CORD-19:5f94b5f7e914cb87d2f607a1b0ef03c135b0a2da JSONTXT 9 Projects

Abstract Background: The regulatory IL-10 and TGF-$1 cytokine gene polymorphisms have been associated with allergic diseases in different populations, like Caucasian, Chinese and Indians. However, no associations between IL-10 and TGF-$1 gene polymorphisms and food allergy (FA) in Japanese children have been evaluated so far. To clarify the relationship of polymorphisms of these 2 regulatory cytokine genes with FA, not atopy itself, polymorphisms IL-10 AV1082G, CV819T and TGF-$1 T+869C, G+915C, CV509T in FA patients were compared with those in non-FA atopic controls. Methods: One hundred-eleven childhood FA patients, with a mean 7.6T4.0 years of age and 115 atopic control children without FA (mean = 8.2T1.5 years of age) were recruited. Most of FA patients and atopic controls were sensitized with house dust mite (92% and 93%, respectively). DNA samples from these subjects were genotyped by using Real Time PCR. Results: The odds ratio (OR) of IL-10 V1082 AA genotype was 2.5 (95%CI, 1.0Y6.4) for food allergy risk when compared with atopic control subjects (p = 0.03). This study had a power of 80% to achieve significance at the 0.05 level for IL-10 V1082A allele when OR is greater than 2.3. Our OR value was 2.4; therefore, we consider this association had enough statistical power to support our finding. There were no significative differences in the frequency of IL-10 CV819T and TGF-$1 gene polymorphisms between both groups. Conclusion: This result indicates that IL-10 AV1082G gene polymorphism is associated with food allergy susceptibility in atopic Japanese children. Our findings should encourage further studies to elucidate the functional relationship of IL-10 AV1082G gene polymorphism in FA pathogenesis. 2 Abstract withdrawn 3 Safety assessment of bacterial choline oxidase protein introduced in transgenic crops Background: Previously, transgenic Brassica juncea expressing bacterial choline oxidase protein did not show enhanced allergenicity as evaluated by WHO/FAO guidelines. In the present study, choline oxidase protein derived from Arthrobacter globiformis was assessed for allergenicity and toxicity. Methods: In-vitro heat stability of the protein was assessed. Choline oxidase induced allergenicity was assessed in Balb/c mice. IgE reactivity of choline oxidase protein was assessed with atopic patients`sera. Acute toxicity of choline oxidase protein was also assessed in mice model. Results: Choline oxidase protein was stable at 90-C for 1 hour and reacted with choline oxidase antibodies on immunoblot. Specific IgE levels were low in choline oxidase treated mice comparable to control, while OVA sensitized mice showed high IgE levels. Intravenous challenge with choline oxidase did not induce any adverse reaction but i.v challenge with OVA led to anaphylaxis in OVA sensitized mice. Choline oxidase sensitized mice demonstrated IL-4 release similar to control in splenic culture supernatant however, OVA sensitized mice showed higher IL-4 levels. Histological analysis of lung tissues from choline oxidase sensitized mice showed intact epithelium with normal airways, whereas OVA sensitized mice showed narrowing of airways with increased eosinophilic infiltration. ELISA with allergic patients`sera (n=45) revealed low specific IgE binding (mean OD 0.281) with choline oxidase protein. Acute toxicity studies of choline oxidase protein in mice model showed no significant difference (p90.05) with control in growth, body weight, food consumption and blood biochemical indices. Histopathology of gut tissues of mice fed with choline oxidase showed normal gastric mucosa lining with normal villi in jejunum and ileum sections. Conclusion: Bacterial choline oxidase protein is heat stable but failed to elicit allergic response in mice model. These data indicates choline oxidase protein may be safe for use in transgenic crops. 4 Identification of amino acids critical for IgE-binding to sequential epitopes of bovine 0-casein and the similarity of these epitopes to the corresponding human 0-casein sequence 9 Abstract withdrawn 10 CXC and CC chemokines produced by human respiratory epithelium Ilja Striz 1 , Eliska Krasna 1 , Libor Kolesar 1 , Antonij Slavcev 1 , Marcela Jaresova 1 , and Jaromir Musil 2 . 1 Institute for Clinical and Experimental Medicine, Department of Immunology, Prague, Czech Republic; 2 University Hospital Motol, Pulmonary Department, Prague, Czech Republic. : Chemokines regulate leukocyte trafficking during their physiological turnover and recruitment to mucosal surfaces in inflammatory reactions. Respiratory epithelium with the ability to respond to locally generated cytokines might be an important source of chemokines attracting diverse cell populations. With this respect, the aim of our study was to compare the capacity of different proinflammatory cytokines to stimulate the gene expression, production and release of multiple CC and CXC chemokines by respiratory epithelial cell line. The chemokine distribution was studied also in bronchoalveolar lavage fluid (BAL) and exhaled breath condensates (EBC) of patients undergoing lung transplantation. A total of 96 chemokines and chemokine receptor genes has been studied using an oligoarray system (Superarray Inc.) in cultured human alveolar type-II like cells A549 stimulated by multiple concentrations of TNF alpha, IFN gamma, IL-1 beta, IL-18, and IL-33. The chemokine levels in culture supernatants, BAL, and EBC were measured using multiplex immunoluminometric assay (Luminex) or by ELISA. In repetitive experiments, epithelial cells constitutively expressed mRNA for CXCL1 (Gro-alpha), CXCL2 (Gro-beta), CXCL3 (Gro-gamma), CXCL5 (ENA-78), CXCL6 (GCP-2), and CXCL8 (IL-8), chemokines attracting preferentially neutrophils. Cells stimulated with IL-1 beta or TNF alpha upregulated mRNA expression of chemokines specific for mononuclear cells recruitment such as CCL2 (MCP-1), CCL4 (MIP 1 beta), CCL5 (RANTES), and CCL20 (MIP 3 alpha). For the induction of of CXCL10 (IP-10) and CXCL11 (I-TAC) which attract activated T lymphocytes, IFN gamma was the most potent stimulus. The induction of epithelial cells by IL-1 related cytokines, IL-33 and IL-18, resulted only in a moderate upregulation of few CC or CXC chemokines compared to a potent effect of IL-1 beta stimulation. Epithelium derived chemokines such as CXCL5, CXCL8, CCL2, CCL4 or CCL5 are abundant in BAL fluids but can be only rarely detected in EBC. We conclude from our data that respiratory epithelial cells are involved in regulating the influx of different populations of inflammatory cells by release of multiple CC and CXC chemokines in a highly coordinated manner. diarrhea are, protein-losing gastroenteropathies. Intestinal lymphangiectasia (InL) . Is a rare entity which can be congenital or acquired it is characterized by abnormal, dilated lymphatics throughout the intestinal tract. The disorder leads to leakage of lymph causing hypoproteinemia, lymphopenia and hypogamaglobulinemia, which can leads to depression of both the humoral and cellular immune systems. Case Report: A 35 year old woman from rural Mexico with a history of asthma for 20 years, presented with a chief complaint of diarrhea for 15 months with up to 10 evacuations a day, she referred colic pain with abdominal distension and flatulence with recurrent symmetrical systemic edema. The patient showed fatigue, weaknesses and muscle spasms as well as myalgias and fasciculations, she lost 22 lbs and developed onychomycosis in all of her toenails. She recalls showing the same clinical signs and symptoms once during childhood. Her medical team did not have a clear diagnosis. Laboratory revealed a lymphopenia, hypoalbuminemia, hypogammaglobulinemia, hypocalcemia and negative HIV by ELISA. The stool Ü 1-antitrypsin level was elevated. The CT of chest and abdomen was normal. An esophagogastroduodenoscopy and colonoscopy were performed showing gastritis, a duodenal biopsy demonstrates dilated lymphatics and white villi spots. Discussion: The lack of elevated levels of protein in the urine along with the depressed serum levels of IgG drew attention to the gastrointestinal tract as the likely portal of protein loss. Detecting an elevated level of "1-antitrypsin in a 24-hour stool collection adds support to the diagnosis of InL. A duodenal biopsy with the characteristic histological findings, demonstrating dilated lymphatics and white spots, confirmed the diagnosis. Treatment of primary InL is symptomatic and only marginally effective. We treat our patient with strict dietary lipid restrictions, pancreatic enzymes, probiotics, diuretics, Mediumchain triglycerides and multivitamins. The patient showed a marked response regained her normal weight, but the diarrhea and edema sporadically presents. To date is the seventh reported case of primary InL in Mexico and to our knowledge the only described in literature of biphasic presentation. Atrial natriuretic peptide receptor (NPRA) signaling in human dendritic cells controls treg development Weidong Zhang 1 , Xueqin Cao 1 , Gary Hellermann 1 , Xiaoyuan Kong 1 , Dongqing Chen 1 , Jia-Wang Wang 1 , Richard F. Lockey 2 , and Shyam S. Mohapatra 1 . 1 University of South Florida, Internal Medicine, Tampa, United States; 2 James A. Haley VA Hospital, Internal Medicine, Tampa, United States. : The natriuretic peptides (NP) are key endogenous factors that control inflammation and immune tolerance, the latter is essential to maintain immune homeostasis, control autoreactive T cells, prevent the onset of autoimmune diseases and achieve tolerance of transplants. The mechanism of regulation of ANP-NPRA signaling in DCs is crucial for the understanding of how ANP regulates innate immunity and how failure of this signaling might contribute to autoimmune disease, chronic inflammation and tissue damage. Herein, the role of the c-terminal NP, ANP and one tolerance inducing NP, NP73-102, in modulating DC function was examined. The results demonstrate that in contrast to ANP, NP73-102 primes DCs to induce regulatory T (Treg) cells. The ANP receptor, NPRA, binds to TLR-2, SOCS3 and STAT3 and affects induction of IL-6, IL-10 and TGF-!, not IL-17. SOCS3 expression is controlled in a MyD88-dependent manner. Also, down-regulation of SOCS3 and TLR2, but not STAT3, affects NPRA expression. These results demonstrate that TLR2, and SOCS3 are key players in integrating ANP-NPRA signaling with innate immunity and provide insight into how inhibition of IRNP affects ANP-NPRA signaling promoting the tolerogenic phenotype of DCs. Thus, we found a tendency towards the prevalence of ever having got an asthma diagnosis, and having current asthma being slightly more common in 2003 compared to 10 years earlier. However, using asthma medication had increased and suffering from asthma symptoms had decreased. Furthermore, self-reported allergy had increased, especially towards furry pets. Discussion: The study population was not a representative sample of the Swedish adult population, and one should be cautious when generalizing from the results. However, apart from gender, school employees should not differ substantially from the general working population. Furthermore, the two study populations were very homogenous to each other, with respect to gender, age, occupation and geographical habitat, and the study methods were identical. Conclusion: The results imply a small increase of asthma and allergy during the 10-year period 1993Y2003 in Swedish adults. Azienda Ospedaliera Universitaria San Martino, Azienda Ospedaliera Universitaria San Martino, Genoa, Italy; 4 GP SIMG Sezione Genova, GP SIMG Sezione Genova, Genoa, Italy; 5 GP ASSIMEFAC, GP ASSIMEFAC, Genoa, Italy. Background: Despite asthma control and treatment adherence have been largely investigated in current literature, the appropriateness of general practitioner in prescribing for patients with asthma is less studied. At the beginning of a continuing professional development (CPD) program on rhinitis and asthma, part of ARGA (Allergopatie Respiratorie studio di monitoraggio linee-guida GINA e ARIA), a study supported by Italian Drug Agency (AIFA), the drugs prescription and the healthcare resource utilization of physicians adhering to the study were performed. Materials and Methods: We observed 70.147 patients followed by 60 GP during 2006. 709 patients had diagnosis of asthma. 68 patients were hospitalized for asthma exacerbations, mean age 51 yrs. Among these patients only 16 were treated with association of beta 2 agonist/inhaled steroid coupled with antileukotrienes (9/16), antihistamines (8/16), theophylline (4/16) . Systemic steroids (prednisone, betamethasone) were prescribed to 10 patients, among them, 3 received no other therapy, other 3 patients received only oral antihistamines. In 4/16 patients were prescribed tiotropium although asthma indication is not approved. The total amount of prescribed asthma drugs for these patients was low, it could last for a maximum of two months. The most prescribed drugs were beta 2 short acting bronchodilators (up to 22 drug boxes for a patient). No patient was evaluated by a specialist (allergist or pneumonologist) before hospitalization; only 6 were visited by a specialist after their discharge from the hospital and 11 underwent a spirometric evaluation. 7 patients were hospitalized twice in a year. Conclusion: Regardless of the efforts of national and international societies for the diffusion of asthma diagnosis and treatment guidelines, asthma is still undervalued and untreated by general practitioners; this could be the principal cause of hospitalization for asthma exacerbation in the observed group of patients. It could be interesting to evaluate in a CPD blended program (5 residential plus 4 distance learning courses) in which the contents are tailored on physicians knowledge, which healthcare resource utilisation and drugs prescription would be able to improve the asthma management. elicits rhinitis and conjunctivitis especially in younger generations. The twodimensional IgE-binding spectrum of C. japonica pollen allergens demonstrated that many allergens remain to be identified. Here we present the molecular cloning and immunochemical characterization of a novel C. japonica pollen allergen belonging to aspartyl protease family. Methods: TOF-MS analysis of a high IgE-binding protein, termed CPA63, revealed its internal amino acid sequences. Based on these sequence information, cDNA-encoding CPA63 was cloned by RACE-PCR. The allergen was produced as a recombinant protein using baculovirus-insect cell culture system, and purified by double chromatographic technique using HisTrap and HiTrap Q columns. Putative mature recombinant CPA63 (r-CPA63) was produced upon autolysis by incubation in acetate buffer (pH 3.3) , and used for ELISA experiment. Its proteolytic activity was tested using FITC-casein as a substrate at different pHs, and substrate specificity was evaluated by using series of protease inhibitors. Results: CPA63 cDNA encoded a 472 amino acid polypeptide with calculated molecular weight and isoelectric point of 51.1 kDa and 4.69, respectively. Homology search revealed that CPA63 polypeptide sequence showed about 40% identity with plant aspartyl protease/nucleoid DNA binding protein family members. ELISA demonstrated that purified r-CPA63 was recognized by pollinosis patient IgE at a frequency of 58% (18/31). The r-CPA63 also showed an aspartyl protease-like proteolytic activity, demonstrating its enzymatic maturation upon autolysis. Conclusion: CPA63 is the first plant aspartyl protease identified as an allergen. That might well open new investigations for other plant aspartyl protease allergens. The availability of CPA63 sequence and recombinant allergen production could be useful to develop future diagnostic technique and therapeutic approaches. Background: Hereditary angioedema (HAE) is characterized by recurrent skin swelling, abdominal pain attacks, and potentially life-threatening upper airway obstruction. The classic HAE types (I and II) are caused by mutations in the complement C1 inhibitor gene, resulting in a quantitatve or qualitative deficiency of C1 inhibitor. In contrast, in a novel HAE type, affecting mainly women, C1 inhibitor concentration and activity in plasma are normal (HAE type III, HAE with normal C1 inhibitor); we hypothesized that an abnormal coagulation factor XII molecule may lead to inappropriate activation of the kinin-forming cascade and, therefore, performed a search for mutations in the F12 gene. Methods: Twenty unrelated index patients from families with hereditary angioedema and normal C1 inhibitor activity were screened for mutations in the coagulation factor XII gene by sequencing of the 14 exons and splice junctions of the F12 gene; subsequently, beside 235 control individuals, another six such index patients as well as 90 patients with idiopathic angioedema were selectively sequenced for exon 9 of the F12 gene. Results: Two different non-conservative missense mutations, both located in exactly the same position within exon 9, namely in the second position of the codon (ACG) encoding Thr309 of the mature protein, were identified. Five of the twenty patients screened showed a heterozygous CYA transversion (1032CYA), predicting a threonine-to-lysine substitution (Thr309Lys); one additional patient showed a heterozygous CYG transversion (1032CYG), resulting in a threonine-to-arginine substitution (Thr309Arg). Sequencing of exon 9 in another six index patients revealed one further patient heterozygous for the Thr309Lys mutation. Thus, in 7 of 26 unrelated patients the wild-type threonine is substituted by a basic amino acid residue. The mutations were not found in healthy control individuals (n = 235) and co-segregated with the phenotype in seven families with altogether 23 affected women, providing strong support that they cause disease. Finally, the Thr309Lys mutation was also identified in 2 out of 90 patients with idiopathic angioedema; thus, this mutation may also play a role in a subgroup of these singular angioedema cases with no affected family members. Conclusion: These findings provide strong support that the coagulation factor XII gene is a new angioedema gene. Subcutaneous Background: Disinhibition of kallikrein results in excess of bradykinin and is responsible for signs and symptoms of hereditary angioedema (HAE). The highly potent, specific plasma kallikrein inhibitor, DX-88 (ecallantide) has been shown to ameliorate symptoms of HAE. Methods: Patients 10 years of age or older with documented HAE (12 naBve patients and 48 patients who were treated for a single acute HAE attack in the double-blind, placebo-controlled stage of EDEMA3) participated in this openlabel extension. Patients were given 30 mg DX-88 to treat acute HAE attacks at all anatomic site locations-abdominal, peripheral, and laryngeal. Outcome measures at 4 and 24 hours were the HAE-specific Treatment Outcome Score (TOS) and change from baseline in Mean Symptom Complex Severity (MSCS) score. TOS and MSCS are composite scores measuring all attack sites. Time to onset of response and time to significant improvement in overall response were recorded. Safety was assessed by treatment-emergent adverse events (TEAEs) and serious adverse events (SAEs). Results: Interim data are presented for 59 patients treated for 145 acute HAE attacks (20 men, 39 women, median age 34.0 years, range 12Y77 years). Mean TOS was 75.3 T 39.64 at 4 hours and 81.8 T 36.55 at 24 hours (Q 50= improvement). Mean MSCS change from baseline was j2.3 T 0.6 at both 4 and 24 hours (j1 to j3 = clinically-significant improvement). Median time to onset of overall improvement was 37.5 minutes, and time to onset of significant overall improvement was 165.0 minutes. Safety data for 51 patients included 166 TEAEs (87% mild or moderate in severity and 89% resolved without sequelae), with 30 related TEAEs in 15 patients. Related events experienced by more than 1 patient include diarrhea, nausea, injection site reaction (3 patients each) , HAE, myalgia, headache, cough, and pruritus (2 patients each). A total of 7 SAEs occurred in 7 patients; 6 were unrelated to DX-88, and 1 SAE of anaphylactic reaction reported previously, was assessed as related, and resolved without sequelae. Conclusion: Repeat dosing of DX-88 (ecallantide) for acute HAE attacks resulted in onset of overall improvement in G40 minutes and significant overall improvement in G3 hours. Ecallantide was generally well tolerated with the majority of adverse events being mild or moderate in severity and resolved without sequelae. Background: A new type of hereditary angioedema was described recently. It was characterized by recurrent bouts of angioedema in various organs and normal C1 inhibitor and was observed mainly in women. Our aim was to conduct a detailed study of the clinical features of this condition. Methods: A total of 138 patients with hereditary angioedema and normal C1 inhibitor who belonged to 43 unrelated families were examined through the use of standardized questionnaires. Results: A majority of patients with hereditary angioedema and normal C1 inhibitor had skin swellings (92.8%), tongue swellings (53.6%), and abdominal pain attacks (50%). Laryngeal edema (25.4%) and uvular edema (21.7%) also were frequent, whereas edema episodes of other organs were rare (3.6%) . Facial swellings and tongue involvement occurred considerably more frequently compared with hereditary angioedema due to C1 inhibitor deficiency. The number of patients with recurrent edema of only one organ was higher than in classic hereditary angioedema. The number of patients with disease onset in adulthood was significantly higher in hereditary angioedema with normal C1 inhibitor compared with classic hereditary angioedema. Erythema marginatum was not observed. A subgroup of patients from families with coagulation factor XII mutations showed the same symptoms as the other patients. Long-term treatment consisted in tranexamic acid and attenuated androgens. Conclusion: Hereditary angioedema with normal C1 inhibitor levels shows a characteristic pattern of clinical symptoms. The main clinical features include skin swellings, tongue swellings, and abdominal pain attacks. There are many differences in the clinical symptoms and course of disease between this type of hereditary angioedema and classic hereditary angioedema due to a genetic C1 inhibitor deficiency. Skin reactivity to autologous serum and safety of COX-2 inhibitors in NSAID-intolerant patients with urticaria and/or angioedema Mauro Cancian, Raffaele Bendo, Elena Ossi, and Giuseppe Realdi. Clinica Medica 1 -University of Padova, Medical and Surgical Sciences Department, Padova, Italy. Background: Pseudoallergic reactions (PAR) to non-steroidal antiinflammatory drugs (NSAID) are very common in chronic urticaria (CU), as aspirin and related drugs impair symptoms in about 20Y30 % of patients. Moreover, a group of otherwise normal subjects who develop urticaria and/or angioedema (UA) only after the intake of NSAIDs also exists. Although the inhibition of cyclo-oxigenase (COX) pathway seems to play a central role in NSAIDintolerance, a very high prevalence of skin reactivity to autologous serum (AS) has also been reported in NSAID-induced UA, pointing to a possible involvement of the immune system in the pathogenesis of these forms. Methods: 50 patients who had experienced NSAID-induced UA were recruited and gave their informed consent. 28 had a history of CU exacerbated by NSAIDs, whereas the remaining 22 reported acute UA after NSAIDs intake. In 26 cases, UA had been elicited by a single NSAID, whilst 24 patients signalled adverse skin reactions to two or more COX-inhibitors. None of the patients presented nasal polyps, nor had evidence of asthma or sinusitis. Oral tolerance tests were conducted on different days by single-blind, placebo controlled challenge with increasing amounts of both conventional (nimesulide, 100 mg whole dose; meloxicam, 15 mg), and highly selective (celecoxib, 400 mg; eterocoxib, 90 mg) COX-inhibitors. The test was continued until the whole dose was reached, or UA appeared. Furthermore, skin reactivity to AS was assessed in all subjects in basal conditions, as well as whenever challengeinduced adverse reactions occurred. Results: No patients developed skin symptoms on the placebo day, and 28 tolerated each of the four COX-inhibitors. 16 experienced UA with nimesulide, 8 with meloxicam, 4 with celecoxib and 2 with eterocoxib. Only 4 patients with CU showed a positive cutaneous response to AS, whilst in the other subjects skin tests with both basal serum and samples collected during challenge-induced UA always elicited negative results. Conclusion: Our findings confirm that highly selective COX-inhibitors represent the safest anti-inflammatory drugs in NSAID-induced UA, although it is well known that coxib intake is not recommended in subjects with renal or cardiovascular diseases. Moreover, these results strengthen the concept that cutaneous reactivity to AS is almost exclusively confined to chronic urticaria, and that autoimmunity is not involved in the pathogenesis of NSAID-induced PAR. Treatment of skin swellings with pasteurized C1 inhibitor concentrate in patients with hereditary angioedema Background: Gum arabic is used as an emulsifier, a thickening agent and as stabilizer in foods, in pharmaceutical industry, lithography and cosmetics. Exposure to gum arabic has been reported to cause occasional cases of occupational asthma, but ingestion of it does not commonly cause immediate allergic symptoms. Methods: In 2005 workers of a candy factory were examined for suspected occupational allergy. Hard boiled candies were covered with spray dried gum arabic in rotating drums, where the workers poured the gum arabic powder. 7/11 had respiratory and skin symptoms and 4 had skin symptoms. Spirometry, histamine challenge test, exhaled breath NO and PEF measurements, skin prick tests (SPT), gum arabic IgE measurements and cutaneous or bronchial challenge tests with gum arabic were carried out. Patch tests were carried out to a patient with eczematous skin disease. In addition, SPT data from 2997 subjects examined for food allergy symptoms from 1998 to 2006 were evaluated. Subjects with positive SPT to gum arabic received a postal questionnaire. Results: Four workers had occupational asthma caused by gum arabic. All of them had a positive SPT with gum arabic and specific IgE to gum arabic (0.60-5.6 kU/l). Three patients had a positive bronchial challenge test with gum arabic and one of them had a positive skin challenge test. One patient had significant PEF decrease when exposed to gum arabic at work, a positive skin challenge and immediate oral symptoms associated with ingested gum arabic. None of them had other significant allergies. One worker had contact dermatitis caused by thiuram chemicals and another with positive SPT to house dust mites had work related allergy to carmine red used in candies. Five workers had no occupational disease. Of patients tested for food allergies 17/2997 (0,005%) had positive SPT to gum arabic and most of them also had positive SPT to pollen and foods. 11/17 (65%) returned the questionnaire. At the time of skin testing gum arabic associated allergic symptoms were suspected in three, and two reported gum arabic ingestion associated oral symptoms in the questionnaire. One gum arabic SPT positive patient without respiratory symptoms had an occupational history of work in a candy factory for years ago. Conclusion: Occupational allergy caused by IgE mediated allergy to gum arabic is a risk in exposure occupations. Ingested gum arabic causes symptoms in few patients. Determinants of elevated exhaled nitric oxide (eNO) among bakery workers in South Africa Background: Severe asthma and COPD are characterized by clinical exacerbations. GPs are often required to decide between hospitalisation and home treatment for these patients. Aim: To evaluate if the ambulatory measurement of Oxygen Saturation (SO2) could be safe and effective, to verify the correlation between SO2 measurement during scheduled visits an during exacerbations, to define the patient pattern that requires hospitalisation, to assess the possibility of SO2 database registration in Gps setting. Methods: 450 GPs and 50 pneumologists have been enrolled in the study with the target to evaluate 16000 ptz (10000 by Gps; 6000 by pneumologists). Each physician has been provided with a pulse oxymeter. The SO2 measurement and the fill in of a specific questionnaire have been required to the outpatients attended for disease exacerbation. Results: A descriptive analysis of results will be provided. The patients will be divided in two population according to the necessity of hospitalisation and a comparison of all the parameter registered in GPS and specialists setting will be performed. The relative importance of SO2 measurement for GPS and pneumologists will be evaluated. Discussion: The present ongoing study should provided important information about the utility and safety of SO2 measurement in obstructive patient evaluation during exacerbation. The results should be useful in defining guidelines for exacerbation management. Total IgE in urban black south african teenagers: the influence of atopy and helminth infection Michael Levin. Red Cross War Memorial Childrens Hospital, Allergy Clinic, Cape Town, South Africa. Background: IgE levels are usually elevated in allergic diseases, being highest in atopic eczema, followed by atopic asthma and allergic rhinitis. Genetic factors are believed to play a role in total IgE levels, with higher levels seen in Black African subjects. Total IgE is also raised in parasite infection. Thus, the higher total IgE levels in Black Africans could be due to environmental rather than genetic factors. Few studies investigate the usefulness of total IgE levels in the assessment of atopy in Black Africans. Aim: The objective of this study was to determine the total IgE levels in unselected urban Black African high school children and to correlate this with atopy and ascaris sensitization. Methods: Two hundred and twenty Urban Xhosa children (mean age 17 years) attending a school in Cape Town were studied. Atopic status was assessed by means of specific allergen sensitization (ALK\ skin prick tests to 8 inhalant and 4 food allergens), self reported asthma and bronchial hyperresponsiveness measured by methacholine challenge. Total IgE and Ascaris specific IgE were measured by CAP-RAST (Pharmacia). Total IgE levels were subanalysed according to atopic status and ascaris sensitisation in order to establish ranges of total IgE in atopic subjects, ascaris sensitised subjects, and non-atopic, non ascaris sensitised subjects. Results: Total IgE levels were markedly skewed toward the left and were not distributed in a Gaussian or a log-normal distribution. Mean total IgE was 307.7 kU/L. The median was 106 kU/l and the interquartile range 50.4 kU/l to 288 kU/l. Skin prick tests were positive for aeroallergens in 32.3 % of subjects. Thirty four percent had elevated ascaris IgE. Total IgE was higher in atopic versus non-atopic subjects and correlated with the number of positive skin prick tests (Kruskall Wallis ANOVAR p G0.0001), self reported asthma (p = 0.025) and bronchial hyperresponsiveness (p = 0.0002). In addition total IgE correlated with ascaris IgE (p G0.0001). Subjects with no ascaris sensitisation had median total IgE of 77.1 kU/l, similar to the levels seen in people of other genetic origins. Conclusion: This study has demonstrated that total IgE is correlated on one hand with atopy, bronchial hyper responsiveness and self reported asthma and on the other hand with ascaris sensitisation. The likelihood of helminthic infection rather than genetic differences, is thus the major factor determining a population`s specific IgE range. Alfred Hospital, Allergy, Immunology and Respiratory Medicine, Melbourne, Australia. Background: Elevated levels of total IgE (tIgE) in serum are characteristic of allergic diseases. Levels of tIgE are influenced by genetic predisposition, age, sex and helminth infections. However, the association between tIgE and allergic diseases in children living in areas endemic for helminth infections is not clear. The aim of our study was to assess the relationship between tIgE, allergic diseases and geohelminth infections in children. Methods: A total of 640 schoolchildren 9Y11 years of age was selected by stratified random sampling. Data regarding allergic diseases (asthma, rhinitis and eczema) were collected by a standard questionnaire given to the parent or guardian. Screening for helminth infections was done by examining their fresh stool samples by modified Kato-Katz technique. Serum tIgE was measured by Fluoroenzymeimmunoassay in 67 geohelminth-positive subjects and in a comparable group of geohelminth-negative subjects. Results: The mean age in the study population was 10 years (SDT0.3). The prevalence of geohelminth infection was 15.5%. Trichuris trichiura (14.3%) was the most common followed by Ascaris lumbricoides (4.2%) and hookworm (0.2%). Mixed infection was detected in 20.3% of infected children. Infection intensity was light in 68.9% of infected children while 28.4% and 2.7% showed moderate and heavy infection respectively. The cumulative prevalence of allergic diseases was 33.7%. Prevalence of asthma, rhinitis and eczema was 17%, 21.4% and 5% respectively. Serum tIgE concentrations showed a positively skewed distribution. Geometric mean (GM) for tIgE for the geohelminth infected group (1039.9kU/L) was significantly higher than that of the non-infected group (575.4kU/L) (p = 0.004). It was also higher in the allergic group (933.3kU/L) than in the non-allergic group (639.7kU/L) but the difference was not statistically significant (p = 0.068). The GM for tIgE for non-allergic children in the study population was much higher than that seen in non-allergic children in developed countries. Conclusion: Serum tIgE concentration was strongly associated with the presence of geohelminth infections in children. Serum tIgE may not be a useful marker for allergic diseases in children living in areas endemic for geohelminth infections. Evidence of eosinophil activation in adenoid and tonsil tissues from atopic children Seung-Youp Shin 1 , Chang-Ki Hong 1 , Sung-Jin Choi 2 , Kun-Hee Lee 1 , Joong-Saeng Cho 1 , and Hae-Sim Park 2 . 1 KyungHee University Hospital, Department of Otorhinolaryngology, Seoul, Republic of Korea; 2 Ajou University Hospital, Department of Allergy and Rheumatology, Suwon, Republic of Korea. Background: Adenoid and tonsil have been considered as immune organs. They may present different cellular and cytokine profiles according to the atopic status. The objective of this study was to find the status of eosinophil activation and compare between atopic and non-atopic children. Materials and Methods: Adenoid and tonsil tissues obtained from 40 children (18 atopics and 22 non-atopics) were enrolled. Seven atopic and 9 non-atopic children had chronic inflammatory diseases such as chronic rhinosinusitis (CRS) and/or otitis media with effusion (OME). Atopic status was classified as a positive result ( 92+) on MAST test to common inhalant allergens. Serum total IgE and eosinophil count were measured. ECP level and cellular activation markers, including sCD23, sIL-2R and IL-6 within the supernatant of adenoid and tonsil tissue were measured by CAP system (Pharmacia, Sweden) or ELISA. Results: The ECP level in adenoid and tonsil tissue from atopic children was 451.06g/L, 514.26g/L, which were significantly higher than those from nonatopic children (353.2 6g/L, 374.4 6g/L, respectively). The ECP level in adenoid and tonsil tissues from atopic children with CRS showed the highest (564.6 6g/L and 718.1 6g/L), followed by those from non-atopic children with CRS (449.3 6g/L and 430.7 6g/L), those from atopic children without CRS (371.5 6g/L and 371.6 6g/L), and those from non-atopic children without CRS (291.5 6g/L and 338.2 6g/L). And the ECP level in adenoid were significantly correlated with locally produced total IgE and cellular activation markers, including sCD23, sIL-2R and IL-6. Conclusion: This findings suggest that activated eosinophil could be involved in inflammatory response occurring in the adenoid and tonsil from both atopic and non-atopic children. The more activated eosinophils, the more CRS can be combined. The activation of T cell, especially Th2 cell could lead to the activation of eosinophil in adenoid and tonsil tissues. Plant homeodomain finger protein gene polymorphisms are associated with plasma total IgE and exhaled nitric oxide levels in Chinese children Hing Yee Sy 1 , Ting Fan Leung 1 , Chung Yi Li 1 , Iris H. S. Chan 2 , Edmund Yung 1 , Nelson L. S. Tang 2 , Christopher W. K. Lam 2 , and Gary W. K. Wong 1 . 1 The Chinese University of Hong Kong, Department of Paediatrics, Hong Kong, Hong Kong; 2 The Chinese University of Hong Kong, Department of Chemical Pathology, Hong Kong, Hong Kong. Background: Single-nucleotide polymorphisms (SNPs) of the gene encoding plant homeodomain finger protein (PHF11) were shown by positional cloning to be associated with severe asthma and elevated circulating total IgE level in Caucasians. These SNPs were also associated with childhood atopic dermatitis. Such genetic association has not been studied in Chinese. The objective of this study is to investigate the association between PHF11 SNPs and asthma phenotypes in Chinese children. Methods: 269 asthmatics and 165 non-allergic children were recruited from our paediatric clinics. Their lung function was assessed by spirometry, and exhaled nitric oxide (FeNO) levels were measured online by chemiluminescence analyser at expiratory flow rate of 50 ml/s. Plasma total and specific IgE were quantified by immunoassays. Ten SNPs in PHF11 were genotyped by multiplex SNaPshoti reaction with an ABI-310 Genetic Analyser. The linkage disequilibrium (LD) pattern of these SNPs was analysed by Haploview, and the association between asthma traits and PHF11 was analysed by multivariate regression. Results: The mean (SD) log-transformed plasma total IgE and FeNO levels in cases and controls were 2.62 (0.61) and 80.5 (59.2) ppb and 1.81 (0.71) and 40.6 (43.2) ppb, respectively (P G 0.001 for both). All 10 SNPs followed Hardy-Weinberg equilibrium (P 9 0.05). There were significant interethnic variations in PHF11 minor alleles (up to 19%) in our children when compared with Caucasians. PHF11 polymorphisms were not associated with asthma diagnosis (P 9 0.5), asthmatics with FEV 1 G 65% of predicted (P 9 0.3), or atopy (P 9 0.09). Multivariate linear regression showed that PHF11 T167A and G491A were associated with plasma total IgE (P = 0.048 and 0.051, respectively); and C507T and C732T were associated with FeNO (P = 0.019 for both). Two haplotype blocks could be constructed -one with T79C, G491A, C507T and T661A (D' 9 0.9) and the other with T395C and C732T (D' = 0.95). Three major haplotypes from these 6 SNPs (CTGA CT [53%], TTAT TT [29%] and TCGT TC [18%]) were identified, but which were not associated with asthma diagnosis (P90.3). Conclusion: Our results suggest that PHF11 polymorphisms are associated with plasma total IgE and FeNO levels but not asthma diagnosis in Chinese children. Ma Anunciación Martin Mateos 1 , Montserrat Garriga Badía 2 , Ana Plaza Martin 3 , Maite Giner 3 , and Mónica Piquer 3 . 1 Hospital Sant Joan de Déu-Hospital Clinic., Universidad de Barcelona, Barcelona, Spain; 2 Fundación Altahia, Hospital de Manrresa, Barcelona, Spain; 3 Hospital Sant Joan de Déu-., Sección de Inmunología y Alergia, Barcelona, Spain. : Following a bronchiolitis caused by respiratory syncytial virus (RSV), later recurrent wheezing are common. Objective: The aim of this study was to establish the relationship between repiratory syncytial virus and the modifications in immunity that cause appearance of childhood asthma. Patients and Methods: The prospective study Two groups of study: CHILDREN WITH BRONCHIOLITIS. (Group A). The problem group of children was originally made up of 65 children (with an average age of 3 months, 1 day to 7 months) who were admitted to this Hospital for their first episode of VRS positive bronchiolitis, from December 1997 to February 1998. 50 of them completed follow-up for 2 years. In the tests done at the moment of admittance :hemogram, and Ig G, Ig M, Ig A, Ig E, lymphocyte count, eosinophil cationic protein,IL-4 IFN-g. were at 6, 12 and 18 months, At 18 months an adhesion molecules count was requested: sVCAM.1, sLselectin. The second group (group B), was made up of 80 children, sub-divided into 4 groups of 20 according to age: IL-4, IFN-g and adhesion molecules. Results: In the acute stage of the bronchiolitis, the immunological response to the viral infection was anomalous, in that they presented a humoral immunity response (with an increase of immunoglobulins G, A, M, the later with p = 0,014) and a decrease in T, T4 and T8 cellular immunity. The immunolological response to the viral infection must be of cellular predominance, in order to be effective. Among all children suffering from VRS bronchiolitis, we found a statistically significant increase (p = 0,001) of sL-Selectine at 18 months of age in comparison with healthy children of the same age. Conclusion: the characteristic patterns that lead us to believe that a child who suffers from bronchiolitis will develop asthma are: Increase in humoral immunity response at the acute stage of the bronchiolitis Decrease in cellular immunity response, from the acute VRS infection until 6 months of age Increased IL-4 values mainly at 6 and 12 months of age. Decrease in INF-g between 6 and 18 months of age. Association of polymorphism of the mast cell chymase gene promoter region (-1903 G/A) and (TG)n(GA)m repeat downstream of the gene with bronchial asthma in children Elham Hossny 1 , Nermine Amr 1 , Shereen Elsayed 2 , Rasha Nasr 2 , and Eman Ibraheim 1 . 1 Ain Shams University, Pediatrics, Cairo, Egypt; 2 Ain Shams University, Microbiology and Immunology, Cairo, Egypt. Background: Mast cell chymase is an important mediator of inflammation and remodeling in the asthmatic lung. Various studies examined the association between -1903 G/A single nucleotide polymorphism (SNP) of the mast cell chymase gene (CMA1) and allergic phenotypes with inconsistent results. The (TG)n(GA)m repeat polymorphism 254 bp downstream of the chymase gene was previously found to associate asthma in adults. We sought to investigate the association of the -1903 CMA1 SNP as well as the (TG)n(GA)m repeat polymorphism with childhood asthma and its associated traits in a case-control study aiming to identify local data from our country. Methods: The study comprised 15 children (6Y10 years old) with bronchial asthma and a family history of atopy enrolled consecutively during asthma exacerbation and 15 age and sex matched children with negative personal or family history of allergy as a control group. They were subjected to clinical evaluation and serum total IgE estimation as well as search for polymorphism in the CMA1 gene promoter region 1903 G/A using PCR restriction fragment length polymorphism based genotyping (RELP) and the (TG)n(GA)m repeat polymorphism 254 bp downstream of the CMA1 gene. Results: Investigating the -1903 G/A SNP revealed significant difference in the distribution of CMA1 A and G alleles between the patients and controls. Allele G was detected in 70% of patients compared to 16.7% of controls, while allele A was overexpressed in the control group (83.3%). Our data showed for the first time a positive association between -1903 G/A SNP and asthma. Concerning the (TG)n(GA)m repeat, a significant difference was obtained in the allele counts among patients and controls in the current study. The largest difference was observed in allele 39 which was only represented in the asthmatic children and allele 37 which was overexpressed among the controls. A positive association for the allele 39 of the (TG)n(GA)m repeat with serum IgE levels was also detected in our series. The findings are limited by the sample size. Conclusion: We report the association of -1903 G/A SNP and (TG)n(GA)m repeat polymorphism downstream of the CMA1 gene with bronchial asthma in a group of Egyptian children. The findings suggest that the alleles, genotypes and haplotypes investigated are possible important determinants of asthma susceptibility and are probably involved in regulating IgE levels in atopic asthma. Efficacy of mometasone furoate nasal spray-MFNS-in the treatment of allergic rhinitis. Meta-analysis of randomized controlled trials Martin Penagos, Enrico Compalati, Francesco Tarantini, Giovanni Passalacqua, and Giorgio Walter Canonica. Allergy & Respiratory Diseases Clinic, Genoa University, Department of Internal Medicine, Genoa, Italy. Objective: To assess the efficacy of MFNS in the treatment of allergic rhinitis. Study Selection: Randomized, placebo-controlled and double blind trials that studied the effects of MFNS in patients with allergic rhinitis. Comprehensive searches of the EMBASE, LILACS, Cochrane Library and MEDLINE databases from 1966 up to January 2007 and references of identified articles and reviews. Outcomes: Different outcomes measured in the active treatment and control groups were considered. Review Manager 4.2.8 Program (Cochrane Collaboration) was used for data synthesis. Outcomes were extracted from original articles. If information was not available, authors of each trial were contacted. Some graphics were digitalized. The analysis included the calculation of standardized mean difference (SMD). There was significant heterogeneity among the study results, because of differing study methodologies. Randomeffects model was used. Results: The initial scanning identified 91 articles, 26 of which were potentially relevant trials on the use of mometasone for the treatment of allergic rhinitis. 14 studies were randomized controlled trials and met inclusion criteria for the meta-analysis. All randomized clinical trials included 23 comparison groups and more than 20 different outcomes (individual and global). For efficacy assessment, 2163 patients were analyzed; 1113 received mometasone and 1050 placebo. MFNS was associated with significant reduction in total symptoms scores (SMD j0.59; 95% CI j0.80 to j0.39; P G 0.00001; heterogeneity I2 = 78.3%.). Safety: For adverse events report, 1758 patients were included. 884 received MFNS and 874 placebo. No significant difference between MFNS and placebo was revealed (SMD 1.02; 95% CI 0.83 to 1.25; P = 0.86; heterogeneity I2 = 1.0%) Conclusions: First among nasal CSs, MFNS efficacy achieved Evidence Ia in the treatment of allergic rhinitis.The frequency of adverse events is similar in both groups, when it was compared with placebo. Studies on the role of tumor necrosis factor alpha gene polymorphisms in the therapeutic response to intranasal UV phototherapy Márta Boros-Gyevi, Edina Garaczi, Andrea Koreck, Márta Széll, and Lajos Kemény. University of Szeged, Dermatology and Allergology, Szeged, Hungary. : Tumor necrosis factor alpha (TNF") is one of the key cytokines in UVinduced immunosuppression. We have shown that intranasal phototherapy with mixed UVA-UVB-visible light (mUV/VIS) is a new therapeutic tool for the management of allergic rhinitis. The aim of this study was to investigate whether the efficiency of rhinophototherapy is related to specific TNF" polymorphisms. Fifty-nine patients with a history of at least 2 years of ragweed-induced allergic rhinitis were treated during ragweed seasons in Szeged, Hungary. Rhinophototherapy was carried out 3 times a week for 2 weeks and continued by a one-week follow up period, or 3 times a week for 3 weeks with increasing doses of mUV/VIS. The total dose of mUV/VIS therapy was the same at the end of the study in both therapeutic regimens. The patients graded the following 4 symptoms of allergic rhinitis on a standard severity scale: nasal itching, nasal obstruction, rhinorrhea and sneezing referred to as total nasal score (TNS). The average improvement in TNS of all patients was evaluated using repeated measures ANOVA. Patients with more than 50 percent decrease of the average change in the TNS were considered as responders and with less than 50 percent decrease as non-responders. According to these parameters, 38 patients were considered to be responders (64%) and 21 patients were non-responders (36%). Blood sample was taken from all patients for single nucleotide polymorphism (SNP) analysis of TNF". Three SNPs of the promoter region of TNF" gene were selected for polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. The genotypes and allele frequencies of selected SNPs' were compared between the responders and non-responders for rhinophototherapy. Allergic rhinitis: occurrence of sinusitis in Bsneezers-runnersâ nd BblockersŜ andeep Sahay, Danish Jamal, and Ashok Shah. Vallabhbhai Patel Chest Institute, Respiratory Medicine, Delhi, India. Background: Patients with allergic rhinitis can be categorized as Bsneezers-runners^and Bblockers^. The presence of sinusitis is often overlooked in these patients. The occurrence of sinusitis was assessed in these two clinical presentations of allergic rhinitis. Methods: Consecutive patients with allergic rhinitis, with skin allergy test positivity were enrolled and categorized into Bsneezers-runners^and Bblockers^as per their predominant symptoms based on ARIA guidelines. All were subjected to computed tomography (CT) of the paranasal sinuses and extent of sinusitis was staged as per Lund and Kennedy criteria. The presence of sinusitis was then correlated with their clinical symptoms. Results: Of the 338 patients (220 males, 118 females), 134 (39.6%) were Bsneezers-runners^and 204 (60. 4%) were Bblockers^. Sinusitis was present in 257 (76%), 78 (58.2%) were Bsneezers-runners^while 179 (87.7 %) were Bblockers^. Sinusitis was significantly higher in Bblockers^(78/134 vs 179/204, P = 0.019). Furthermore, the occurrence of sinusitis was significant in Bblockers^(179/204 vs 25/204, P = 0.000) but not so in Bsneezers-runners( 78/134 vs 56/134, P = 0.12). The mean sinus CT score was significantly higher in Bblockers^as compared to Bsneezers-runners^(8.9 T 7.6 vs 6.1 T 5.6, P = 0.0014). The mean number of sinuses involved too was significantly more in Bblockers^(6.54 T 4.53 vs 4.5 T 3.2, P = 0.0001). In clinical practice, caring for patients generates many questions about diagnosis, prognosis, and treatment that challenge health professionals to keep up to date with the medical literature. One approach to meeting these challenges is to learn how to practise evidence based medicine (EBM). According to EBM, conclusions from meta-analyses of randomized trial represent the most solid evidence source to assess the efficacy of a treatment. Meta-analysis is a statistical procedure that integrates the results of different independent studies pooled together, thus allowing a more objective appraisal of the evidence than traditional narrative reviews can do. Metaanalysis provides a quantitative estimate of treatment effects. Pharmacological treatment of allergic rhinitis represents a bench mark of therapeutic management. Practical guidelines need high quality proves of evidence in order to provide reliable recommendations for therapeutic management. Meta-analysis may represent the answer to this need, as a source of solid evidence. Cochrane Collaboration recommendations and Quorom Statements represent the gold standard of quality assessment. At present only few systematic reviews of high methodological quality are available or in progress: evidence Ia of efficacy for antihistaminic treatment is provided by desloratadine which on 3108 patients showed a significant reductions in total symptoms scores, total nasal symptoms score and nasal blockage relief when compared with placebo; a meta-analysis confirms that also ebastine induces a greater decrease from baseline in mean rhinitis symptom scores than placebo (749 patients). Mometasone furoate is at present the only one nasal steroid which may benefit an evidence of effect Ia by means of an in-progress meta-analysis, whose preliminary results show in 2163 patients a significant reduction in total symptoms scores. According to a recent systematic review with pooled analysis, leukotriene receptor antagonists produce a small but statistically significant improvement in nasal symptom when compared with placebo (3924 patients). All drugs showed good safety profile. Antihistamines, nasal steroids and antileukotrienes demonstrated evidence of efficacy in the treatment of allergic rhinitis; these tools should also demonstrate a good benefit/cost ratio in order to satisfy the future methodological approaches of guidelines formulation (GRADE). Leukotriene receptor expression in allergic rhinitis: the effect of allergen exposure and treatment Teet Pullerits 1 , Carina Malmhäll 2 , and Jan Lötvall 2 . 1 Lung Pharmacology Group, Department of Allergology and Lung Medicine, Gothenburg, Sweden; 2 Lung Pharmacology Group, Department of Allergology and Lung Medicine, Gothenburg, Sweden. Background: Cysteinyl leukotrienes are mediators contributing to the symptoms of allergic rhinitis and exerting their effect through specific receptors. We aimed to evaluate the effect of a naturally occurring grass pollen season and the anti-rhinitis treatment on the expression of cysteinyl leukotriene receptor 1 (CysLT(1)R) in the nasal mucosa of allergic rhinitis patients. Methods: In a double-blind randomised study, 59 patients with allergic rhinitis received treatment with either nasal steroid (fluticasone propionate; FP, 200 microg/day), oral antileukotriene montelukast (ML, 10 mg/day) alone or in combination with antihistamine loratadine (LT, 10 mg/day), or placebo over the grass pollen season. Nasal biopsies for immunohistochemical CysLT(1)R analysis were taken before and during the peak of the grass pollen season. Results: The grass pollen season induced a significant increase in the CysLT(1)R expression in placebo (from 0.097 T 0.005 to 0.178 T 0.012 cells/ field), ML (from 0.113 T 0.011 to 0.148 T 0.009), and ML + LT (from 0.122 T 0.008 to 0.154 T 0.016), but not in FP (from 0.123 T 0.013 to 0.128 T 0.016) treated patients. FP treated patients had significantly lower pollen-season induced change in the CysLT(1)R expression compared to other groups. Also, both ML and ML+LT treatment groups had significantly lower increase in the CysLT(1)R expression compared to placebo treatment. Conclusion: We conclude that the inhibitory effect on CysLT(1)R expression is one of the mechanisms by which nasal steroids and antileukotrienes alone or together with antihistamines exert their anti-inflammatory effect in allergic rhinitis with nasal steroids demonstrating the strongest inhibition. separation. The protein signatures of 79 inflammatory mediators from these paired serum samples were analyzed using RayBio\ Human Cytokine Antibody Array V. Serum levels of leptin, being differentially expressed in umbilical cord blood (UCB) from those with maternal asthma, and C-reactive protein (CRP) were measured by ELISA assays. The respective lower detection limits for these markers were 125 ng/L and 0.1 mg/L. Results: Six pregnant mothers with asthma and 6 non-allergic controls were recruited, with respective mean (SD) age being 30.7 (5.9) years and 35. 1 (4.9) years. Five of 6 UCB samples from newborns with maternal asthma had increased leptin levels when compared with newborns with non-allergic mothers. Other mediators with altered levels in two or more UCB samples with maternal asthma included IL-3, IL-8, IL-12, IL-15, PDGF-BB, NAP-2, GRO and MIG. Serum leptin and CRP levels from asthmatic mothers were significantly higher than those in UCB from their newborns (P G 0.001), so were serum CRP levels in non-allergic mothers as compared with their newborns (P G 0.01). Leptin levels showed significant positive correlation with CRP levels (r = 0.712, P G 0.001) in sera from asthmatic mothers. On the other hand, CRP was below the detectable limit in most UCB serum samples. Serum leptin levels in UCB did not differ in those with and without maternal asthma. Background: Cysteinyl leukotrienes (cysLTs) are potent inflammatory mediators acting through at least two receptors CysLTR 1 and CysLTR 2 . CysLTR 1 mRNA and protein expression is upregulated in patients with severe asthma phenotypes: frequent severe exacerbations, chronic airflow limitation or steroid-resistant asthma. Changes in the CysLTR 1 promoter might influence its transcriptional activity and therefore might modulate airway inflammation. Aim: Investigate an association of polymorphisms in the CysLTR 1 gene promoter with severe and non-severe asthma phenotype. Methods: The study groups consist of 100 patients with severe asthma, 70 patients with non-severe asthma, diagnosed according to GINA 2005 criteria and 100 healthy subjects. Genomic DNA was isolated from the peripheral blood leukocytes. Fragments of the CysLTR 1 promoter region were amplified by PCR and sequenced directly. Results: Three single nucleotide polymorphisms (SPSs) -634 T/C (rs321029), -475C/A (rs2637204), -336G/A (rs2806489) were identified in the promoter region of CysLTR 1 gene. All SNPs were in strong linkage disequilibrium both in patients and in control group. Two most common haplotypes were identified: ht1 [C-A-A] and ht2 [T-C-G], among which ht1 was observed in 71.43% of male patients and 90.48% of male controls. Significant differences were observed in allele frequencies of -336G/A in male subjects. -336G allele occurred significantly more often in men with severe asthma comparing with male control group (p cor = 0.04; OR = 3.800 C.I. = [1.112Y12.983] ). Conclusion: Genetic variants of CysLTR 1 gene promoter might be associated with severe asthma phenotype in male asthmatic patients. Further elucidation of this association might improve understanding of heterogeneous condition in severe asthma. Tatjana Caparoska 1 , Biserka Kjaeva 1 , Jagoda Stojkovic 1 , and Olivera Caparoska 2 . 1 Clinical Center-Skopje, Clinic for Pulmology and Allergology, Skopje, Macedonia, Fyrom; 2 University Health Network, IMG Ontario, Toronto, Canada. Background: Patients with aspirin-sensitive asthma often have continious inflammation of their upper sinuses complicated by chronic rhinitis, chronic sinusitis and nasal polyposis which frequently leads to asthma attacks. These patients often are not aware of the problems in their upper respiratory tract. Our goal with this study was to evaluate the inflammatory changes in upper respiratory tract in patients with aspirin sensitive asthma (ASA). Methods: We followed 88 patients with paranasal x-ray, CTscan, skin prick tests, pulmonary function test, rhynomanometry and chest x-ray. Patients were divided in 2 groups. 39 pts with NSAIDs intolerance of~15yrs were included in the first group (mean age 45, 8 Q 4,7yrs) . In the second group were 49 aspirin intolerant asthma pts (mean age 44, 8 Q 4, 1yrs) . The diagnosis of asthma was made based on clinical evaluation, skin prick test and pulmonary function test including histamine provocation test. Endoscopic methods were used for assesment of paranasal sinuses and nasal passages. Results: Endoscopic examination showed in group 1 nasal polyposis in 8, deviation of nasal septum in 16, hypertrophy of nasal mucosa in 6, and chronic tonsilitis in 4 pts. In 16 pts we have observed changes in x-ray of paranasal sinuses and in 12 changes in CTscans of PNS. In 13 pts-rhynomanometric evaluation showed decrease in of the nose. pathways Bacteriological analysis of nose and throat showed predominance of bacteria species Moraxella influenze in 33,3%, Moraxella cataralis in 28,2% and Stahylococcus aureus in 15,3%. Conclusion: Our study and results confirm the coexistence of chronic inflamation in upper respiratory tract in pts with bronchial asthma regardless of it's type. Hopkins, Allergy, Baltimore, United States; 2 National Institute of Aging, Immunology, Baltimore, United States; 3 University of Missouri-Columbia, Surgery and Molecular Immunology, Columbia, Missouri, United States; 4 Johns Hopkins University, Allergy, Baltimore, United States. Rationale: Posttranscriptional regulation (PTR) is a key yet ill-defined area of gene expression in T cells. We investigated the PTR of interleukin (IL)-13 and the role the RNA-binding protein HuR plays in this process. HuR associates with adenylate-uridylate-rich elements (ARE) in the 3'untranslated regions (UTR) of mRNAs, promoting mRNA stability and translation. Methods: IL-13 mRNA decay was monitored in human peripheral bloodderived Th2 cells using the transcriptional inhibitor Actinomycin D. The IL-13 3'UTR was subcloned into a tetracycline (Tet)-off $-globin reporter construct and transiently expressed in the absence or presence of overexpressed HuR in H2 cells. Association of HuR with IL-13 mRNA was detected by immunoprecipitation (IP) of messenger ribonucleoprotein complexes (mRNPs) and by biotin pull-down. Results: IL-13 mRNA half-life increased in cells treated with PMA and ionomycin compared to unstimulated cells (6.5 T 2.0h vs 1.6 T 0.1h, p G 0.05, n = 3). The decay of the $-globin transcript was faster in cells transfected with pTet-BBB-IL13 compared to those transfected with the ARE-less pTet-BBB (2.1 h vs 11.4 h, p G 0.05, n = 6). Overexpression of HuR increased $-globin mRNA half-life in cells transfected with pTet-BBB-IL13 (22.7 h vs 2.3 h, p G 0.01, n = 4). Enrichment of IL-13 mRNA was detected by PCR following IP of Jurkat cell mRNPs with anti-HuR vs a control antibody. HuR binding to IL-13 3'UTR was confirmed by pull-down assay using biotin-labeled RNA probes spanning the 3'UTR or the coding region of IL-13. Conclusion: PTR plays a relevant role in expression of IL-13. HuRmediated stabilization of IL-13 mRNA could be a relevant therapeutic target in asthma. Functional study of prostaglandin D2 receptor (chemoattractant receptor molecule expressed in th2 cells, CRTH2) promoter polymorphism with aspirin-intolerant asthma Nami Palikhe 1 , Seung-Hyun Kim 1 , Eun-Mi Yang 1 , Young-Min Ye 2 , Gyu-Young Hur 1 , Eun-Joo Nam 1 , and Hae-Sim Park 1 . 1 Ajou University, School of Medicine, Department of Allergy, Suwon, Republic of Korea; 2 Yonsei University, School of Medicine, Department of Pharmacology, Seoul, Republic of Korea. Background and Objective: Prostaglandin D2 (PGD2) is an important lipid mediator in the pathogenesis of eosinophilic airway inflammation via its receptor CRTH2. The human CRTH2 gene encodes a G protein-coupled chemoattractant receptor which is expressed on Th2 cell, eosinophil, basophil and monocyte. The main objective of the study is to investigate the association of CRTH2 promoter polymorphism in patients with aspirin intolerant asthma (AIA). Subjects and Methods: The case control study was performed in three groups of patients, 106 AIA, 115 aspirin tolerant asthma (ATA) and 133 normal controls (NC). Two promoter polymorphism of CRTH2 gene were genotyped by a primer extension method, which was performed with the SNaPshot ddNTP primer extension kit (Applied Biosystems, CA, USA). Results: Two novel single-nucleotide polymorphisms (SNPs), CRTH2-466T9C and -129 C9A were identified in the promoter region of gene. Among two SNPs, the polymorphism of CRTH2 at positions of -466T9C showed a significant difference in genotype frequency between AIA and ATA; AIA patients showed significantly higher frequency of homozygous TT genotype than combined homozygous CC and heterozygous CT genotype of CRTH2 -466T9C (p = 0.037, multiple logistic regression analysis controlling for age and sex, recessive model). The luciferase reporter assay showed that the reporter plasmid carrying the -466C allele showed significantly higher promoter activity than -466T allele in human monocytes cell line (U937) and human mast cell line (HMC-1) (p G 0.001and p G 0.001 respectively). Similarly, the promoter activity of CRTH2 -466C allele cotransfected with GATA-3 was found to be increased dose dependently from the baseline promoter activity in HMC-1 cells. Conclusion: These results suggest that GATA-3 may be directly or indirectly associated with CRTH2 -466C allele by stimulating mast cell activation with the release of Th2 cytokines and other proinflammatory mediators contributing the increased susceptibility of AIA in CRTH2 -466T9C promoter polymorphism. The relationship between early infections due to RSV and the specificities of subsequent development of asthma : Clinical studies of RSV-infected patients indicated increased levels of Th2 cytokines and IgE, suggesting that an allergy-like condition developed during infection and skew the Th1/Th2 balance toward Th2 9 Th1. Comparative retrospective analysis of 76 children, who been admitted between winter seasons 2001 and 2005 for a first attack of brochiolitis before the age of 12 months, was performed in order to elaborate and correlate possible specificity of asthma in term of pathogenesis (A, NAA), immune response and compliance to treatment. In the 3 groups of children, divided on the basis theirs phenotypes: I group (n = 21) IgE associated persistent wheezier, II group (n = 47) nonatopic recurrent wheeze, III (n = 8) viral associated wheeze, RSV infection was responsible for the first attack of wheezing in 98%, 2% has early adenovirus infections in the II group. Family history of asthma and multiple asthmas triggers was performed in 95% in I (bilateral genetic risk was funded in 18%) comparing with 3% from II group. In opposite, many risk-factors in prenatal, peripartal and early neonates period of life in the present of serious another illness, was found in the II group (p G 0,05): oligohydramnion, placentas abruptions and infarct, small for date with extremly LBW 800 T 57g, gigant baby with extremely higher birth weight: 5250 T 50g, low Apgar score and reanimation/adaptation problems after birth. Patients in I group demonstrated higher levels of Neu, Eo, moderate degree of diseases and recurrent wheezing and/or asthma. Severity of the illness, many immune, endocrine, metabolic and genetics deregulations, ly-cytosis and Mo-cytosis were typical for II group. Compliance to ICS was significantly better in the I group. On the basis these data we may indirectly conclude implicate excess type 2 and/or deficient type 1 immune responses in the pathogenesis of RSV bronchiolitis and subsequent asthma development in I group. In contrast, II group had probably higher Th1 immune responses during RSV infections, on the basis maternal and intra-uterine factors, and has more severe disease and had shown higher percentage of immune deregulations. Patients in II group have attributes specific for NAA and complex immunoregulatory pattern followed by inappropriate response to treatment. These results indicate that immunological background via CD8+T cells may play an important role in the regulation of the differentiation and activation of the Th2CD4+Tcells during RSV infection. 109 Indoor air quality and dyspnea, mucosal, dermal and general symptoms in relation to room temperature and ventilation in university computer classrooms -an experimental study Dan Norbäck, and Klas Nordström. Uppsala University, Dept. of Medical Science, Uppsala, Sweden. : The aim was to study effects of increased ventilation and temperature changes in computer classrooms on measured indoor air quality and medical symptoms in university students. Technical university students in four classrooms participated in a blinded study. Two classrooms had higher air exchange (4.1Y5.2 ac/h); two others had lower air exchange (2.3Y2.6 ac/h). After one week, ventilation conditions were shifted between the rooms. The students reported medical symptoms last hour, on a seven step rating scale. Room temperature, RH, CO2, PM10, and ultra fine particles were measured simultaneously during 1 hour. In addition, illumination, air velocity, operative temperature, supply air temperature, formaldehyde, NO2, O3 and airborne levels of bacteria, moulds and common allergens was measured. Totally 355 students participated at least once in the study during the two weeks , 121 participated twice in the longitudinal analysis. In total, 31% were females, 2.9% were smokers, 3.8% had asthma, 18.1% pollen allergy (hay fever), and 14.0% furry pet allergy. Mean CO2 was 993 ppm (674Y1450 ppm), temperature 22.7 C (20Y25 C), and RH 24% (19Y35 %). Low and high air exchange rate corresponded to a personal outdoor air flow of 7 L/s*p and 10Y13 L/s*p, respectively. Mean PM10 was 20 microgram/m3 at low and 15 microgram/m3 at high ventilation flow. In the crude analysis, ocular, nasal and throat symptoms, dermal symptoms, dyspnea, sinusitis, headache, tiredness and nausea was significantly more common at higher CO2 levels and higher room temperature. At higher air exchange rate dyspnea and dermal symptoms were significantly less common. Similar results were obtained in the multivariate analysis, controlling for potential confounders such as gender, smoking and a history of atopy. In the longitudinal analysis of a subset participating twice, increased temperature was significantly related to increase of tiredness. In conclusion, computer classrooms may have CO2 levels above the current ventilation standards (more than 1000 ppm) and temperatures above 22 C, due to crowdedness and high thermal load. Increased temperature and CO2-levels may affect both dermal and mucosal membrane symptoms, as well as headache and tirdness. It is recommended that the personal ventilation flow is at least 10 L/s. Allergenicity of the recombinant and native blomia tropicalis allergens among atopic subjects John Donnie Ramos 1 , Nge Cheong 2 , and Kaw Yan Chua 2 . 1 University of Santo Tomas, Research Center for the Natural Sciences, Manila, Philippines; 2 National University of Singapore, Department of Paediatrics, Singapore, Singapore. Background: Blomia tropicalis (Bt), a predominant house dust mite (HDM) species in tropical and subtropical regions of the world, is a source of multiple allergens causing allergic sensitization among atopic individuals. Isolation and characterization of the allergenicity these Bt allergens are essential to evaluate the clinical relevance of this HDM species. Methods: Eight recombinant Bt allergens were expressed as GST-fusion allergens in E. coli or secreted allergen in P. pastoris while two native Bt allergens were purified by immuno-affinity chromatography. Specific-IgE reactivity of the Bt allergens were determined using 192 Bt-allergic and 85 nonatopic sera. The panel of Bt allergens were likewise used for skin prick test (SPT) in 110 Bt-allergic pediatric patients. The ability of the recombinant and native Blo t 11 allergens to inhibit the IgE-binding activity in Bt extract was determined by absorption studies. Results: Blo t 1, Blo t 3, Blo t 5, Blo t 10, Blo t 11-fD, Blo t 12 and Blo t 19 were expressed as soluble GST-recombinant proteins in E. coli while Blo t 4 was expressed in P. Pastoris. Native Blo t 5 and Blo t 11 were purified from Bt extracts using monoclonal antibodies. Of the 192 Bt-allergic patients`sera tested, 68% and 53% reacted positively to recombinant Blo t 5 and Blo t 11-fD, respectively, while 5-30% IgE reactivity was observed with the other recombinant Bt allergens. Interestingly, the IgE reactivity recombinant Blo t 5 and Blo t 11-fD compares with the IgE reactivity of their native counterparts at 70% and 54%, respectively. SPT results showed comparable results with ELISA. Absorption assays, showed that up to 80% inhibition of IgE reactivity in Bt extract can be obtained with the different Bt allergens. Conclusion: Results obtained from this study suggest the clinical importance of Blomia tropicalis as major source of allergens causing allergic sensitizations in tropical regions. The incorporation of Bt allergens in the panel of diagnostic and immunotherapeutic allergens for HDM allergy are highly recommended. Phleum pratense alone is representative for pooideae grass pollen species due to high immunochemical similarity between homologous grass pollen allergens Jorgen N. Larsen, Charlotte Hejl, Henrik Ipsen, Peter A. Wurtzen, and Niels Johansen. ALK-Abello, Research, Horsholm, Denmark. Background: The Pooideae grasses constitute a large taxonomical subfamily with thousands of species and worldwide occurrence. Extensive homology between amino acid sequences of grass pollen allergens occur, and grass pollen counts are published together due to a large immunochemical similarity between the grass pollen allergens. Objective: Study the immunochemical similarity of grass pollen allergens from different Pooideae species. Methods: Amino acid sequences representing grass pollen allergens were obtained from the Uniprot database. Structural similarity of the group 1 allergens was illustrated by mapping identical amino acids on the surface of the x-ray structure of Phl p 1, PDB: 1N10. A large number (913,000 data points) of grass pollen allergic patients' IgE was measured by Magic Lite solid phase immunoassay to eight Pooideae grass pollen extracts, respectively. IgE to eight Pooideae grass pollen extracts was measured by ADVIA Centaur solid phase immunoassay with or without inhibition by 2 mg Phleum pratense pollen extract. Standard T cell stimulation assays using T cell lines from grass pollen allergic donors were applied to assess T cell cross-reactivity. Results: Sequence alignment of Pooideae grass pollen allergens showed high homology. Mapping of identical amino acids on the surface of the x-ray structure of Phl p 1 clearly showed the presence of identical surface structures large enough to harbor IgE binding epitopes. A high correlation (0.86Y0.98 Spearman rank correlation coefficient) was observed when comparing levels of IgE to Phleum pratense with those of pollen extracts from individual grass species. Phleum pratense pollen extract inhibited IgE to other grass species more than 95% in most patients. T cell lines specific for Phl p 1 and Phl p 5 both showed similar stimulation indices when stimulated with different grass pollen extracts indicating extensive T cell cross-reactivity between individual grass species. Conclusion: Phleum pratense alone covers most of the immunochemical reactivity of all tested Pooideae grass species with respect to allergic patients' IgE. T cell lines specific for the group 1 and 5 major allergens showed extensive cross-reactivity between grass species. One specie alone, e.g. Phleum pratense, therefore seems adequate for specific management of allergy to Pooideae grass pollens. Gradual shift in the aeroallergen index affecting the allergy patients in recent years in Texas panhandle Nabarun Ghosh 1 , Mandy Whiteside 1 , Kyla Kersh 1 , Constantine Saadeh 2 , Michael Gaylor 2 , and Don W. Smith 3 . 1 West Texas A&M University, Life, Earth, Environmental Sciences, Canyon, United States; 2 Allergy Aarts, Pathology, Amarillo, United States; 3 University of North Texas, Biology, Denton, United States. : Aeroallergens cause serious allergic and asthmatic reactions. We have been analyzing the Aeroallergen data of the Texas Panhandle region for the last 7 years. We determined the aeroallergen index regularly by analyzing the coated Melinex tape from the Burkard Volumetric Spore Trap. Samples were examined, counted and photographed every 24 hours using a BX-40 Olympus microscope with a DP-70 Olympus Digital Camera. Data were correlated with daily temperature, precipitation, peak wind speed and clinical studies established from Allergy A.R.T.S. Clinical Research Laboratory in an effort to aid in diagnosis of mold and pollen-related allergies. Aeroallergens that we recorded were Alaternaria conidia, Pezizales ascospores, Curvularia, Cladosporium, Dreschlera, Stachybotrysand pollen from short ragweed (Ambrosia artemisiifolia), grass (Poaceae), hairy sunflower (Helianthus hirsutus), buffalo bur (Solanum rostratum), purple nightshade (Solanum elaeagnifolium) and lamb`s quarters (Chenopodium album). Due to severe drought conditions in the previous years (2002Y5), the pollen concentration was significantly low, that reached the highest peak from April onwards this year (2007) with plenty of rainfall. We observed this pick from April to July in last year 2006. We noticed a gradual shift in the aeroallergen index in the Texas Panhandle in recent years. Global warming with increased CO2 concentration exerted widespread impacts on the biotic system. Many regions are currently experiencing warming effect associated with global climate change including longer growing seasons and early arrival of spring. Short ragweed (Ambrosia artemisiifolia) released 54.8% more pollen with an ambient CO2 release (Rogers et al, 2006) . In Japan a 21-year study showed a gradual shift in Cryptomeria japonica pollen season. These results support the steadily increasing number of reports indicating a global warming trend. The temperature change affecting the start dates of the C. japonica pollen season is particularly relevant in the context of human health. From the clinical data from the AARTS clinic it is clearly evident that there were more patients suffering from allergic rhinitis during the months of March to June. 4 years back the peak pollen and mold season was between May to September (Ghosh et al., 2006) that has gradually shifted in 2007 to March-June so as the frequency of the patients visiting the Allergy Clinic. Background: $-(1,3)-glucan is pro-inflammatory and has been associated with airway inflammation and respiratory symptoms. Methods: This study assessed $-(1,3)-glucan levels from upper body clothing (jerseys) of 55 subjects. $-(1,3)-glucan levels were estimated with a modified Limulus amoebocyte lysate kinetic assay. Results: $-(1,3)-glucan levels ranged widely from 2,697 to 162,690 ng/g. $-(1,3)-glucan levels were significantly lower from cotton jerseys, and from warm water washed jerseys. Background: We have developed a natural CACR in which subjects will be exposed to consistent levels of Fel d 1, the major cat allergen, in a natural setting, such that there is transient exposure to disturbance of reservoir cat allergen. Objective: To develop the CACR in which Fel d 1 levels range from approximately 500Y2500 ng/m 3 . This concentration range of Fel d 1 was selected as it is representative of higher levels of Fel d 1 in homes with cats, and is known to induce an allergic and/or asthmatic response. Methods: The CACR is fully carpeted, contains a couch covered with a cotton sheet, and a chair. The CACR was preloaded with Fel d 1 using a proprietary aerosol generator and maintained by allergen from two CACR inliving cats. Fel d 1 levels were transiently increased by vigorously shaking the sheet to aerosolize Fel d 1. Fel d 1 airborne concentrations were measured using personal air samplers. Total airborne particulate was collected using a Button Sampler for 1h. Samples were analyzed using ELISA and particle sizes were independently measured with a laser particle counter. Results: Background measures indicated that Fel d 1 was undetectable in the CACR before aerosolization. Following one hour of 1g cat allergen aerosolization and 11 days cat housing, the average maximum total particle count was maintained over 4 consecutive days to be 9.01 T 3.65X105 ppm 3 and the Fel d 1 concentration 538 T 192 ng/m 3 . After an additional 2h aerosolization of 3.2g Fel d 1 and a total of 15 cat housing days, the average maximum total particle count maintained over an additional 4 consecutive days was 9.8 T 3.65X105 ppm 3 and the average Fel d 1 concentration was 621 T 135 ng/m 3 . Following an additional two days the concentration of Fel d 1 was 1046 T 982 g/m 3 . Overall, these two periods of aerosolization of Fel d 1 and a total of 20 cat housing days resulted in the mean concentration of Fel d 1 to be 686 T 455 ng/m 3 . Conclusion: These data indicate that aerosolized Fel d 1 is effective to obtain natural allergen levels rapidly and that live cats can maintain these levels over time within the targeted range. This work demonstrates that we have developed a CACR model that can be used towards the study of the etiology and putative therapeutics for cat allergy and asthma. Funding provided by Allied Research-Cetero Research, CA. Drug-induced T regulatory cells in asthma: Toll like receptors 2 and 9 represent distinct markers of glucocorticoid versus 1?25dihydroxyvitamin D3 action on human CD4+ T cells (1?25VitD3) alone, or together with glucocorticoids, induces an IL-10 secreting T regulatory phenotype (IL-10-Treg) in human CD4+ T cells, and these cells inhibit naBve T cell, Th1 and Th2 responses. Glucocorticoid induced IL-10 production is defective in patients with clinically insensitive asthma and 1?25VitD3, either in vitro or following oral ingestion by these patients, restores this impaired IL-10 response. Aim: To identify the distinct phenotypic and functional effects of the two drugs on human CD4+ T cells. Methods: In vitro based tissue culture, quantitative RT-PCR, flow cytometry and antibody capture assays (ELISA, cytometric bead array) were used to study human CD4+ T cell phenotype and function following polyclonal activation (anti CD3, in the presence or absence of dexamethasone and/or 1?25VitD3. T cell phenotype following 1?25VitD3 ingestion was also analyzed. Results: We show that 1?25VitD3 alone inhibits Th1 and Th2 cytokine production, enhances the generation of IL-10-Treg, but also the percentage of FoxP3 positive cells. However expression of 1?25VitD3-induced IL-10 and FoxP3 appears mutually exclusive. Furthermore, optimal effects of 1?25VitD3 on IL-10, Foxp3 and effector cytokine production are observed at distinct drug concentrations. Dexamethasone selectively induced expression of TLR2 in vitro whilst 1?25VitD3 selectively induced expression of TLR9 on CD4+ T cells both in vitro and following patient ingestion of 1?25VitD3, in comparison to other T cell populations (naBve, CD25+ Treg, Th1, Th2). Ligation of these TLR on the drug-induced IL-10-Treg resulted in loss of Treg function in both cases, but by distinct mechanisms. Conclusion: Glucocorticoids and 1?25VitD3 promote IL-10 production and regulatory function in human CD4+ T cells alone and in combination. TLR2 and TLR9 can be used to distinguish glucocorticoid and 1?25VitD3 exposure of human CD4+ T cells and ligation of these receptors downregulates Treg function. Sublingual immunotherapy to inhalant allergen sensitisation and the effect of chitosan P. T. Cunningham, C. E. Elliot, P. G. Holt, and Wayne R. Thomas. Telethon Institute for Child Health Research, Molecular Biotechnology, Perth, Australia. Background: Immunotherapy is an established form of treatment but it is necessary to develop more efficacious methods. We have developed a model for respiratory sensitisation induced by intranasal (i.n.) administration of the cysteine protease papain, without adjuvant. It produces high, boostable IgE titres. Upon challenge pulomonary eosinophilia is evident as is the release of Th2 cytokines into the bronchoalveolar lavage (BAL) fluid. The model has been used to optimise sublingual desensitisation in mucosally sensitised animals. We have previously demonstrated its ability to suppress allergen specific IgE. In these studies lymph nodes were examined and the effect of the excipient chitosan. Methods: Allergic sensitisation was achieved by the i.n. administration of low doses of papain, a homologue of Der p 1. Desensitisation was examined after the sublingual administration of the antigen with chitosan, antigen alone, or chitosan alone. Serum antibodies were examined and high dose challenge followed by BAL was used to study cellular infiltrates and cytokine production in the lung. Lymph nodes were cultured and proliferation and cytokine production in response to allergen stimulation measured. Results: All treatments, even chitosan alone, suppressed IgE and IgG1 antibodies. Chitosan delivery at desensitisation resulted in a significant reduction of cells infiltrating the lungs that was not seen in the other treatment groups however both the chitosan alone and chitosan with papain significantly suppressed the neutrophilia seen. The Th2 cytokines in BAL fluid were not reduced in the treatment groups although all groups receiving chitosan had elevated levels of IL-10 and IFN-,. Cultured lymph node cells had lower levels of proliferation after sublingual therapy with a soluble antigen but when chitosan was used cells proliferated to similar levels to controls. Cytokine levels in lymph node cell supernatants showed significantly increased levels of TGF-$ and significantly decreased IL-5, IL-10 and IFN-, in all treatment groups. Conclusion: Sublingual immunotherapy decreased IgE and IgG antibodies but not lung inflammatory responses. It is however shown that lymph node proliferation and cytokine production was decreased and this could have a long-term effect. Chitosan caused a non-specific reduction that was not greater than allergen alone. Analysis of epitope-specific immune responses induced by vaccination with structurally folded and unfolded recombinant Bet v 1 allergen derivatives in man Background: Previously we have constructed recombinant derivatives of the major birch pollen allergen, Bet v 1, with a more than hundredfold reduced ability to induce IgE-mediated allergic reactions. These derivatives differed from each other because the two recombinant Bet v 1 fragments represented unfolded molecules whereas the recombinant trimer resembled most of the structural fold of the Bet v 1 allergen. Methods: Here we analyzed the antibody (IgE, IgG subclass, IgA, IgM) response to Bet v 1, recombinant and synthetic Bet v 1-derived peptides in birch pollen allergic patients who had been vaccinated with the derivatives or adjuvant alone. Furthermore, we studied the induction of IgE-mediated skin responses in these patients using Bet v 1 and Bet v 1 fragments. Results: Both types of vaccines induced a comparable IgG1 and IgG4 response against new sequential epitopes which overlap with the conformational IgE epitopes of Bet v 1. This response was much higher than that induced by immunotherapy with birch pollen extract. Trimer more than fragments induced also IgE responses against new epitopes and a transient increase in skin sensitivity to the fragments at the beginning of therapy. However, skin reactions to Bet v 1 were reduced one year after treatment in both actively treated groups. Conclusion: We demonstrate that vaccination with folded and unfolded recombinant allergen derivatives induces IgG antibodies against new epitopes. These data may be important for the development of therapeutic as well as prophylactic vaccines based on recombinant allergens. Materials: Group of 34 subjects included: 19 individuals with allergy to grass pollen confirmed by medical history, positive skin prick tests (SPT) and specific IgE (sIgE) min. 2 class, treated with SIT before pollen season in 2007 (gr. A), 11 sex and age matched control subjects with confirmed as above allergy to grass pollen untreated with SIT (gr. B) and 4 control healthy nonallergic to grass pollen subjects (gr. C). Methods: i) subcutaneous SIT to grass pollen with allergen extract absorbed on aluminum hydroxide according to manufacturer's quidline in the gr. A; ii) Evaluation with questionnaires of clinical rhinoconjunctivitis symptom score during 2006 and 2007 pollen season (the endpoint) in groups A and B; iii) The flow cytometry analysis of blood samples was performed at the beginning of SIT treatment, after reaching a maintenance dose and at the end-point in gr. A, and at the end-point in gr. B and C. Results: 17 subjects from gr. A reached the end point phase of SIT without significant adverse events. Reduction of symptom score 950% in 6 subjects, 26Y50% in 9 subjects and G25% in 3 subjects after SIT treatment was shown. In gr. B the symptom score in grass pollen season was comparing to Group A before the SIT treatment. The flow cytometric analysis showed: i) lower percentage of CD4+ CD25+ regulatory T cells in all allergic patients (gr. A-2,1%, gr. B-1,1%) versus the control (gr. C-3,4%) at the end point; ii) statistically nonsignificant percentage upward trend in CD4+ CD25+ regulatory T cell population during the course of SIT in treated with SIT groups. Conclusion: Lower count of CD4+ CD25+ regulatory T cells in both groups of allergic patients suggests a dysfunction within this population might contributes to the pathology of allergy. The percentage upward trend in Treg population during the SIT treatment, accompanied by the improvement in symptom score, suggesting the maintenance of balance between Th1, Th2 and Treg populations is crucial for peripheral tolerance. The project will be continued to cover broader population and investigate other immunological parameters. Early suppression of basophil activation during allergen-specific immunotherapy by upregulation of histamine receptor 2 Wroclaw University, Medical Faculty, Wroclow, Poland. Background: Most of the primary effects of allergen specific immunotherapy (SIT) are exerted on effector cells. However, there is surprisingly few information about the mechanisms by which SIT modifies and suppresses immune responses of basophils and mast cells in particular during the repetitive administration of increasing allergen doses in the built-up phase. One of the main mediators released by effector cells upon allergen challenge is histamine. The diverse immunoregulatory functions of histamine are based on the differential distribution of histamine receptors on immunocompetent cells. While histamine receptor (HR)1 decreases humoral immunity and increases cellular immunity, HR2 displays opposing functions, decreasing cellular immunity and mediating tolerogenic immune mechanisms. Therefore, the early desensitizing effect of SIT due to reprogramming of effector cell responsiveness to histamine via the modulation of the histamine receptor repertoire represents an exciting and plausible strategy. Methods: Honey bee and wasp venom SIT was applied to 10 patients according to the rush protocol. The mRNA level of HR1, HR2 and HR4 in PBMC isolated from the peripheral blood of patients during the built-up phase of SIT (5 days) was evaluated by real time PCR. Histamine receptor mRNA levels in basophils enriched from the peripheral blood, which were triggered via IgE receptor were determined. Additionally, the activation state of basophils after IgE receptor cross-linking and treatment with ligands selective for HR2 as well as cAMP inducer forscolin, a second messenger of HR2, was analysed by flow cytometry and sulfido-leukotriene release assays. Results: Significant increase of the HR2 mRNA level as well as the HR2/HR1 ratio was observed in PBMC of patients from the 3rd day of SIT. IgE receptor cross-linking of basophils from the peripheral blood with increasing anti-IgE doses led to the upregulation of HR2. The selective triggering of HR2 with specific agonist dimaprit or forscolin strongly suppressed IgE-receptorinduced activation of basophils in a dose dependent fashion. Conclusion: HR2 mediates immunosilencing functions on IgE-receptor activated basophils. The up-regulation of HR2 expression on basophils in response to repetitive IgE-receptor cross linking as well as allergen challenge during the built-up phase of SIT might represent a key mechanism and early desensitization effect of SIT on the level of effector cells. *these authors contributed equally. Jean-Pierre Allam 1 , Gligor Stojanovski 1 , Nikolaus Friedrichs 2 , Thomas Bieber 1 , Reinhard Büttner 2 , and Natalija Novak 1 . 1 University Hospital Bonn, Dermatology, Bonn, Germany; 2 University Hospital Bonn, Pathology, Bonn, Germany. Background: Sublingual immunotherapy (SLIT) has been proven to be a safe and efficient alternative to subcutaneous immunotherapy (SCIT) in the treatment of allergic rhinitis. While antigen-presenting cells such as Langerhans cells (LC) are thought to play a major role in the effectiveness of SLIT, mast cells (MC) most likely account for the observed adverse reactions such as oral itching and sublingual edema. Since only little is known about LC and MC within the oral cavity, we investigated their distribution in search for alternative mucosal application areas with the highest density of LC and lowest presence of MC. Methods: Different biopsies were taken simutaneously from mucosal tissue of the vestibular, buccal, palatinal, lingual, sublingual and gingival region (n = 10). Tissue was further processed for immunohistochemistry and flow cytometry. MC were detected by chymase, and LC by CD1a expression. Results: The highest density of MC could be detected within the gingiva while the lowest density of MC was found within the palatinum and lingua. However, in the sublingual region MC were located within the lobe and duct of sublingual glands in a substantial number of individuals, which might explain swelling of sublingual caruncle in some SLIT patients. Considering LC, we could detect the highest density within the vestibular region followed by the region of bucca, palatinum and lingua. Interestingly, the lowest density of LC was located in the sublingual region. By flow cytometry, we investigated the expression of the high affinity receptor for IgE (Fc(RI) which might play a central role in allergen uptake during SLIT. Thereby, we could detect the highest expression of Fc(RI on LC of the vestibular region. Conclusion: In view of our data, different mucosal application sites such as the vestibulum might represent an alternative region with potent allergen uptake especially in SLIT patients suffering from sublingual edema. Furthermore our data might serve as a basis in the development of new application forms for SLIT such as tablets or stripes. Because probiotics were proposed to decrease the prevalence of allergy in susceptible individuals, we investigated whether they could affect IgEinduced mast cell secretory responses. To this end, mouse Bone Marrowderived Mast cells (BMMC) were exposed to lactic acid bacteria. BMMC express high-affinity receptors for IgE (Fc&RI) . When sensitized with IgE antibodies and challenged with specific antigen, they release and secrete a variety of inflammatory molecules including granular mediators and enzymes (among which "-hexosaminidase), lipid mediators, cytokines (among which TNF-!) and chemokines. We checked first that none of the bacteria tested activate mast cells. Indeed, an incubation of BMMC for 20 min or 3 hr with bacteria (at a ratio of 1000 bacteria/cell) did not induced "-hexosaminidase release or TNF-! secretion, respectively. We then investigated whether a previous exposure of BMMC to bacteria would affect the subsequent IgE-induced biological responses of mast cells. We found that an overnight incubation of BMMC with one strain of bacteria (at a ratio of 1000 bacteria/cell), prior to sensitization with IgE antibodies, inhibited Ag-induced "-hexosaminidase release and TNF-! secretion. Live and gamma-irradiated bacteria were equally inhibitory. These bacteria did not decrease the expression of Fc&RI and did not affect sensitization of BMMC with IgE. Inhibition required a direct contact between cells and bacteria. Inhibition were similar in BMMC from TLR-2/4-, MyD88-, and NOD2-deficient mice as in BMMC from WT mice, excluding TLR1, 2, 4, 5, 6, 7, 8 and 9 , as well as NOD2 as being the responsible receptors. Preliminary works aiming at analyzing how bacteria interfere with Fc&RI signaling indicated that: 1) several intracellular signaling molecules had a reduced expression, 2) both early and late Fc&RI-dependent phosphorylation events were decreased, 3) I.B degradation was prevented, thus interfering with the nuclear translocation of NF-.B, 4) the IgE-induced increase of the intracellular concentration of Ca2+ was profoundly inhibited. Altogether, our results indicate that probiotics, especially lactic acid bacteria, can exert direct inhibitory effects on mast cell activation. They support the idea that probiotics can protect from allergies, by preventing IgE-induced mast cell activation. Induction of allergic airway inflammation by house dust mite allergen specific Th2 cells in mice C. H. Huang 1 , L. M. Liew 1 , I. C. Kuo 2 , H. M. Wen 1 , D. L. Goh 1 , D. L. Goh 2 , B. W. Lee 1 , and K. Y. Chua 1 . 1 National University of Singapore, Paediatrics, Singapore, Singapore; 2 A*STAR, Singapore Institute for Clinical Sciences, Singapore, Singapore. Background: It is known that allergic inflammatory diseases such as asthma are Th2 cells-mediated, however, the pivotal roles of allergen-specific Th2 cells in the induction of allergic lung inflammation have not been fully elucidated. The aim was to elucidate the imunopathological roles of allergenspecific-Th2 cells in allergic airway inflammation. Methods: Mice were epicutaneously sensitized with a major dust mite allergen, Blo t 5 and a well-characterized Blo t 5-specific-Th2 cell line was subsequently established from the splenocytes of the sensitized mice. The immunopathological roles of the cell line were assessed in vivo by adoptive cell transfer approach. NaBve mice received Blo t 5 specific-Th2 cells intravenously followed by intranasal challenge with Blo t 5. The responses of recipient mice were analysed by immunological and histochemical methods. Results: A long term TCRV$3 + Blo t 5 specific Th2 cell line producing high levels of IL-4, IL-5, IL-13 and IL-10 but not IFN-, was established. These CD44 high CD62L -Th2 cells showed up-regulation of CTLA-4, ICOS, OX40, 4-1BB, CD27 but not CD40L upon stimulating with Blo t 5. After intranasal challenge with Blo t 5, Th2 cells recipient mice developed Blo t 5-specific IgG1 and IgE, airways eosinophilia and mucus production of the Goblet cells. In addition to the donor Th2 cells, the cellular infiltrate consisted of CD4 + , CD8 + T cells and NK cells of the recipient mice. Such cellular inflammation could be suppressed by dexamethasone intervention. The pathological results were not observed in the PBS challenged recipient mice. Conclusion: Blo t 5-specific Th2 cells played a central pathological role in mediating allergic airway inflammatory responses resembling those seen in humans. This animal model is particularly useful for screening of novel therapeutics for asthma and allergy. Differences in Foxp3 + CD4 + ratio between symptomatic atopic patients and asymptomatic atopic controls with similar levels of Th1/Th2 markers Kanami Orihara 1 , Masami Narita 2 , Akira Akasawa 3 , Yukihiro Ohya 2 , Kenji Matsumoto 1 , and Hirohisa Saito 1 . 1 National Research Institute for Child Health & Development, Department of Allergy and Immunology, Tokyo, Japan; 2 National Center for Child Health and Development, Division of Allergy, Tokyo, Japan; 3 National Center for Child Health and Development, Department of Interdisciplinary Medicine, Tokyo, Japan. Rationale: Foxp3, a responsible gene for IPEX syndrome in which both Th1 and Th2 responses are hyperreactive, is known as a master gene of naturally occurring regulatory T cell and maybe of some other regulatory T cell subsets. Therefore, the regulatory roles of Foxp3 + cells in allergic disorders are highly expected; however, the precise role of circulating Foxp3 + cells in common allergic diseases remains unclear. Then in order to evaluate its diagnostic potential, we examined intracellular Foxp3 protein expression in circulating peripheral blood CD4 + T cells from allergic patients and healthy controls at the single-cell level. Methods: The ratio of Foxp3 + CD4 + cells in the CD4 + cell fraction as well as various laboratory data were measured for 64 donors, including 37 patients with atopic dermatitis and/or bronchial asthma. None of the subjects was treated with oral corticosteroids. All the patients were judged positive for specific IgE antibodies against at least one of Dermatophagoides pternyssinus, Dermatophagoides farinae, and Japanese cedar pollen using Pharmacia's CAP system fluorescence enzyme-linked immunoassay (CAP-FEIA). Twenty-three of the 27 age-matched healthy volunteers also showed at least one positive reaction to those allergens by CAP-FEIA, although they never had subjective symptoms related to allergic diseases. Results: Foxp3 + CD4 + ratio in the total population was correlated inversely with the levels of total serum IgE (p = 0.021), % eosinophil count (p = 0.022), and serum IFN-, (p = 0.048). By performing a case-control study by matchedpair analysis to eliminate the influence of the Foxp3 + CD4 + level correlations with the total IgE level and eosinophil ratios, Foxp3 + CD4 + ratio was significantly lower in active atopic patients compared to asymptomatic donors having similar levels of IgE, eosinophils and IFN-,. Conclusion: Our findings imply that circulating Foxp3 + CD4 + regulates both Th1 and Th2 responses in vivo. Moreover, symptomatic atopic patients had a lower Foxp3 + CD4 + ratio than asymptomatic atopic controls having similar levels of Th1/Th2 markers. Measurement of Foxp3 + CD4 + ratio has the potential to aid in evaluating the presence of active inflammation in patients with allergic diseases, which can not be evaluated by known Th1 and Th2-related markers in patients with allergic diseases. Molecular biomarkers of eosinophil-lineage commitment: Multiplex Q-PCR analysis of GATA-1, MBP and IL-5 receptor mRNA expression kinetics in both peripheral and umbilical cord blood Background: We examined practices of physicians in India, China, Sri Lanka, Australia, Singapore, Indonesia, Philippines and Taiwan in monitoring and treating childhood asthma. Methods: Our 6-page standardized questionnaire was sent to doctors via post. The questionnaire is made up by questions on: a) methods of monitoring of childhood asthma, b) practices in managing acute asthma exacerbations, and c) choice of therapy in maintenance treatment. Results: Our study reflects mostly the practices of doctors who practice in urban regions. Of respondents, 41.4% were general pediatricians, while 26.3% were general practitioners. A small fraction of doctors used score cards or diaries to monitor asthma, ranging from 0% (Philippines and Australia) to 15.9% (India). Only 8.1% (Sri Lanka) to 51.7% (Australia) use either a peak flow meter and/or spirometry to monitor asthma. However, for frequency of usage, 35% (China) to 94% (Indonesia) never or seldom make use of a peak flow meter, and 33% (China) to 97.6% (Indonesia) never or seldom use spirometry for monitoring. Nebulised salbutamol given every 20 minutes was the treatment of choice in treating acute asthma attacks by the majority of doctors in each country, ranging from 29% (China) to 85.7% (Sri Lanka). A minority (4% (China) to 18.8% (Taiwan)) administer systemic corticosteroids in an outpatient setting (ER or polyclinic) in acute asthma. 53.6% (India) to 93.9% (Sri Lanka) of doctors also indicated that their duration for treatment with corticosteroids would be for 3Y5 days, and at the same dosage throughout the duration. A good number (32.7% (Sri Lanka) to 80.2% (Taiwan)) use antibiotics in the treatment of acute asthma, but only when pneumonia, otitis media or sinusitis was likely. Few doctors favoured high-dose inhaled corticosteroids in acute management, ranging from 0% (Indonesia and Sri Lanka) to 8. 4% (Philippines) . For maintenance treatment, a significant fraction of doctors chose a long-acting beta agonist (LABA) monotherapy as a firstchoice treatment for asthma maintenance. For infants the percentage ranged from 1.3% (Australia) to 76.3% (Indonesia),in preschoolers 0%(Australia) to 61% (China), and in older children, 0% (Philippines) to 61% (China). Conclusion: There is much room for improvement in increasing doctors' awareness to guidelines for more effective management of paediatric asthma in Southeast Asia, especially regarding the use of LABA monotherapy. Efficacy of inhaled heparin is effective in the treatment of acute exacerbation of asthma Nancy Mahmoud Abd-Elaty 1 , Mahmoud Elprince 1 , and Magdy Abd El-salam 2 . 1 Faculty of medicine, Suez Canal University, Chest Department, Ismailia, Egypt; 2 Faculty of medicine, Ain Shams University, Chest Department, Cairo, Egypt. Background: Inhaled Heparin was found to be highly protective against methacholine induced bronchospasm in bronchial asthma possibly via a direct effect on smooth muscle or may have potential as anti-inflammatory activity. Objective: The aim of the study was to determine the additional therapeutic benefit of inhaled heparin in the treatment of hospitalized patients for acute asthmatic exacerbation and treated with inhaled bronchodilators and glucocorticoid therapy. Methods: Thirty patients (20 male, 10 female), mean age( 31 +/j 10 years), admitted for acute exacerbation of asthma, participated in a prospective, randomized, double-blind, placebo-controlled study. All the subjects received hydrocortisone, administered intravenousl, and nebulized salbutamol. The treatment group received inhaled heparin therapy (20,000 U in 4 mL) every 4 hr. and the placebo group received 0.9% saline solution for 24 hours. Baseline respiratory parameters such as oxygen saturations, respiratory rates, and peak Background and Aims: Recently, we could demonstrate that mucosal targeting with Aleuria aurantia lectin (AAL) coated Poly(D,L-lactide-co-glycolic acid, PLGA) microspheres modulated a specific immune response in birch pollen allergy. Using grass pollen as a novel allergen encapsulated in AAL-coated particles we aimed to further evaluate the immune modulating functionality of microspheres in a murine asthma model. Methods: BALB/c mice were sensitized intraperitoneally with grass pollen extract followed by aerosol challenges inducing acute allergic asthma. Thereafter, animals were treated repeatedly with grass pollen loaded PLGA microspheres coated with AAL for M-cell targeting. Mice immunized with microspheres coated with glycine, with further intraperitoneal allergen injections and naBve animals served as control. The outcome of the treatment was evaluated by measurements of grass pollen specific antibodies (IgG1, IgG2a, IgE and IgA) in sera and bronchio-alveolar lavage (BAL) and by determination of cytokine profiles (IL-5, IL-10, TGF-$) in spleen supernatants. Results: In the animals treated with AAL-coated mircrospheres allergenspecific IgG1, IgG2a and IgE levels were unaffected by treatment. However, allergen-specific IgA levels increased during therapy. Interestingly, in cytokine evaluations IL-10 levels did not change, whereas reduced amounts of IL-5 and high levels of TGF-$ were measured in the mice treated with AALfunctionalized microspheres compared to the other groups. Conclusion: These data indicated that in the murine model grass pollen allergy was beneficially influenced by targeting M-cells via AAL-coated, allergenloaded microspheres. Thus, the potential of functionalized microparticles for specific immunotherapy for acute respiratory allergy was confirmed. Oral sulforaphane safely and effectively induces antioxidant phase II enzymes in the human airway Substantial evidence implicates particulate air pollution exposure as an important factor for asthma exacerbations and the increasing prevalence of allergic respiratory disease. As the inflammatory effects of particulate air pollution are mediated by the induction of cellular oxidative stress, strategies to reduce oxidative stress may potentially reduce the harmful effects of particulate air pollution. Endogenous Phase II enzymes abrogate oxidative stress through metabolism of particulate-associated reactive chemicals and the scavenging of reactive oxygen species. We conducted a placebo-controlled dose escalation trial to investigate the in vivo effects of sulforaphane, a naturally occuring potent inducer of Phase II enzymes, on the expression of GSTM1, GSTP1, NQO1, and HO-1 in the upper airway of human subjects. Sixty human study subjects consumed oral standardized broccoli sprout homogenate (BSH) doses containing sulforaphane once daily for 3 consecutive days. An escalating block design was used with 25, 50, 75, 100, 125, 150, 175, and 200 grams (g) of BSH to ensure safety and tolerability. Additional subjects were subsequently enrolled at doses of 125, 150, 175, and 200 g to examine dose-response effect. Five subjects completed the protocol with non-sulforaphane containing alfalfa sprout homogenate dosing at 200 g as a control group. RNA expression for selected Phase II enzymes was measured in nasal lavage cells by RT-PCR ( † Wilcoxon) Nebulized single dose formoterol powder (24 mcg) is as effective as three doses of albuterol and at one fifth the cost in children's acute asthma; is well tolerated and patients of all ages can benefit. Nebulized Formoterol single dose is non -inferior to three doses of albuterol and underscores many possibly substantial care savings, given the heavy load that acute asthma care represents in our country and many other developing nations (3) . * PARI \ nebulizer (Pronet Turbo) 1 before and after BSH dosing. All subjects tolerated oral BSH dosing without significant adverse events. Increased Phase II enzyme expression in nasal lavage cells occurred in a dose dependent manner with maximal enzyme induction observed at the highest dose of 200 g BSH. At 200 g BSH daily GSTM1, GSTP1, NQO1, and HO-1 expression was 219, 201, 299, and 221% of baseline expression respectively (p e 0.001 for all enzymes). Phase II enzyme induction was not seen with ingestion of non-sulforaphane containing alfalfa sprouts. Thus, oral sulforaphane safely and effectively induces mucosal Phase II enzyme expression in the upper airway of human subjects. Based on this work and previous in vitro/animal studies, sulforaphane may represent a novel therapeutic strategy for the treatment of allergic respiratory conditions. Additional studies are underway to determine whether Phase II enzyme induction is effective in reducing the inflammatory effects of particulateinduced oxidative stress in the human airway. Background: Increasing incidences of Aspergillus fumigaus-induced allergic asthmatics all over the world necessitate identification of a catalogue of allergens of clinical importance. In the present study, immunoproteomic approach was used to establish a set of candidate allergens, which could be explored further for diagnostic application in allergic aspergillosis and asthmatics including ABPA. Methods: Immunoproteomics combined with mass spectrometric analysis was used to identify proteins of three-week culture filtrate (3wcf) responsible for inducing A. fumigatus-specific IgE immunoreactivity, using pooled sera from A. fumigatus-sensitised asthmatics. Their diagnostic potential was also examined against patients with allergic bronchopulmonary aspergillosis (ABPA), by 2-DE immunoblotting with individual sera from such patients. Results: Peptide mass fingerprint (PMF) using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and/or de novo sequencing by MS/MS analysis of the protein spots from 2-D gels led to identification of a total of sixteen allergens of A. fumigatus. Eleven of them are being reported as allergens for the first time and five were reported earlier. Putative isoforms of the proteins Asp f 13 and chitosanase have been observed for the first time. When studied for immunoreactivity of these proteins among patients with ABPA using their individual sera, these patients exhibited sensitisation although the pattern was varying. Taken together, these proteins could thus be considered as potential allergens even among patients with ABPA. Three of these proteins viz. the hypothetical protein, extracellular arabinase and chitosanase could be major allergens. Conclusion: The immunoproteomic approach applied to the analysis of culture filtrate proteins resulted in identification of several candidate allergens, many of them novel, contributing to the catalogue of A. fumigatus allergenic proteins. These allergens may facilitate improved serodiagnosis for allergic aspergillosis. In addition, the immunoreactivity of these proteins observed among the patients with ABPA may have potential for serodiagnosis and opens up scope for evaluation and development of personalized immunotherapeutics. Laboratory testing and evaluation of olive pollen allergy and cross reactivity in Jordan Hani Ababneh 1 , Hani Ababneh 1 , and Ghada Maaytah 2 . 1 President of the Jordanian Society of Allergy Immunology, Immunology Queen Alia Hospital, Amman, Jordan; 2 King Hussein Medical Center, Immunology, Amman, Jordan. Background: The extensive and unorganized planting and cultivation of the holly olive trees in our country is the first cause of allergy. Olive tree (olea europaea) pollen is a major cause of seasonal allergy-making it the first outdoor pollen calendar in Jordan. To date, ten allergens, named Ole e 1 to Ole e 10, have been isolated and characterized from this pollen Ole e 10, a small protein, has been recently described as a major allergen from this pollen, since it affects more than 55% of allergic patients. Few studies regarding olive allergy have been reported in our area. The olive pollination season lasts two months approximately from the middle of May to the middle of July. Objective: The main aim of this study was to address the performance of olive screen testing by total and specific IgE and the cross reaction with other pollens. Methods: Total and specific serum IgE to olive pollen was tested and evaluated in 400 allergic patients before, during and after the peak pollen season using immunoblot Allergy Screen-Respiratory pannel. Results: Out of the 400 patients serum specimens 132 (30%) had allergy to IgE olive pollens. 125(94%) of the allergic patients had elevated total serum IgE 9100 IU/ml). 10 samples from healthy subjects were used as negative controls. Cross-reactivity with orange, Banana, Grass pollens and Ambrosia has been noticed in 10 cases (15%). Conclusion: Olive tree pollen is one of the main causes of allergy in Mediterranean region included Jordan. It is worth to mention the importance of the total and specific IgE testing for olive pollen allergy detection and cross-reaction with other pollens and fruits Suggesting the presence of Ole e 10-like proteins in many sources which may be related to the development and exacerbation of allergic and asthmatic process, but these results could be interpreted with caution regarding clinical symptoms, signs, and skin prick testing. In vivo reactivity to grass pollen in correlation to specific IgE antibody levels in patients with allergic rhinoconjunctivitis Johannes Huss-Marp 1 , Ulf Darsow 2 , Florian Pfab 1 , Ingrid Weichenmeier 1 , Carl Johan Petersson 3 , Magnus Borres 3 , Johannes Ring 2 , and Heidrun Behrendt 1 . 1 ZAUM -Center for Allergy and Environment GSF/TUM, Division of Environmental Dermatology and Allergy, Munich, Germany; 2 Department of Dermatology and Allergy Biederstein, Technical University Munich, Munich, Germany; 3 Phadia AB, Phadia AB, Uppsala, Sweden. Background: Previous investigations established dose-response curves between the severity of clinical symptoms and specific IgE antibody levels to both indoor and food allergens in allergic individuals. We questioned whether this also holds true for outdoor allergens e.g. grass pollen allergens. Methods: 100 patients with allergic rhinoconjunctivitis (ARC) and allergy to timothy grass pollen as well as 40 healthy controls were recruited during the grass pollen season in 2006 and 2007. To investigate the correlation between IgE antibody levels and threshold in vivo reactivity we determined specific serum IgE antibodies to timothy grass pollen (ImmunoCAPi, Phadia AB, Uppsala) at three time points during one year (June/July; October; December) and performed conjunctival and nasal provocation tests with grass pollen extracts at different concentrations (Phleum pratense, HAL Allergy, Duesseldorf, Germany). In addition, skin prick tests were performed. The investigations were paralleled by grass pollen counts using a Burkard trap performed at the University Campus Biederstein, Munich (G50 m distant from the laboratory, 1.80 m above the ground). Additionally, the patients scored their symptoms with visual analog scales and exhaled nitric oxide was analysed as measure for inflammatory lung reaction. Results: Preliminary results from 41 patients with ARC revealed a significant relationship between specific IgE concentration in serum and the titrated level of the skin prick test (p G 0.0096, Jonckheere-Terpstas trend test). However, no correlation between specific serum IgE to timothy grass and thresholds of nasal and conjunctival provocation tests were found. Conclusion: Measurement of specific serum IgE levels seems to accurately predict the in vivo reactivity with regard to skin prick test in patients with ARC to grass pollen allergen. The preliminary results indicate a good correlation between indicators of sensitization, but not between serum IgE levels and threshold allergen concentrations that induce symptoms Further data analysis concerning conjunctival and nasal provocation tests and symptom scores in a greater number of patients is in progress. The diagnosis of Pollen-Food Syndrome (PFS) through the use of a structured questionnaire PFS, a manifestation of Oral Allergy Syndrome (OAS), is a common food allergy. The characteristic symptoms, speed of onset and the typical foods involved suggest PFS could be diagnosed though clinical history alone. Subjects with reported springtime hay fever completed a PFS diagnostic questionnaire (PFSDQ) and were allocated to either Group 1 (has PFS), Group 2 (No PFS, ? food allergy) or Group 3 (no PFS or food allergy). All subjects then had a consultation with an allergy specialist who made a provisional PFS diagnosis. This was followed by skin prick testing to fresh foods using the prick by prick test method (PPT) and oral food challenge (OFC). The PFSDQ was measured against the gold standard of positive OFC, and a second standard, the final diagnosis accorded to each subject by the medical allergy specialist, using PPT and OFC results to modify his original diagnosis. 119 subjects completed the study; 58 (49%) were allocated to Group 1 (PFS+ve), and 61 (51%) to Groups 2 (30) and 3 (31) (PFS-ve). 47 subjects had a positive OFC, 41 of whom were from Group 1. Although there was a significant difference between the PFSDQ and OFC results (p G 0.05), this was not the case when the PFSDQ was compared to the final diagnosis standard (p = 0.5). The sensitivity, specificity, positive and negative predictive values of the PFSDQ were all Q 90% when tested against the final diagnosis standard. The reliability of the PFS-DQ was high with a standardised alpha score of 0.819 (Chronbachs Alpha Test). Factor analysis showed the key components of the PFSDQ to be symptom type, raw or cooked foods, speed of onset of symptoms and number of reported symptoms (Varimix rotation). Logistic regression analysis showed the only predictive factors of PFS diagnosis were reactions to raw plant foods (p G 0.01), onset of symptoms within 5 minutes of eating (p G 0.01) and number of reported symptoms to foods (p G 0.05). Diagnosis of PFS using the PFSDQ was not significantly different to that made by a specialist allergist using clinical history, PPT and OFC. The most important predictive variables for PFS were reported reactions to raw plant foods, rapid onset of symptoms and multiple symptoms. The PFSDQ could be a useful diagnostic aid for those working in primary care to screen adults with reported food symptoms for the presence or absence of PFS. Laure Rouanet-Bousquet 1 , Philippe Jean Bousquet 2 , Bernard Arnoux 3 , Pascal Demoly 1 , and Antonino Romano 4 . 1 University Hospital Arnaud de Villeneuve, Exploration des Allergies, INSERM, Montpellier, France; 2 University Hospital of Nîmes, Département de l`Information Médicale, Montpellier, France; 3 INSERM, U 454, Montpellier, France; 4 Complesso Integrato Columbus, Unita di Allergologia, Roma, Italy. Introduction: Drug hypersensitivity to beta-lactam was often skin tested using benzyl-penicillin, major (PPL) and minor (MDM) penicillin determinants, amoxicillin, ampicillin and any other culprit beta-lactam. However, PPL and MDM were removed from the marked. The aim of the study was to assess the importance of benzylpenicillin before and after the withdrawal. Methods: Using our Drug Allergy and Hypersensitivity Database (DAHD) and data coming from an Italian centre, we conducted an historico-prospective cohort study. All patients who consulted between 1996 and 2007 for a suspected beta-lactam hypersensitivity reaction and who had at least positive skin test to benzylpenicillin were included. Diagnosis and skin tests followed the European Network on Drug allergy (ENDA) recommendations. Benzylpenicillin, PPL, MDM, Ampicillin, Amoxicillin were always skin tested. Results: 133 patients (48Y36.1% men), 15 (11.3%) asthmatics and 41 (30.8%) atopics were included. 13 (9.8%) were only positive to benzylpenicillin, 32 (26.5%) were also positive for PPL and 66 (55.5%) for MDM. Without skin testing for PPL and MDM, the number of positive to benzylpenicillin increased to 20 (15.0%). No difference was observed for asthma and atopic between subjects only positive to benzylpencillin and those also positive for another penicillin (p = 0.84 and p = 0.34 respectively). Anaphylaxis and anaphylactic shock were more common in subjects positive to several penicillins (76Y67.3% vs 7Y35%, p = 0.001). Conversely, only 26 (23.0%) vs 8 (40.0%) presented an urticaria / angiodema. Conclusion: Since the withdrawal of PPL and MDM, benzylpenicillin skin test appeared to be mandatory, 15% of the patients being only positive to this drug. This should significantly reduce oral challenges. Zenro Ikezawa, Chieko Watanabe, Kazuko Nakamura, Yumiko Yamane, Yuki Shigehira, Yuko Ikezawa, and Michiko Aihara. Yokohama City University Graduate School of Medicine, Enviromental Immuno-Dermatology, Yokohama, Japan. Background: (BDrug-induced hypersensitivity syndrome (DIHS) is a unique severe adverse drug reaction, which is well known to be accompanied with reactivation of human herpesvirus 6 (HHV-6) in many cases 2Y3 weeks after development of drug reactions. The clinical features of this syndrome are acute widespread maculopapular, polymorphous, eczematous and/or erythrodermic erythema, fever, lymph node swelling, liver dysfunction, eosinophilia and leukocytosis with atypical lymphocytes. After first reports of DIHS with HHV-6 reactivation in Japan in 1998, 118 cases have been reported with data of HHV-6 reactivation up to now. Methods: We retrospectively analyzed recent reports of suspected adverse drug reactions submitted to medical journals from Japan, in order to define the presenting characteristics of these diseases in Japan. Results: Ages ranged from 0 to 89 years old with a mean of 48.6 years old and the ratio of M/F was 64/54. 63.7 % of causative drugs were anticonvulsants (carbamazepine, 63.3% of the anticonvulsant drugs). The other common causative drugs were sulfonamides, mexilletine (an antiarrhythmic drug) and allopurinol. The symptoms developed 3 weeks or more after the beginning of causative drug administration in 80.5% of the patients. Recurrence of the symptoms was observed in 40% of the patients. Four of them died and the mortality rate was 3.6%. Reactivation of HHV-6 was detected in 83.9% of the patients by the increase of IgG against HHV-6 and/or increase of HHV-6 DNA in the peripheral blood and sera. On the other hand, reactivation of cytomegalovirus was observed in 4 patients without HHV-6 reactivation. In treatment of 118 patients with DIHS, steroid without puls was in 71 cases (60.2%), steroid puls with mPSL in 15 cases (12.7%; 1g/day in 40.0% of 6/15, under 1g/ day in 33.3% of 5/15, unclear in 26.7% of 4/15), intravenous injection of high dose immunoglobulin (IVIG) with steroid in 5 cases (4.2%) , and plasmaapheresis (PA) with steroid in 3 cases (2.5%). The mortality in DIHS was unexpectedly high as 3.4% (4/118) , as compared to that in SJS. Discussion: A probable role of HHV-6 in DIHS was discussed in comparison of that of Epstein-Barr (EB) virus in infectious mononucleosis and Mosquito bite hypersensitivity. Drug provocation test in patients`\drug hypersensitivity reaction Background: It has bndeen confirmed that, drug hypersensitivity are common and life threatening. This diagnosis can be vigorous and based on clinical history and a physical examination, possibly under skin tests and drug provocation test. We intend to describe the out come of drug provocation test in analyzing patients with histories suggesting drug allergy, using retrospective analysis of clinic case series. Methods: Some 416 consecutive patients with suspected immediate drug allergy referred to the hospital between, January 1999 to February 2005. Patients with served skin reaction and those with positive result on skin test for B-lactams were excluded. it should be noted that single-blinded administration of increasing doses of the suspected drug up to the usual daily dose, under struck hospital surveillance. Results: 920 drug provocation tests were performing using various drugs, including aspirin (11.4%) nonsteroidel anti-inflammatory drugs (7.5%) macrolides (8.5%) paracetamole (10.1%) and other non-steroidal antiinflammatory drugs (12.2%). We had 192 (15.6%) positive provocation test results which also reproduce the same symptoms, albeit milder and of a shorter duration -in the following patients.10 (4.2%) with history of anaphylactic shock, 15(5.5%) the history of anaphylaxis without shock. 70 (46.4%) with a history of urticaria and 5(2.4%) with a history maculopapular eruption. All reactions were completely refers by prednisone and epinephrine. However, we assume or accept that false negative results on drug provocation test may have occurred because of sanitation, rare cofactors not included in the diagnostic procedure and tolerance introduction during provocation. Conclusion: It can be certain that drug provocation tests in individuals with suspected drug allergy performed in carefully control settings can confirm drug hypersensitivity. 142 Antituberculous drug rechallenge: success rate among patients with cutaneous hypersensitivity reactions Lara Theresa Aleta. University Of The Philippines-Philippine General Hospital, Allergy, Quezon City, Philippines. Background: This is the first study to systematically document the proportion of cutaneous hypersensitivity reaction (CHR) in patients with adverse drug reactions (ADR) to antituberculous medications and their excipients. Objective: 1.To determine the proportion of CHR due to specific antituberculous medications; 2. To investigate the success rate of rechallenging by shifting to a different brand of antituberculous medication. Methodology: All patients with CHR to antituberculous medications from January 1 to August 31, 2001 were included. After at least 48 hours off antihistamines, the patient was rechalleged with white or light yellow colored drugs to minimize excipients the patient may be sensitized to. Rechallenge was considered successful if there was no recurrence of symptoms while on the multiple antituberculous drug regimen. Results: Sixty-six patients were referred for ADR to antituberculous medications. Forty-five had CHR. Forty-two patients were rechallenged. Fourteen were successful. Sixteen had documented cutaneous adverse reactions to one or more drugs with the highest incidence occurring with ethambutol and pyrazinamide. The success rate of rechallenge was 14/42. Conclusion: 1. Skin allergy is common (68%) with first line anti-TB drugs. The most common drugs associated with CHR were ethambutol and pyrazinamide; 2. The success rate of drug rechallenge when shifting to a different brand of anti-TB medication is 14/42 (33%) 143 Meta-analysis: Risk of angioedema with angiotensin receptor blockers (ARBs) in patients with prior angioedema associated with angiotensin converting enzyme inhibitor (ACE-I) Bret Haymore 1 , Jiun Yoon 1 , Cecilia Mikita 1 , and Kent DeZee 2 . 1 Walter Reed Army Medical Center, Allergy-Immunology, Washington DC, United States; 2 William Beaumont Army Medical Center, Internal Medicine, El Paso, United States. Background: Patients who experience angioedema (AE) after taking ACE-I have been reported to develop AE when taking an ARB, but few studies describe the risk. We sought to answer this question by performing a systematic review of the medical literature. Methods: A literature search was performed in MEDLINE, EMBASE, BIOSIS, and Current Contents with no limitations from Jan 1990-May 2007. SCISEARCH was also used to identify additional citations of key articles. Reference lists of retrieved studies and review articles were evaluated for additional citations. Two authors independently evaluated studies for inclusion and abstracted relevant data according to pre-defined parameters. Any article that described a cohort of patients who had experienced AE after taking an ACE-I, were subsequently exposed to an ARB and followed for at least one month were included. The percentage of patients experiencing AE was abstracted from each article and confidence intervals were calculated using the exact binomial method. The pooled percentage was calculated using the inverse variance method. Results: From the 238 unique articles initially identified, three articles describing 71 patients met inclusion criteria. One was a randomized controlled trial and two were retrospective cohorts. The mean age of patients was 63 years with 56% male and 44% female. Two of the studies were predominantly Caucasian subjects with the third study being 69% African-American. The mean time of follow-up was 20.3 months. These articles described both confirmed and possible cases of AE secondary to ARB. For possible cases the risk of AE was 9.4% (95%CI 1.6Y17); for confirmed cases it was 3.5% (95% CI 0Y9.2). There were no fatal events. There was no statistical heterogeneity between trials (p 9 0.3). Conclusion: Limited evidence suggests that for patients who developed AE when taking an ACE-I, the risk of developing any AE when taking an ARB is between 2Y17% and for confirmed AE the risk is 0Y9.2%. This information will aid clinicians in counseling patients regarding therapy options after developing AE due to ACEI. nervous systems as well as the immune and hematopoietic systems. IL-31 was secreted in larger amounts preferentially in Th2 than in Th1 cells. IL-31 functiones by the heterodimeric receptor composed of IL-31RA and OSMR that is expressed constitutively on epithelial cells and keratinocytes. These cells are likely to be involved in the dermatitis and pruritis of patients upregulating IL-31. Objective and Methods: Peripheral blood mononuclear cells were obtained from 22 patients with atopic dermatitis allergic to hen-egg ranging from 1 to 13 years in age, as well as from 9 healthy individuals who had no allergic symptoms. The patients had recurrent eczema, pruritis and positive skin reactions to egg white. Patients also had positive responses to the oral provocation test to raw hen egg. Diagnostic criteria for atopic dermatitis was based on the criteria of Hanifin and Rajk. We evaluated expression of IL-31-pruducing cells by the methods of newly developed FCAS analysis. Results and Discussion: IL-31 was preferentially expressed in CLA (skinhorming cutaneous lymphocyte antigen) as well as CD45RO-positive cells in atopic children complaining of skin lesions and pruritis compared to normal subjects. Severity of these symptoms were correlated with amount of IL-31 in the cells. Conclusion: These result suggests larger amounts of expression of IL-31 is likely to be involved in the pruritis as well as skin lesions in atopic children. Dysregulation of TLR-2 induced effects in monocytes from patients with atopic dermatitis: Impact of the TLR-2 R753Q polymorphism Margarete Niebuhr 1 , Jens Langnickel 1 , Christian Draing 2 , Alexander Kapp 1 , and Thomas Werfel 1 . 1 Hannover Medical School, Department of Dermatology and Allergology, Hannover, Germany; 2 University of Konstanz, Biochemical Pharmacology, Department of Biology, Konstanz, Germany. Background: Atopic dermatitis (AD) is often complicated by an enhanced susceptibility to bacterial skin infections, especially with Staphylococcus aureus (S. aureus). Toll-like receptors, especially (TLR)-2 recognizes cell wall components of S. aureus, e.g. lipoteichoic acid (LTA) and peptidoglycan (PGN) . A heterozygous TLR-2 R753Q polymorphism occurs in a frequency of 11.5% in adult AD patients and has been shown to be associated with a severe phenotype. Objective and Methods: The aim of this study was to investigate the impact of TLR-2 agonists (LTA, PGN and Pam3Cys) on cytokine production and cell surface marker expression in human monocytes from AD patients with the TLR-2 R753Q polymorphism compared to AD patients with Bwild typeT LR-2 and control individuals to elucidate the functional role of the TLR-2 R753Q polymorphism. Results: We could show that AD patients with the TLR-2 R753Q mutation produced significantly more IL-6 and IL-12 and significantly less IL-8 compared to AD patients with non-mutated TLR-2 upon stimulation with TLR-2 agonists. Expression of CD86 was significantly higher upon Pam3Cys stimulation in TLR-2 R753Q polymorph patients. Expression of CD80 was unaffected after stimulation with TLR2 agonists. Conclusion: We show for the first time functional differences in TLR-2 responsiveness of monocytes from AD patients with the TLR-2 R753Q mutation compared to wild type AD patients in a ligand dependant manner. Clinical implication: Our data stratify the emerging concept that AD patients have a dysbalance in innate as well as aquired immunity. TLR-2 might be essential in the pathogenesis of AD and involved both in the enhanced susceptibility to skin infections with S. aureus and in a higher inflammatory response in patients with the TLR-2 polymorphism. 147 Impact of atopic dermatitis on the quality of life in Japanese patients Yukihiro Ohya 1 , Yasuo Kubota 2 , Akira Hoshioka 3 , Ritsuko Hosoya 4 , Namiko Kojima 1 , Masami Narita 1 , Akiko Miyazaki 1 , and Nahoko Sakamoto 5 . 1 National Center for Child Health and Development, Division of Allergy, Tokyo, Japan; 2 Kagawa University, Department of Dermatology, Kagawa, Japan; 3 Chiba Children's Hospital, Division of Allergy, Chiba, Japan; 4 Hosoya clinic, Dermatology, Tokyo, Japan; 5 Juntendo University, Department of Public Health, Tokyo, Japan. Background: Prevalence of atopic dermatitis (AD) in Japanese population has been rapidly increased for these decades. Impact of AD on their quality of life (QOL) is difficult to be assessed by using a translated QOL scale developed in western nations because of the influence of different culture. The aim of this study is to develop an original QOL assessment scale for Japanese patient with atopic dermatitis. Methods: We have collected our own data from patients who visited Japanese hospitals and clinics by using an anonymous open questionnaire and some by interviews. One hundred and eighty items were extracted from whole data and based on them, secondary questionnaire comprising 68 items was made by expert's discussion. It was applied to 200 patients with atopic dermatitis for psychometric item reduction. Results: Through the process of factor analysis and item analysis, sixty eight items were converged to 9 items comprising of four factors such as influence to daily activities, burden of medication, itchiness generated annoyance, anxiety about future. Internal consistency of each factor was excellent such that Cronbach" of each factor was 0.0772, 0.843, 0.865 and 0.789. Item content and factor loading score of each item is weak enthusiasm o.818, influence of sleep disturbance 0.617, burden of house keeping 0.589, burden of topical therapy 0.824, unwillingness of topical application 0.817, annoyance to itchiness 0.821, annoyance to scratching 0.756, anxiety about remission 0.824, and anxiety of fluctuating symptoms 0.684. Conclusion: This study has revealed impact of atopic dermatitis on the QOL of patient is not only associated with dermatological symptoms but also adherence burden and anxiety about prognosis. The compact 9 time questionnaire developed in Japan this time would be useful to detect compre-hensive quality of life of patients with atopic dermatitis even in a busy situation of our outpatient clinics. Eosinophil proteins in serum and urine of children with atopic dermatitis Magdalena Czarnecka-Operacz, Dorota Jenerowicz, and Wojciech Silny. Poznan University of Medical Sciences, Dermatology, Poznan, Poland. : Atopic dermatitis (AD) is an inflammatory skin disease characterized by chronic and recurrent course, beginning primarily in an early childhood. The aim of the project was an evaluation of the usefulness of selected eosinophil proteins in serum (ECP, EPX) and urine (EPX) of children suffering from AD, as markers of disease severity. The study also aimed to analyze correlations between the level of examined proteins and such parameters as: skin prick tests (SPT) results, serum concentration of total IgE and coexistance of symptoms of other atopic diseases. We examined 40 AD children attending Allergic Diseases Diagnostic Center and hospitalized in the Department of Dermatology. As control group we selected 23 healthy children without any symptoms of allergic diseases. Mean level of eosinophil proteins measured in serum (ECP, EPX) and urine (EPX) of children suffering from AD was higher than in controls and statistically significant difference was detected for serum level of EPX. Patients presenting with very severe/severe AD had higher level of eosinophil proteins than patients presenting with mild/moderate AD, although no significant difference was found. AD children with positive SPT results and detectable serum specific IgE presented with higher mean level of serum and urine eosinophil proteins than in compared groups of patients with negative SPT results and undetectable serum specific IgE, although with no statistically significant difference. Total IgE level in AD children representing allergic form of the disease was statistically significantly higher than in children with non-allergic AD. In children with very severe/severe AD, total IgE level was significantly higher than in children with moderate/mild disease. Results presented above indicate singnificant role of eosinophils in etiopathogenesis of AD. Measurements of serum and urine level of selected eosinophil proteins may be considered as an important part of diagnostic approach in children suffering from AD, especially in differentiating of allergic and non-allergic form of disease. Among eosinophil proteins, EPX has always been the subject of particularly thorough investigations, because of easy and non-invasive way of measurement in urine, which is an important feature in diagnostics of children. EPX is a marker significantly better differentiating AD patients from healthy individuals in comparison to other factors, such as ECP and peripheral blood eosinophilia. Quantitative analysis of nerve growth factor (NGF) in the horny layer of atopic dermatitis and effect of treatment on NGF Zenro Ikezawa 1 , Junko Yamaguchi 1 , Michiko Aihara 1 , Takeshi Kambara 1 , and Yusuke Kobayashi 2 . 1 Yokohama City University Graduate School of Medicine, Environmental Immuno-Dermatology, Yokohama, Japan; 2 Shiseido CO, R&D Planning Division, Yokohama, Japan. : The expression of nerve growth factor (NGF) is known to increase in the skin of patients with atopic dermatitis (AD) and to be related to disease aggravation. In the present study, we measured skin NGF levels in AD patients and assessed the possibility of a relationship to AD severity, as well as effects of treatments. Methods: NGF in the horny layer (horn NGF) of skin lesions on the cubital fossa was collected via tape stripping and measured using ELISA in AD patients before and after 2 and 4 week treatments. Itching and eruptions on the whole body and on lesions measured for NGF were evaluated. Peripheral blood eosinophil count, serum LDH level and serum total IgE level were also examined. Results: The level of NGF was significantly higher in AD patients than in healthy controls, and correlated with the severity of itch, erythema, scale/ xerosis, eosinophil count, and LDH level. The NGF level decreased significantly after treatments with olopatadine and/or topical steroid for 2 and 4 weeks. The decrease of NGF correlated with the decrease in the severity of itching, severity of AD on the whole body, erythema, papule, scale/xerosis and lichenification of the lesion, eosinophil count, and LDH level. Conclusion: The level of the horn NGF was found to reflect the severity of itching and eruptions in AD. Therefore, measurement of the NGF via this harmless method appears to be useful in assessing severity and therapeutic effects in AD. Association between the allelic variant delta 32 of CCR5 receptor and the prevalence of urticaria, angioedema and atopic dermatitis symptoms Jorge Molinas 1 , Rut Aguero 1 , Ledit Ardusso 2 , Cintia Crudelli 1 , Rodolfo Navarrete 1 , Natalia Pujato 1 , Guillermo Mujica 1 , and Nora Figueroa 1 . 1 Rosario National University, Human Physiology, Rosario, Argentina; 2 Rosario National University, Allergy Department, Rosario, Argentina. Introduction: C-C chemokines, including RANTES, MIP-1, MCP-2, and the CCR5 specific receptor play an important role in the genesis of allergic inflammation. The presence of homozygosis for the delta32 (d32) polymorphism of CCR5 produces the absence of this receptor from the cell surface and heterozygosis promotes a very low expression of this receptor, hence reducing the availability of receptors capable of interacting with chemokines to induce the inflammatory response. We have previously informed a reduced risk of developing asthma and rhinitis in adults that present the d32 allelic variant. We could not find any report about the relationship between d32 and atopic dermatitis (AD), urticaria (URT) or angioedema (ANG). Objective: To evaluate association between d32 polymorphism and AD, URT and ANG in adults from Rosario city. Materials and Methods: 111 caucasian subjects, 36 males (32.7%) and 74 females (67.3%), aged 19 to 46 years old (x = 23.88 T 4.45), answered questionnaires for AD (ISAAC), URT and ANG. DNA was extracted from blood samples and d32 polymorphism was determined by PCR. Oligonucleotide sequences were: sense: 5 ¶-GTCTTCCATTACACCTGCAGCTCT-3 ¶, antisense: 5 ¶-CACAGCCCTGTGCCTCTT-3 ¶. Results: 6.3% of the subjects presented the heterozygotic polymorphism and 0.9% the homozygotic polymorphism. The risk of suffering from AE, URT or ANG was significantly lower in the subjects that presented the d32 polymorphism. Among the subjects that referred for AD symptoms at least once in their life (15.3%), no one presented heterozygosis or homozygosis for d32, while in the subjects that did not refer these symptoms the frequency of the same genotype was 8.5% (RR = 0.83 IC:0.77Y0.91; p G 0.05). Among the subjects that referred for URT symptoms at least once in their life (36.9%), 2.4% presented heterozygosis or homozygosis for d32, while in the subjects that did not refer these symptoms the frequency of the same genotype was 11.1% (RR = 0.70 IC:0.52Y0.95; p G 0.05). Similar results were obtained in subjects that referred for ANG symptoms at least once in their life (13.6%), since no one presented heterozygosis or homozygosis for this polymorphism, while in the individuals that did not refer these symptoms the frequency of the mentioned allele was 8.4% (RR= 0.85 IC:0.79Y0.92; p G 0.05). Conclusion: The results obtained in this study allow us to suggest that the reduced risk to develop AD, URT and ANG could be influenced by the presence of the CCR5-d32 allelic variant. Background: In contrast to the advancement of knowledge on the pathogenesis of atopic dermatitis (AD), establishing international diagnostic criteria standards remains a challenge. The criteria of Hanifin and Rajka, though extensively mentioned in research settings, rely on features with significant variation due to age, ethnicity or other factors and may be unsuitable for epidemiological studies when clear-cut case/control definitions are needed. The validity of the UK Working Party criteria has been reported in population and hospital-based studies, mostly in children. Data concerning adults are however very limited. Objective: We aimed to test the validity of the UK Working Party criteria in adult patients referred to a university hospital in Bulgaria. We also tested the relevance of six clinical signs commonly associated with AD without being criteria, namely, fine hair, periorbital and periauricular dermatitis, infraauricular fissures, cheilitis angularis and Hertoghe sign. Methods: 118 patients aged 18Y60 years were recruited for the study. All patients were examined independently by a senior dermatologist and a resident, both blinded to the study aim. The results were evaluated by a third independent and blinded observer using contingency tables. Results: We found higher sensitivity (94,03%), specificity (92,16%), positive (94,03%) and negative (92,16%) predictive value for the UK Working Party diagnostic criteria than for the ones of Hanifin and Rajka (88,06%;82,35%; 86,76;55,26 respectively). Family history for atopy and erythroderma were significant factors for false positive results. Of the additionally examined features, the highest sensitivity and specificity was found for Binfraauricular fissures [ (59,70% and 84,31%, respectively) . Conclusion: Our results confirm the validity of the UK Working Party criteria in an adult hospital population in Bulgaria. Although the cases may represent a more severe disease end spectrum, there are only few systematic publications on AD features in adulthood and our study provides valuable data on the performance of the diagnostic criteria and relevance of the individual clinical signs in this age group. Is atopic dermatitis really atopic? Mariana Mandazhieva-Pepelanova, and Nevena Berova. Military Medical Academy, Department of Dermatology and Allergology, Sofia, Bulgaria. Background: There is little data on the correlation between atopic dermatitis and other atopic diseases (asthma, rhinitis) and atopy markers (positive skin prick test and specific IgE to Dermatophagoides pteronissinus /Dpt./). This is why the comorbidity of atopic dermatitis and allergic rhinitis and bronchial asthma and the presence of atopy markers should be further investigated as well as the correlation between the positive atopy markers and a positive patch test with Dpt. Methods: The correlation between patch-test and skin prick test with Dpt. and specific IgE was evaluated in 32 patients (8 men), with atopic dermatitis, 27 had rhinitis as a concomitant disease (17 with allergic rhinitis) and 16 had bronchial asthma. All of them were tested for specific IgE and skin prick test and patch test with Dpt. were performed. Results: 17 patients had a positive patch test with Dpt., 12 had positive specific IgE antibodies to Dpt. and 14 -positive skin prick test with Dpt. Conclusion: Cases with a positive patch test prevail when compared to positive skin prick test and specific IgE. Positive patch test does not always correlate with atopy, evaluated as positive skin prick tests and specific IgE to Dpt. The patch test could be used as a routine and reliable method in practice for evaluating the role of Dpt. in the pathogenesis of atopic dermatitis. Viktoriia Klymenko. Kharkiv State Medical University, Department of Propedeutic Pediatrics N2, Kharkiv, Ukraine. Background: Immune cells play a main role in development of atopic dermatitis (AD); however, so far only limited data are documented on the distribution on these cells in the skin during cutaneous inflammation. Objectives: To gain better insight into the number and phenotype of immune cells in lesional skin of AD patients. Methods: The lesional skin biopsies of patients with different forms of atopic dermatitis (an exudative, an erytemato-squamouse, a lichenoid) has been investigated. It is carried out 38 skin's biopsies in the children aged 1,5Y15 years. Biopsies were made by needles (diameter -1,6MM). Avidin-biotin immunoperoxidase staining of paraffin-embedded skin sections, with quantitative counting of cells labeled by anti-CD4, anti-CD8, anti-CD16, and anti-HLA-DR (Human Leukocyte Antigen) anti-IgE monoclonal antibodies was used. Results: The results of immunomorphologycal analysis showed a great infiltration of T-lymphocytes and a high intraepidermal expression of IgE. The amount of main immune cells were (per 100 cells): CD4 = 47 T 5,8; CD8 = 17,3 T 3,2; CD4:CD8 = 2,8 T 0,5; CD16 = 5,7 T 0,7; HLA-DR = 1,1 T 0,1. Conclusion: Immunophenotyping was found to be a useful diagnostic method in AD patients. These data help to elucidate the pathogenesis of AD. Undiagnosed alllergic dermatitis -a prospective study Vipul Shah 1 , and D Tripathi 2 . 1 Allergy Clinic, Allergy, Surat, India; 2 Bombay Hospital, Allergy, Mumbai, India. Introduction: Undiagnosed cases of dermatitis were referred to our clinic after treated by number of dermatologists in our city. These cases were treated with different types of symptomatic drugs for last 7 to 10 yrs, but did not yield any significant result and these were labeled as difficult dermatitis. Materials and Methods: Ten such patients who were referred to our clinic were subjected to detailed medical history & clinical examinations. We decided to perform skin prick test (modified SPT) as to determine possible role of airborne allergens. These patients showed positive skin prick test reactions to many allergens such as Asp. flavus, Asp.Tarmare, Alt Alternaria, D. Farinae, D.Pter., House dust, Hay dust, Parthenium & Paltophorm among airborne allergens, while reactions were observed significantly positive to certain food allergens also ( Milk, Almond, Yeast, Hazel nut ) .Since there patients showed strong reactivity to certain allergens (IgE mediated hypersensitivity), an attempt was made to administer allergen immunotherapy course. Allergens which were strongly positive and showed airborne dominance in the vicinity of the patient were selected for allergen immunotherapy mixture. The allergen immunotherapy was initiated on this subject as per guidelines laid down by WHO. Results and Discussion: The improvement was observed & symptomatic relief was recorded within 3 to 4 month of initiation of allergen immunotherapy. Conclusion: Allergen immunotherapy could play a major role in the treatment of airborne contact dermatitis. Prognosis of allergic contact dermatitis in patients patch tested in a university hospital in Beijing Lin-feng Li. Peking University Third Hospital, Dermatology, Beijing, China. Background: Although it is believed that the prognosis of allergic contact dermatitis (ACD) is good, long-term prognosis of ACD has not been reported. Objectives: To investigate the 1-year outcome of ACD in a university dermatology setting. Patients and Methods: In a two-year period, 655 consecutive patients were patch tested in Peking University Third Hospital for suspected ACD. At one year after patch testing, all patients were asked a revisit and prognosis of ACD was evaluated by the rate of clearance (complete free of dermatitis without recurrence for more than 3 month before the final evaluation). Results: Of 599 patients finished study, 167 cases were ACD. The rates of clearance and relapse were 49.7% and 41.9% respectively. Concurrence of atopic diathesis, ichthyosis, history of drug allergy as well as disease location had no effect on the clearance rate. Significant lower clearance rate (26.8%) was found in patients with longer disease duration (over 6 months) before diagnosis. The clearance rate in patients with non-continuous ACD was also lower than that of continuous ACD (28.3% vs. 59.6%, chi square test). Ignorant reexposure to contact allergens was the mean reason for relapse. Conclusion: 1-year outcome of ACD is not as good as expected. Longer disease duration before diagnosis and non-continuous ACD are main risk factors for poor prognosis. Patients with suspected ACD should be patch tested as early as possible. Mandatory ingredient labeling and sufficient patient education are necessary to improve the prognosis of ACD. Fishing for allergens hiding as prohaptens: cinnamic aldehyde but not cinnamic alcohol identified as potent sensitizer Skin is a major target organ of allergic reactions to small molecular weight compounds such as allergic contact dermatitis or allergic drug allergy. It has been generally accepted that most small molecular weight compounds must be bound to high molecular weight compounds in order to become immunogenic. For this binding of the hapten to a protein the hapten must be a highly reactive chemical such as intermediate metabolites of cytochrome P450 (CYP) dependent metabolism of prohaptens. Recently we demonstrated the usefulness of an organotypic CYP cocktail to identify prohaptens by combining such a CYP cocktail with a dendritic cell-based in vitro model to identify sensitizing small molecular weight compounds (Bergström et al., JID 127 (2007 Mukhtar et al., JID (2007) 992Y993). Cinnamic alcohol is oxidised to cinnamic aldehyde by a CYPdependent metabolism. Therefore we were interested whether cinnamic aldehyde is the nominative antigen in allergic reactions to cinnamic alcohol. We generated immature DCs from human peripheral blood monocytes by depletion of CD2, CD7, CD19, CD56, CD16, CD235a positive leukocytes and consecutive incubation with GM-CSF and IL-4. At least 6 million cells were stimulated with the following test compounds for 30 hours respectively: 2,4,6-trinitrobenzene sulfonic acid -TNBS (200 mg/ml), sodium dodecyl sulfate -SDS (5 mg/ml), dimethyl sulfoxide -DMSO (0.1%), CAld (0,1 mM) and CAlc (0.1 mM). After RNA isolation of the treated cells, reverse transcriptase-PCR analysis was performed using a primer/probe set specific for the detection of IL-8 mRNA expression. A sensitizing effect of CAld could be shown for all 6 donors as indicated by a median IL-8 upregulation of 34.3 fold normalized relative to mediumtreated controls. In contrast, the median enhancement of IL-8 mRNA expression was not significantly greater after incubation with CAlc (2.13) than after stimulation with SDS (2.22), DMSO (1.2) . In conclusion, cinnamic aldehyde possesses a stronger allergenic activity than cinnamic alcohol which is CYP-dependently metabolised to cinnamic aldehyde. These results further supports the concept that the combination of prohapten modified organotypic CYP cocktails with a dendritic cell-based in vitro model of contact allergens may streamline and increase the predictive efficiency of current safety test methods in allergic contact dermatitis. The profile of patch test reactions to common contact allergens is related to sex Jochen Brasch 1 , Axel Schnuch 2 , and Wolfgang Uter 3 . 1 University Clinics of Schleswig-Holstein, Campus Kiel, Department of Dermatology, Kiel, Germany; 2 Institute at the University of Göttingen, Information Network of Departments of Dermatology, Göttingen, Germany; 3 University of Erlangen-Nürnberg, Dept. of Med. Informatics, Biometry and Epidemiol., Erlangen, Germany. Background: It is a worldwide observation that patients exposed to diagnostic patch tests are predominantly women. This is commonly explained by distinct sex-related patterns of allergen exposure that cause different patterns of sensitization in men and women and by a higher health awareness of women. Nevertheless, additional sex-related differences in the responsiveness to contact allergens should be taken into consideration. Methods: Sex-related reaction profiles of 16 common patch test allergens that are used in standard patch test series were retrospectively analysed based on data of 47.000 patients filed by the Information Network of Departments of Dermatology in Germany within a period of 10 years. All patch tests had been done by use of identical methods in patients with suspected contact allergy but no history of atopic dermatitis. The proportions of weak positive reactions and of questionable and irritant reactions were evaluated by calculating the positivity ratio and the reaction index separately for men and women for each allergen. Results: Out of the 16 allergens evaluated, 8 had a slightly higher positivity ratio and a slightly higher reaction index in women than in men. 4 allergens had a lower positivity ratio and another 4 had a lower reaction index in women than in men, but no allergen had a lower positivity ratio plus a lower reaction index in women than in men. In particular for allergens with similar rates of positive reactions in men and women female sex was significantly related with a higher positivity ratio and reaction index (pG0.01). Conclusion: There is a small but statistically significant disparity in the reactivity of men and women to common patch test allergens. Women in general have a higher rate of weak positive reactions but less questionable or irritant reactions to an allergen than men. This marginal difference is probably not relevant for patch testing but adds a new element to the pathogenetic puzzle of contact allergy. Reduced allergy rates to contact allergens/haptens with both oral and cutaneous exposure in atopic eczema subjects. Atopy and the "hapten hypothesis" Background: Whilst allergy to food proteins is almost exclusively seen in atopics, it has been our impression that this does not hold true for haptens/ contact allergens which are present in our diet. Diallyl disulfide is the major haptenic allergen in garlic and allergic contact dermatitis to this has been observed in people who handle garlic whilst preparing food. Parabens and lanolin are both contact allergens commonly used in many cosmetic and medicament creams. However, unlike lanolin, the preservative parabens is used in some processed foods. We retrospectively reviewed data from our patch test clinic, comparing the frequency of allergy rates (patch test positive, PT+) to these haptens between atopic eczema/dermatitis AD and non-AD dermatitis patients. Results: Between 1980 and 2006 in total population of 36658 patients with eczema/dermatitis were patch tested, of whom 10326 (28.2%) had AD; in contrast 13/83 (12.5 %) patients PT+ to diallyl disulfide/garlic had AD (AD/total population vs AD/diallyl disulfide PT+ p = 0.011). 54/239 parabens PT+ had AD (22.6%), whilst 181/608 lanolin PT+ had AD (29.8%) (pG 0.05) Discussion: Frequency of contact allergy to haptens with both oral and skin exposure is reduced in AD patients compared to non-AD patients in direct contrast to food protein allergy. This decrease was not observed in lanolin, which is used only on the skin. Possible reasons for these results could be 1) confounding factors e.g. AD patients handle garlic less than non-AD patients, or 2) AD patients are very efficient at tolerising haptens, and this is in some way secondary to their atopic status, or 3) oral tolerance of haptens may in some way antagonise tolerance of proteins and contribute to the development of atopy (the hapten hypothesis). We note that the increase in atopy over the last 50 years also coincides with a period where processed food and milk, with chemical/hapten content, is used in western diets. Incidence and prevalence of para-phenylenediamine allergy in an adult Thai population: a public health problem 3 Kirikkale University Hospital, Pathology, Kirikkale, Turkey. Hypersensitivity to sex hormones has long been recognized as cyclic and recurrent rushes. Progesterone sensitivity is termed as autoimmune progesterone dermatitis (APD) which is shown with a positive skin test . Our patient was a 33year-old female who presented with cyclic skin eruptions of two years'duration, right after she delivered her baby. At the beginning a fixed single maculopapular and itchy skin lesion of 1-2 cm was occuring two days prior to her monthly cycles and resolving by the beginning of mens. However, new lesions occured in other fingers, lately. She had no prior exogenous hormone use. Her menstruel cycles were irregular since she was 15 years old. Progesterone levels were found to be high before her pregnancy, but wasn_t treated. The possibility of APD was suspected because of the cyclic nature of her lesions. Serum levels of sex hormones were normal, as well as her gonadotrophins, and prolactin. Skin biopsy showed a non-spesific dermatitis with a moderate perivasculer infiltration of lenfocytic cells in the upper dermis. The direct immunofluorescence study was negative. Challenge test with aqueous progesterone was performed. Prick and patch tests were negative, whereas intradermal test with 50 mg/ml of progesterone was positive confirming the diagnosis of APD. Despite many localizations reported, there wasn_t any case appearing only in fingers, as it was seen in this case.Exposure to exogeneous progesterone as oral contraceptives (OC) has been suggested as a stimulus for the production of autoantibodies reacting with endogeneous progesterone in APD. However, in some cases OC use isn_t necessary as was observed in our patient. There is a possibility of autoimmune damage to progesterone containing ovarian tissue which could be resulted with premature ovarian failure. In our patient there was no evidence of ovarian disfunction as she had a child. In this case, APD was diagnosed with patient_s typical history of recurrent lesions during the luteal phase of her menstrual cycle and this was confirmed by intradermal test with progesterone. As the endogenous progesterone is produced during ovulatory cyclus, the goal of therapy is the suppression of ovulation by a combination OC. If this is ineffective, danazol, gonadotropin releasing hormone analogs, tamoxifen, and oophorectomy may be tried. Our patient didn_t accept any treatment as she wanted another baby. Analysis of discoidin domain receptor-1 expression by tissue-infiltrating eosinophils in allergic skin diseases Yukari Nishimura, Hiroyuki Murota, Norihisa Kotobuki, Shun Kitaba, Mika Terao, and Ichiro Katayama. Course of Integrated Medicine Graduate School of Medicine, Osaka University, Dermatology, Oasaka, Japan. Background: Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase that is activated on binding to its ligand-collagen a component of the extracellular matrix (ECM) in the organs and is known to be constitutively expressed in the normal tissues such as the lungs, kidneys, colon, and brain. It has been reported that the tissue-infiltrating eosinophils expressed DDR1 and interaction between endogenous DDR1 on eosinophils and collagen in the ECM might affect eosinophil survival in the tissue microenvironment through NFK-B activation in Churg-Strauss syndrome (CSS) characterized by necrotizing granulomatous angitis with massive eosinophil infiltration, asthma, and hypereosinophilia. Patients and Methods: DDR1 expression on peripheral blood eosinophils from asthma patients, and healthy volunteers has been reported to be significantly lower than that from CSS patient. However little is known about the DDR1 expression and its significance in tissue infiltrating eosinophils in allergic diseases. Therefore, we analysed DDR1 expression by tissueinfiltrating eosinophils in allergic skin diseases such as atopic dermatitis, drug eruption and Kimura's disease with eosinophil infiltration. Results and Conclusion: DDR1 was significantly expressed by eosinophils in the lesional skin of CSS, but not atopic dermatitis, drug eruption or Kimura's disease. Our current results indicate that DDR1 expression by eosinophils in the local inflammatory sites is specific for CSS and could be the disease marker in allergic skin diseases with eosinophil infiltration.Mechanisms of eosinophil survival and activation might be different between CSS and allergic skin disease such as atopic dermatitis. Gene-gene interaction between IL-13 and IL-13 receptor !1 is associated with total IgE in Korean children with atopic dermatitis Objective: We evaluated if serum IgE levels correlate with the symptomatology and plasma chemokine levels in children with AD. Methods: AD patients younger than 18 years old were recruited from the paediatric dermatology clinic of a university teaching hospital and AD severity evaluated with the SCORing Atopic Dermatitis (SCORAD) index. Concentrations of serum total IgE, eosinophil count and plasma AD-associated chemokines (cutaneous T cell attracting cytokine, CTACK; thymus and activation regulated chemokine, TARC) were measured. Results: One hundred and seventeen Chinese children with AD (64 boys and 53 girls), with mean (SD) age of 10.7 (4.4) years, were recruited. Their mean (SD) overall SCORAD was 51. 1 (22.8) . % (defined as total serum IgE level divided by age-specific upper limit) correlated well with the extent and intensity of AD except for oozing/crusting which was significant only in the males. There was significant correlation for % with pruritus or sleep loss only in the females. 82 (70%) of patients reported history of allergic rhinitis and 35 (30%) reported history of asthma or hyperactive airway disease. In a subgroup of patients who had never had allergic rhinitis, asthma or urticaria (n = 26), log-transformed % correlated well with the objective SCORAD (Spearman's rho = 0.554, p = 0.003).Levels of IgE, CTACK, and TARC and eosinophil count differed significantly among patients with mild, moderate and severe disease. % correlated well with TARC (r = 0.50, p G 0.001) and eosinophil count (r = 0.41, p G 0.001) but not CTACK (r = 0.11, p = 0.270). The prediction of moderate-to-severe eczema by % gave an area under the receiveroperating characteristic curve of 0.76 (95% CI 0.65Y0.86, p = 0.004). An optimum positive predictive value of 94.2% was achieved with a cut-off point of % of 2.95, sensitivity of 75.0% and specificity of 66.7%. Conclusion: % correlates well with the objective clinical and serum TARC level, and may serve as an overall marker for disease severity. Furthermore, % of 2.95 predicts moderate-to-severe disease. Unlike the chemokines, Serum IgE measurement is inexpensive and readily available in most clinical service. IgE is not only a laboratory marker for atopy but also a parameter of disease severity. Soy intake and reduced prevalence of skin allergic deseases symptoms Jorge Molinas 1 , Ledit Ardusso 2 , Cecilia Torrent 1 , Paula Daneri 1 , and Sara Molinas 1 . 1 Latinoamerican Center University, Physiopatology, Rosario, Argentina; 2 Centenary Hospital, Allergy Department, Rosario, Argentina. Introduction: Soy is one of the few foods that contain important levels of linoleic acid, an omega-3 polyunsaturated fatty acid (O3FA). It is known that O3FA reduces the production of arachidonic acid derivates which are able to reduce chronic inflammatory response. Objective: To evaluate the association between dietary soy intake and the presence of skin allergic symptoms in adults. Materials and Methods: We conducted a cross-sectional study of 765 students from Centro Educativo Latinoamericano University in Rosario, randomly chosen, 638 females (83.4%) and 127 males (16.6%), aged from 17 to 65 years old (x = 21.42 T 4.18) . They answered questionnaires about urticaria/angioedema, contact dermatitis, atopic dermatitis and about the frequency of soyfood intake. Results: Differences were found when the food intake was classified into two groups: A (never or less than once a month) and B (once or more than once a month, week or day). For soy-based mayonnaises and margarines intake, 48.1% of the subjects were found in group B and the prevalence of skin diseases within this group was significantly lower than in group A; for example atopic dermatitis: .5% and in this group the prevalence of angioedema symptoms (1.3%) was lower than in the rest of groups (5.9%) (OR = 0.22; IC:0.04Y0.96; pG0.05). The soyfood ingestion was significantly higher in male and men presented significantly less prevalence of the symptoms studied. The statistical significance remained the same after adjustment for sex and ingestion of histamine-releasing food. No differences were found between soy intake and urticaria symptoms. Conclusion: The results obtained allow us to estimate that the intake of a soy rich food, at least once a month, could protect against some skin disorders. Is house dust mite sensitivity a factor in chronic urticaria? Mojgan Safari. Hamedan University of Medical Sciences, Pediatric Ward, Hamedan, Islamic Republic of Iran. Introduction: chronic urticaria is the most common cutaneous disorder seen in outpatient allergy clinics. There are a few reports associating house dust mite sensitivity with chronic urticaria. This study investigates the possible association between house dust mite sensitivity and chronic urticaria. Methods: In this study four groups of patients were enrolled. Group I: Chronic urticaria (35 subjects). Group II: allergic rhinitis(135 subjects). Group III: asthmatic patients (14 subjects).Group 4: allergic rhinitis +asthma(12 subjects). Group II, III and IV considered as positive controls. All of the patients underwent skin prick testing with antigens of the house dust mite, Dermatophagoides pteronyssinus (DP) and Dermatophagoides farinae (DF), with positive and negative controls. Results: In Group I, 13/35 (37.1%): In group II 64/135(47.4%); In group III 5/ 14(35.7%) and in group IV 4/12(33.3%) patients had skin sensitivity to house dust mites .There were no statistical differences between prevalence of positive skin test to mite in four groups. Conclusion: we suggest a possible association of house dust mite sensitivity with chronic urticaria. Sensitisation and colonisation status of Malassezia in patients with atopic dermatitis 168 Frequency of selection MRSA-strains in composition of a microbiocenosis of a skin the patient_s atopic dermatitis Yuriy Turin, Dmitry Dolbin, Sergey Kulikov, and Olga Tupkina. Kazan scientific research institute of epidemiology and microbiology, Rospotrebnadzor, Kazan, Russian Federation. Background: to Define a share of allocation MRSA-strains and structure of a microbiocenosis of a skin of patients an atopic dermatitis. To define properties staphylococcus with skin of patients. Methods: 63 patients an atopic dermatitis are examined at the age from 3 till 35 years. Degree of a gravity of the patients defined with use of the international standard -index SCORAD (Scoring of Atopic Dermatitis). Research of microbiocenosis of skin is carried out bacteriological method. 155 strains staphylococcus with skin of patients are investigated. Are checked up on stability to antibiotics by method of disks on cups Petri. Activity catalases in lysates staphylococcus by a colorimetric method on Sinha A. K. The statistical analysis of results is executed by means of statistical programs Microsoft Excel 2000 and ?Statistica 6X StatSoft. Results: As a part of a skin microbiocenosis in 84, 2% was Staphylococcus aureus, in 15, 8% was coagulase negative staphylococcus. High frequency of occurrence MRSA-strains (75%) in structure microbiocenosis skin of patients is revealed. High frequency has been found out among MRSA, repeatedly from steady pressure to antibiotics. Conclusion: As a part of a microbiocenosis with skin of patients sick an atopic dermatitis dominate MRSA-strains. Level of activity for pressure MRSA correlates catalases with their ability to survive in an organism. More poisonous MRSA also are more often allocated from patients with heavy degree (SCORAD 87, 6 T 1, 4) . Phenotypic features of coccal microflora of skin in the norm and with atopic dermatitis Sergey Kulikov, Yuriy Turin, Lira Bayazitova, Dmitriy Dolbin, Rustem Fassakhov, and Elena Sukmanskaya. Kazan Scientific Research Institute of Epidemiology and Microbiology, Rospotrebnadzor, Kazan, Russian Federation. Background: Phenotypic features of coccal microflora of skin and disease severity are correlated in children with atopic dermatitis (AD). Methods: Forty eight AD patients (age 0, 1Y16 years) were recruited. The SCORing Atopic Dermatitis (SCOARD) we used for clinical score for assessing AD symptomatology. The skin analyzed for Staphylococcus aureus and coagulase-negative staphylococci detecting. Ig-protease, collagenase, lysozyme and DNAase activities of staphylococcal isolates were studied. Methicillin-resistant Staphylococcus aureus isolates were determined by disk diffusion method. Results: S.aureus colonization increased in skin with AD with increasing SCORAD -5,2 T 1,2 log10 CFU/sm2 (SCORAD = 35,5 T 5,5) and 5,7 T 0,8 log10 CFU/sm2 (SCORAD = 91, 9 T 5, 8) . 41% staphylococcal isolates from skin with moderate AD (SCORAD = 35,5 T 5,5) and 100% isolates from skin with severe AD (SCORAD = 91, 9 T 5, 8) had DNAase activity. Also Igprotease, collagenase activities of staphylococcal isolates from skin with atopic dermatitis were higher then from skin in the norm. Conclusion: Virulence factors of staphylococcal isolates correlated with SCOARD index and disease severity. And staphylococcal isolates from skin with AD were essentially aggressive in comparison with isolates from healthy skin The role of fecal microflora in children with allergic eczema/dermatitis syndrome Alla Nakonechna 1 , and Tatiana Umanetz 2 . 1 National Medical University, Clinical Immunology and Allergology, Kiev, Ukraine; 2 Institute of Pediatry, Obstetrics and Gynaecology, Pulmonology, Kiev, Ukraine. Rationale: The role of intestinal microflora in children with allergic eczema dermatitis syndrome (AEDS) is still disputable, although a beneficial influence on AEDS course of certain intestinal bacteria, administered as probiotics has been described in intervention studies. The purpose was to investigate the relationship between gut microbiota and sensitization and cytokines production in children with AEDS. Materials and Methods: A case-control study with 57 children (4Y7 years) having AEDS and 55 age-matched healthy control subjects was conducted. For differential diagnosis we used Skin Prick Test (SPT) with common allergens, total and specific IgE. IL-5, IL-10, +-IFN levels were assessed by ELISA. All patients had investigation of gut microflora. Results: In children with AEDS were found significantly low counts of Bifidobacterium comparing to healthy control subjects (10 5 Y10 6 vs 10 9 Y10 10 CFU/g) as well as Lactobacillus (10 5 vs 10 7 Y10 8 CFU/g) (pG0.05). At the same time atopic children presented high level of Escherichia coli, Staphylococcus aureus, Clostridium dificile, Candida albicans in investigated microflora profile. Furthermore AEDS children demonstrated positive SPT to common allergen (especially food allergen); increased level of total and food specific IgE (mostly -egg, milk, peanut, hazelnut). In this group were shown increased production of IL-5 to 192,3 T 6,1 pkg/ml (N = 74,3 T 3,3 pkg/ml); IL-10 to 153 T 9 pkg/ml (N = 5,8 T 0,25 pkg/ml) and decreased production of +-IFN to 241 T 6 pkg/ml (N = 331 T 35 pkg/ml) (p G0,05). Conclusion: This investigation demonstrated the relationship between perturbation in intestinal microbiota and IgE sensitization in children with AEDS. There is an augmentation of Th-2 type lymphocyte function reflected by increased IL-5 and IL-10 production in AEDS children. Disorder of the intestinal microflora might play a role in the onset of atopic eczema. self-healing blistering disorder of the skin caused by a small insect belonging to genus Paederus, family Staphylinidae, order Coleoptera. It appears on the skin by hemolymph of Paederus beetles. This study was undertaken to identify the dominant species of paederus beetles and to report the expose parts of the body in the study area. Methods: Peaderus beetles were collected by hand and UV black rays every month for entomological studies. For medical studies questionnaires were completed by physicians in health centers in the studied areas. Results: By this investigation, it was revealed that Paederus ilsae Bernhauer and paederus iliensis Coiffait are two species that were identified for the first time in the studied area. This research has revealed the 27.02%, 23.42%, 18.01%, 17.11% and 14.41% cases of linear dermatitis occurred in face, neck, eyes, hands and legs respectively in kazeron area. Furthermore, 42.6% of cases also had secondary infections later. Conclusion: Public awareness of how to avoid contact with the adult beetles is considered to be the best way to reduce the cases of linear dermatitis. A new cause of facial eczema: the ferret Barthelemy Splingard, Jean Pol Dumur, Jean-Luc Schmutz, Annick Barbaud, and Nancy Barbaud. CHU Nancy, Dermatology, Nancy, France. Observation: a 36 year old patient has had for one year a face eczema, mainly on eyelids and lips. She had no history of allergy and no medication. She was a beautician and suspected a professional sensitization but the eczema persisted during her holidays. She had been exposed to numerous perfumed products in her institute for six years. For many years, she had had horses and pets: dogs, cats, and for two years a ferret. Patch-tests of the standard, cosmetic product, methacrylate and fragrance series, with the products brought by the patient (creams, perfumes, household and professional products) were negative at 48 and 96 hours. Pricktests with aeroallergens, molds, latex and 28 foodstuffs were negative. Specific IgE for ferret were undetectable with normal total IgE. The ferret was given to one of her friend for two months and the eczema dramatically improved. When she had a short new contact with the ferret the eczema relapsed. Conclusion: a few cases of asthma or urticaria caused by ferrets have been reported but there is no former case of eczema. The ferret belongs to the fissiped family (mink, marten, ermine and polecat). Theorically a cross reactivity between them could occur but no case has been yet reported. The culprit allergens are not well known (albumin?). Ferrets, like felides, lick themselves and the salivary proteins could be the allergens (protein contact dermatitis?). We emphasize that sensitization to ferret, that is a new pet, has to be evoked in case of eczema or asthma, even when specific allergological tests are negative. Allergic contact dermatitis due to Propolis Alejandro Joral 1 , Jose Antonio Navarro 1 , Jose Francisco Garmendia 1 , Olga Villarreal 2 , Ascensión Aranzabal 3 , and Susana Lizarza 1 . 1 Hospital Donostia, Allergy Section, San Sebastian, Spain; 2 Hospital de Mendaro, Allergy Unit, Mendaro, Spain; 3 Hospital de Zumárraga, Allergy Unit, Zumárraga, Spain. Propolis is a product made by bees from various vegetal sources, mainly from the resin of the poplar; it is well known as an inducer of occupational dermatitis in apicultors. Its use as a natural cosmetic is widespread. given its antiinflammatory and antiseptic properties. We present the case of a 24 years old woman without an atopic background who presented an eczematous, itchy rash in the areas where she had used a cosmetic that had Propolis among its ingredients. Patch test were performed with True Test series, the cosmetic series and Propolis. The tests were positive for Propolis (+++ at 48 and 96h), and were negative for the other contact allergens, including Balsam of Peru. The sensitizing capacity of Propolis is mainly attributed to the methylbutenil cafetate and to phenylethil cafetate mainly. It has crossreactivity with the Balsam of Peru with which it shares at least 13 components. We therefore present an allergic contact dermatitis to Propolis in a "natural" cosmetic in a patient who is not an apicultor and who has no relationship with this hobby. Adverse reactions to dark chocolate -a case report Christina Oberhuber 1 , Sonja Gaier 2 , Eva Untersmayr-Elsenhuber 2 , and Karin Hoffmann-Sommergruber 2 . 1 Biomay, Vienna, Austria; 2 Medical University of Vienna, Department of Pathophysiology, Vienna, Austria. Background: Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by pruritus, inflammation, lichenifications, and typically distributed eczematous lesions. Environmental factors as well as allergens from various sources may trigger skin inflammation. The exact cause of AD is unknown, but immunological and psychological factors should be taken into account. We report a case of a 28-year old female suffering from atopic dermatitis with a history of recurrent episodes of pruritus, urticaria and facial angioedema starting 6-12 hours after eating dark chocolate pralines. With the elimination of milk chocolate and dark chocolate from the diet no more episodes occurred. White chocolate is well tolerated and consumed frequently. Methods: Skin prick tests to commercial extracts of common aeroallergens and food allergens were performed. Furthermore prick to prick tests using white chocolate, milk chocolate, dark chocolate (65% cocoa powder and 85% cocoa powder), and cocoa powder were carried out. Specific IgE, IgG1 and IgG4 levels to a large panel of purified food allergens from milk, egg, peanut, hazelnut, soy, peach, apple, celery, and chocolate extracts were determined by ELISA experiments. Results: Skin prick tests with commercial extracts were negative. IgE and IgG1 levels to various food allergens and chocolate extracts were not increased compared to non atopic individuals, whereas IgG4 levels to all tested allergens and chocolate extracts were significantly higher than in control sera. Prick to prick tests with chocolate brands revealed erythema with increasing intensities according to higher contents of cocoa powder. White chocolate (without cocoa powder) did not elicit any skin reactions. Conclusion: This case report illustrates the complexity of diagnosing adverse reactions to foods. Ingestion of dark chocolate elicits angioedema and AD, while white chocolate is tolerated. Even in the absence of allergen-specific IgE, an allergen-specific Th2 reactivity (high IgG4 levels) can be observed. Case report-contact dermatitis to perfume Nicoleta Cimpean, Cristina Barbinta, Adriana Bujor, and Diana Dumitrascu. 3rd Medcial Clinic Cluj-Napoca, Allergology, Cluj-Napoca, Romania. Background: Dermatitis is an inflammation of the skin. Contact dermatitis is a localized rash or irritation of the skin caused by contact with a foreign substance. Substances that cause contact dermatitis in many people include "poisonous" plants such as poison ivy, certain foods, some metals, cleaning solutions, detergents, cosmetics, perfumes, industrial chemicals, and latex rubber. Allergic contact dermatitis is a type IVa1 T-cell-mediated hypersensitivity. Case Report: A 26-year-old female with clinical signs of persistent allergic rhinitis (sneezing, itching and rhinorrhea). She presented a severe eruption with intense itching after using her new perfume. Clinical aspect of skin lesions were: erythema, papules and vesicles on the neck and chest with augmentation of symptoms after solar exposure. The manifestations have begun after 24 hours of exposure to the new perfume. Results: Skin prick tests revealed an atopic patient with sensitization to Alternaria. Skin patch tests with european standards to cosmetic fragrances (fragrance mix, balsam of Peru), cosmetic preservatives (paraben mix, quaternium) and other cosmetic ingredients (colophony, thiomerosol, ethylenediamine, formaldehyde) and her own perfume revealed late sensitization (72 hours) to her perfume and fragrance mix. We recomanded to avoid contact with known allergens and treatment with H1 anihistamine and topical steroids. Conclusion: The patient with contact dermatitis may be very uncomfortable and have poor quality of life. The best treatment is to identify and avoid the substances that may have caused the allergic reaction. Influence of Psidium guajava tea for atopic dermatitis Itsuo Suzuki, and Masaru KIsida. International University of Health and Welfare, Pediatrics, Tokyo, Japan. I previously reported the antihistaminic effect and anti-leukotriene effect of Psidium guajava(PG) which was a native plant of North America. Thus I made PG tea and perviewed an effect to atopic dermatitis. Methods: Seventy-five patients with atopic dermatitis who were treating in Sanno Hospital,were enrolled in this study. Ages of them 7 to 34 years old. PG tea made 0.5g with hot water of 200ml and drank it three times per day. After one week observation without any treatment, I used PG tea for fours weeks and skin condition. And I measured eosinophil cationic protein (ECP) values and histamine levels of blood. Results: Forty-two (56%) improved itching of skin and Thirty (40%) improved skin condition. ECP valued in serum were decreased from 82.4 T 42.4ng/ml to 21.8 T 23.4ng/ml. The Histamine levels in serum decreased from 211.0 T 50.4ng/ml to 98.6 T 52.4ng/ml. Conclusion: I investigated the effects of PG tea for atopic dermatitis. This study showed it was able to improve the clinical features of this disease significantly. Furthermore,the ECP values and histamine levels in serum which are indexes of allergic inflammation were decreased. I concluded from these results that Pg tea is an Effects method for the treatment of atopic dermatitis. The role of contact sensitization in contact dermatitis patients among the seaweed farmers in Bantaeng, South Sulawesi, Indonesia Atopic dermatitis remains one of the serious problems of pediatrics as prevalence of disease among children is quite high (10Y20 % according data of different countries) and have increasing tendency. The aim of the study was to assess the effects of probiotics on clinical course of atopic dermatitis in children. We studied 38 patients aged 1Y6 year, who admitted at M. Guramishvili Pediatric clinic (2006Y2007). All patients fulfilled the criteria (at least 3 major and 3 minor) for diagnosis of atopic dermatitis (Hannifin and Rajka). The children were divided into 2 groups. The first group (20 children) for 8 weeks received probiotic (Lactobacterine containing Lactobacillus acidophilus and BifidumBacterine containing Bifidumbacterin,) and emollient ointment the second group (18 children) only emollient ointment. The SCORAD index was used to evaluate severity and extent of AD at the end of the study of disease. Objective signs -spread of lesions, intensity (erythema, edema, oozing, excoriation, lichenification, and xerosis) and subjective signs (pruritus and sleeping disorders) were assessed. The mean SCORAD index before treatment was 45, 9. After the 4 week as well as 8 week treatment was founded improvement of clinical (SCORAD index decreases) in both groups. At the same time the reduction in the SCORAD index was significant in the probiotic group. After the 8 week of study 65% of children had a better SCORAD index then baseline, comparing to the second group improvement was in 43%. So, we can conclude that supplementation with probiotics is beneficial in improvement severity of AD in children. Influence of peeling procedure in allergic contact dermatiitis Jun Young Lee, and Jung Eun Kim. St. Mary's Hospital, The Catholic University of Korea, Dermatology, Seoul, Republic of Korea. Background: The prevalence of allergic contact dermatitis (ACD) in patients previously undergone peeling has been rarely studied. Objectives: We compared the frequency of positive patch test (PT) reactions in a patient group with a history of peeling, to that of a control group with no history of peeling. Patients and Methods: The Korean standard series and cosmetic series, was performed on a total of 262 patients. Sixty-two patients had previously undergone peeling, and 200 patients did not. Results: The frequency of positive PT reactions on Korean standard series was significantly higher in the peeling group compared to that of the control group (p G0.05, Chi-square test). However, the most commonly identified allergens were mostly cosmetic-unrelated allergens. The frequency of positive PT reactions on cosmetic series in the peeling group was higher than that of the control group, but lacked statistical significance. There was no relationship between the frequency of peels and the frequency (%) of positive PT reactions or the total number of positive PT results. Conclusion: It appears peeling may not generally affect the development of contact sensitization. Further work is required focusing on the large scale prospective studies by performing a PT before and after peeling. The therapeutic efficacy & safety profile of allergenic extracts(EA) in in treating patients with labial lichen planus M. Ishaq, I. Khan Sameera, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Introduction: Lichen planus an uncommon skin complaint, thought to be due to an abnormal immune reaction provoked by a viral infection (such as hepatitis C),or a drug. Inflammatory cells seem to mistake the skin cells as foreign and attack them. In 80Y85% of cases it clears from skin surfaces within 18 months but when affecting the mouth may persist longer. Methods: The classical lichen planus is characterized by shiny, flat-topped, firm papules varying from pin point size e.g.guttate to larger than a centimeter. The lesion had shinny flat-topped, firm papules varying from pinpoint to much larger sized lesions. In the study concerned 2 individuals were included. Both had been medicated with different empirical therapies. There was a remitting/ relapsing course patients were treated with a combination regimen of anti histaminic & gradual lowered dilutions of dust mite therapeutic extracts over a period of 4 months. The remission of the lesions were evident in the form of a reduction in the intensity of itchiness, ooze & redness. The final outcome was evident in the form of scar formation. Results: In the ongoing trials the therapeutic response was dramatic in female than in the male patients. Conclusion: Lichen planus a chronic intractable condition with multiple causes some times with a diagnostic dilemma. A detailed drugs/dietary history supplemented by immunological assessment provides diagnostic clue. *1Pre-treatment *2Skin prick test profile *3Post treatment presentation. Tacrolimus ointment is effective and well tolerated for the treatment of atopic dermatitis in asian countries 9 Changi General Hospital, Dermatology, Singapore, Singapore; 10 Astellas Pharma Inc., Department of Asia International, Tokyo, Japan. Background: Tacrolimus ointment is used for the treatment of atopic dermatitis (AD) in adult and pediatric patients worldwide. However, no studies have been reported on the efficacy and safety in a large population of Asian patients with AD. The aim of this overview is to survey the assessments of the efficacy and safety conducted in the patients across 8 Asian areas, China, Indonesia, Korea, Malaysia, Philippines, Singapore, Taiwan and Thailand. Methods: We analyzed the combined data of patients with AD from studies conducted in the Asian areas. Adult and pediatric patients with moderate to severe AD were enrolled. Adult patients applied 0.1% or 0.03% tacrolimus ointment and pediatric patients applied 0.03% twice daily for 3 to 4 weeks. The primary efficacy end point was Physician's Global Evaluation of Clinical Response (PGE). Other evaluations included Eczema Area Severity Index (EASI), Percent Body Surface Area affected (%BSA), Patient's Assessment of Itch and Patient's Assessment of Overall Response, and each category of Dermatology Life Quality Index (DLQI) and children's DLQI (CDLQI). Results: More than 800 patients were included in these Asian studies. Success based on PGE was observed in 83.6% of all patients. The improvement in Patient's Assessment of Overall Response was similar to PGE. Percent BSA affected, EASI and Patient's Assessment of Itch improved at weeks 1, 2, and at the end of treatment. The decrease of Patient's Assessment of Itch was greatest compared with decrease of %BSA affected and EASI. All DLQI subscales (Symptoms and feelings, Daily activities, Leisure, Work and school, Personal relationships, and Treatment) improved, and also the subscales of CDLQI (Symptoms and feelings, Leisure, School or holidays, Personal relationships, Sleep and Treatment) improved at the end of treatment. In particular, Symptoms and feelings showed a marked decrease in both age groups. In subscales of CDLQI, Sleep also had marked improvement. Improvement in the PGE was associated with the decrease in %BSA, EASI and Patient's Assessment of Itch, and also with DLQI/CDLQI. Adverse events frequently reported were skin burning and pruritus at the application site. The incidence of skin burning was lower in pediatric patients than that in adults. No serious adverse events were reported from the studies. Conclusion: Tacrolimus ointment is effective and well tolerated in the treatment of patients with AD in Asian countries. Downmodulatory effect of epinastine on Th2 chemokine production by epidermal Langerhans cells Kazunari Sugita, Miwa Kobayashi, Tomoko Mori, Kenji Kabashima, and Yoshiki Tokura. University of Occupational and Environmental Health, Department of Dermatology, Kitakyushu, Japan. Background: Epinastine belongs to the second generation of antihistamines and also possesses anti-inflammatory or anti-allergic properties in cutaneous as well as systemic allergy, including mast cell stabilization, suppression of costimulatory molecule expression, inhibition of eosinophils chemotaxis and granulocytes accumulation, and suppression of cytokine and chemokine production. In Japan, systemic preparation of epinastine has been approved for not only atopic dermatitis but also pruritic psoriasis vulgaris. Skin is a wellorchestrated immune organ where epidermal Langerhans cells (LCs) function as antigen-presenting cells and keratinocytes serve as producers of various cytokines and chemokines. We have previously reported that epinastine suppresses the production of Th1 chemokines and RANTES/CCL5 by keratinocytes. Since keratinocytes and LCs are the main epidermal sources of Th1 and Th2 chemokines, respectively, the effect of epinastine on the Th2 chemokine production by LCs is an issue to be clarified. Methods: We prepared epidermal cell suspensions from BALB/c mice and enriched them for LCs. The LC-enriched epidermal cells were cultured with epinastine for 48 h, and the expression levels of Th2 chemokines, TARC/ CCL17 and MDC/CCL22, were assessed by real-time PCR analysis. Results: The expression of CCL17 was depressed by epnastine as low as 10-6 M, and CCL22 expression was also inhibited by epinastine 10-7 M or more in a dose-dependent manner. Conclusion: Our study suggests that epinastine exerts not only an antihistaminic action but also a downmodulatory effect on Th2 cell migration toward the epidermis by inhibiting LC production of Th2 chemokines. Taken together with the previous finding that epinastine downregulates Th1 chemokine production by keratinocytes, we conclude that this antihistamic drug has an inhibitory potential for migration of both Th1 and Th2 cells in the skin. Study of efficacy of epinastine hydrochloride in patients with pruritic dermatosis and chronic urticaria and its effects on QOL of the patients Akiko Miyata, Mika Yamamoto, Yoshiaki Hayashi, Mariko Nanbu, Noboru Nakagawa, Tatsuya Tsuda, Shoichiro Minami, and Kiyofumi Yamanishi. Hyogo College of Medicine, Dermatology, Nishinomiya, Japan. Epinastine hydrochloride (Alesion\, Nippon Boehringer Ingelheim Co., Ltd.) 20 mg once daily was administered to patients aged 15 years and older with pruritic dermatosis or chronic urticaria for 4 weeks, and its efficacy and effects on the patients' QOL were assessed. This study enrolled 57 patients in total, 35 patients with eczema/dermatitis, 6 patients with cutaneous pruritus, 7 patients with urticaria, and 9 patients with other pruritic dermatosis, and their mean age was 56. The endpoints were the severity of pruritus, the severity of skin eruption, visual analog scale (VAS) for pruritus, and QOL of the patients (symptoms, emotional status, daily living, and leisure, work/school, and treatment). Concerning general improvement, remarkable and moderate improvement was achieved in 43% and 33% of all patients, respectively. The severity of pruritus indicated on VAS was significantly decreased, and QOL of the patients was also improved. As a possible adverse drug reaction, an increase in AST was observed in 1 patient. These results show that oral epinastine hydrochloride once daily appears to be safe and beneficial for symptoms and QOL of patients with pruritic dermatosis or chronic urticaria. A randomised, double-blind, placebo-controlled study of the efficacy and tolerability of a Chinese herbal medicine concoction for atopic dermatitis Background: There has been considerable interest in traditional Chinese herbal medicine (TCHM) as a treatment for atopic dermatitis (AD). A twicedaily concoction of an ancestral formula containing five herbs has been found to be beneficial in an open study. The five herbs include Flos lonicerae (Jinyinhua), Herba menthae (Bohe), Cortex moutan (Danpi), Rhizoma atractylodis (Cangzhu) and Cortex phellodendri (Huangbai). We demonstrated that there was no corticosteroid (CS) or CS related compound in this formulation. Objective: To assess the efficacy and tolerability of the concoction in children with AD. Methods: Following a 2-week run-in period, children with longstanding moderate-to-severe AD were randomised to receive a 12-week treatment with twice daily dosing of three capsules of either TCHM or placebo. The SCORing Atopic Dermatitis (SCORAD) score; Children Dermatology Life Quality Index (CDLQI), allergic rhinitis score, requirement for topical corticosteroid and oral antihistamine were assessed before, and at week 4, 8, 12 and 16 after treatment. Adverse events, tolerability, haematological and biochemical parameters were monitored during the study. Results: Eighty-five children with AD were recruited. Over 12 weeks, the mean SCORAD fell from 58.3 to 49.7 in the TCHM group (n = 42; p = 0.003), and 56.9 to 46.9 in the placebo group (n = 43; p = 0.001). However, there was no significant difference in the scores at the corresponding time points between the two groups. The CDLQI in TCHM treated patients were significantly improved compared with patients receiving placebo at end of the 3-month treatment and 4 weeks after stopping therapy (p = 0.008 and 0.059, respectively). The total amount of topical corticosteroid used was also significantly reduced by one-third in the TCHM group (p=0.024). No serious adverse effects were observed between the groups. Analysis of biochemical data also revealed no significant change in the IgE levels, haematological (complete blood counts, eosinophil counts) and biochemical (electrolytes, renal and liver functions) parameters monitored. No patient complained that the capsule was unpalatable. Conclusion: The TCHM concoction is efficacious in improving quality of life and reducing topical corticosteroid usage in children with moderate-to-severe AD. The formulation was palatable, well tolerated and can probably be used as an adjunct treatment for children with refractory AD. Histamine (H1R) blocker inhibits histamine-induced collagen synthesis in dermal fibroblasts Shun Kitaba, Hiroyuki Murota, and Ichiro Katayama. Course of Integrated Medicine Graduate School of Medicine, Osaka University, Dermatology, Osaka, Japan. Background: Mast cell-derived histamine is known to act on dermal fibroblasts and contribute to formation of an intractable chronic allergic dermatitis. Although this fibrotic event may also occur in other organs such as nasal mucosa, no direct evidence has been reported as to whether responsiveness to histamine by fibroblasts derived from different organs is of the same intensity. Furthermore, while histamine (H1R) blocker has been proved to be effective for alleviation of the symptoms of allergic diseases, its ability to affect histamine-induced tissue remodeling has not yet been clarified. Objective: Our aim was to study the effect of H1R-blockers on histamineinduced tissue remodeling. Methods: A macro array assay was used for a comprehensive analysis of histamine-induced gene expression by normal human fibroblasts. Fibroblasts derived from skin or nasal mucosa were cultured in the presence of various concentrations of histamine, and the synthesis of type 1 collagen was measured by means of semi-quantitative RT-PCR and ELISA assay. To determine the effect of the H1R blocker, diphenhydramine hydrochloride and emedastine difumarate were investigated in this assay. Results: Histamine induced various kinds of fibrogenic molecules from fibroblasts. Increased type 1 collagen expression was observed in fibroblasts treated with high-dose (j4 to j6 logM) and low-dose (j12logM) histamine. This histamine-induced type 1 collagen synthesis was effectively diminished by emedastine difumarate. Conclusion: We found that the expression of these fibroblast-derived genes are regulated differently by different concentrations of histamine, and that the robustness of the inhibitory action of H1R blockers is different for skinderived and nasal mucosa-derived fibroblasts. We believe that our findings may contribute to a better understanding of the mechanisms of histamineinduced tissue remodeling, and provide information useful for the management of refractory allergic dermatitis. Attenuating effect of fexofenadine hydrochloride on the development of cutaneous inflammatory responses through the inhibition of substance P production in mice Kazuhito Asano 1 , Ken-Ichi Kanai 2 , Atsuko Furuta 2 , and Harumi Suzaki 2 . 1 Showa University, School of NRS, Division of Physiology, Yokohama, Japan; 2 Showa University, School of Medicine, Department of Otolaryngology, 1-5-8 Hatanodai, Shinagawa-Ku, Tokyo, Japan. Background and Purpose: Fexofenadine hydrochloride (FEX), a secondgeneration antihistamine, is used for the treatment of both allergic diseases and inflammatory skin reactions and successful results are reported. The primarily therapeutic mode of action of FEX is generally believed to be owing to its suppressive effect on effector cell activation, which responsible for the development of allergic inflammation. Recently, neuropeptide such substance P (SP) attracts attention as the essential factors that trigger, exacerbate or modulate allergic skin reactions. However, the influence of FEX on neuropeptide-induced inflammatory skin reaction is not fully understood. Methods: NC/Nga mice (8-weeks of age, female) were received repeated local application of 2,4,6-Trinitrochlorbenzene (TNCB) to provoke chronic cutaneous inflammation. FEX at a single dose of 1.0 mg/kg was orally administered into mice once a day for 2 weeks from either 1 day before or 1 week after the application of TNCB. The number of scratching for 5 min was counted 30 min after the final FEX administration. After counting the number of scratching, the SP content in skin tissues and serum IgE levels were examined by ELISA. Results: Repeated local application of TNCB induced itching skin lesions, together with an increase in the levels of SP and IgE. Treatment of mice with FEX caused significant decrease in the levels of SP, but not IgE. Scratching behavior was also decreased significantly by the treatment of mice with FEX: the number of scratching in non-treated mice was 80 T 13 and that in FEXtreated mice was 48 T 9. Conclusion: These results may suggest that FEX inhibits the scratching behavior observed in chronic cutaneous inflammation through suppression of SP production in the skin lesions. The efficacy and safety of levocetirizine 5mg vs. cetirizine 10mg in patients with dermatitis or eczema with pruritus: A multi-center, double-blind, double-dummy, radomized, active-controlled study Background: Levocetirizine is a lately developed selective H1 antagonist. The purpose of this study is to assess the efficacy and safety of levocetirizine 5 mg comparing to cetirizine 10 mg in patients suffering from dermatitis or eczema with pruritus (moderate-severe). Method: This study was conducted in randomized, 6 centers, parallel, doubleblind, and double-dummy. After screening for 3 to 7 days, subjects were randomly assigned to either levocetirizine or cetirizine treatment group and each were administered for 2 weeks with 1% topical hydrocortisone. Patients visited at screening (visit 1), randomization (visit 2), 7 days after randomization (visit 3) and 14 days (visit 4). The pruritus severity score was assessed by patients using the 4-point score scale (0; none, 1; mild, 2; moderate, 3; severe) and responders were defined as patients who reported a post treatment score rating of "None" or "Mild" at the visit 4 or at the study completion visit. Results: 506 patients were screened and 466 patients of these were randomized. 423 patients completed the study and 340 patients were adequate for efficacy evaluation. The proportion of responders was 77.98% (131/168) in the levocetirizine treated group and 77.91% (134/172) in the cetirizine treated group. Three patients (1.44%) in the levocetirizine treated group and six patients (2.80%) in the cetirizine treated group had drug related adverse events. Unexpected adverse drug reaction was not reported in both groups. Conclusion: Levocetirizine was non-inferior to cetirizine for the improvement of pruritus in patients with dermatitis or eczema with pruritus. Levocetirizine is safe and efficient treatment in patients with dermatitis or eczema with pruritus. Background: The aim of study was to investigate the sensitization to house dust mite (HDM) and S. aureus in atopic dermatitis patients with recurrent pyoderma (AD RP patients) and investigate the efficacy and safety of specific immunotherapy with staphylococcal and HDM allergovaccines in these patients. Materials and Methods: The group of patients consisted of 156 AD RP patients. Intradermal tests with Staphylococcal allergen and prick tests with Der. pteronissinus and Der. farinae allergens were carried out in all AD RP patients. Skin testing was carried out in all AD RP patients in remission period. Specific IgE levels to Staphylococcal enterotoxin B as well as to Der. pteronissinus and Der. farinae in sera of AD RP patients were measured with UniCap 100 system, Phadia. 28 AD RP patients with positive skin tests and positive specific IgE to HDM and staphylococcal enterotoxin B received specific immunotherapy (SIT) with staphylococcal and HDM allergovaccines. Subcutaneous injections of allergen and vaccine were made daily, in increasing doses. The duration of the main course of specific immunotherapy was 10Y14 days and then one maintenance dosage every 2 weeks; further observation period lasted 12 months. Results: We observed sensitization to HDM in 83,9% AD RP patients, to S. aureus in 66,6% patients, both to HDM and S. aureus in 63,5% patients. There was positive effect of SIT with HDM and staphylococcal vaccine. After 1,6,12 months of the further observation period index SCORAD decreased in 26,1%, 58,3%, 44,8% accordingly. There were no exacerbations of RP in 42,8% of patients. The topical corticosteroids and antibiotics decreased significantly. Index of respiratory symptoms decreased significantly too. While treatment mild local reactions, such as redness, edema, and flare were observed in 47,6% patients during Exacerbations of AD Y in 9,5% and RP Y in 4,8% patients were observed. Inspite of adverse events none of the patients were excluded from the study. The levels of specific IgG4 in sera against HDM and S.aureus increased significantly. The levels of specific IgE in sera to HDM and S.aureus did not change. Conclusion: According to our data HDM and S. aureus are the most common offending allergens in AD RP patients. Thus SIT application in these patients is needed. SIT in AD RP patients with HDM and staphylococcal vaccine was effective and safe. Topical application of epigallocatechin-3-gallate improves prolonged atopic dermatitis by suppressing macrophage migration inhibitory factor Background: There are some growing evidences that epigallocatechin-3gallate (EGCG) exerts anti-inflammatory effects on the chronic inflammatory skin conditions such as atopic dermatitis (AD). Macrophage migration inhibitory factor (MIF) has been recently suggested to be one of the most crucial immunoregulatory cytokines of Th1/Th2 imbalance in AD. In this study we assessed the anti-inflammatory effect of EGCG by its topical application to the AD skin lesions in NC/Nga mouse model, and then we determined whether this effect of EGCG is mediated by immunoregulatory cytokines including MIF. Methods: Induction of AD skin lesions was made by painting Dermatophagoides pteronissinus extract (DPE) onto the surface of each ear of NC/Nga mice. One group was then treated with EGCG solution, another group with vehicle, and the other group sham-treated for 4 weeks. To compare the clinical signs of the 3 groups, we measured ear thickness every week. At the last day, all the ears were excised and prepared for the following procedures; 1) hematoxyline and eosin stain, 2) immunohistochemistry for MIF, TNF-! and IFN-+ 3) quantitative reverse transcription polymerase chain reaction for MIF, TNF-!, IFN-+, IL-2, IL-4 and IL-12. Blood sampling was also performed for enzyme-linked immunosorbent assay for MIF and total IgE in serum. Results: As compared to vehicle treatment, topical application of EGCG significantly reduced the ear thickness (pG0.05). Likewise, the histological grading of chronic inflammation including epidermal hyperplasia and mononuclear cell infiltration into the dermis was also significantly decreased on the skin lesions of EGCG group. On the immunohistochemical staining, in comparison with vehicle treatment, EGCG treatment significantly diminished the expression of TNF-!, IFN-+ as well as MIF on the lesions (p G 0.05). Similarly, the mRNA expression of MIF, TNF-!, IFN-+, IL-2 and IL-12, but not IL-4 of the lesions was significantly down-regulated by EGCG treatment (p G 0.05). Serum MIF and total IgE production was also significantly reduced with the EGCG. treatment (p G 0.05). Conclusion: These results demonstrated that topical application of EGCG could reduce the chronic inflammation of AD lesions by suppressing a series of immunoregulatory cytokines including MIF. Taken together, it is suggested that topical application of EGCG can be a potential therapeutic modality in AD. Anti-IgE treatment in a patient with hyper-IgE syndrome and hepatitis c virus Infection S. Sánchez-Ramó n, A. Paravisini, R. Aviles, M. Aldeguer, and E. Fernandez-Cruz. Hospital General Universitario Gregorio Marañon, Immunology, Madrid, Spain. Background: Hyper-IgE syndrome (HIES) is a primary immunodeficiency characterized by recurrent infections, dermatitis and elevated serum IgE. The underlying cause of HIES is unknown and the management of the patient is difficult, the general goals are to control pruritus, eczema, and to prevent infections. We report a patient with typical features of HIES and hepatitis C virus (HCV) infection, in whom anti-IgE monoclonal therapy for allergic manifestations could be an effective therapeutic alternative. Methods: A 29-year-old woman of non-consanguineous parentage presented with moderate asthma and severe atopic dermatitis since early infancy. She had a history of recurrent respiratory infections, delay of shedding of primary teeth and slight facial dysmorphia. She had persistently elevated serum IgE levels (3000 kU/L). She suffered relapses of multiple discrete and confluent erythematous scaly papules and plaques with follicular prominence over face, trunk and limbs, with lichenification and warm hypersensitive skin, which limited her activities of daily living. In 1999, she was diagnosed of HCV infection. Later on, she developed unusual autoimmune associations: haemolitic anaemia, alopecia universalis and autoinmune hypothyroidism. The rash had been treated with antihistamine drugs and topical steroids, with poor response. Given the high levels of serum IgE, she was given Omalizumab therapy in high dose (450 mg sc) and every 15 days schedule, monitoring IgE, IgG4 levels, hemogram and biochemical analyses. Results: The patient improved her current symptoms of asthma since the first dose of Omalizumab and two months after the eczema severity decreased to a mild form. The patient cleared almost all her skin lesions although light lichenification areas persisted in flexures, improved xerosis, lowered itching and suffered no relapses. Serum IgE level persisted elevated and there were no change in the biochemical analyses. She did not present any side effects. Conclusion: Therapy with anti-IgE monoclonal antibodies was found to be well-tolerated and effective. Although, current knowledge and therapeutic options in HIES are limited and with variable success, anti-IgE monoclonal therapy for allergic manifestations seems to be an effective and safe therapeutical alternative, even more as a steroid sparing agent considering her HCV infection. Efficacy of Omalizumab in the treatment of 3 patients with severe refractory atopic dermatitis A. Paravisini 1 , S. Sánchez-Ramón 1 , R. Suarez 2 , J. Gil 1 , and E. Fernandez-Cruz 1 . 1 Hospital General Universitario Gregorio Marañon, Immunology, Madrid, Spain; 2 Hospital General Universitario Gregorio Marañ on, Dermatology, Madrid, Spain. Background: Atopic dermatitis is a chronic cutaneous inflammatory disease mediated by type 1 hypersensitivity reaction. High IgE serum levels are commonly found and correlated with the severity of AD. Treatment of moderate-severe cases included phototherapy, cyclosporine, azathioprine, mycophenolate mofetil and intravenous immunoglobulin with variable results. Omalizumab, humanized monoclonal antibodies bind to free serum IgE and decrease the levels of its high affinity receptor. It has been successfully used in allergy asthma, rhinitis,allergic reactions to certain foods and drugs,latex and insect bites. The experience in the treatment of AD is limited and controversial. We report the preliminary results on 3 AD patients who received anti-IgE treatment. Methods: 3 adult patients(aged 29Y44) with severe refractory AD and high IgE levels9700kU/L(5.720Y12.310kU/L), were treated with omalizumab (Xolair, 450mg/kg subcutaneously every 2 weeks), 2 patients during 6 months and one patient is now on the third month of treatment. All patients had allergic rhinitis, one of them has current symptoms of asthma and other one has history of asthma. They were previously treated with topical and systemic steroids, topical calcineurin inhibitors and cyclosporine, with partial control of the symptoms. All patients were informed this was an offlabel use and the side effects profile. Results: All the patients experienced any improvement during the treatment. After two months the eczema severity decreased in all of them, from severe to moderate-mild forms. Two patients stopped all medications either antihistaminic drugs and only one of them continues taking cyclosporine. One patient obtained partial response, with clearing of lesions, lower itching and was able to restart sport; other patient suffered autolimited relapses and did not require other therapies, and finally reach complete clearance at 6-months; the last one improves her skin lesions and asthma symptoms at 2-months. This later patient diminished the dose of cyclosporine (200 to 50 mg/day). All the biochemical analyses remained normal. None of them suffered adverse reactions. Conclusion: Omalizumab did not systematically diminished serum IgE levels, but achieve significantly clinical improvement even on monotherapy at 6 months. On the other hand, Omalizumab showed a safe profile and might be a promising treatment in those patients in whom systemic therapy has not been successful. Atopic dermatitis (AD) is a chronic inflammatory skin disease whose pathophysiology is the product of complex interaction between susceptibility genes, host environments, infectious agents, defects in skin barrier and immunologic responses. Basic treatment of AD consists in topical emollients, topical and systemic CS and immunosuppressive drugs, antihistamines, phototherapy. Adjunctive HdIVIg for resistant AD may offer a useful approach, as suggested by small number of uncontrolled trials and case reports, via a number of immunomodulatory effects. Most patients with AD are colonized with S aureus and experience exacerbation after infection with this organism. In these cases treatment with antibiotics can result in reduction of skin disease. We signal the case of a 46-y-old man affected by pollens-mitesallergic rhinitis from adolescence, suffering from severe AD for 7 years with typical signs of lesional chronicity with lichenified plaques and fissuration on face and neck, upper arms and back, palm and dorsa of hands. To our attention, typical phenomena of microbial colonization were evident. The subject confirmed high sensitization to mites, grass; total IgE sum of 979 KU/L. Negative for celiac disease and food allergy, normal common haemato-biochemical parameters. Cultural microbiology of lesions demonstrated S aureus, sensible to common antibiotics. First line treatments resulted not efficacious. Short courses of oral antibiotics and CS associated to topical antibiotics obtained a decrease of flare for just few weeks; relapse occurred frequently justifying a not respondent disease to common treatments. Further analysis demonstrated a total gammaglobulins reduction related to a selective decrease of IgG class (325 mg/dl), normal lymphocyte fenotipe and proliferative response to common antigens, a mild deficit of granulocytes chemotaxis to IL8. Diagnosis of common variable immunodeficiency was formulated and HdIVIg instaured. After the first administration of 200 mg/kg the severe lesions of AD significantly improved with evident resolution of infections. First line treatment alone resulted efficient in controlling the disease in the following 7 months. Subjects affected by severe not respondent to conventional therapy AD, especially with evident infections, may reasonably be investigate for possible immunodeficiency. In these cases HdIVIg treatment may be a useful support for both immunomodulatory effect and neutralization of cutaneous pathogens. The influence of laser therapy on the functional activity of nuetrophiles While carrying out NBT-tests, we have found that in 67 patients with atopic dermatitis in the original state the number of spontaneously activated neutrophiles was 17.8 + 1.3% which is considerably higher than in healthy people (6.4 + 0.9% pG0.001). While studying the neutrophiles activation index which reflects their reactivity to extrinsic stimulation, we have found that, in the studied group, the general drop in the index was over 2.5 times larger than compared to the norm of 3.4 + 0.7 and 8.2 + 0.6 (pG0.001). Along with this, in patients with atopic dermatitis we have also registered a considerable decrease in phagocytic activity of nuetrophiles, which amounted to 59.8 + 3.3, compared to the norm of 72.0 + 3.4% (pG0.05). All the examined patients were divided into 2 groups, comparable by all criteria. Group I Y 34 patients who were treated with laser and traditional therapy, and a control group of 33 patients, treated with the traditional drug therapy. The positive dynamic of the studied parameters were registered with the application of different treatment methods; however, it was at its highest under the influence of laser therapy. Thus, in the main group the content of neutrophile leucocytes recovered to the normal quantity and dynamic of the above mentioned parameters was on average 48% higher than in the control group. Hence, this newly developed combined method of treating atopic dermatitis has been proven to have a desensitizing and anti-allergic effect, which is indicated by the recovery of the functional activity of neutrophiles. Background: Because of chronic course and highly disfigured skin lesions, atopic dermatitis (AD) has a substantial impact on quality of life (QOL). Moreover, previous studies assessing the financial cost of AD have shown that it represents a notable economic burden. However, little has been known about QOL and economic burden in Korean patients with AD. Methods: A questionnaire specifically designed by our clinical experience was used to determine how AD affects the lives of the patients and/or their parents. 66 and 39 parents completed the self-administrated questionnaires, including the questions on multidimensional aspects of QOL. They were also surveyed to find how much money they have spent in the treatment of AD during the last 6 months. All of the QOL scores are linearly transformed to 0Y100 scale, with 100 indicating the worst QOL, and 0 the most favorable. In addition, for each patient the disease severity was evaluated by using SCORAD index. Results: The QOL data revealed that AD had significantly negative effects on QOL through all aspects of life including daily living activities (37.4), psychological status (29.7) and social functioning (23.1). The average of the total QOL score was 30.1. The QOL scores significantly increase in proportion to the increasing severity of AD (pG0.0001). With regard to daily living activities, a major stress-causing factor was restriction on cosmetics use (44.5) and choice of foods (40.6) or clothes (36.7) as well as concern about exposure of their skin (41.2). For psychological status, 89% of patients reported that they feel deeply nervous about the likelihood of their offspring having AD in future (54.5). Regarding social functioning, it was told that AD patients had difficulties in making relationship with someone (26.4) and were less interested in the opposite sex (24.7). AD parents also had high level of stress associated with modifying lifestyles (33.8) and keeping their children from scratching (28.2). The estimated cost related to AD has the positive correlation with the severity of AD, widely ranging from 60,000 won to 1,990,000 won per patient a month. Of importance, the average direct medical cost was 77,000 won, corresponding to only 29% of the total, while the indirect cost represents 71% of the total with average cost of 189,000 won. Conclusion: These results suggest that AD has a significant impact on the various components of QOL as well as economic burden in Korean patients and their parents. Background: Extensive evidence has been accumulated on the role of Nerve Growth Factor (NGF) in allergic diseases. Although physical exercise has been associated, in animal models, with an increased NGF expression, no data are available about its effects on NGF in humans. Aim: To study NGF serum levels in top athletes, a population sample in which allergic diseases and neuroimmune disorders Y such as amyotrophic lateral sclerosis (ALS)Yare reported with a significant increased prevalence. Methods: 103 male soccer players (mean age 26.1) from five first division teams were studied for allergic diseases trough an original specific questionnaire (AQUAc2007), history, physical examination, skin prick tests and/or Phadiatop. Sera were taken in all athletes, at different time periods, and stored at j70-C for NGF determination trough a double-antibody ELISA. Data from a pilot sample of 36 athletes and 15 non-allergic, sedentary, matched controls are presented. Results: The prevalence of allergy in soccer players was 47.8%. In spite of a high intra-and inter-individual variability of values, possibly due to the different training loads during the soccer season, mean NGF serum levels were significantly higher in soccer players than in controls (384.19 ng/ml +/j 298.41 ng/ml vs 128.0 ng/ml +/j 10.0 ng/ml; p G0.05). The NGF serum levels in athletes were increased independently from the presence of allergy. Conclusion: Our study confirms the high prevalence of allergic diseases in athletes. The increased NGF serum levels in allergic soccer players are consistent with the role of this neurotrophin in allergic inflammation. This is the first report of increased serum levels of NGF related to intense physical activity in humans. Special features of allergy testing in elite sportsmen Nataliya Shartanova. The Institute of Immunology, Asthma, Moscow, Russian Federation. Sport is inevitable part of modern society. However up today the precise and convincing data on prevalence and features of current allergic diseases in elite Russian sportsmen are absent. The purpose of the present research is to study prevalence, features of allergen spectrum and structure of allergic diseases in elite sportsmen. The skin prick-tests with different groups of allergens, the estimation of total IgE end IgA, IgM, IgG in blood serum, the challenge bronchial tests with physical exercise are used in modern clinical, laboratory and allergic research methods including gathering allergic, the pharmacological and food anamnesis. Screening of 319 elite sportsmen was carried out (fencing, sport and art gymnastics, dzudo, softball, water polo, volleyball, figure skating, hockey on a grass, rowing on a canoe, freestyle, biathlon, a youth soccer team, football). The analysis of carried out research has shown the following. 23,5 % (75) sportsmen have clinical attributes of allergic disease: allergic dermatitis -16%, allergic urticaria -17,3%, allergic rhinitis -30,7%, pollen-allergic rhinitis -17,3%, bronchial asthma -5,3%, drug allergy -0,94%, food allergy -0,63%. Increase of total IgE level in blood serum has been revealed in sportsmen with allergic diseases (23%) from 130 to 1000 ME/ml. Immune status analysis demonstrated the decrease of IgG and IgA level and increase of IgM level in elite sportsmen without any connection to atopy. In 94,5% of sportsmen allergy was diagnosed for the first time. Obtained data testifies widespread of allergic diseases among elite sportsmen and allergic diseases hypo-diagnostics. The hypo-diagnostics of allergic diseases results in late prescription of adequate therapy and restricts sport achievements. Changes in immune status indexes require further investigation. 197 Antiproliferative and anti remodelling effect of beclomethasone dipropionate, formoterol and salbutamol alone or in combination in primary human bronchial fibroblasts Desideria Descalzi 1 , Chiara Folli 1 , Gabriele Nicolini 2 , Anna Maria Riccio 1 , Cinzia Gamalero 1 , Francesca Scordamaglia 1 , and Giorgio Walter Canonica 1 . Background: Bronchial asthma is characterized by lower airway inflammation and remodelling. Anti-inflammatory treatment with inhaled corticosteroids provides the mainstay of asthma therapy together with bronchodilation induced by short and long-acting inhaled $2-agonists. Lower airway fibroblasts may play a critical role in airway inflammation and remodelling suggesting they might represent an important target for the major antiasthmatic drugs. The aim of our study was to investigate the effects of beclomethasone dipropionate, salbutamol and formoterol either alone or in combination on in vitro cultures of human bronchial fibroblasts. Methods: Fibroblasts were cultured in the presence of proinflammatory and proliferative stimuli, beclomethasone dipropionate, salbutamol and formoterol. The effects of drugs on cell proliferation were ascertained by 3Hthymidine incorporation. CD90 and CD44 expression were detected by flow cytometry and fibronectin secretion using an ELISA technique. Results: This study showed that beclomethasone dipropionate alone has significant anti-proliferative effects on lung fibroblasts treated with bFGF while LABAs or SABAs by themselves did not show any significant effect in the different cultures. The combination of BDP with formoterol or salbutamol strengthens these effects. CD44 and CD90 expression and fibronectin production were modulated by proinflammatory and proliferative stimuli; the addition of the drugs brought them back near to the basal level. Conclusion: From this in vitro study we can conclude that beclomethasone dipropionate, when combined with salbutamol or formoterol, exhibits enhanced anti remodelling activity in bronchial fibroblasts, providing new insights on the additive effects of inhaled corticosteroids and $2-agonists for asthma therapy. Galectin-9 inhibits CD44-hyaluronan interaction and suppresses a mirine model of allergic asthma Shigeki Katoh 1 , Akira Yamauchi 1 , and Mitsuomi Hirashima 2 . 1 Faculty of Medicine, Kagawa University, Department of Cell Regulation, Kagawa, Japan; 2 Faculty of Medicine, Kagawa University, Department of Immunology and Immunopathology, Kagawa, Japan. Background: belongs to the galectin family that exhibits affinity for $ galactosides. Gal-9 has a variety of biological activities, however its role in allergic inflammation is unknown. We evaluated the effect of a stable form of the human protein on allergic airway inflammation in a mite allergeninduced asthma model. Methods: Human stable gal-9 was given by intravenous injection to mice during antigen challenge. The effect of gal-9 on airway inflammation and airway hyper-responsiveness (AHR) was then evaluated. Results: Gal-9 reduced AHR as well as T helper type 2 (Th2)-associated airway inflammation. Furthermore, administration of gal-9 as well as anti-CD44 monoclonal antibody inhibited the infiltration of peripheral blood Th2 cells into the airway. Interestingly, gal-9 directly bound the CD44 adhesion molecule and inhibited interactions with hyaluronan (HA). Consistent with the concept that CD44-HA interactions mediate the migration of T cells into the lung, gal-9 blocked CD44-dependent adhesion of BW5147 mouse T cells to HA. Conclusion: We conclude that gal-9 inhibits allergic inflammation of the airway and AHR by modulating CD44 dependent leukocyte recognition of the extracellular matrix. Suppressive effects of a water-soluble fraction from Artemisia capillaris on the ovalbumin-induced allergic asthma model Background: Asthma is a chronic inflammatory disease of the airways characterized by reversible airway obstruction and hyperreactivity and remodeling of the airways. Even though a number of medications or treatments for the disease are available, the demand for a rather mild and preventive medication using the natural products such as Chinese traditional medicine is increasing, especially, in a number of Asian countries. Artemisia carpillaris is a perennial herb easily found around the temperate Asian regions including the entire Korean peninsula. It is called BInjin[ in Korea and BYin Chin[ or BYin Chen Hao[ in China, which is traditionally used for the treatment of liver diseases and also applied to some allergic symptoms such as hives or rash. In order to identify an anti-allergic principle of the plant, we have isolated a water-soluble glycoside fraction of smaller than 1kDa and determined its biological activities on an allergic asthma model animal. Methods: We have investigated suppressive effects of the fraction on an ovalbumin-induced allergic asthma in BALB/c mice and studied the cellular and molecular mechanisms of its anti-allergic activity. Results: A water-soluble fraction of Artemisia carpillaris significantly reduced the pulmonary eosinophilia and the Th2 cytokine expressions such as IL4 and IL5 in the lungs as well as serum IgE levels. The surface expression of the CD11c and class II MHC on the lung dendritic cells was also reduced by the treatment of the fraction, indicating that the fraction also modulates the dendritic cell development in lung tissues. Conclusion: Considering that the pulmonary dendritic cells are crucial in the differentiation of Th2 cells and the production of IL4 and IL5 in lung tissues and that the cytokines are important in the IgE antibody production and eosinophil infiltration, the fraction appears to include an anti-allergic principle modulating the Th2 differentiation and the resulting allergic asthma development. Protective effect of Glu27 allele of $2-adrenergic receptor gene in thai asthmatic patients Background: Genetic polymorphisms involving a variation at the 16th (ArgY Gly) and 27th (GlnY Glu) amino acid positions of the adrenergic receptor-$2 (ADRB2) possibly associated with various asthma related phenotypes including the adverse effect on lung function after regular use of albuterol in asthmatic patients who are homozygous Arg16 and a protective effect of homozygous Glu27 on asthma severity. Objective: To determine association between ADRB2 polymorphisms and asthma phenotypes in Thai patients. Methods and Materials: One hundred and thirty asthmatic patients were genotyped for ArgGly16 and GlnGlu27 polymorphisms. Patients' demographic, disease severity, pulmonary function test results and medication used were collected. Haplotype with unknown phase was inferred using EM algorithm method. ANOVA was applied to compare means among groups. Results: Prevalence of Arg16 and Gln27 alleles were 56.9% and 91.2%, respectively. Linkage disequilibrium coefficient between the two SNPs was 0.36. Three haplotypes were inferred, which were Arg-Gln, Gly-Gln, and Gly-Glu with frequencies of 148 (56.9%), 89 (34.2%), and 23 (8.6%). Mean percentage of predicted FEV1 for these corresponding haplotypes were 73.5 (SD = 16.3), 72.4 (SD = 17.4), and 80.7 (13.1), respectively. Although the means were not statistical different (p = 0.258), the predicted FEV1 of Gly-Glu was 6%Y9% higher than the rest haplotypes. Number of hospitalization and emergency visit were also lower in GlnGlu27 than GlnGln27 genotypes (i.e., 0% versus 11.9%, p = 0.089 for hospitalization; 4.5 % versus 18.8 %, p = 0.084 for visiting emergency). Finally, inhaled corticosteroid/long-acting $2-agonist (ICS/LABA) usage was significantly lower in patients with GlnGlu27 than patients with GlnGln27 (i.e., 50% versus 76.6%; p = 0.042). Conclusion: Presence of Glu27 allele in Thai asthmatic patients is associated with better parameters of asthma severity, including higher percentage of predicted FEV1, less hospitalization and emergency department visit during the past year and significant lower amount of ICS/LABA usage. Clinical implication of vascular endothelial growth factor in children with asthma and eosinophilic bronchitis Kyu-Earn Kim, Kyung Won Kim, Hye Mi Jee, and Myung Hyun Sohn. Yonsei University College of Medicine, Pediatrics, Seoul, Republic of Korea. Background: Eosinophilic bronchitis (EB) is a condition characterized by a corticosteroids responsive cough and sputum eosinophilia, without bronchial hyperresponsiveness. Vascular endothelial growth factor (VEGF) is an important mediator of airway inflammation and remodeling in asthma. We aimed to explore whether VEGF is expressed at elevated levels in airways with asthma or EB and associated with pulmonary function and bronchial hyperresponsiveness in children. Methods: One hundred seventeen asthmatic children, 77 children with EB and 84 healthy controls (mean age, 8.9 years) were enrolled in the study. Sputum supernatants were collected, and VEGF and eosinophil cationic protein (ECP) levels were measured. We performed pulmonary function tests and methacholine challenge tests, while measuring total eosinophil count, serum total IgE and ECP in all subjects. Results: Asthmatic children had significantly higher levels of VEGF in induced sputum (1.82 T 0.55 log pg/mL) compared to children with EB (1.50 T 0.53 log pg/mL; p = .00165) or healthy controls (1.48 T 0.48 log pg/mL; p = .00034). A positive significant correlation was found between FEV1/FVC and sputum VEGF (r = 0.257; p = .006), whereas no significant correlations were found between sputum VEGF and bronchial hyperresponsiveness, sputum eosinophil count or sputum ECP. Sputum eosinophil count showed higher levels in children with asthma or EB than in controls (p G 0.001). Sputum ECP also demonstrated higher levels in children with asthma or EB than in controls (p = .044, p = .001, respectively). Conclusion: Our findings suggest that sputum VEGF might be a valuable marker to distinguish asthma from EB in children with chronic cough. It is also suggested that VEGF would affect the airway inflammation contributing to airway remodeling which may reflect in FEV1/FVC. Proportion of eosinophilic and non-eosinophilic inflammation phenotypes in patients with mild to severe bronchial asthma, its influence on main clinical and functional outcomes and efficacy of inhaled steroid treatment Maris Bukovskis 1 , Normunds Jurka 2 , and Immanuels Taivans Background: Bronchial asthma is probably not a single disease, but a complex of separate syndromes. One of the methods to standardise asthma phenotypes is identification of inflammatory pattern. Inflammation subtypes may influence treatment efficacy with ICS. Objective of our study was to determine the percentage of patients with eosinophilic and non-eosinophilic asthma phenotype, sputum inflammatory cell spectrum in each patient group, its influence on main asthma control parameters and ICS treatment efficacy. Methods: Lung function, PD20 of Mch, induced sputum cell count, asthma symptom score and $2-agonist use was investigated in non-smoking, steroid naive and ICS treated patients with mild to severe persistent bronchial asthma. The cut-off point between patient groups was defined as sputum eosinophilia 93%. Steroid naive patients were allocated to the treatment with FP 250 6g twice daily for 12 weeks and treatment efficacy on previously mentioned parameters was evaluated. Results: 21 steroid naive and 77 patients previously treated with ICS were included in our study. In steroid naive patient group 72% of patients were classified as asthmatics with eosinophilic phenotype and 28% with noneosinophilic phenotype. In ICS treatment group 39% were classified as eosinophilic and 61% as non-eosinophilic asthmatics. Patient demographic data and ICS dose was similar. In patients with non-eosinophilic asthma we found significantly higher relative and absolute sputum neutrophil count, asthma symptom score and $2-agonist use (p G 0.05). There was no significant difference between FEV1 83.1 T 18.1% vs. 87.6 T 25.6% and 90.1 T 20.7% vs. 96,3 T 12.5% in steroid naive and ICS treated patients respectively; p90.05) and lgPD20 Mch (j 1.343 T 0.765 vs. j 1.894 T 0.733 mg and j 0.729 T 0.702 vs. j 1.462 T 1.023 mg in steroid naive and ICS treated patients respectively; p90.05). 11 patients with eosinophilic and 10 with non-eosinophilic asthma were treated with FP. We observed statistically significant better improvement of FEV1 and morning PEF in patients with initial airway eosinophilia compared to non-eosinophilic asthma (p G 0.05). Change of lgPD20 of Mch, asthma symptom score and $2-agonist use was similar in both patient groups. Conclusion: Non-eosinophilic asthma phenotype is associated with increased sputum neutrophilia, more severe symptoms, higher rescue medication use and less effect of ICS treatment on lung function. Influence of anti-IgE antibody omalizumab on airway remodeling and the expression of interleukins in asthma Oleksii Korzh, Sergiy Krasnokutskiy, and Elizaveta Lavrova. Kharkov Medical Academy of Postgraduate Education, Internal Diseases and Clinical Pharmacology, Kharkov, Ukraine. Background: To study the relation between interleukin-4 (IL-4), IL-5, IL-13, transforming growth factor-beta (2) (TGF-beta (2)) and airway remodeling and to investigate the effects of omalizumab on airway inflammation and airway remodeling of asthma. Methods: Thirty five female BALB/c mice were randomly divided into a remodeling group and a treatment group (omalizumab group), with 10 BALB/ c mice in each group. The mice were sensitized by ovalbumin (OVA), and only the omalizumab group was treated with omalizumab. The number of total cells and eosinophils in bronchoalveolar lavage fluid (BALF) were counted. Light and electronic microscope were used to detect the pathologic histology and morphologic change. In situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR) were used to measure IL-4, L-5, IL-13, and TGF-beta (2) mRNAs in the lung. Results: The numbers of total cells and eosinophils in BALF of the remodeling group were (5.7 T 1.3) x 10(5)/ml and 2.43 T 0.18, while those of the treatment group were (4.1 T 1.4) x 10(5)/ml and 1.67 T 0.23, respectively, the difference being significant (P G 0.05). Histological and electronic microscopic examination showed extensive airway inflammation, notably accumulation of significant numbers of eosinophils and lymphocytes in the remodeling group. Other features including prominent proliferation of airway epithelial cells protruded like fingers, increased thickness of smooth muscle, hyperplasia of connective tissue, goblet cell hyperplasia and a marked increase in airway mucus secretion with mucus plugging and extensive collagen deposition around the airways were also noted in the remodeling group. In the treatment group, the inflammation was significantly decreased, with decreased production of mucus, decreased collagen and granule of mucus around airway, less proliferation of airway epithelium, smooth muscle hypertrophy and airway spasm. In situ hybridization showed that the expression of IL-13 mRNA and TGF-beta (2) mRNA in the lung oQf the remodeling group were 22 T 9 and 18 T 3 respectively, while those of the treatment group were 16 T 5 and 9 T 4. Conclusions: Omalizumab could effectively inhibit airway remodeling and decrease in the expression of IL-13 mRNA and TGF-beta (2) mRNA as well as IL-4 mRNA and IL-5 mRNA in the lung in asthma. IgE and IgG4 autoreactivity in bronchial asthma Anna Konischeva, and Valentina Gervasieva. Mechnickov Research Institute for Vaccines and Sera, RAMS, Moscow, Russian Federation. Currently, it pays much attention to the phenomenon of autoreactivity in the immunopathogenesis of allergic diseases. The chronic allergic inflammation in the bronchial tree leads to the tissue injury, which results in the modification of its structure and releasing intracellular autoantigens. It was discovered IgG -autoantibodies to DNA and bronchial endothelium cells antigens in patients with bronchial asthma (BA). Goal: to determine the content of IgE and IgG4-autoantibodies (Ab) to some tissue antigens in patients with atopic BA different degrees of severity. Materials and Methods: We identified IgE-autoAb and IgG4-autoAb with ELISA to the following tissue antigens: epithelial keratin, III and VI collagen types, myelin basic protein (MBP), elastin and myosin (Sigma, USA) using specific IgE and IgG4 reference sets BDr. Fooke[ (Germany). It was studied serum samples of 64 adult asthmatic patients and 25 healthy individuals the same age. Results: The level of IgE-autoAb determined from control subjects was taken us for the average homeostatic norms and it was 1.5 T 0.07 IU/ml. Compared to them, in patients with easy and middle BA was noted reliable elevation of IgE-autoAb to keratin (13.7 T 4.3 IU/ml), collagen III and VI types (4.4 T 2.1 IU/ml, 6.8 T 3.2 IU/ml), elastin (4.9 T 2.4 IU/ml) and myosin (18.21 T 11.5 IU/ml). In patients with severe asthma were raised autoAb only to elastin (2.2 T 0.7 IU/ml), collagen type III (1.7 T 0.2 IU/ml) and myosin (7.5 T 3.1 IU/ ml). At the same time, increased total IgE correlated with IgE-autoAb to elastin and myosin (r = 0.68 and r = 0.45, respectively). IgG4 auto-Ab were also determined in the 35 serum samples of severe and middle asthmatic subjects and in the serum of 25 healthy people. Its normal average value was 2380 T 89.5 ng/ml. The level of IgG4-autoAB to all tissue antigens in asthmatic patients was a 1.5 times higher than in healthy controls. We also revealed significant negative correlation between IgG4 and IgE-autoAB to elastin (r = j0,7), III collagen type (r = j0,56), MBP (r = j0, 9) . So the data indicate that IgE and IgG4-autoreactivity contributes to the development of chronic inflammation in patients with atopic BA and needs to be further investigated. Changes of internal diameter of large airway during and after exacerbation of bronchial asthma Sang-Hoon Kim, Byoung Hoon Lee, and Jae Hyoung Lee. Eulji Hospital, Internal medicine, Seoul, Republic of Korea. Background: Exacerbation of bronchial asthma was characterized by increased airway responsiveness induced by lower airway inflammation and resultant airway obstruction. Small airways are known to be the main area of obstruction which causes limitation of airflow, but inflammatory change during exacerbation occurs in entire airway including large airway. We evaluated the changes of internal diameter of large airway during and after exacerbation. Methods: High resolution computed tomography (HRCT) and pulmonary function test were done as soon as possible after admission in patients with exacerbation of asthma and follow up studies were done after improvement of symptoms and discharge. With HRCT scanned images during exacerbation, internal diameters of large airway were measured consecutively from right main to right subsegmental bronchus and measurement was repeated after discharge at the same level and angle of scanned bronchus. Results: Ten patients (male 5, mean age 43 [range 19Y64]) completed studies. Pulmonary function was markedly improved after treatment of exacerbation; mean FVC from 2.93 T 0.82 L to 3.72 T 0.64 L (p = 0.001) and mean FEV1 from 1.55 T 0.55 L to 2.61 T 0.56 L (p = 0.001). Mean internal diameter on HRCT during exacerbation was 3.38 T 0.81 mm and after treatment 3.41 T 0.80 mm (p = 0.80). There was one patient with significant increase of internal diameter and it decreased in 2 patients. Improvement of FEV1 was not associated with changes of internal diameter on HRCT (r = 0.39, p = 0.26). Conclusion: Before and after treatment of asthma exacerbation, there was marked improvement of pulmonary function but changes of internal diameter measured by HRCT were not consistent. TNF-" is an important pro-inflammatory cytokine that is also a well known inducer of the inflammatory response and a regulator of immunity. A strong argument exists for TNF-" being a critical cytokine in the pathogenesis of chronic inflammatory disorders of the airways. Once released in the airways, TNF-" acts by inducing a general inflammatory response mainly through enhanced release of pro-inflammatory/chemotactic mediators and upregulation of adhesion molecules such as E-selectin, VCAM-1, and ICAM-1, thus facilitating the migration of neutrophils and eosinophils. To date there is very limited information on the use of TNF-" blocking agents in asthma. The present study investigated the effect of soluble TNF-" receptor on the airway inflammation in mice model of bronchial asthma. Mice were treated with intraperitoneal soluble TNF-" receptor during the OVA challenge. Mice exposed to OVA developed sustained eosinophilic airway inflammation and sustained AHR to methacholine compared with control mice. Intraperitoneal administration of soluble TNF-" receptor inhibited the development of AHR and eosinophilic inflammation. Moreover, soluble TNF-" receptor treatment reduced IL-4, IL-5, IL-13 and IL-10 level in bronchoalveolar lavage fluid. These results suggest that soluble TNF-" receptor can modulate the airway inflammation and AHR via inhibition of inflammatory cytokine production. Detection of multi-locus genetic interaction in aspirin-intolerant asthma with multifactor-dimensionality reduction analysis Background and Objective: Aspirin-intolerant asthma (AIA) is a common phenotype of aspirin hypersensitivity and affects about 10~20% of asthmatic patients. Recently, the single gene polymorphism associated with the AIA susceptibility has been investigated, but identification of multi-locus single nucleotide polymorphism (SNP) set in association with the susceptibility has not been investigated. Subjects and Methods: In this study, we selected 23 SNPs in 13 candidate genes for 94 asthmatics with aspirin hypersensitivity (AIA) and 152 asthmatics without aspirin hypersensitivity (aspirin-tolerant asthma, ATA) and genotyped each SNP by a primer extension method. Multi-locus genetic interactions were examined with multifactor-dimensionality reduction (MDR) to test all multi-locus SNP combinations for the efficient prediction of AIA. Results: Through a MDR analysis, we identified four-locus gene-gene interaction models that predict AIA disease risk among asthmatic patients with 64.2 % balanced accuracy. Conclusion: These results suggest that significant epistatic effect of fourlocus genetic interaction may exist in the susceptibility for AIA in asthmatic patients which may be a useful in vitro method to diagnose the AIA with acceptable sensitivity. IL-5 production in response to Candida Albicans secretory aspartic protease 2 is the marker of isolated late-phase bronchial responses upon inhalation challenge for nonatopic asthma To delineate the mechanisms of nonatopic asthma, peripheral blood mononuclear cells (PBMC) obtained from atopic asthmatics, nonatopic asthmatics, and healthy controls were incubated with various allergen molecules. IL-2, IL-4, IL-5, IL-13, and IFN-, productions were measured by specific ELISAs. T cell proliferation was assessed by 3H-thymidine uptake. IgE and IgG antibody were assayed by RAST and ELISA, respectively. Intradermal and bronchial inhalation challenges of the antigens were performed according to the standard procedures. Histamine releasing tests (HRT) were performed using peripheral blood leucocytes. Proliferative response to crude Candida albicans (CA) extract was not statistically different among the three groups, indicating a common sensitization against CA antigen. Significant amount of lL-5 was produced by PBMC obtained from several nonatopic asthmatics upon incubation with crude CA extract and a purified antigen, secretory aspartic proteinase 2 (SAP2). IL-5 production was undetectable for the PBMC obtained from healthy control subjects in response to SAP2. Upon intradermal and bronchial challenge of SAP2, late but not immediate skin and bronchial responses were observed for the IL-5-producing asthmatics, respectively. Neither IAR nor LAR was detectable for the IL-5-nonproducing asthmatics, indicating the specificity of the responses. LAR was not induced for the IL-5-nonproducing, IL-13-producing asthmatics. IgE-dependent mechanism was ruled out by negative RAST, HRT, or immediate skin reaction. Anti-SAP2 IgG antibody (precipitin) was not detectable in the serum of either the asthmatics or the control subjects. Nonatopic asthma may be caused by an IgE-independent, T celldependent immune-recognition, and in vitro cytokine synthesis become a reliable diagnostic test for BT cell allergens[. Correlation between serum immune markers and bronchial hyperreactivity Background: Asthma is characterised by an underlying allergic inflammation resulting bronchial hyperreactivity. The aim of the examination was to study the correlation between serum biological markers (IL-13, RANTES, total IgE) and bronchial hyperreactivity. Symptom-free adults with childhood bronchial asthma and their symptom-free children were studied (n:76). Methods: Bronchial hyperreactivity was proved with metacholine airway provocation in 51 symptom-free patients (30 adults, 21 children), but it was not in 25 patients (21adults, 4 children). Serum IL-13, RANTES and IgE detection was perfomed by ELISA. Results: Significant higher 53 T 20, 24 pg/ml vs. 5, 64 T 4, 24 pg/ml, pG0 .01), RANTES (XTSD:1098 T 543, 1 pg/ml vs. 901, 8 T 322, 8 pg/ ml, pG0.01) and IgE (X T SD:360, 7 T 352, 9 KU/l vs. 130, 0 T 117, 8 KU/l pG0.0001) concentrations were proved in symptom-free patients with bronchial hyperreactivity compared to non-hyperreactives. In symptom-free patients with bronchial hyperreactivity significant positive linear correlation was proved between the serum levels of IL-13 and RANTES, IL-13 and IgE. Conclusion: The serum IL-13 level correlated with the IgE concentration in non-hyperreactive patients as well. In bronchial hyperreactivity IL-13 shows a correlation with the chemokine RANTES and total IgE. A persisting inflammation can be detected in bronchial hyperrectivity even in symptomfree patients. The local and serum level of IL-6 and its soluble receptor (srIL-6) at teenagers with bronchial asthma The analysis of dynamics of system and local content SR-IL-6 at teenagers with various clinical forms of allergic diseases has revealed higher levels of systemic content SR-IL-6 in comparison with its local maintenance in nasal lavage. Despite of different levels of IL-6 and SR-IL-6 the correlation of these markers was found in studied biological liquids estimation the results of the research. So, bronchial asthma (BA) SR-IL-6 level was 23,09 + 0,27 ng/ml in serum of blood and 2,24 T 0,2 ng/ml locally. However, IL-6 research revealed the opposite direction in the present group of patients: higher levels in nasal lavage in comparison with serum (1,97 + 0,4 pg/ml and 3,97 + 0,61 pg/ml accordingly). Combined SR-IL-6 level in serum of blood BA and the atopic dermatitis (AD) patients tended to increase in comparison with group of patients BA and made 27,24 + 0,32 ng/ml, and the SR-IL-6 level in nasal a secret was below, than at BA (1,02 + 0,04 ng/ml). IL-6 research at BA patients in combination with AD revealed authentic (8G0,05) increase of serum level of present cytokine (3,31T0,22 pg/ ml) and decrease local one (0,97 T 0,06 pg/ml). Content SR-IL-6 practically did not differ comparing BA and atopic rhinitis (AR) patients and BA patients making 23,64 T 0,38 ng/ml in serum of blood and 2,03 T 0,14 ng/ml in nasal secret. However, quantity of IL-6 in the present group was the highest both in serum of blood, and in nasal secret (9,68 + 1,11 pg/ml and 4,73 + 0,56 pg/ml accordingly) (8G0,05). Combining clinical forms of AR and AD levels IL-6 and SR-IL-6 cytokines were the least of the studied clinical variants of atopic march, making 21,19 T 0,11 pg/ml in serum and 0,49 T 0,01 pg/ml locally for IL-6 and 0,18 T 0,001 ng/ml and 0,17 T 0,02 ng/ml for SR-IL6 accordingly. Thus, the greatest increase of IL-6 level in serum of blood and in nasal secret occured while combination BA? and A?R with children and teenagers having all forms of atopic march. The level of soluble receptor of IL-6 with children also increased while combining BA? and A?R, whereas with teenagers -while having clinical evidences of BA? and AD more essentially. Zinc iron ratio in allergic asthma and allergic rhinitis Vojislav Djuric. Institute of Allergology and Immunology, Functional Diagnostics Unit, Belgrade, Serbia and Montenegro. Background: Zinc was shown to have protective effect on the respiratory system exposed to oxidative stress, soit may play a role in allergic airway diseases. Iron can have a role in oxidative stress in allergic respiratory diseases. Iron, as well as copper, can transfer electrons and produce reactive oxygen species which can damage epithelium in allergic diseases. Both trace elements also have significant influence on immune system. Methods: Using atomic absorption spectrophotometry, we have measured concentration of zinc and iron in serum and in supernatant of induced sputum in 21 patients with allergic asthma, in 13 patients with allergic rhinitis and in 10 control subjects. Results: Postivie corelation was found between serum zinc and iron concentrations of patients with allergic asthma. There was no correlation between zinc and iron concentration in sputum. Conclusion: Zinc and iron trace elements with antagonistic action on respiratory epithelium are positively correlated in serum of alergic asthmatics. Immunohistochemical co-localization of transient receptor potential vanilloid-1 in the trachea using a guinea-pig asthma model Aim: We investigated the distribution of transient receptor potential vanilloid-1 (TRPV1) in the airway of guinea-pigs. In this study, we compared the changes in TRPV1 in the trachea using a guinea-pig asthma model. Methods: We created an asthma model of 5 guinea-pigs by the administration of ovalbumin (OA) and 5 controls (Sham) according to a double blind study. Each trachea was removed from them and sectioned (306m) in a cryostat. The slidemounted sections were incubated in 10% normal donkey serum for 1 hr followed by 0.3% H2O2 for 30 minutes. In addition, they were blocked by avidin/biotin blocking kit. And the subsequently sections were incubated in polyclonal anti-TRPV1 antibody (1:30,000) for 40 hr. Next, they were incubated with biotinylated donkey anti-rabbit immunoglobulin G (1:400) for 90 minutes. Then they were incubated in streptavidin biotin-peroxidase complex for 1 hr followed by fluorescein tyramide (1:75) for 7 min. Finally, we observed the TRPV1 activity of each section by confocal microscopy and then compared the results. Results: The TRPV1 immunoreactive axons were localized to the fine axons within the epithelium and around the smooth muscle area. The TRPV1 axons were found to be stronger and more frequent in the OA group than in the Sham group. Conclusion: These findings suggest that the TRPV1 immunoreactive axons of the trachea increase in number under allergic inflammatory conditions. Water soluble chitosan attenuates mite allergen-induced macrophage activation in allergic asthmatic patients Chitin and chitosan have versatile anti-tumor, anti-fungal, and antimicrobial biological properties. Oral intake and intranasal administration of chitin attenuated allergen-induced airway inflammation in sensitized mice, which may be due to its Th1 adjuvant properties. However, the detailed mechanism of action is not clear. In this report, we demonstrated that water soluble chitosan had specific immunomodulatory effects on dust mite allergen Dermatophagoides farinae (Der f)-stimulated, monocyte-derived macrophages (MDM) in the shifting of Th2 cytokine polarization, decreasing inflammatory cytokine production of IL-6 and TNF-", down-regulation of CD44, CD14, TLR14, and PAR2 receptor expressions, and inhibiting T-cell proliferation in the presence of allergen-stimulated MDM. Under scanning electron microscope (SEM) examination, chitosan reduced cellular change of pseudopodia formation in Der f-stimulated MDM from allergic asthma patients. The effect of chitosan on allergen-stimulated MDM may occur through inhibited PKCUa phosphorylation and NF-kB pathway activation. In a murine model of asthma, we found that intranasal application of chitosan attenuates Der f-induced lung inflammation by reducing infiltration of inflammatory cells, epithlieal damage, and goblet cell hyperplasia. The production of Arg1, iNOs and thymic stromal lymphopoietin (TSLP) in the bronchial epithelium of allergen-challenged sensitized mice was markedly decreased in chitosan-treated mice. Therefore, we believe that these results of examination of the anti-allergic effect of chitosan may provide a new therapeutic modality for allergic asthma. Basic FGF2 plays a key role in the allergic sensitization of airways and the pathogenesis of asthma Background: Asthma is characterized as a chronic inflammatory disorder of the airways associated with airway hyperresponsiveness (AHR), mucus production, and airway remodeling such as smooth muscle hypertrophy and subepithelial fibrosis. Basic FGF (FGF2) is a member of the large FGF family and plays roles in the proliferation of fibroblast, and the migration and proliferation of airway epithelial cells during wound healing. However, the role of FGF2 in the allergic sensitization is totally unknown, although FGF2 levels are thought to be elevated in patients with asthma. Objective: To evaluate the immunological roles of FGF2 on airway allergen sensitization, and pathogenesis of asthma. Methods: 6-week-old female BALB/C mice were sensitized intranasally with LPS-depleted ovalbumin (OVA) in the presence or absence of 106g of rFGF2 at days 0, 1, 2, 7, and then challenged intranasally with OVA at days 14, 15, 21 and 22. AHR was measured by whole both plethysmography after 24 hours from last challenge. Mice were sacrificed at day 24, and then BAL cellularity, lung histology, and several immunologic parameters were assessed to evaluate the effects of rFGF2 co-treatment. Results: AHR against methacholine challenge was significantly increased in mice sensitized with OVA plus rFGF2 compared with other groups. And number of total cell, macrophage, eosinophils, and lymphocytes in BAL fluids were also makedly increased in rFGF treated mice, accompanied by severe peribronchial and perivascular eosinophilic inflammations from histologic findings. Mucus secretion and subepithelial fibrosis were also increased in these mice, respectively. IL-4, IL-13, and TGF-$1 levels in BAL fluids were markedly enhanced in mice sensitized with OVA plus rFGF2. Conclusion: These finding suggested that FGF2 plays important roles in the pathogenesis of allergic asthma, especially promoting allergic sensitization via production of inflammatory cytokines, such as IL-4, IL-13, and TGF-$1. The role of VEGF and its receptors on airway allergic sensitization and pathogenesis of asthma diseases such as asthma and chronic bronchitis. It has been reported that the levels of VEGF in tissues and biologic samples are increased in patients with asthma. VEGF has been assumed to contribute to asthmatic tissue edema through its effect on vascular permeability. The recent study using lung specific VEGF over-expressing transgenic mice showed that VEGF induced TH2 lung inflammation. However, it still remains whether the VEGF affects during allergen sensitization or adaptive immune responses by allergen specific T cells. Objective: To assess the roles of VEGF and its receptor (Flt-1/KDR) in allergic sensitization of airways. Methods: C57BL/6 wild type mice were sensitized intranasally with LPS depleted with ovalbumin (OVA) in the presence of 10 lšg of LPS on days 0, 1, 2 and 7 and then challenged intranasally with OVA on days 14, 15, 21 and 22. Before sensitization, the mice were intraperitoneally injected with SU5416 as the inhibitor of VEGFR1 and VEGFR2 at the only sensitized period. On the proper times (days 4, 23), we sacrificed mice and analyzed phenotypes including BAL cellularity, lung histology and evaluation of immunologic parameters such as the production of cytokines in BAL fluid. Results: On day 4, the number of total cells, macrophage, lymphocytes and neutrophils increased in BAL fluid from the mice sensitized with OVA in the presence of LPS compared to only OVA treated mice. Interestingly, SU5416 blocked the recruitment of inflammatory cells in sensitization period. At this time, the production of TNF-" and IL-12 also decreased in BAL fluid from SU5416 treated mice. After OVA challenge (day 23) the number of total cells was decreased in BAL from mice treated SU5416 at sensitized period. And histological finding showed that infiltration of inflammatory cells was also significantly decreased in SU5416 treated mice. Conclusion: VEGF induced during allergic sensitization and its signaling pathway play a key role in allergen specific T cell priming. Cytokine responses during exacerbation compared with stable phase in asthmatic children Background: Bronchial asthma is a chronic inflammatory disorder of the airways. T lymphocytes are crucial for the initiation and maintenance of the allergic inflammatory response, particularly T helper 2 (Th2) cells. Balancing in Th1 and Th2 response is targeted in treatment. Recent studies show that interleukin-10 (IL-10) have an important role in the regulation of Th2 and allergic responses and decreased in asthmatic patients. Objective: To examine cytokine responses, including interferon-gamma (IFN-,), IL-4, IL-10 in asthmatic children during acute exacerbation. Methods: Fourteen asthmatic children were included in this study. Fresh whole blood obtained from patients at two phases: exacerbation and stable phase, were separated to peripheral blood mononuclear cells and were stimulated with phytohemagglutinin (PHA) and mite allergen (Der p1) for 72 hours. ELISA assays were applied to measure cytokine concentration (IFN-,, IL-4, IL-10) of supernatant. : IL-10 level (PHA-stimulated) was significantly decreased in acute asthma exacerbation (464 T 628.25 pg/mL) compared with stable phase (859.5 T 796 pg/mL) (p = 0.03). IL-4, IFN-, levels and cytokine ratios for all stimulations were not significantly different among episodes. For PBMC with PHA-stimulated, IL-10 levels were slightly decreased in moderate persistent asthma compared with mild persistent asthma (737 T 691.25 vs. 1,375 T 870.75, p = 0.055) . There were no correlations between severity of exacerbation, asthma score, size of skin prick test and cytokine levels. Conclusion: The decrease of IL-10 production in asthmatic children during acute exacerbation may have a role in asthma exacerbation. The decrease of IL-10 production in more severe asthma (moderate persistent) supports the idea of defective immune regulation of IL-10. Further studies in IL-10 producing cells and treatment to enhance IL-10 responses might be useful in prevention and treatment of asthma. Alternative transcripts of cysteinyl leukotriene 1 receptor (CysLTR 1 ) in patients with bronchial asthma Introduction: Cysteinyl leukotrienes are lipid mediators that have been implicated in pathogenesis of several inflammatory processes, including asthma. They cause bronchoconstriction, mucous hypersecretion, increased microvascular permability, bronchial hyperresponsiveness, and eosinophil infiltration. The biological action of cysLTs is mediated via CysLT 1 and CysLT 2 receptors. Human CysLTR 1 gene consists of five exons variably spliced. Transcript I, composed of 1, 4 and 5 exons is a major transcript present in human leukocytes, smooth muscles. Transcript II, composed of 1 and 5 exons is less abundant but find in blood leukocytes, smooth muscles, hart, brain. The role of alternative transcripts in asthma pathogenesis is still not known. Aim: The goal of our study was to investigate differences in expression of CysLTR 1 alternative transcripts I and II in patient with bronchial asthma and healthy control group. Methods: PBMCs were obtained from peripheral blood of 20 patients with severe and non-severe asthma and 15 healthy volunteers. Total RNA was obtained from mononuclear cell using TotalRNA and cDNA was synthesized using Reverse Transcriptase. We designed specific primers spanning exonexon junction in the transcript I (exons 1Y5) and transcript II (exon 1Y4) and common primer in the CysLTR 1 coding region according to published CysLTR 1 transcripts sequences. Real-time PCR reactions were performed on RotorGene using Evagreen dye. Results: We found that CysLTR 1 transcripts I and II are present in patients with asthma and in control group. The CysLTR 1 transcripts II/I ratio was 26,02 % in patient with severe asthma, 25,99 % with moderate asthma, 56,44 % with mild asthma and 44,20 % in control group. In addition, we found significant difference in CysLTR 1 transcripts II/I ratio in severe asthmatic subgroup in comparison to controls [p = 0,004], severe asthmatic in comparison to mild [p = 0,018] and in CysLTR 1 transcripts II/I ratio in moderate asthmatic in comparison to control group [p = 0,0004] and in mild asthmatic in comparison to control group [p = 0,007]. Conclusion: Differences in CysLTR 1 alternative transcripts expression might possibly contribute to airway inflammation in patient with bronchial asthma. Prostaglandin production in different asthma patients Valeriya Nemtsova, Inga Fedotova, and Volodimir Fedotov. Kharkov Allergological Center, Allergology, Kharkov, Ukraine. Background: Special regulatory role of prostaglandin E2 has been postulated in aspirin-induced asthma. The aim if this study was to investigate the effects of aspirin on systemic production of prostaglandin E2 and cysteinyl leukotrienes in patients with asthma. Methods: We determined urinary concentrations of two main prostaglandin E2 metabolites: 13,14-dihydro-15keto-PGE2 using commercial enzyme immunoassay and 9,15-dioxo-11alpha-hydroxy-2,3,4,5-tetranor-prostane-1,20-dioic acid using gas chromatography/mass spectrometry; and leukotriene E4 using immunoassay. Determinations were performed at baseline and following oral aspirin and celecoxib challenges, in two well-defined asthma phenotypes: aspirin-sensitive and aspirin-tolerant patients. Results: Aspirin precipitated bronchial reactions in all aspirin-sensitive, but in none of the aspirin-tolerant patients. Celecoxib 400 mg was well tolerated by all patients except for one with aspirin-induced asthma. At baseline mean prostaglandin E2 metabolites values did not differ between the groups. Following different aspirin provocation doses, the two main prostaglandin E2 metabolites were decreased in the aspirin-tolerant group, but their mean level remained unchanged in the aspirin-sensitive group. The dose of aspirin had no effect on the magnitude of the response on the prostaglandin E2 metabolites and its duration. In both groups urinary prostaglandin E2 metabolites decreased following celecoxib challenge. No correlation was found between prostaglandin E2 metabolites and leukotriene E4. Conclusion: Aspirin-precipitated asthmatic attacks are not associated with changes in the systemic prostaglandin E2 production. In contrast, prostaglandin E2 systemic production becomes depressed by aspirin in nonsensitive patients. This different response might indicate COX-1 dependent prostaglandin E2 control of inflammatory cells in AIA. Thus, PGE2 is released during the clinical reactions to aspirin through an alternate COX-2 pathway. Clinical implications of this finding are in line with current observations of good tolerance of the selective COX-2 inhibitors in sensitive patients. Mast-cell activation in aspirin-induced asthma Inga Fedotova, Irina Feclina, and Irina Tikhonova. Kharkov Allergological Centre, Allergology, Kharkov, Ukraine. Background: There is increasing evidence of the importance of cysteinyl leukotrienes (LT) as mediators of aspirin-induced bronchoconstriction in aspirin-sensitive asthma but the cellular origin of the LT is not yet clear. Methods: Urinary concentrations of leukotriene E4 (LTE4), 11-dehydrothromboxane B2, 9alpha,11beta-prostaglandin F2, and Ntau-methylhistamine were measured during the 24 h following cumulative intravenous administration of increasing doses of lysine aspirin to asthmatic patients. In addition, the urinary concentrations of these metabolites were measured on 10 consecutive days in a patient who suffered an asthma attack after percutaneous administration of nonsteroidal anti-inflammatory drugs. Results: In aspirin-induced asthma patients (AIA, n = 18), the basal concentration of urinary LTE4, but not the other metabolites, was significantly higher than that in aspirin-tolerant asthma patients (ATA, n = 16). After intravenous aspirin provocation, the AIA group showed a 12.6-fold (geometric mean) increase in excretion of LTE4 during the first 6 h, and 9alpha,11beta-prostaglandin F2 also increased in the AIA group during the first 0Y6 h and the 6Y9 h collection period. Ntau-methylhistamine excretion was also increased, but to a lesser degree. Administration of aspirin caused significant suppression of 11-dehydrothromboxane B2 excretion in both the AIA and ATA groups. When the percentage of maximum increase of each metabolite from the baseline concentrations was compared between the AIA group and the ATA group, a significantly higher increase in excretion of LTE4, 9alpha,11beta-prostaglandin F2, and Ntau-methylhistamine was observed in the AIA group than the ATA group. An increased excretion of LTE4 and 9alpha, 11beta-prostaglandin F2 has been detected in a patient who suffered an asthma attack after percutaneous administration of nonsteroidal anti-inflammatory drugs. Conclusion: Considering that human lung mast cells are capable of producing LTC4, prostaglandin D2, and histamine, the results of our study support the concept that mast cells, at least, may participate in the development of aspirininduced asthma. Zlatica Goseva, Angelko Gjorcev, Dejan Dokic, Tatjana Caparoska, and Zoran Arsovski. Clinic of Pulmology and Allergy, Department of Allergy, Skopje, Macedonia, Fyrom. Background: Bronchial asthma is characterized by airway inflammation, hiperresponsiveness to variety of stimuli and airway obstruction, which can be reversible. Different cells are involved in the pathogenesis of asthma, like: T-lymphocytes, mast cells, eosinophils, macrophages etc. A number of different inflammatory cells infiltrate the airways. The trigger can cause the release of inflammatory mediators from the cells, which could be the markers of inflammation. Methods: We have investigated the role and the importance of inflammatory cells and mediators like eosinophils, ECP, IL-4 and IL-5. We studied 77 subjects divided in three groups as follows: 1. asthma patients; 2. patients with obstructive bronchitis and 3. control group. Results: The number of eosinophils was significantly increased in the group of asthma patients versus second and third group. The eosinophils have their very important role in allergic inflammation. We found that the presence of ECP demonstrate an ongoing inflammation, with or without clinical symptoms in asthma patients. There was significant difference between the values of ECP of asthma patients versus second group with bronchitis and healthy controls. The values of IL-4 were not significantly increased between the groups. We found the explanation in the fact that IL-4 is a marker for early allergic inflammation. Our results have shown that IL-5 is involved in the pathophysiology of asthma. The values of IL-5 were significantly increased versus second group and controls. We also found the decrease of the values of inflammatory markers after the treatment with corticosteroids. Conclusion: We concluded that eosinophils, ECP and IL-5 could be useful markers for selecting allergic patients and they could be used together with other examinations. The markers of inflammation could have their important role in the measurement of the allergic inflammation and they could be the monitors of treatment effects. Mucosal immunotherapy with CpG oligodeoxynucleotides increases local IL-10 concentration in a murine model of chronic asthma Tahereh Mousavi, Alireza Salek Moghadam, and Reza Falak. Iran university, Immunology, Tehran, Islamic Republic of Iran. We examined CpG motif effects on a previously developed murine model of asthma in which chronic airway inflammation was induced by repeated allergen [Chenopodium album (Ch.a)] inhalation. Using this model, we examined the responses to mucosal administration of CpG DNA (oligonucleotides) and specific antigen immunotherapy. CpG-based immunotherapy significantly reversed both acute and chronic markers of inflammation compared with specific Ag immunotherapy. IL-10 levels were also measured both in splenocyte and lung culture supernatants. The results showed that antigen recall responses of lung culture from mucosal treated mice demonstrated an antigen-specific enhanced release of IL-10, but the concentration of this regulatory cytokine had no significant changes in splenocyte culture medium. These results suggest that mucosal immunotherapy with CpG DNA may induce local production of cytokines without systemic effects on Th responses. We suggest that mucosal co-administration of CPG /Ag may provide the basis for a more efficient form of immunotherapy in allergic asthma. Indeed local induction of cytokines by this procedure may diminish potential toxicity of systemic antigen administration. Assessment of airway remodeling by HRCT in asthmatics: correlation with age, smoking, disease duration and severity Aliae Mohamed-Hussein 1 , W Hasan 1 , G Agmy 1 , S Abdel Aziz 2 , and E Abou Elhamd 2 . 1 Assiut University Hospitals, Assiut, Egypt, Chest, Assiut, Egypt; 2 Assiut University Hospitals, Assiut, Egypt, Radiology, Assiut, Egypt. Background: To date, airway remodeling is usually assessed using histological examination of airways. However, now it is possible to assess and quantify the extent of airway remodeling in vivo using high-resolution CT (HRCT). The aim of this study was to prospectively evaluate airway wall thickness as indicator of remodeling by using thin section HRCT in asthmatics, and to correlate these findings with pulmonary function tests (PFTs) results and other clinical indices in asthmatics. Methods: 41 patients with asthma and 20 healthy controls participated in the study. Remodeling as measured by whole airway wall thickness was assessed with HRCT. Thickness-to diameter ratio (TDR) and the percentage wall area (PWA) were determined. Spirometric tests were also performed. Results: TDR and PWA were significantly higher in asthmatics than in controls. Both TDR and PWA were strongly correlated with disease severity and duration. Also, TDR and PWA were inversely correlated with the percentage of predicted forced vital capacity (FVC %), forced expiratory volume in 1st second (FEV1%) and FEV1/FVC and post-bronchodilator reversibility in asthmatics. Conclusion: These findings indicate that HRCT is useful non-invasive method for assessment of airway wall thickness. Airway wall thickening occurs more in patients with moderate and severe asthma and its degree is related to the duration and severity of asthma and degree of airflow obstruction. Clinical and immunologic findings of methylene diphenyl diisocyanate -induced occupational asthma in a single car upholstery Gyu-Young Hur 1 , Dong-Hee Koh 2 , Gil-Soon Choi 1 , Han-Jung Park 1 , Sung-Jin Choi 1 , Young-Min Ye 1 , Kyoo-Sang Kim 2 , and Hae-Sim Park 1 . 1 Ajou University School of Medicine, Allergy and Rheumatology, Suwon, Republic of Korea; 2 Korea Occupational Safety and Health Research Institute, Incheon, Republic of Korea. Background: Although methylene diphenyl diisocyanate (MDI) has been widely used in many industries, there have been few studies of MDI-induced occupational asthma. Objectives: We investigated to present the clinical and immunologic findings in methylene diphenyl diisocyanate-exposed workers in a single industry of car upholstery. Methods: Fifty-eight exposed workers in a single industry were enrolled. Work related respiratory symptoms were screened using a respiratory questionnaire. Serum specific IgE and IgG antibodies to MDI-human serum albumin conjugate were measured by ELISA. Atopy status was evaluated using allergy skin prick test. For confirmation of MDI-induced occupational asthma, methacholine bronchial challenge test and MDI-specific inhalation test with changes of sputum eosinophil counts were performed in symptomatic workers. Results: Thirteen (22.4%) subjects had complained of respiratory workrelated symptoms. The prevalence of MDI-occupational asthma was noted in 5 (8.6%) workers and occupational eosinophilic bronchitis was noted in 2 (3.45%) workers. The prevalence of specific IgG (20.7%) was higher than that of specific IgE (8.6%). The prevalence of MDI-occupational asthma/ eosinophilic bronchitis was strongly associated with the presences of workrelated respiratory symptoms and MDI-specific IgG antibodies (P G 0.01, P G 0.05, respectively). Conclusion: MDI may be a causative agent of occupational asthma among MDI-exposed workers. The prevalence of MDI-occupational asthma was 8.6%, and MDI-eosinophilic bronchitis was confirmed in 2 workers. The presence of work-related lower respiratory symptoms and serum specific IgG to MDI-human serum albumin conjugate may be useful to predict MDIoccupational asthma/eosinophilic bronchitis in MDI exposed workers. Results: The average duration of employment of workers was 10 years and 12% were pesticide crop sprayers. Work-related wheeze (26%), ocular-nasal (24%), urticaria/skin symptoms (14%) were more prevalent in the orchards. The prevalence of sensitization (SPT) was the highest to TU (22%) followed by house dust mite (16%), with 25% being atopic. The prevalence of allergy to TU (skin reactivity and work-related symptoms) was 9.5%, with respiratory (6%) more common than skin allergy (3%). Work-related ocular-nasal (OR = 4.9) and skin (OR = 3.7) symptoms were more commonly reported by pesticide crop sprayers. Workers with TU-allergic rhino-conjunctivitis and probable asthma were more likely to be atopic, spray pesticides and have low (G30 U/g Hgb) AChE levels. Background: Toluene diisocyanate (TDI), a highly reactive industrial chemical, is one of the leading causes of occupation-related asthma in industrialized countries. The pathogenesis of TDI-induced asthma remains not fully understood, in part due to lack of appropriate animal models. In our study, we established a novel TDI asthma mouse model by epicutaneous sensitization and intranasal challenge. Methods: Thirty two 6Y8 week BALB/c mice were randomly divided into four groups (n = 8): TDI, acetone olive oil (AOO), ovalbumin (OVA) and saline (sal). On days 0, 7 and 14, the mice were epicutaneously sensitized with 1% TDI in 100 2l acetone: olive oil (3:2, AOO), 100 2l AOO, 100 2l 0.1% OVA in sal and 100 2l sal respectively, every allergen or solvent was placed on a patch of sterile gauze (1Â1 cm) which was taped to the skin of shaved mouse back and kept for 3 days per sensitization. On days 21, 23 and 25, mice were challenged intranasally with 25 2l identical allergen or solvent respectively once a day. All mice were sacrificed on day 27. 24 hours before first intranasal challenge and after last intranasal challenge, airway responsiveness to methacholine (Mch) was measured in an unrestrained whole body plethysmogragh and was expressed as enhanced pause (Penh). Mice were exposed to nebulized phosphate-buffered saline as the baseline and then increasing dose of Mch, namely 5%, 10%, 20% and 40% followed by measurement of Penh values for 5 minutes. TDI-and OVA-specific IgG1, IgG2a and IgE in sera were measured by ELISA and expressed as optical density (OD) mean value's. Results: 1. Histology: TDI-exposed mice exhibited neutrophil-dominant pulmonary inflammation in the peri-bronchial and peri-vascular regions by lung tissue haematoxylin and eosin (HE) staining, and increased mucus secretion by PAS staining, while OVA-treated mice exhibited an eosinophildominant inflammation and mucus production. 2. After challenge in TDItreated mice airway responsiveness was significantly augmented, but not in OVA-and solvent-treated mice. 3. Significant increased production of sera TDI-specific IgG1, IgG2a and IgE in TDI-treated mice was observed as well as the increased OVA-specific IgG1, IgG2a and IgE in OVA mice. Conclusion: Our study showed that allergic asthmatic responses to a chemical sensitizer such as TDI may occur after dermal sensitization and airway challenge via IgE-mediated mechanism. Remei Guspí Bori 1 , Cinta Castellà Valldeperez 2 , and Miquel Baltasar Drago 1 . 1 Hospital of Tortosa Verge de la Cinta, Unit of Allergy, Tortosa, Spain; 2 Hospital of Tortosa Verge de la Cinta, Internal Medicine, Tortosa, Spain. We report a case of a patient male of 33 years old with recurrent symptoms of rhinitis, conjunctivitis, and dermatitis related with his job. He hasn_t had family nor personal antecedents of atopy. His symptoms were recurrent pruriginous eczema located to body and legs, accompanied of rhinitis and conjunctivitis, without symptoms of bronchial reactivity. These symptoms appeared while he was in the place of work and improved at the weekend and holidays. He was working for eighteen months in a pipe_s factory as support agent. They used polyvinyl for made the pipes and was in contact with resins, cobalt, styrene and glass fibre in this factory. We preformed a skin prick test with a battery of common aeroallergens and a standard patch test (True test\) with negative result. The basal espirometry performed had a normal pattern and the bronchodilatation test was negative. The blood analysis showed a normal biochemical levels and normal haematological values, with antinuclears and antitissue antibodies negatives. The specific IgE determination against anhydride phthalic was 25,7 KU/l and against formaldehyde was G0.35kU/l. Our diagnosis_ suspicion was sensitization to anhydride phthalic as the cause of the symptoms that he presented. The patient_s evolution after the job_s change was very favourable. He has remained asymptomatic until nowadays and the specific IgE levels to anhydride phthalic has gone lowering, being of 0.42 KU/l eighteen moths later. The use of anhydride phthalic as hardener of epoxy resins forms polymers that are useful to make materials as glass fibre. Sometimes there are too many substances in the occupational environment then it can difficult the causal diagnosis. The acid anhydrides are substances with a low molecular weight that can cause rhinitis, asthma and contact dermatitis associated to an IgE mediated mechanism. The airborne exposition to anhydride phthalic can happen more frequently in the plastic industry. Predictors of work-related symptoms, allergic sensitisation and occupational asthma among supermarket bakery workers in South Africa Background: A recent study reported a high risk of developing work-related asthma among supermarket bakers. This study aimed to determine the predictors for work-related symptoms, allergic sensitisation, non-specific bronchial hyper-responsiveness (NSBH) and baker's asthma in small bakeries of a supermarket chain store in South Africa. Methods: A cross-sectional study of 517 (current and previously employed) bakers was conducted in 31 Cape Town bakeries using a modified European Community Respiratory Health Survey (ECRHS) questionnaire, skin prick tests to common aeroallergens and cereal flours, as well as serum specific IgE to wheat flour, rye flour and fungal alpha amylase by ImmunoCAP-system (Phadia, Sweden). NSBH was assessed using the Medic Aid Pro Nebulizer Dosimeter method. Exposure-response modeling was conducted to identify significant determinants. Results: The mean age of bakers was 32 years and 47% were current smokers. The prevalence of atopy (positive SPT to Q1 common aeroallergen) was 42%. Common work-related symptoms were ocular-nasal (31%) and chest tightness/ wheezing (17%). One third of bakers were sensitised to bakery allergens with 25% sensitised to cereal flours such as wheat and rye. There were 22% of the workers who demonstrated evidence of bronchial responsiveness with 2/3 of these having airway obstruction. Doubling the employment duration was associated with an increased odds for specific IgE reactivity to wheat (OR: Background: Occupational allergy in cutlers is a very strange phenomena even though they handle a lot of potencially allergenic substances. Case Report: A 40-year-old man, cutler since he was 14, active smoker of 20 cigarettes/day. He was referred to our deparment because he presented during the last 4 months daily dysnea, wheeze, cough and thoracic oppression accompanied by rhinitis and conjunctivitis of diurnal predominance and clearly related to his work place. He had been treated with formoterol 9 mcg/ budesonida 320 mcg every 12 hours since 45 days before his visit. He got worse throughout the week and specially on Fridays, when he swept the floor of the factory, forcing him this fact to go to the emergency deparment in 3 occasions. He improved during the weekend and referred to work with metals, wood of olive tree, oak (Bwith resin[) and an artificial wood, although other workers next to him handled deer antler (since 4 years before) and bull horn. In his previous work in a cutlery (16 years before), he used to work with deer antler and wood; during the last three months of working there he presented daily cough, dysnea and wheeze which were related to wood dust. Once he changed the company to the present one, he remained asymptomatic during 15 years until now. Results: Skin prick test (SPT) with a standard battery of inhaled and epithelial allergens and a battery of woods and mites: all of them were negative. SPTwith an extract of deer antler gave positive results (4Â4 mm) with 10 negative controls (5 were atopic patients). SPT with bull horn: negative. Respiratory functional tests after remaining 48 hours without treatment: Forced spirometry: mild obstructive pattern (FVC: 82%, FEV1: 75%) with positive bronchodilator test. Total IgE: 75 UI/ml. Specific IgE was measured by EAST method (Enzyme AllergoSorbent Test) obtaining the following results: deer antler, 0.7 kU/l (class 1); fur and dander from cow, 1.6 kU/L (class 2); epithelium from dog, cat and horse, all of them less than 0.35 kU/L (class 0). The serum specific IgE levels to serum albumins from bovine, cat and chicken (alpha livetin) was also measured, and all of them were below 0.35 kU/L (class 0). The molecular mass of the IgE binding proteins was determined by SDS-PAGE inmunoblotting: two bands of 63 and 52 kDa were detected in deer antler extract. Conclusion: We report the first case in the literature of allergic rhinitis, conjunctivitis and asthma caused by deer antler. Acute allergic reaction and chronic pulmonary effects of occupational inhalation exposure to talc dust The main purpose of this study was to assess and characterize the pulmonary reactions associated with occupational exposure to talc dust. Methods: Ninety-seven talc workers and 110 unexposed employees as the reference group were randomly selected from a local rubber industry. Standardized respiratory questionnaires were administered to the subjects, they underwent chest X-ray and were examined by a specialist for any possible respiratory abnormality to be diagnosed. Furthermore, Pulmonary Function Tests (PFTs) were measured just before and after the work shift. Moreover, to assess the extent to which workers had been exposed to talc dust, using standard methods, inhalable and respirable dust concentrations were measured in different dusty worksites. Results: The average (mean T SD) age (years), weight (kg), height (cm) and duration of exposure to talc dust (years) for the exposed group were 35.8 T 6.75, 73.1 T 9.2, 172.3 T 5.9 and 11.79 T 5.3, respectively. The corresponding values for the non-exposed group were 36. 1 T 6.87, 73.36 T 8.1, 173.2 T 5.7 and 0 T 0, respectively. Atmospheric concentrations of inhalable and respirable talc dust were estimated to be 41.8 T 23.52 and 19.8 T 8.04 mg/m3, (mean T SD), respectively. Talc exposed subjects had a significantly higher prevalence of respiratory symptoms. Similarly, PFTs revealed that exposure to this lubricating agent was associated with significant decreases in the mean percentage predicted of vital capacity (VC), forced vital capacity (FVC) and forced expiratory volume in the first second (FEV1). Likewise, there was a general tendency for VC, FVC and FEV1 to decrease as estimated cumulative exposure (years worked) increased. Moreover, there was an acute reduction in some parameters of pulmonary function such as VC, FVC and FEV1, over the work shift. Chest radiographs of exposed workers showed that pneumoconiosis profusions were between p 0/0 and p 2/1 according to the ILO 1980 chest X-ray classification. Conclusion: These results support the notion that occupational inhalation exposure to talc is associated with chronic respiratory disorders and induces bronchitis and interstitial lung disease. Additionally, they support the hypothesis that inhalation exposure to talc dust induces acute aalergic reversible adverse effect on PFTs. Change of lung function and airway hyperresponsiveness in occupational asthma exposure to causative agents. Significant improvement or disappearance of symptoms or AHR at follow up period is frequently used for the parameter of OA outcome measurement. However, few studies have longitudinally evaluated the change of lung function and AHR by serial measurement. To evaluate the significance of the lung function or AHR as outcome parameter, we serially measured it and compared its change with the clinical condition of OA. Methods: Pulmonary function test, methacholine bronchial provocation testing, asthma symptom and medication scores were evaluated at regular interval in 35 patients with OA confirmed by specific challenge tests. Results: AHR continuously resolved during the follow-up period (mean: 7.5 years, range: 1Y16 years) in 5 (14.3%) of 35 patients. AHR transiently disappeared, however, subsequently re-developed in an additional 5 patients. The level of AHR improved (increase in methacholine PC20 by 3.2-fold or more) in 9 (25.7%) patients, but fluctuated in remaining 16 (45.7%) patients. The level of FEV1 was fluctuated along with the severity of asthma symptom, but not in proportion to the level of AHR. In 20 of 24 patients with reduced lung function at initial examination, lung function did not recover during the follow up period. Significant improvement of symptoms and maintenance of nearly asymptomatic state were achieved in 5 patients. Three of them had disappearance of AHR, but 2 patients showed fluctuated levels of AHR. Lung function and severity of asthma at diagnosis, and the interval between the development of OA symptoms and avoidance was associated with functional impairment. Conclusion: In spite of medical treatment and avoidance, many patients with OA had persistent and fluctuated AHR and lung function impairment even in the absence of respiratory symptom. Intermittent or single assessment of lung function or AHR as outcome measure might lead to incorrect clinical evaluation, and more comprehensive approach will be required. Background: Allergic diseases constitute a global health problem, as they have an increasing economic and social impact and, especially, they can deeply interfere with patients_ daily life, being a cause of physical and emotional discomfort. This is why health related quality of life (HRQoL) has become increasingly important in healthcare research. Aim: To examine the scientific literature of the last three years dealing with the impact of allergic rhinitis (AR) treatments suggested by ARIA guidelines on patients_ HRQoL, and to identify the unexplored or not fully investigated areas concerning this issue. Methods: Studies were sought from MEDLINE (1 January 2004 to 31 December 2006 using as key words the ARIA suggested drugs [AND] allergic rhinitis [AND] and quality of life. Only randomized, DBC trials published in peerreviewed journals, using validated questionnaires were selected. Results: Our research has lead to a selection of 34 DBPC trials, 4 in paediatric and 30 in adult population. HRQoL has been assessed in a total of 10957 patients. The RQLQ questionnaire was used in 27 studies. The Rhinasthma was applied in one study involving patients with both rhinitis and asthma. SF-36 was used together with the specific questionnaire in 3 trials. Only a paper on sublingual immunotherapy (SLIT) evaluated health status by means of SF-36 and subjective satisfaction by means of Satisfaction Profile (SAT-P). The duration of the studies varied from 2 days to 3 years. 19 studies were about SAR, 3 about PAR, 1 about seasonal and perennial rhinitis. 6 studies investigated rhinitis according to ARIA classification: 3 of them were about persistent allergic rhinitis, and 3 considered persistent allergic rhinitis and its comorbidity with asthma. 3 studies investigated rhinitis without providing any classification of the disease. Out of 34 trials, only 8 evaluated the possible presence of comorbid asthma. Discussion: On the basis of our research, it is possible to identify some unexplored areas in HRQoL of patients affected by allergic rhinitis: the new ARIA classification (intermittent and persistent rhinitis) should be carried out for all the treatments in order to fully appreciate the effect of drugs used in AR on aldult and children patients_ HRQoL; the effect of comorbidity on HRQoL should be extensively investigated; HRQoL study should include some other aspects of patient_s perspective life (i.e. satisfaction, adherence). Disturbances in nocturnal sleep, excessive daytime sleepiness and sleep-specific quality of life impairment in patients with allergic rhinitis Danish Jamal and Ashok Shah. Vallabhbhai Patel Chest Institute, Respiratory Medicine, Delhi, India. Background: Nocturnal sleep disturbances, excessive daytime sleepiness and impairment in quality of life (QOL) in allergic rhinitis are often overlooked both by patients and physicians alike. Methods: Consecutive skin allergy test positive patients with allergic rhinitis and matched controls were subjected to computed tomography of paranasal sinuses (CT-PNS) Patients were categorized as allergic rhinitis (group 1), allergic rhinitis with sinusitis (group 2) and controls (group 3). They were evaluated for nocturnal sleep, excessive daytime sleepiness and sleep specific QOL disturbances using the Pittsburgh Sleep Quality Index (PSQI), Epsworth Sleepiness Score (ESS) and Nocturnal Rhinoconjunctivitis Quality of Life Questionnaires (NRQLQ) respectively. They were also categorized as Bsneezer-runners[ or Bblockers[. Results: Of the 207 patients (141 males, 66 females), 75(36%) were in group 1, while 132(64%), in group 2 and 23 in group 3. Patients in group 2 had significantly higher mean scores for PSQI, ESS and NRQLQ questionnaires as compared to group 1. Seasonal allergic rhinitis (SAR) is a hypersensitive immune response to allergens that causes nasal and ocular symptoms and leads to significant decline in the sufferers' Quality Of Life (QOL). In the Environmental Exposure Chamber (EEC) model of allergen exposure, the relationship between SAR scores and their QOL has not been investigated. To this end, a specialized Rhinoconjunctivitis QOL (EEC-RQOL) Questionnaire has been developed to evaluate the QOL problems experienced by patients while they are in the chamber. The questionnaire contains 16 questions in 4 domains: Non-nose/Eye symptoms (NNE), practical problems (PP), emotional problems (EP), and global assessment (GA). Aims: To examine whether correlations exist between QOL and instantaneous scoring of SAR symptoms in study subjects at the EEC. Secondly, to determine the effect of sequential EEC visits on QOL. Methods: A group of 208 Ragweed-sensitive study subjects were exposed to Ragweed pollen (3500 T grains/m3) in the EEC for 3h in which they recorded instantaneous TSS every half-hour on 4 consecutive days. The relationship between EEC-RQOL scores and TSS over the 4 visits was examined using Pearson's correlation and linear regression. Results: During each of the four visits, all individual domains as well as the total EEC-RQOL scores significantly correlated with the symptom scores (see Table 1 ). As subjects participated in subsequent EEC visits, the line of linear regression from 1st to 4th visit shows a clear rightward shift and a gradual increase in slope. Conclusion: The QOL measured in the EEC using EEC-RQOL Questionnaire significantly correlated with the SAR symptoms of subjects on all visit days. The change in linear regression over the four visits indicates an increasing SAR severity, and that is accompanied by a subsequent decline in QOL. Moreover, subjects appear to experience a greater deterioration in QOL over visits as its rate of decline is faster than the worsening of SAR symptoms. Our findings suggest the negative effect on QOL associated with EEC confinement and thus confirm the importance of developing an EEC-specific RQOL questionnaire. Furthermore, we have demonstrated its cross-sectional construct validity. Quality-assessment of disease-specific quality of life questionnaires for rhinitis and rhinosinusitis: a systematic review Caroline van Oene 1 , Ellen V Reij 1 , Mirjam Sprangers 2 , and Wytske Fokkens 1 . 1 Academic Medical Centre, ENT, Amsterdam, Netherlands; 2 Academic Medical Centre, Medical Psychology, Amsterdam, Netherlands. Background: In recent years multiple HRQL questionnaires have been developed specifically for rhinitis and rhinosinusitis. Several reviews described HRQL questionnaires concerning rhinitis and/or (rhino) sinusitis. However, little attention has been paid to the quality of the psychometric properties of these questionnaires. Objective: The aim of this systematic review is to give a quality-assessed review of the existing disease-specific health related quality of life questionnaires concerning rhinitis and rhinosinusitis for adults. Methods: We reviewed the literature up to May 2007 in Pubmed, EMBASE and Medline, to identify studies of interest. Additionally the database of the AAAAI Quality of Life Resources and the Patient-reported Outcome and QOL Instruments Database were searched. The quality is assessed by defining the characteristics of a quality of life questionnaire with assessment criteria. Results: The results of the construction, description, feasibility, and the psychometric performance of the instruments are provided. We finally provide a clinician_s guide to choose a questionnaire based on the measurement goals, the discriminant validity, responsiveness and the points obtained in the quality assessment. Of the top scoring instruments regarding the overall quality assessment, only 4 health related quality of life The cause of a disorder of ventilation and drainage of the middle ear is mainly a disorder opening mechanism of the Eustachian tube.The purpose of the study is to evaluate the persistent symptoms of serous otitis media in children with adenoid hypertrophy and allergic rhinitis. Materials and Methods: The group study is made up of 128 patients with adenoid hyperplasia and serous otitis media, examined and treated in the Pediatric Hospital of Oradea. From all of this, 72 patients were diagnosed with allergic rhinitis. The objective endocavitary E.N.T. examination was systematically performed on all patients. Evaluation criteria: degree of hearing loss, serous rhinorrhea, nasal obstruction and sneezing. We perform adenoidectomy under general anesthesia in all the patients. In the second phase, 3 and 6 month later, patients were re-evaluated clinically and audiological. Results: In the case of patients group just with adenoid hypertrophy, serous otitis media was remitted in 89% of cases at 3 month re-evaluation. In the case of patients with allergic rhinitis and adenoid hypertrophy, serous otitis media was remitted just at 39% of cases, with fluctuant evolution. At 6 month reevaluation, after treatment with desloratadine, mucolitic agents, the serous otitis media was remitted completely. On second evaluation, in the case of the first group, the study showed the reduction of nasal obstruction in 78% of patients as compare to the second group, where recovery was present in only 38% of patients, certified by hearing tests and clinical aspects of tympanic membrane at otoscopy. Conclusion: As a result of adenoidectomy, improvement in permeability of the Eustachian tube, diminish the effusion into the middle ear, but they persist in the case of patients with associated allergic rhinitis. Antihistaminic drugs, desloratadine, have an important role to prevent recurrent attacks of serous otitis media in children. Allergic rhinitis and asthma survey: clinical and psychological perspectives Fulvio Braido, Ilaria Baiardini, Silvia Brandi, Anna Porcu, and Giorgio Walter Canonica. University of Genoa, Allergy and Respiratory Diseases, Genova, Italy. Background: The existence of a close link between asthma and rhinitis, which brings consequences both in therapy and disease management, has been demonstrated by many studies. Objectives: The trial we performed had the following aims: to assess physicians_ knowledge on rhinitis/asthma comorbidity, to evaluate patients_ management behaviour and their experience about symptoms and expectations, to investigate the clinical and psychological meaning of GPs_ and patients_ knowledge. Methods: 101 general practitioners and 504 asthmatic patients were involved in the study. They were asked to fill in two different multiple-choice questionnaires about the association between asthma and rhinitis and its impact. Results: 34.7% of general practitioners are aware of the asthma-rhinitis link, and 43.6% of them assume the comorbidity on the basis of their clinical experience. 21.8% of physicians make the diagnosis autonomously. 27.8% of asthmatic patients experience three or less rhinitic symptoms, 41% from 4 to 6 symptoms, and 31.2 more than 6 symptoms. The symptoms have a deep impact on daily life, as they are cause of sleep problems (87.3%), lack of concentration (78.9%), difficulties in spare time (71.8%) and sport (71.7%). Rhinitis symptoms are responsible for the worsening of asthma, with an increase of dyspnoea (86.3%), cough (73.9%) wheezing (59%). 93% of patients state they are interested in a combination therapy approach. Clinical efficiacy of antileukotriene threapy in allergic rhinitis: changes in subjective and objective parameters, and quality of life measures Cemal Cingi 1 and Kivanc Gunhan 2 . 1 Eskisehir Osmangazi University, ENT, Eskisehir, Turkey; 2 Celal Bayar University, ENT, Manisa, Turkey. Background: Leukotrienes and histamine are thought to be quantitatively the most prominent mediators in the final pathways of allergic rhinitis (AR). Intranasal corticosteroids and oral antihistamines are the cornerstones of therapy. Antileukotriene therapy combined with antihistamine showed a synergistic effect in treating AR, however, there is little data on objective parameters and quality of life measures. Methods: This multi-centered, prospective, randomized and plabebo controlled study randomized 275 patients with mild or moderate intermitant AR into three groups as: fexofenadine (120 mg/day), fexofenadine plus montelukast (10 mg/day) or fexofenadine plus placebo. The nasal endoscopic examinations, objective upper airway measurements (anterior rhinomanometry), were evaluated before and at end of 21 days treatment period. Daily symptom scored by a standard visual analoge scale (VAS) and by a daily diary of quality of life measures. Results: Nasal endoscopic examination showed an additional effect on turbinate congestion with combination therapy. Objective nasal airway assessment revealed that total nasal resistance decreased from 0,42 Pa/cm3/s to 0,32 Pa/cm3/s with monotherapy, and from 0,43 Pa/cm3/s to 0,27 Pa/cm3/s with combination therapy (p G 0,05) in average. Mean symtom scores of nasal congestion, nasal itching and sneezing pointed out a significant decline in the first three days, however nasal congestion scores were better with the antileukotriene add-on therapy. The positive effect on quality of life measures mainly in sleep, daily life activities and performance were increased, but combination therapy revealed significantly better results at the end of 21 days. It is significantly more effective than placebo. No side effects were encountered. Conclusion: These data provide a basis for optimism in the control of AR with antileukotriene-antihistamine combination therapy. The control on nasal congestion was more pronounced subjectively and objectively comparing to antihistamine alone. The effect might be due to the additional antiinflammatory activity as provided by reduction of inflammatory infiltrate and cytokine levels. More long-term studies are needed to evaluate the clinical effectiveness of antileukotrienes, especially as add-on therapy. Available data suggest it is reasonable and safe to add these agents to standard therapy if nasal symptomatology remains unresolved. The role of montelukast on perennial allergic rhinitis and associated sleep disturbance and daytime somnolence Background: One of the main manifestations of perennial allergic rhinitis is congestion. Nasal congestion can predispose to sleep apnea and microarousals. This sleep disturbance can result in daytime somnolence and fatigue. The use of montelukast may be effective at reducing congestion, but objective and subjective studies are lacking that demonstrate if the reduction of congestion will result in improved sleep and reduced somnolence. It was the purpose of this research to determine if montelukast is effective at improving sleep and daytime somnolence in patients with perennial allergic rhinitis. Methods: The study was a crossover, double-blinded, placebo-controlled study using montelukast 10 mg tablet or matched placebo and was approved by the IRB. Active phase of the study included a 2-week run-in followed by a treatment period of 2 weeks. After a wash out of 2 weeks, subjects were crossed over to the alternate arm of the study for 2 weeks. Subjective instruments to assess sleep and daytime somnolence were utilized. SAS was used for statistical assessment. Results: Montelukast 10 mg as compared to placebo statistically improved daytime sleepiness (p = 0.0089) and daytime fatigue (0.0087). With our small cohort we were unable to significantly demonstrate decreased congestion; however; congestion was reduced by 0.52 on a scale of 0Y3, while placebo reduced congestion by 0.16. Conclusion: Montelukast can reduce daytime somnolence and fatigue and may be a suitable alternative to topical nasal corticosteroids in those unwilling to use or are intolerant to nasal steroids and have daytime impairment from perennial allergic rhinitis. Intranasal phototherapy is more effective than fexofenadine hydrochloride in the treatment of seasonal allergic rhinitis Edina Garaczi 1 , Zsolt Bella 2 , Márta Boros-Gyevi 1 , Emese Tóth 1 , Zsanett Csoma 1 , Lajos Kemény 1 , and Andrea Koreck 1 . 1 University of Szeged, Dermatology and Allergology, Szeged, Hungary; 2 University of Szeged, Otolaryngology, Head and Neck Surgery, Szeged, Hungary. We recently showed that intranasal phototherapy represents an efficient therapeutic modality in the treatment of allergic rhinitis. The aim of this study was to compare the efficacy of intranasal phototherapy with that of a second generation antihistamine fexofenadine HCl in allergic rhinitis. A randomized open study was conducted in patients with history of moderate to severe ragweed-induced allergic rhinitis. Thirty-one patients were randomly assigned to receive either intranasal irradiation (low doses of UV-B, UV-A and visible light, referred to as mUV/VIS) 3 times a week for 2 weeks (n = 18), or 180 mg fexofenadine HCl per day for 2 weeks (n = 13). Each patient kept a diary of symptoms for nasal obstruction, nasal itching, rhinorrhea, sneezing and palate itching during the treatment. Total nasal score (TNS), a sum of scores for nasal symptoms was also calculated. In the mUV/ VIS group the individual scores significantly decreased compared with baseline for all of the parameters, sneezing (p = 0.0001), rhinorrhea (p = 0.0004), nasal itching (p = 0.0005), nasal obstruction (p = 0.009) and palate itching (p = 0.0001). In the fexofenadine HCl group none of the scores improved significantly at the end of the treatment except sneezing (p = 0.03). TNS was significantly decreased in the mUV/VIS group (p = 0.00003), but no significant change was observed in the fexofenadine HCl group. In conclusion, we found that intranasal phototherapy is a more efficient therapeutic tool than fexofenadine HCl in reducing clinical symptoms for allergic rhinitis. The burden of illness of allergic rhinitis in Canada Background: The objective was to assess the burden of symptoms in Canadian adults with allergic rhinitis (AR). Methods: A cross-sectional, random-digit-dialing telephone survey of 30,987 Canadian households was conducted in July 2006 to identify adult AR patients. After screening 3671 adults, structured interviews were done with 1001 respondents (patients diagnosed by a physician as having AR or taking medication for AR). Results: About 45% of Canadian adults report suffering from nasal symptoms due to allergies unrelated to colds. Less than half (45%) have been diagnosed by a physician. Half only have seasonal symptoms, with spring and summer being the worst seasons. Of those with AR, 27% had asthma, 17% chronic or recurrent sinusitis and 5% nasal polyps. More than onequarter cannot tolerate their symptoms without treatment. Most (83%) have sought medical attention for their symptoms at one time and one-quarter have done so in the past year. The most bothersome symptoms include stuffy nose, runny nose, repeated sneezing and watering eyes. In their worst month, two-thirds of patients reported having a stuffed nose either daily or several days per week. Almost one-quarter reported headaches and sleep loss. One-fifth describe symptoms as poorly controlled or not controlled during the worst month of the year. One-half use only OTC products, 12% use only prescriptions while one-quarter use both. Conclusion: Despite treatment, many Canadians experience allergic rhinitis symptoms that could be better evaluated and controlled. Asthma, sinusitis and nasal polyposis are common concomitant conditions. Radiofrequency tissue ablation treatment in persistent allergic rhinitis: effects on quality of life and objective parameters Kivanc Gunhan, Halis Unlu, Ali Vefa Yuceturk, and Murat Songu. Celal Bayar University, Otorhinolaryngology and Head-Neck Surgery, Manisa, Turkey. Background: In patients who suffer from persistent allergic rhinitis (PAR), a severe drug-resistant hypertrophy and increase in glandular structures of the inferior turbinates may develop, which leads to constant nasal obstruction and rhinorea. Radiofrequency tissue ablation of the turbinates can reduce nasal obstruction and secretions. Methods: This prospective, single-sited study randomized 50 patients with mild or moderate PAR who had substantial bilateral hypertrophy of the inferior turbinates to mometasone furoate monohydrate nasal spray (MFMNS), (2 sprays per nostril [total dose 200 2g] once daily), or radiofrequency inferior turbinate ablation (RIFA), (2 or 3 punctures on each site, total dose of 1510 T 110 joule, plateau temperature of 75-C, energy of 10 watt) treatment groups. Both objective outcomes evaluated by total nasal resistance at anterior rhinomanometry and subjective outcomes assessed with the Quality of Life Questionnaire were analyzed before and at least 12 months after treatment. Results: The median total nasal resistance in patients treated with MFMNS decreased from 0,49 T 0,17 Pa/cm 3 /s to 0,39 Pa/cm 3 /s (p = 0,42), and 0,51 T 0,18 Pa/cm 3 /s to 0,29 Pa/cm 3 /s in patients with RIFA (p = 0,003) 1 year postoperatively. Compared with preoperative scores, the postoperative scores of these patients significantly improved in both 7 separate domain scores and overall Quality of Life Questionnaire scores (p = 0,004). Nasal symptomatology was markedly reduced 1 month after radiofrequency application. No adverse reactions including bleeding, infection, adhesions or worsening of allergic symptoms were encountered. The patients experienced a lasting benefit from this procedure. Conclusion: These results suggest that topical mometasone reduces the volume of inferior turbinate at some point while improving the quality of life in patients with PAR. Radiofrequency inferior turbinoplasty is also improving quality of life and is more effective for decreasing nasal resistance in patients with PAR who have substantial nasal congestion. This effect might be due to the switch between the inflammatory cells and the fibrotic tissue. Histopathologic and longer term studies are required to enlighten the potential of RIFA in the management of allergic rhinitis. The role of allergic rhinitis in suppurative chronic otitis media, prelaminary reports Background: Eustachian tube dysfunction has a significant effect on pathogenesis of ear diseases especially chronic otitis media (COM). In the other hand, allergic rhinitis as a prevalent disease is a well-known condition which has some effects on eustachian tube function and its role in serous otitis media has been studied from several aspects, but its effects on chronic suppurative otitis media with & without cholesteatoma has not been studied yet. Methods: In a prospective case control study, 40 patients with suppurative chronic otitis media who were candidates for surgery and 38 healthy, sex and age-matched persons were evaluated for allergy using the standard questionnaire, prick test and serum IgE. Results: Fifteen of patients (37.5%) and six of controls (15.8%) have allergic rhinitis. The results showed the higher incidence of allergy in the patients' group (P G 0.05). Conclusion: To the authors' knowledge, this is the first documented survey showing the correlation between allergy and chronic otitis media. More extensive studies on the effect of allergy especially allergic rhinitis on chronic otitis media even postsurgical prognosis strongly recommended. Nasal mucosa remodeling and re-activation of epithelial mesenchymal trophic unit in human allergic rhinitis Tong Wang. Tongren Hispital Beijing, China, ENT Department, Beijing, China. Objective: To investigate the histopathologic characteristic of a marked chronic inflammatory reaction and remodeling, and that the role of reactivation of epithelial mesenchymal trophic unit in this marked structure change of the nasal mucosa in allergic rhinitis. Methods: Our study will be divided into two parts. The first part is to investigate whether remodeling of the airway mucosa are present in nasal mucosa of allergic rhinitis. The second part is to investigate the possible role of epithelial mesenchymal trophic unit in the pathogenesis of inflammation and remodeling in allergic rhinitis. The tissue blocks were embedded in paraffin, and stained with hematoxylin and eosin (HE), alcian blue periodic acid-Schiff (AB-PAS), Masson's Trichrome(MT), and immunohistochemical staining. The infiltrating eosinophils in nasal mucosa were examined, AB-PAS-positive cells in the surface epithelium in nasal mucosa were counted. The percentage area of MT stained extracellular matrix in mucosa and conchae and damage of epithelium were determined by an image analyzer. immunohistochemical staining revealed EGF, EGFR, TGF-", and !-SMA immunoreactivity in nasal mucosa. Results: Epithelial damage (shedding), basement membrane thickening, submucosa fibrosis, mucosal goblet cell hyperplasia, submucous gland hyperplasia, and strikingly eosinophil infiltration, lymphocyte infiltration was more prominent in patients with AR compared to the non-AR group (p G0.05). Compared with the control group, immunohistochemical staining revealed EGF, EGFR immunoreactivity in the damaged epithelium were decreased in allergic rhinitis. However, stronger TGF-" and !-SMA immunoreactivity was observed in allergic rhinitis mucosa as compared with normal nasal mucosa. Conclusion: The histopathologic findings of asthma, epithelial damage, goblet cells hyperplasia and extracellular matrix deposition, namely, inflammation and features of airway remodeling, are also present in allergic rhinitis. It maybe result from activation of the epithelialmesenchymal unit, leading to myofibroblast activation and propagation of remodeling responses into the submucosa. The concept of epithelialmesenchymal unit maybe contribute to the further understanding the pathogenesis of allergic rhinitis and may provide potential targets for novel therapy of allergic rhinitis. Zsolt Bella 1 , Edit Kadocsa 1 , Laszlo Szekely 2 , and Jozsef Jori 1 . 1 University of Szeged, ENT, Szeged, Hungary; 2 MediCenter Medical Institute, ENT, Szeged, Hungary. Objectives: Our previous randomised double blind clinical study proved that intranasal phototherapy using a combination of UVB (5%), UVA (25%) and visible light (70%) (mUV/VIS)(Rhinolight\) is an effective modality to treat allergic rhinitis (AR). The aim of this study was to show our clinical experiences on grass pollen sensitized allergic patients. Methods: We have treated AR patients (N = 243) with intranasal mUV/VIS between 2003Y2006. 79 patients had moderate/sever intermittant grass pollen induced AR. The pollen counts were over 40/m3 continuously. Rhinophototherapy was performed in monotherapy or combined with oral antihistamines (once a day) or/with nasal steroids (maximum 400 2g/die). Each intranasal cavity was irradiated 2Y3 times a week, for 2 weeks with mUV/VIS. The dose of irradiation was raised step by step from 1.6 to 2.7 J/cm2/nasel cavity. Nasal symptoms of each patient were registered on the treatment days with a scale of 0 to 3 and the total symptoms on a visual analog scale (VAS: 0Y10). Results: 3 patients dropped out (3,8%) . Finally 75 patients got full therapy. Monotherapy mUV/VIS: got worse: 6% of the patients, didn't change 19%, improved: 75% by VAS. mUV/VIS+drugs: got worse 5%, didn't change 8%, improved 87%. The nasal symptoms were significantly improved in each group. Rhinophototherapy was tolerated well. Using vitamin A oil 3Y4 times a day the development of painful xeromucosa could be avoided. Only 2 patients (2,53%) needed to have extra (more than one) brake days between two treatment occasions. Conclusion: These results suggest that intranasal phototherapy alone or with drug combined is an effective modality to treat grass pollen induced AR. Clinical characteristics of children with allergic rhinitis (AR) vs those with nonallergic rhinitis (NAR) from a university pediatric allergy clinic Chanthana Suratannon, Nualanong Visitsunthorn, and Piyarat Lerdbannapong. Mahidol University/ Siriraj Hosital, Pediatrics, Bangkok, Thailand. Background: Proper differentiation of chronic rhinitis into AR and NAR is essential since avoidance of offending allergen in AR could rapidly lead to alleviation of symptoms whereas NAR is usually a chronic and non-remitting condition. Unfortunately, allergy skin test is not feasible in general practice in several countries around the world. The aim of this study is to determine specific characteristics among pediatric patients with AR and NAR. Methods: Three hundred and two children with symptoms of chronic rhinitis presented to a Pediatric Allergy Clinic between January to December 2006 were categorized into 2 groups, i.e, allergic rhinitis (AR) and nonallergic rhinitis (NAR) according to their skin prick test results. Nasal cytology was performed to further subcategorize NAR into nonallergic rhinitis with eosinophilia syndrome (NARES). Results: There were 222 (73.5 %) patients with AR and 80 (26.4 %) with NAR. Nineteen (23.7%) NAR patients had NARES. The median age of onset in patients with AR and NAR were 4 and 3 years old, respectively. Symptoms of rhinorrhea, postnasal drip and nasal congestion did not differentiate the two groups apart. Interestingly, nasal itching, sneezing and eye symptoms were more common among AR than in NAR group (p G 0.01) while snoring and sinusitis were more common in NAR than in AR group (p G 0.01). Seasonality, severity of symptoms and the proportion of asthma did not differ between both groups. Prevalence of food allergy were more common in AR than NAR group (p G 0.05). Conclusion: AR was about threefold more prevalent than NAR among children presented to our allergy clinic with chronic rhinitis. Nasal itching and sneezing were more common in AR than NAR group (p G 0.01) while snoring and sinusitis were more common in NAR than AR group (p G 0.01). Eye symptoms and history of food allergy were more common in AR than NAR group (p G 0.01, p G 0.05). Allergic diseases are the increasing global health problem either in all over the world or in Hungary. In 2006 the incidence of allergic rhinitis was 25429 new cases (252%oo ), and the prevalence 263 925 (2619 %oo), and incidence of allergic asthma was 12 693 (126,0%oo) and prevalence 141 561 (1404,,9%oo). The basic treatment of the allergic disease has been H-1 receptor antagonists, and the levocetirizine is a new oral, non-sedating H-1 antihistamine, that has been shown to be effective against allergic symptoms, and offers good tolerability. Objective: To collect data concerning the treatment of allergic disease in general practice. to measure the effectiveness of levocetirizine in the allergic disease. to report the side effects during the levocetirizine therapy. Methods: A nationwide survey was organized in 2005 in Hungary with using the same questionnaire, filled two times ( at the start and at he end of treatment period) in.The patients participated in survey had allergic disease and was treated by levocetirizine during 6 months.Data of 17800 questionnaires were analysed. 57,8% of patients were female and 42,2 % male (p G 0,001). 57,2% of all patients has been aged 20Y49 y, 19 % was younger than 19 y. Results: 14 637 patients ( 82%) had allergic rhinitis, 3335 (19%) had urticaria, 3660 (20%) had allergic asthma, 1048 (6,8%) had atopic dermatitis, and 365 (2%) had food allergy. 57,1% of patients had symptoms of allergic disease longer than 4 years. Cutan test was performed in diagnosing of allergic disease in 78,5 % of patients, and in 92% was positive. The allergic disease influenced the way life of patients in 76,3%, but 88% of patients had clinically important symptoms (serious 40,4 %, or intermedient 47, 4%) . At the end of the survey 6,7% of patient had clinically significant symptoms, so in 93,3 % of patients_s symptoms improved. The 58,5% of allergic rhinitis and 51,0% persistent rhinitis patients become symptomless, and the ratio of symptom free cases was 51,3% of allergic asthma, 43,8% of atopic dermatitis, 51,9% of food allergy and 47,0% of urticaria. 52,4% of patients were treated by combination therapy. Side effects occured in 2,1% of patients during the treatment period. Summary: According the results of 6 months treatment of allergic diseases by levocetirizine it has been established that the levocetirizine was effective, decreased the clinical symptoms of allergic diseases, was well tolerated without important side effects. Frequency of adenoid hypertrophy and atopy in childhood Elena Korovkina. Mechnikov's research institute for vaccines and sera, Allergological department, Moscow, Russian Federation. Background: The adenoid is a part of Waldeyer's ring, the basic function of which is antibody formation, that react later against a great variety of antigens. Pediatric adenoidal obstruction of the nasal airway is associated with significant morbidity and is a frequent indication for surgery, but its relationship to allergy has not been often studied. Methods: To examine the influence of atopy on the adenoid hypertrophy we studied 35 children 1Y11 years old. In the analysed group all children suffered from difficulty in nose breathing, rhinorrhea, recurrent upper respiratory tract infections, hearing disorders, sleep disturbance. For the diagnostic of adenoid hypertrophy we had used the anamnesis, anterior rhinoscopy, endoscopy rhinoscopy. All patients had been consulted by the allergist-immunologist. For the diagnosis of allergic rhintis we had used interview, skin prick-test, presence of sIgE (RAST) in the serum, nasal provocation test with dust and polen allergens. Results: All children had had adenoid hypertrophy 2 or 3Y4 degree, 3 of them had got the relapse of adenoids after the surgery. In the analised group 21 children (60%) had had the positive skin prick-tests and/or positive reactions to specific IgE, mostly to different kinds of pollen or house dust mites; eosiniphils in nasal secrete; positive nasal provocation tests. In this group 9 patients (25%) had had the bronchial asthma, and 6 children (17%) Y atopic dermatitis. All patients with adenoids and allergic rhinitis had got the complex therapy Y topical nasal steroid and antihistaminic in the ages doses. The children with brochial asthma and atopic dermatitis also received a pharmacological treatment. In a during of 3 weeks we observed the most significant decrease of clinical symptoms and endoscopic adenoid size. Conclusion: The study shows that allergy and sensitivity to different kinds of allergens is an important risk factor for a greater degree of adenoid hypertrophy in children. Chronic allergic inflammation of the upper airway is the causes of lymphoid hypertrophy with prominence of adenoidal and tonsillar tissue. So, all patients with the difficulty in nase breathing and reccurent upper respiratory tract infections need in the consultation by allergist-immunologist for exception of allegic diseases. The quality of life in patients with atopic asthma associated with allergic rhinitis Ion Gabriel Stoica. Spitalul de Pneumologie, Ambulatoriul TBC Sector 3, Bucharest, Romania. Objectives: The aim of the study was to assess the effect of allergic rhinitis on quality of life in patient with atopic asthma. Methods: We examined 45 atopic asthmatics patients with persistent allergic rhinitis and 36 patients with atopic asthma without allergic rhinitis. Quality of life was measured by the Asthma Quality of Life Questionnaires (AQLQ). Clinical symptom score and use of rescue medications were recorded in diaries for 4 weeks before the assessment of quality of life. Lung function was measured on the same day as the study of quality of life. Results: Clinical symptom score and use of shortYacting $ 2Yagonists were similar in both groups and there was no difference in FEV1 (63.00 3.75 vs. 65.84 3.58, p 9 0.05). Asthma symptom scores (2.83 0.14) and activity limitation (3.47 0.22) within AQLQ were significantly lower for patients with asthma and allergic rhinitis compared with those asthma patients without allergic rhinitis (4.01 0.21 and 4.23 0.25, p G 0.05). Conclusion: Persistent allergic rhinitis may have a negative effect both on asthma symptoms and on quality of life in patients with atopic asthma. Background: Allergic rhinitis is the most common cause of rhinitis affecting approximately 20% of the population. While allergic rhinitis is not a lifethreatening condition, complications can occur and the condition can significantly impair quality of life, which leads to a number of indirect costs. In susceptible individuals, exposure to certain foreign proteins leads to allergic sensitization, which is characterized by the production of specific IgE directed against these proteins. This specific IgE can be tracked with Standard Skin Prick Test. Methods and Materials: During 6 months of study, 334 cases of allergic disorders referred to our allergy clinic, among them 295 were allergic rhinitis. Standard Skin Prick Test with ALUSTAL prick allergens, aero-allergens and food allergens, were performed for all of them and compared with negative and positive standard control. Wheals of 3-mm and flares of 10-mm larger than negative control regarded as positive test. Results were analyzed with SPSS 11.5 and prevalence of allergens was calculated in different groups of patients in regard to age, sex and type of allergic rhinitis (perennial or seasonal). Results: 150 males with mean age of 25.6 years (SD: 14.31) and 145 females with mean age of 28.32 (SD: 12.93) were studied among them 85 patients (30%) had seasonal Allergic Rhinitis (SAR), 117 patients (41.3%) had Perennial Allergic Rhinitis (PAR), 65 patients (23%) had PAR with seasonal aggravation and 16 patients (5.7%) had only episodic symptoms. 116 patients had less than 5 years, 117 patients had 5Y10 years and 102 patients had more than 10 years history of rhinitis. Aeroallergen sensitization were detected in 256 (86.5%) of patients and botanical allergens`sensitization were shown to be present in 225 (76%) of patients. Chenopodiacea including Chenopodium alba and Rough pigweed, trees especially ash, platanus, willow and birch, 12 grasses including artemisia, rye-grass and timothy were among the most prevalent botanical allergens. Conclusion: Allergic rhinitis is the most prevalent allergies which cause signs and symptoms in genetically susceptible patients confronting environmental allergens. In our study, aeroallergens were studied. As in other studies, aeroallergens especially botanical allergens were shown to be the most common allergens both in Seasonal and perennial allergic rhinitis with 86.5% sensitive to one or more aeroallergens and 76% sensitive to one or more botanical allergens. Rhinitis: is it allergic or not? Azita Hekmatdoost, Nader H. Rad, and Meran Ghoreishi. Tehran University of Medical Sciences, Faculty of Health, tehran, Islamic Republic of Iran. Introduction: there are no acceptable criteria for distinguishing between allergic and non-allergic rhinitis. The aim of this study was to compare the clinical and para-clinical features of allergic and non-allergic rhinitis. Methods: in a retrospective study, we compared the clinical and para-clinical features of 442 allergic and 103 non allergic rhinitis patients (18Y50 year-old). The evaluations included respiratory signs and symptoms, lab tests, and respiratory function tests. Results: patients with allergic rhinitis had more signs and symptoms within their season in terms of cough, sneezing, nasal congestion, rhinorrhea, itchy eyes and nose, and respiratory function test (p G 0.001), whereas those with non-allergic rhinitis had more persistent symptoms, and headaches (p G 0.01). Food allergy, OR = 1.8, (1.4Y2.1), P G 0.01, serum Ig E, OR=2.1, (1.9Y2.19), P G 0.001, and airway hyperresponsiveness, OR=1.7 (1.4Y1.9), P G 0.01 were more common in patients with allergic rhinitis. Conclusion: our results indicate that the signs and symptoms have more fluctuation in allergic rhinitis, and more sever within their season. More studies with respect to both adults and children with rhinitis is recommended. Allergic and non-allergic rhinitis in adolescent and adults Levan Bolokadze, Eugeniya Bolokadze, and Volodimir Fedotov. Kharkov Allergological Centre, Allergology, Kharkov, Ukraine. Background: The aim of this study was to describe differences between allergic rhinitis (AR) and nonallergic rhinitis (NAR) in a large communitybased sample of adolescents and adults. Methods: A total of 354 subjects, 10Y50 years of age, who in a screening questionnaire had reported a history of airway symptoms suggestive of asthma and/or allergy, or who were taking any medication for these conditions were clinically examined. All participants were interviewed about respiratory symptoms and furthermore skin test reactivity, lung function and airway responsiveness were measured using standard techniques. Results: A total of 64% of the subjects with rhinitis had AR, whereas 46% had NAR. Subjects with NAR were more likely to be females, OR = 2.13 (1.24Y3.22), P = 0.004, to have persistent symptoms within the last 6 weeks, OR = 1.87 (1.23Y2.65), P = 0.002, and to have recurring headaches, OR = 1.94, (1.34Y3.45), P = 0.021. On the other hand, subjects with NAR were less likely to have airway hyperresponsiveness, OR = 0.34, (0.32Y0.46), P G 0.001, food allergy, OR = 0.32, (0.18Y0.33), P = 0.007 and to have been treated with antihistamines in the last 6 weeks, OR = 0.23, (0.22Y0.35), P G 0.002 compared with subjects with AR. Subjects with AR were symptomatically worse within their season in terms of sneezing (P G 0.002) and itchy eyes (P G 0.002), compared to subjects with NAR, whereas nasal congestion and rhinorrhea were equally frequent in the two groups (P = 0.32 and P = 0.44, respectively). Conclusion: The proportion of subjects with NAR in an adolescent and adult population with rhinitis is around one-fourth. Women have NAR twice as often as men. In general, subjects with NAR have more persistent but equally severe symptoms compared to subjects with AR. However, subjects with allergic rhinitis have more sneezing and itchy eyes within their particular season of allergy compared to subjects with nonallergic rhinitis. Relationship between histamine H1 receptor occupancy (RO) and reduction of symptoms of seasonal allergic rhinitis (SAR) by levocetirizine (L) in subjects exposed to pollen in an environmental exposure unit (EEU) Background: the estimation of in vivo RO, which takes into account the affinity of an H1 antihistamine for the histamine H1 receptor and its free plasma concentration, is a far better predictor of human pharmacodynamics evaluated by the inhibition of histamine or allergen-induced wheal & flare than considering in vitro affinity and plasmatic half-life only (Gillard et al. Inflamm Res 2005 , Frossard et al. Br J Clin Pharm 2007 . The purpose of this study is to assess whether this is valid also for other pharmacodynamic models such as that scoring the symptoms of SAR in ragweed sensitive subjects exposed to ragweed pollen in an EEU. Methods: 119 subjects (mean age 34 yrs, mean bodyweight 81 kg) were exposed to pollen in the EEU at 9 am of Day 1. At 11 am, they received a single dose of 5mg L and remained in the EEU until 4 pm. On Day 2, subjects were exposed again to pollen in the EEU from 8 am until noon. The primary efficacy variable was the reduction from baseline in the Major Symptoms Complex (MSC) score (calculated as the sum of the following 6 symptoms: runny nose, itchy nose, sniffles, nose blows, sneezes and watery eyes). Baseline value was the MSC score before drug administration (9Y11 am of Day 1). The MSC score was evaluated at different times after L administration (see table) . Plasma concentrations at different times after administration of a single dose of 10 mg L were obtained from a study in 24 healthy volunteers (mean age 35 yrs, mean bodyweight 67 kg) (Baltes et al. Fund Clin Pharm 2001) , normalized to the therapeutic dose of 5 mg (see table) . The plasma protein binding data of L come from Bree et al. (Fund Clin Pharm 2002) . The RO has been calculated as described by Gillard et al. (Inflamm Res 2005) . Results: The kinetics of change from baseline in MSC score fit much better with those of RO than with those of free plasma concentrations (see table) . It is possible however that the free plasma concentrations of L do not represent those at the receptor at very short times and that the RO values calculated are over-estimated at these times. It is also possible that there is a lag time between the RO and the relief of some of the symptoms. Conclusion: the EEU model confirms previous results that the kinetics of RO by L are a better predictor of the efficacy of the drug than are plasma pharmacokinetics. Expression of matrix metalloproteinase 2, 9 and 13, TIMP 1, 2 in the allergic nasal mucosa inhibitors of MMPs known as TIMP regulate MMP function. Allergic rhinitis (AR) and asthma share many similarities in their inflammatory response but epithelial shedding, basement membrane thickening and fibrosis are not seen in AR. By contrast, nasal polyps share some features with asthma. As in asthma, we demonstrated an increased expression of MMP-9 and low TIMP in nasal polyps. We hypothesized that the differential expression of MMPs may at least in part contribute to the differences between AR and asthma. Methods: We performed nasal allergen challenge (NAC) in AR patients with mite allergen disks and control disks and analyzed the number of MMP 2, 9, 13 and TIMP 1,2 in nasal biopsies at 30 min, 6 hrs and 12 hrs post challenge. By ELISA, we examined the levels of MMP-2, 9 and 13, and TIMP1,2 in the nasal mucosa of AR patients. Results: At 30 min post NAC, MMP-2 and 13 were increased. At 6 hrs post NAC, MMP-2 and at 12 hours post NAC, MMP-2 and 13 were significantly increased. TIMP-1 was increased at 30 min. At 30 min and 12 hr post NAC, the MMP 2 : TIMP1,2 and MMP-13 : TIMP1,2 ratio was high. The levels of MMP-2 and 13 but not MMP-9 were high in the allergic nasal mucosa. Conclusion: These results suggest that MMP-2 and 13 may play an important role in the pathomechanisms of allergic rhinitis and that this differential expression of MMP may contribute at least in part contribute to the differences between AR and asthma. CRTH2 plays an essential role in the pathophysiology of Cry j 1-induced pollinosis in mice Tokyo Medical and Dental University, Human Gene Sciences Center, Tokyo, Japan. Background: Prostaglandin (PG) D2 is the major prostanoid produced during the acute phase of allergic reactions. Two PGD2 receptors have been isolated, DP and CRTH2, but whether they participate in the pathophysiology of allergic diseases remains unclear. We investigated the role of CRTH2 in the initiation of allergic rhinitis in mice. Methods: First, we developed a novel murine model of pollinosis, a type of seasonal allergic rhinitis. Pathophysiological differences in the pollinosis were compared between wild-type and CRTH2-gene deficient mice. An effect of treatment with ramatroban, a CRTH2/T-prostanoid dual antagonist, was also determined. Results: Repeated intranasal sensitization with Cry j 1, the major allergen of Cryptomeria japonica pollen, in the absence of adjuvants significantly exacerbated nasal symptoms, Cry j 1-specific IgE and IgG1 production, nasal eosinophilia, and Cry j 1-induced in vitro production of IL-4 and -5 by submandibular lymph node cells. In addition, CRTH2 mRNA in nasal mucosa was significantly elevated in Cry j 1-sensitized mice. Following repeated intranasal sensitization with Cry j 1, CRTH2-gene deficient mice had significantly weaker Cry j 1-specific IgE/IgG1 production, nasal eosinophilia, and IL-4 production by submandibular lymph node cells than wild-type mice. Similar results were found in mice treated with ramatroban. Conclusion: These results suggest that the PGD2-CRTH2 interaction is elevated following sensitization and plays a proinflammatory role in the pathophysiology of allergic rhinitis especially pollinosis in mice. The clinical course of allergic rhinitis and asthma and changes in skin prick test and spirometry in the aftermath of hurricane Katrina Prem Kumar, Annette Fiorillo, Sandhya Mani, and Douglas Barstow. LSU, Allergy/Immunology, New Orleans, LA, United States. Background: Allergic rhinitis and asthma are both prevalent diseases seen in everyday practice. In the aftermath of hurricane Katrina several of our patients reported exacerbations of allergic rhinitis, allergic conjunctivitis and either exacerbations or development of asthma. As a result of Katrina many homes were flooded, causing significant increased mold levels. We sought to investigate if these changes in reported symptoms correlated with changes in Skin Prick Test (SPT) reactivity and/or changes in spirometry. Methods: At the start of the study, we looked at patients who reported increased allergic symptoms and in whom we had previously performed SPT. We then repeated SPT and compared the results to earlier findings. In addition, patients who had a history of asthma or those complaining of respiratory symptoms had spirometry done and their results were compared with those done before Katrina. Results: Four of five patients were reactive to many antigens to which they were previously anergic. Reactivity to molds was increased, most commonly Alternaria, Aspergilus fumigatus and Cladosporium. In addition changes in spirometry were also noted. In one patient who initially had normal spirometry with negative methalcholine challenge, developed abnormal spirometry. This patient's FEV1 was markedly decreased and improved by more than 15% after bronchodilators. Conclusion: There has been much concern about the effect Katrina may have had on patient health. Many patients have demonstrated increased symptoms relating to allergic rhinitis and asthma. These cases demonstrate that atopic patients in New Orleans have been sensitized to new allergens. Patients that are experiencing increased symptoms of rhinitis, conjunctivitis or asthma should be re-evaluated with SPT, spirometry and a thorough history to environmental exposure. With the large number of people affected by Katrina, further studies should be performed as more patients return to the New Orleans area. Mechanisms of mast cell migration into the allergic nasal epithelium Chika Ozu, Ruby Pawankar, Takizawa Ryuta, Nonaka Manabu, and Yagi Toshiaki. Nippon Medical School, Departoment of Otorhinolaryngology, Tokyo, Japan. Objective: Mast cells are increased in the nasal epithelium of patients with allergic rhinitis (AR). Yet, the precise mechanisms of this increase are unclear. We previously reported that nasal mast cells express CCR3 and exhibit increased chemotaxis to RANTES suggesting a role for RANTES in mast cell migration. To further confirm this, in the present study we examined the levels of RANTES, eotaxin and SCF in the epithelium and lamina propria of patients with AR and the kinetics of RANTES+, tryptase+, and CCR3+ cells in the epithelium and lamina propria after nasal allergen challenge. Methods: By ELISA, we examined the levels of RANTES, eotaxin and SCF in homogenized nasal scrapings and lamina propria of AR patients. In AR patients, we performed nasal allergen challenge with house dust mite, took biopsies at 30 min, 6 hrs, and 12 hrs and by immunohistochemistry, we examined the number of Tryptase+ RANTES+, eotaxin+ and CCR3+ cells as compared to control. Results: The levels of RANTES, but not Eotaxin and SCF was greater in the epithelium than in the lamina propria. At 30 minutes after nasal allergen challenge, Tryptase+, RANTES+ and CCR3+ cells were increased in the epithelium. At six hours post challenge, Tryptase+ and RANTES+ cells were increased in the epithelium but at 12 hrs only an increase in Tryptase+ cells was detected. Conclusion: Migration of the mast cells in the allergic nasal epithelium occurred as early as 30 min with a parallel increase in CCR3+ and RANTES+ cells. These results further confirm that RANTES is one of the critical factors regulating mast cell migration into the allergic nasal epithelium. (4) :389Y96) and explained as a consequence of competition between the two drugs for renal excretion mediated by the transporter proteins organic cation transporter 2 (hOCT2) and P-glycoprotein. The effect of levocetirizine and dextrocetirizine, the eutomer and the distomer of cetirizine, respectively, on the transport of the hOCT2 substrate tetraethylammonium (TEA), was investigated in vitro in chinese hamster ovary (CHO) cells stably transfected with hOCT2. Methods: CHOhOCT2 cells were seeded in 12 well plates and grown to confluence. Once confluent, transport experiments were conducted. Transport buffer containing [ 3 H]TEA, and levocetirizine, dextrocetirizine or positive control (cimetidine) were added to the wells. At intervals, the transport buffer was removed, and each well rinsed three times with 1 mL of ice cold buffer to stop transport. Cells were solubilized and aliquots removed for scintillation counting. Results: Levocetirizine and dextrocetirizine inhibited TEA uptake in CHOhOCT2 cells with IC50 values of 197 and 714 6M, respectively. In comparison, the positive control cimetidine inhibited TEA uptake with an IC 50 of 266M. Discussion: Levocetirizine is a weak inhibitor of hOCT2. The IC 50 value for inhibition of tetraethylammonium transport by levocetirizine is more than 300 fold the C max (0.66 6M) at steady state following the therapeutic dose of 5 mg. It is unlikely that levocetirizine can cause drug interactions by interference with renal elimination through inhibition of renal OCT2. As dextrocetirizine is a weaker inhibitor of hOCT2 than levocetirizine, the racemate cetirizine is also unlikely to cause drug interactions through inhibition of hOCT2. Expression of glucocorticoid receptor-$ in glucocorticoid-resistant allergic rhinitis Akihiro Ishida, Nobuo Ohta, and Masaru Aoyagi. Yamagata Univ., School of Medicine, Department of Otolaryngology, Yamagata, Japan. Background: Glucocorticoid (GC) has been commonly used as an antiinflammatory reagent in the treatment of chronic allergic diseases including allergic rhinitis. The effects of GC are mediated by glucocorticoid receptor-" (GR-"). Upon binding GC, activated GR-" can not only enhance transcription of anti-inflammatory genes but also interact with other protein regulating inflammation, such as nuclear factor-0B (NF-0B). However, these clinical benefits are sometimes limited because some patients demonstrate persistent tissue inflammation despite treatment with high doses of GC. It is generally considered that the interference of non-functional GR variants in immune cells may result in GC-resistance. GR-$ is a well-known natural spliced variant consisting of 742 amino acids including exon 2-8 and part of 9$ as coding region, but cannot bind to GC. It has been reported that there was an increase of GR-$ expression in patients with bronchial asthma and in ulcerative colitis patients who did not respond to GC administration. We have tested this hypothesis by investigating correlation between the expression of GR-", GR-$, and NF-0B proteins in patients with allergic rhinitis and the responsiveness to GC treatment. The patient group consisted of 20 subjects with allergic rhinitis showed persistent GC-resistance, resulting in a required surgical removal after the GC treatments for over 6 months. As normal control, nasal tissues were obtained from 10 subjects underwent maxillofacial surgery. They had not been exposed to GC treatment. We have performed immunohistochemical analysis to detect GR-", GR-$ and NF0-B proteins in nasal tissues. Results and Conclusion: Compared to normal subjects, whereas the number of GR-"-positive inflammatory cells was decreased, the number of GR-$positive cells was significantly increased in nasal tissues from patient group. The number of NF-0B-positive cells was at a similar level both patients and normal control. In conclusion, our data suggests that increased over expression of the GR-$ relative to GR-" is associated with GC-resistance. Effect of azelastine on substance P release into nasal lavage from non-allergic rhinitis patients Radoslaw Gawlik 1 , Barbara Jawor 1 , and Lawrence DuBuske 2 . 1 Silesian University School of Medicine, Allergy and Immunology, Zabrze, Poland; 2 IRINE, Allergy and Immunology, Gardner, United States. Introduction: Non-allergic rhinitis is a condition which affects 30 to 50% of patients with perennial nasal symptoms, occurring either alone or in conjunction with allergic rhinitis. Neural mechanisms may have a role in this condition. This study assesses the impact of intranasal azelastine on the concentration of substance P (SP) in nasal lavage fluid. Methods: 28 patients were defined as having non-allergic rhinitis by history, examination in addition to the presence of negative skin prick tests using common inhalant allergens and low total IgE using the Phadia CAP. 16/28 patients were treated with intranasal azelastine two sprays twice daily for 10 days. The control group consisted of 12 patients with non-allergic rhinitis who used a saline nasal spray, 2 sprays twice a day. Nasal lavages were performed before and after a 10 day treatment period with intranasal azelastine or control saline nasal spray. The concentration of SP in nasal lavage fluid was determined by an EIA (Assay Designs Inc., USA) method. All patients recorded their nasal symptoms daily, including rhinorrhea, sneezing, pruritus, and congestion, using a Visual Rating Scale (VRS). The baseline concentrations of SP in the nasal lavage fluid were similar in the azelastine and the placebo group (86.8 T 18.2 pg/mL versus 82.3 T 21.4 pg/mL). Significantly lower concentrations of substance P were noted in nasal lavage fluid after 10 days of azelastine treatment (73.2 T 16.9 pg/mL) when compared with placebo treatment (83.1 T 17.8 pg/mL). The differences between the azelastine and the placebo treated group VRS symptom scores (6.4 T 2.4 versus 8.6 T 3.2) were also statistically significant demonstrating that the reduction in nasal lavage SP levels occurred in conjunction with improvement in clinical symptoms. Conclusion: Intranasal azelastine reduces substance P release into nasal lavage fluid of non-allergic rhinitis patients associated with clinical improvement during a 10 day treatment period. The efficacy of azelastine in perennial non-allergic rhinitis may be related to reduction of release of SP and other non-adrenergic non-cholinergic neural mediators into nasal secretions. A double blind placebo controlled study on the clinical efficacy and in vivo pharmacodynamics of potassium humate in the treatment of hayfever in patients with inhalant allergies Several mechanistic studies were done on potassium humate, derived from bituminous coal, during the last few years. It was established that this product stimulates lymphocyte proliferation by an increased production of the growth factor, IL2. However, no in vivo studies have been done on the anti-inflammatory effects of humate derived from coal in humans. It was proven that humic acid s extracted from brown coal have no toxic or teratogenic effects. Purpose: The aim of this study is to investigate potassium humate`s antiinflammatory properties in patients suffering from exacerbations of hayfever during the grass pollen season in South Africa, using clinical symptoms and signs as well as establishing changes in inflammatory markers using established and new laboratory techniques. A total symptom score over a 12 hour period and a quality of life questionnaire were evaluated. Potassium humate (1.8g in daily divided doses) was randomly assigned to 20 atopic patients presenting with acute symptoms of hayfever. 20 Patients received a placebo resembling the humate. Treatment period was one month, preceded by a one week run-in period. Symptoms were scored on a scale of one to four for nasal and nonnasal symptoms. A global clinical impression was scored by the doctor at end of study: Measurement of surface area of skin prick test was done at baseline and again at the end of the study. It was restricted to the inhalant that indicated the biggest reaction. Results: No differences in the presence and improvement of symptoms were detected between the 2 groups. Results of the quality of life assessment were also the same for the 2 groups. Flare size comparison showed some promising results with a significant difference (p G 0.05) observed when comparing the before and after results of the treated versus placebo groups respectively. Conclusion: Potassium humate shows promise in the treatment of hayfever or allergic rhinitis. The study was also conducted on inflammatory markers and showed positive results, the results presented elsewhere. The duration of the study, one month, might have influenced the results. Evidence that nasal mucosal hyperreactivity in healthy women is induced by high levels of estrogen Karin Toll 1 , Peter Graf 2 , Magnus Backheden 3 , and Par Stjarne 1 . 1 Karolinska Institutet, Division of Otorhinolaryngology, Stockholm, Sweden; 2 Karolinska Institutet, Dept. of Women and Child Health, Stockholm, Sweden; 3 Karolinska Institutet, LIME, Stockholm, Sweden. Background: Pregnancy rhinitis, a common condition, is thought to affect 18Y30 % of pregnant women. It is very annoying for many women, may develop at any time during pregnancy and usually disappears shortly after delivery. The cause is not known and it may be due to hormonal factors. Some data have shown that there may be a relation between high estrogen levels and nasal mucosal reactivity. The knowledge of the etiology of this condition is important since no satisfactory treatment is available. The aim of this study was to determine whether nasal mucosal reactivity and microcirculation change with various levels of estrogen in the blood, but constant levels of other hormones. Methods: 15 women who were undergoing in vitro fertilization (IVF) were included. The examination and measurements of the nasal mucosa were done in the first part when the concentrations of estrogen in the blood are extremely low and then, when the concentrations of estrogen is high. The nasal mucosa was studied with a combination of rhinostereometry and laser-Doppler flowmetry during challenge with histamine. The swelling of the nasal mucosa was recorded with rhinostereometry. This optical, direct, non-invasive method is designed to measure nasal mucosal swelling with a high degree of accuracy. Laser-Doppler flowmetry, a non-invasive method for studying the microcirculation, is providing continuous and instantaneous measurements of nasal mucosal blood flow. The combination of rhinostereometry and Laser-Doppler flowmetry has the advantage of using two non-invasive methods which permit direct and simultaneous measurements of congestion and the microcirculation. Results: With rhinostereometry we found an increase in nasal mucosal swelling after histamine challenge when the estrogen levels in blood were high. With laser-Doppler flowmetry the increases of the microcirculatory parameters velocity of moving blood cells and perfusion were lower when the estrogen levels in blood were high. Conclusion: High levels of estrogen in blood during IVF treatment of healthy women may induce nasal hyperreactivity and changes in microcirculation. This suggests that estrogen plays a role in pregnancy rhinitis. Effect of a Lactobacillus paracasei on grass pollen allergic rhinitis Sophie Nutten 1 , Jacqueline Wassenberg 2 , Régine Audran 2 , Julie Moulin 1 , Irène Corthésy-Theulaz 1 , Annick Mercenier 1 , and Fran0ois Spertini 2 . 1 Nestle Research Center, Nutrition and Health Department, Lausanne, Switzerland; 2 CHUV, Service d'Immunologie et d'Allergie, Lausanne, Switzerland. Background: Lactobacillus paracasei strain NCC2461 has been found to exert anti-allergic effects in animal models. Aim: the aim of the present study was to investigate the effect of NCC2461 in patients with allergic rhinitis to grass pollen. Methods: 31 subjects (18-35 years-old) were enrolled in a randomized double-blind, placebo-controlled cross-over study. The study consisted of a first 4 weeks period in which subjects consumed either fermented milk containing NCC2461 or placebo (acidified milk drink), a washout period of six to eight weeks and a final 4 weeks period cross-over of the first treatment phase. The entire study was performed out of the pollen season. Clinical symptoms were analysed after a nasal provocation test using grass pollen allergens and immunological parameters (specific immunoglobulins in serum, percentage of eosinophils in nasal washes and cytokines secreted by restimulated PBMC) were compared between the two treatment periods. Results: A trend to a decrease of nasal itching favouring probiotic treatment was noticed, and a significant improvement of the nasal blockage favouring probiotic treatment was observed. However, no significant change of the nasal reaction threshold was observed between the end of the first and second treatment periods, whatever the sequence (active product before or after placebo) of product consumption. Immunological parameters' analyses are under investigation. No side effects were reported for both groups during the study period. Conclusion: A decrease of nasal blockage was observed in allergic patients nasally challenged with grass pollens, after NCC2461 consumption. The ongoing analysis of immunological parameters should help understanding the mechanisms leading to the mitigation of respiratory allergy symptoms. Fluticasone furoate nasal spray demonstrates consistent efficacy against both the nasal and ocular symptoms of seasonal allergic rhinitis Robert Naclerio 1 , Tom Toler 2 , and Anna Ellsworth 2 . 1 University of Chicago, Department of Surgery, Chicago, United States; 2 GlaxoSmithKline, Research and Development, Research Triangle Park, NC, United States. Background: Fluticasone furoate (FF) is a novel enhanced-affinity glucocorticoid that has been developed for topical respiratory use. Data from four randomised studies are presented to demonstrate the consistent nasal and ocular efficacy of FF nasal spray (FFNS) in adolescent and adult patients with seasonal allergic rhinitis (SAR). Methods: Patients aged Q12 years with confirmed SAR were enrolled in the four studies (n = 1141, combined total) and received once-daily double-blind treatment with FFNS 110 6g (n = 571, combined total) or vehicle placebo spray (n = 570, combined total) for 2 weeks. Individual nasal and ocular symptoms were scored by patients each on a 4-point categorical scale (0 = none to 3 = severe) each morning and evening. Nasal and ocular efficacy was evaluated by the mean change from baseline over the 2-week treatment period in daily reflective Total Nasal Symptom Score (rTNSS; average of AM and PM score totals for nasal congestion, nasal itching, rhinorrhoea, and sneezing) and daily reflective Total Ocular Symptom Score (rTOSS; average of AM and PM score totals for eye itching/burning, tearing/watering, and redness). The AM pre-dose instantaneous Total Nasal Symptom Score (iTNSS) and Total Ocular Symptom Score (iTOSS) were also assessed as a measure of 24-hour symptom control. Results: Significant and consistently greater improvements in rTNSS were seen with FFNS than with placebo across all four studies ( Background: The safety profile of the novel enhanced-affinity glucocorticoid fluticasone furoate (FF) administered using a unique, side-actuated device has been investigated in children with perennial (PAR) and/or seasonal allergic rhinitis (SAR). Methods: Safety was evaluated in an integrated analysis of data from three randomised, double-blind, parallel-group studies in 1224 paediatric patients aged 2Y11 years with SAR or PAR. Patients received once-daily FF nasal spray (FFNS) 55 6g (n = 369) or 110 6g (n = 426), or placebo (n = 429) for 2 weeks (SAR study) or 6 or 12 weeks (PAR studies). Assessments included evaluation of adverse events (AEs), clinical laboratory tests, nasal examinations, ophthalmic examinations (12 week PAR study only) and electrocardiograms (ECGs). In the 6-week study, the effects of once-daily FFNS 110 6g on 24-hour serum cortisol (SC) was assessed in a domicile setting in patients with PAR. Results: No safety or tolerability issues were identified in paediatric patients with PAR or SAR. In the integrated analysis, the most common AEs with an incidence of 93% and more common in FFNS than in placebo were headache, nasopharyngitis, epistaxis, pyrexia and pharyngolaryngeal pain. The incidences of drug-related AEs were similar across the FFNS 55 6g and 110 6g treatment and placebo groups: epistaxis was reported in 4%, 2% and 3% of patients, respectively, and headache in 2%, G1% and 1% of patients, respectively. Data from the 6-week PAR study demonstrate that SC levels following administration of FFNS 110 6g were similar to those following placebo (least squared means, 0.94 vs 0.97; treatment ratio, 0.97; 95% confidence interval, 0.88, 1.07) which suggests that FFNS 110 6g is not associated with any effect on the HPA axis. In addition, plasma levels of FF were non-quantifiable. Conclusion: Once-daily FFNS 55 6g or 110 6g has a favourable safety and tolerability profile in children aged 2Y11 years with PAR or SAR. Furthermore, FFNS shows low systemic exposure and is not associated with hypothalamic-pituitary-adrenal axis suppression in children aged 2Y11 years with PAR. Long-term safety of fluticasone furoate nasal spray 1106g once daily in adults and adolescents with perennial allergic rhinitis Background: Fluticasone furoate is a novel enhanced-affinity glucocorticoid, with unique pharmacological properties suitable for topical use in rhinitis. This study investigated the long-term safety and tolerability of fluticasone furoate nasal spray (FFNS) over 12 months in adults and adolescents with perennial allergic rhinitis (PAR). Methods: Following a 7-to 14-day screening period, patients aged Q12 years with PAR were randomised in a 3:1 ratio to double-blind treatment with oncedaily FFNS 1106g (n = 605) or vehicle placebo nasal spray (n = 201) for 12 months. Adverse event (AE) data were collected using diary cards and interviews at each study visit; the investigator graded AE severity as mild, moderate or severe, and assessed the relationship of each AE to the administration of study treatment. Safety was assessed by 24-hour urinary cortisol excretion, nasal and ophthalmic examinations, ECGs and clinical laboratory testing. Plasma levels of FF were determined from blood samples. Compliance was measured using diary cards, change in bottle weights, and symptom scores. Results: 592 patients (73%) completed the study. Demographic characteristics were similar in treatment and placebo groups. FFNS was well tolerated: the incidence of most AEs in the FFNS group was similar to that in the placebo group, with the exception of epistaxis (defined as any observation of blood in or from the nose, irrespective of quantity), which was more frequent in FFNS than placebo recipients (20% vs 8%). There were no other clinically meaningful differences between FFNS and placebo in terms of safety assessments, including 24-hour urine cortisol excretion, mean ophthalmic parameters, ECGs and clinical laboratory tests. Plasma levels of FF were not quantifiable in the majority of patients following administration of FFNS. Compliance with study treatment was high based on diary cards (85% of patients had 990% compliance) and bottle weights. Over the treatment period, improvements in reflective Total Nasal Symptom Score were greater with FFNS (j3.37) than with placebo (j2.49). Conclusion: Long-term (12-month) administration of FFNS 1106g once daily in adult and adolescent patients with PAR revealed a safety profile typical of intranasal corticosteroids as a class, with no evidence of clinically relevant systemic corticosteroid exposure. The prevalence of allergic rhinitis in college students at Kenya Medical Training College-Nairobi,Kenya Charles Gathiru 1 , and Isaac Macharia 2 . 1 Malindi District Hospital, Ear Nose Throat Department, Mombasa, Kenya; 2 University Of Nairobi, Department of Surgery-Ear Nose Throat, Nairobi, Kenya. Background: Allergic rhinitis is one of the commonest atopic diseases world wide yet its epidemiology in Kenya remains largely unknown. Currently, there is only one questionnaire based study (International Study of Asthma and Allergies-ISAAC) in children documented in Kenya. Objectives: The primary objective was to determine the prevalence of allergic rhinitis in Kenya Medical Training College students, aged 18Y50 years. The other objectives were; to determine the severity, pattern of symptomatology and the common aeroallergens involved in the study group. Methods: The study was done in two steps. In stage 1, using a stratified random sampling, 423 students were screened for symptoms of allergic rhinitis based on International Consesus Report definition of rhinitis. In stage 2, the positive responders (63 students) were subjected to a physical examination and a skin prick test to confirm presence of allergic rhinitis. Results: A point prevalence rate of 13% was found with no sex or age predilection in the study group. The average age of onset was 15.2 years,seasonal peaks were in January, July and December. 81.8% of the students with allergic rhinitis had their daily activity affected to a certain degree. Sneezing (83.6%) was the commonest smptom and hypertrophied inferior turbinates (70.9%) the commonest physical finding. Patients with intermittent disease (73%) were the most, while 36% of the students with allergic rhinitis had a family history of atopy. The commonest aeroallergen was the house dust mite (76.4%) and the least was Aspergillus Niger (1.8%). Conclusion: Allergic rhinitis affects a significant proportion of the college students and has symptoms which have an impact on the lifestyles of these patients. The common aeroallergens are found within our immediate surroundings e.g, house dust mite, which can be controlled if patients are educated and proper, cheap enviromental control measures are instituted. Eduard Semyatichko, Valeriya Nemtsova, and Irina Tikhonova. Kharkov Allergological Center, Allergology, Kharkov, Ukraine. Background: Nasal polyposis (NP) often coexists with asthma and rhinitis. Polyp histology typically shows chronic, eosinophilic inflammation including eosinophils, lymphocytes, plasma cells and mast cells. We studied mediator levels and leukocyte values in nasal fluids (NFs) and eosinophil cationic protein (ECP), total IgE levels and eosinophils in the blood in both allergic and non-allergic patients with NP and in patients with allergic rhinitis (AR). Methods: Forty-six patients with NP and 37 patients with AR as a control group: 15 patients with seasonal AR to grass pollen, 12 with AR sensitive to Parietaria and 10 with AR sensitive to house dust mite (HDM) entered the study. Twenty-one patients with NP were also allergic patients (11 were sensitive to Parietaria and 10 -to HDM), whereas 25 were non-allergic patients. Tryptase and histamine values were assayed in NFs, total IgE was determined in serum. ECP values were assayed both in NF and serum. Eosinophils were quantified both in the blood and NFs. Results: Tryptase levels were significantly higher in the NFs from patients with NP than in those from patients without NP (3.9 vs. 3.5 U/l, p G 0.001) and correlated with symptom scores (r(s) = 0.36, p G 0.0001). The median levels of histamine in NFs from patients with NP were significantly higher than those in patients without NP (40.0 vs.19.4 ng/ml, p G 0.001), but did not correlate with symptom scores. The median levels of ECP in NFs from patients with NP were significantly higher than those in patients without NP (37.4 vs. 17.2 ng/ml, p G 0.001) and correlated with symptom scores (r(s) = 0.33, p G 0.001). With regard to leukocyte counts in NFs, no significant differences were between rhinitis patients with NP and those without NP. With regard to serum ECP and serum total IgE, no significant differences were detected between the two groups. Blood eosinophil levels in patients with NP were significantly higher than those in patients without NP (5.7 vs. 5.5, p = 0.002). Conclusion: Chronic eosinophil mucosal inflammatory disease in NP involves a self-sustaining mechanism independent of allergen stimulation of nasal mucosa. Increased release of inflammatory mediators contributes to the development of NP, determining oedema and an increased recruitment of inflammatory cells. Eosinophils, mast cells also play a key role in this process. Physicians' compliance with international guidelines in the treatment of allergic rhinitis Ralph Mösges, and Juliane Köberlein. University of Cologne, IMSIE, Cologne, Germany. Background: Allergic rhinitis (AR) is an allergy associated with a high burden of costs. This disease is also considered an important risk factor in the development of asthma. Within a time slot of 10 years, 20Y40% of rhinitis patients develop asthma. The ARIA guidelines, introduced in 2001, address this problem and make treatment recommendations for allergic rhinitis based on the concept Bone airway, one disease.[ Several studies have investigated the implementation process of these guidelines and have stressed their suitability for daily use. Methods: The objective of the present analysis was to estimate the compliance with and the acceptability of international guidelines among ENT specialists and general practitioners when treating patients with allergic rhinitis. For this purpose, we examined data from 122,000 patients using an IPD meta-analysis from seven post-marketing surveys collected from 1998 to 2005. Results: First, we investigated the data pool as a whole, and the results showed that 38% of the patients who were treated by ENT specialists received therapy according to these international guidelines. In contrast, only 16.3% of the general practitioners heeded the guideline recommendations. Next, we examined the time line in general, regardless of the physician`s specialty. We observed that the rate of compliance and acceptability in 2002 was higher than that in previous years. Moreover, in more than 50% of cases the patients with rhinitis and concomitant asthma were treated by their ENT specialists according to the ARIA guidelines. Conclusion: The results are evidence of the well-structured and successful implementation process of the ARIA guidelines. ENT specialists could apply the recommendations in practice more easily as compared to other guidelines. However, the investigation also shows that the ARIA document has not yet found its way into the daily routine of general practitioners. This finding thus supports the goals set by the BInternational Primary Care Respiratory Group[ for implementing special guidelines for general practitioners in the primary care setting. Inhibition of the nasal reaction by second-generation antihistamines in patients with Japanese cedar pollinosis in an artificial pollen exposure chamber (OHIO chamber) Kimihiro Okubo 1 , Minoru Gotoh 1 , and Kazuhiro Hashiguchi 2 . 1 Nippon Medical School, Department of Otolaryngology, Tokyo, Japan; 2 Kitasato Institute Hospital, Department of Otolaryngology, Tokyo, Japan. Background: Pollinosis is seasonal allergic rhinitis due to pollen antigens, and its prevalence is high enough to be called a national disease in Japan. Among the many pollen antigens, Japanese cedar pollinosis is the most common. A pollen exposure chamber (OHIO Chamber) was built in central Tokyo, Japan, in order to study seasonal allergic rhinitis (SAR). Epinastine hydrochloride (epinastine), the second-generation antihistamines, is largely used in the indication of allergic rhinitis in Japan. The purpose of this study was to investigate the protective efficacy of epinastine in patients with Japanese cedar pollinosis in an artificial pollen exposure chamber (OHIO Chamber). Methods: The study was designed as a double-blind study. After preliminary study, 20 volunteers were initially exposed to a low concentration (4500 grains/m3) of JC for at most 1 hour in this chamber, volunteers were randomized into 2 groups (group A and group B) and allocated to receive either epinastine 20 mg tablets or a placebo tablets once a day for 2 weeks. At the end of 1 week interventional period, volunteers were exposed to a low concentration (4500 grains/m3) of JC for 2 hours. And then at the end of another 1 week interventional period, volunteers were exposed to a high concentration (6000 grains/m3) of JC for 2 hours again. Subjective nasal and ocular symptoms were recorded at 0, 15, 30, 60, 90, and 120 minutes using personal computer systems and the amount of nasal secretion was measured during the allergen exposure periods. Results: Total symptom scores (TSS) in epinastine group was significantly lower than in placebo group during low concentration of JC exposed. However during high concentration of JC exposed, significant differences in TSS were not observed. Conclusion: This is the first clinical study using Japanese cedar pollen under well-controlled conditions in the OHIO chamber. This study showed that 20mg of epinastine once daily reduced the severity of allergic symptoms compared with that once in a week or placebo in pollen season. Development of a next-generation delivery system for allergic rhinitis: fluticasone furoate nasal spray Jim Godfrey. GlaxoSmithKline, Research and Development, Ware, United Kingdom. Background: Patients with allergic rhinitis (AR) consider ease of use to be the most important feature of prescription nasal sprays, with formulation-related attributes (e.g. medication that runs down the throat/out the nose and bitter taste) being key barriers to continued use. Thus, improved delivery systems and formulations may help improve adherence to nasal spray treatment. Methods: The fluticasone Furoate nasal spray (FFNS) device was designed for optimal ergonomics. Following a review of published data, key dimensions and operational ranges required for device operation were determined, and device concepts were modelled and evaluated by user groups, whose feedback was used to refine and finalise the design. The reliability of device was tested in vitro, and its ease of use was assessed in a Phase III clinical study in 302 patients with AR. The delivery system was developed with minimal steps for use, to be suitable for use by a wide patient population, for easier third-party administration and to allow determination of medication remaining in the pack. Results: In vitro tests confirmed that the FFNS device is robust, operates reliably when a force of 25Y45N is applied to the side lever, and delivers a consistent dose, even when left unused for 6 weeks. A low-dose volume (50 6L) is delivered as a fine mist which consistently delivers 27.5 6g/spray and minimises the amount of formulation available to run down the back of the throat or drip from the nostrils. In a Phase III clinical study, 84% of 302 patients found the device easy to use, 95% found it easy to carry, and 97% found the drug product comfortable to use. In addition, a review by an independent ergonomist concluded that the FFNS device is more comfortable to hold and easier to operate than current Btop-down[ nasal spray devices and is suitable for use in children as young as 2 years. The unique side-actuated delivery mechanism also allows for easier third-party administration. Conclusion: The novel FFNS device is easy to use, delivers a consistent dose, and addresses patient-reported barriers to the use of existing nasal sprays. Methods: Total 227 patients, visited us for AR without BA (2001~2004), were enrolled in 2006. We reviewed their clinical data and followed them up with a BA-detecting questionnaire, based on International Primary Care Respiratory Group (IPCRG) guidelines 2005. These results were statistically analyzed. Results: Among 227 (215 loss to f/u, 12 regular visitors), 91 responded to the questionnaire (85 by phone, 6 via mail). 37 responders, who answered yes to either doctor-diagnosed asthma or any of 4 pivotal IPCRG questions, were suspected as the group with subsequent development of BA (BAS). BAS showed higher female rate (73%) than non-suspected group (NBAS, 54%). In BAS, males were younger and females had higher BMI than NBAS (p G 0.01, respectively). Familial allergic diseases (36.8%) and nasal polyp/surgery were more frequent in BAS. BAS had longer AR duration (10.7T10.1 yrs) than NBAS (6.3 T 4.5 yrs). According to the increase of AR duration, BAS (%) increased linearly (p = 0.012). Current AR symptoms were more prevalent in BAS (89.2%, NBAS 64.8%). Atopic tendency, skin test results, serum total IgE, and induced sputum eosinophil (%) were not different between both groups. In pulmonary function test, BAS males showed lower initial FEV1 (L) and FVC (L) (p G 0.05, respectively). In initial methacholine challenge test (M-test), airway hyperreactivity (AHR: PC20 G 25 mg/mL) were more common in BAS (93.7%, NBAS 33.3%). Conclusion: 40.7% of AR patients (mean duration 8.0=57.5 years) were suspected to develop subsequent BA. Such progression might be suggested by female gender, younger age in male, higher BMI in female, longer AR duration, and more frequent AHR positive results in initial M-test. AR patients without lower respiratory symptom should be followed up regularly for early diagnosis of subsequent BA. Possible roll of the nitric oxide as an immune marker for the diagnosis of respiratory pathologies in Venezuelan scholar children Background: It has been proposed a clear association between exhaled nitric oxide (NO) and asthma exacerbation. We evaluated the NO in nasal mucus and sera, from 144 Venezuelan scholar children (6Y12 years old), from low to middle socioeconomic background covering different areas from Caracas, Venezuela, with allergic rhinitis and bronquial asthma. Methods: All children were evaluated with a validated modified Graffar's socioeconomic questionnaire and an allergic rhinitis, asthma and atopic dermatitis after ARIA, GINA and Hannifin criteria; skin prick testing (ALK-ABELLO) for common food and inhalant allergens plus total IgE (ELISA), complete blood count (COULTER) and serial feces examination for ova and parasites were performed in all children. Pre and post bronchodilator FEV1 and PEF spirometric measurements values were obtained (MICROLOOP) (we obtained the approval by the ethical committee of the Institute of Biomedicine, ratified by the academic council of the medical Faculty of Central University of Venezuela, After an informed consent was signed from parents or guardians). In nasal mucus and sera from children NO were detected by a simple colorimetric method based on Griess Methodology. Results: According to clinical evaluation children's were classified in four group. Healthy children's, children's with rhinitis and without asthma, children's with asthma and without rhinitis, and children's with asthma plus rhinitis. Healthy children present lower NO value both in sera (X = 34,482M) and nasal mucus (X = 7,992M). Rhinitis children had the higher NO levels both in sera than in nasal mucus, finding statistical differences in nasal mucus levels (21.232M) compare to control group (7,992M). In asthmatic children NO levels were lower than in rhinitis once, but higher than healthy control group. Conclusion: In nasal mucus, the study of immune parameter as nitric oxide by a rapid and simple methodology, could clarify the local inflammatory process related to allergic rhinitis in order to bring better strategies for the diagnosis and also for the evolution of the treatment. Inducement of eosinophils to apoptosis by injection of steroid into nasal polyp Kensuke Watanabe, Tomonori Eguti, and Sigenori Ohde. Dokkyo Medical University Koshigaya Hospital, Department of Oto-Rhino-Laryngology, Koshigaya, Japan. Background: It is wellknown that the basic protein of eosinophils induce respiratory epitherial disorders. The eosinophils which migrate to nasal mucosa do not return again into the circulating blood stream. It has been reported that eosinophils into nasal mucosa and mucus were degranulated by cytolysis. It is preferable treatment of allergy that the eosinophils are promptly excluded nasal mucosa without degranulation. It is aspired that the eosinophils induced to apoptosis not but necrosis. Methods: After the polyp was taken from the right nostril of the patient in whose mucus many eosinophils were observed, corticosteroid was injected into the polyp of left nostril. Two days after the steroid injecion, the polyp of the left nostril was taken. The both side polyps were fixed with 2% glutaraldehyde for 1 hour and postfixed with 2% osmic acid for 40 minutes and embedded in Epon 812. Ultrathin cross sections were prepared and observed under transmission electron microscope. The total numbers of eosinophils and macrophages with and without phagocytosing eosinophils. In some ulrathin cross section, the localizaion of ss-DNAwas examined which was a mark of apoptosis. Results: The nucleus of eosinophils showed typical characteristics of apoptpsis but apoptoic bodies were not observed after the steroid injection. The ss-DNA was admitted in the part of heterochromatin of the nucleus, which proved apoptosis had been caused. It has been known that the cells are phagocytosed by macrophages as soon as they were induced to apoptosis. There was not a significant difference in the number of total eosinophils and macrophages in the polyp before and after the steroid administration, but the rate of eosinophil phagocytosis by macrophage significantly increased after the steroid administration. Conclusion: It was clalified that eosinophils were induced to apoptosis after the steroid administration by the feature of nucleus with electron microscope and proof of ss-DNA. Eosinophils which had induced to apoptosis were promptly phagocytosed by macrophages. Acoustic Rhinometry in children with allergic rhinitis Fernando de la Torre, Eva Perez Rodriguez, Elena Rodriguez Plata, Guacimara Hernandez Santana, and Victor Matheu. Hospital Universitario NS Candelaria, Tenerife, Spain, Allergy Service, Santa Cruz de Tenerife, Spain. Acoustic Rinometry (AR) is a sound-based techinique by which the reflected wave provides information about volume & area of the nasal cavity. By AR the explorer has the abilitiy to assess nasal patency in patients suffering nasal obstruction. Goal of Study: To study basal figures of endonasal measurements in subjects with allergic rhinitis by dust mites during winter season in Canary Islands (subtropical climate with high humidity (65Y80%) and mild temperatures j20Y25-C-) and to compare with subjects without obstruction. Methods: Patients with clinical history of Rhinitis were assessed. Skin Prick Test (SPT) with mites and German cockroach were performed. Acoustic Rinometry was performed with Rinometer RhinoScan with SER 2000 module (Denmark). Three measurements were made for each nose. The mean area distance curve was calculated for each side of the nose. Curves with artifacts were discarded. Patients without blockage have a normal Minimal Cross-Sectional Area (MCA) at C-notch with values around 0.7 cm2 Results: A clinical history of rhinitis and positive SPT with mites was seen in 82% of patients. In most, the history was a mild/severe persistent rhinitis. In 17% of patients SPT were negative. In most of patients 86% of patients MCA was located in first notch (I-notch), which corresponds with Itsmus nasi and the remainder MCA was located in second notch (C-notch), which corresponds to the head of the inferior turbinate. In all the study, the average distance calculated from nostril to the head of inferior turbinate was 22.0 mm. Average of MCA was 0.54 cm2 in allergic patients and 0.57 cm2 in nonallergic rhinitis patients with no significant differences between groups Conclusion Measurements by AR are similar between allergic and non-allergic rhinitis patients. In most of patients MCA was located in first notch (I-notch), which corresponds with Itsmus nasi AR has major advantages of over other methods for assessing nasal patency since it is a very simple method and requires minimal cooperation. Clinical and therapeutic aspects of moderate/severe allergic rhinitis in Transylvania Ioana Adriana Bujor, Diana Dumitrascu, and Luise Horvath. University of Medicine and Pharmacy BIuliu HaSieganu^, 3rd Medical Clinic, Allergology, Cluj Napoca, Romania. Background: Moderate/severe allergic rhinitis (MSAR) is an affection of nasal mucosa induced by allergen exposure and produced by a specific IgE mediated chronic inflammation. Symptoms are present in over 4 days in a week, and over 4 weeks (ARIA guide), and the symptoms interfering with activities and sleep. It is related that MSAR has a 10Y25% incidence from all allergic rhinitis, it is more frequent in adolescent and young people (who, usually don`t have the financial resources for the therapy: topical corticosteroids, immunotherapy). SMAR patients have a reduce quality of life, and by absenteeism and costs this seems to be an important health problem. Methods: The aim of this study is to evaluate 40 patients with SMAR that last over a year, for 8 months (age between 9 and 57 years, medium age 33.85). Our patients have completed questionnaires about their nasal symptoms and the effect of the treatment (antihistamines, topical corticosteroids and immunotherapy. They were evaluated by skin prick tests to 10 inhaled allergens, rhinoscopy for nasal mucosa and questionnaires for the symptoms score. Results: The majority of MSAR have a polisensitization (80%: to dust mites 90.62%, to cat 37.5%, to pollen 37.5%, to cockroach 37.5%, to moulds 37.5%, to dog 18.75%). Only 20% from our patients have monosensitization (to house dust mites). The score of the symptoms was high-over 4 on a day in 4 days from the last 7 days-we evaluated the nasal congestion, rhinoreea, nasal itching and sneeze by 0 to 3. Antihystamines therapy has a lower efficiency-to one patient (3.12%) -in polisensitization patients, and in those with severe congestion has practically no effect, in 26 patients none feels better. The only therapeutic method with higher efficiency was topical corticosteroids, witch make an obvious amelioration, the score of the symptoms decreased between 0 and 2 on a day in 7 days. The financial aspect of the patients with MSAR showed that only a few from them could provide the topical corticosteroid for at least 3 months (12.5%-5 patients). Conclusion: MSAR is hard to be treated and controlled without topical corticosteroids. The effects of olopatadine hydrochloride on rhinitis induced by intranasal instillation of toluene-2,4-diisocyanate in rats Tadafumi Tamura, and Masato Komai. Kyowa Hakko Kogyo Co., Ltd., Pharmaceutical Research Center, Shizuoka, Japan. Background: The main symptoms of allergic rhinitis are sneezing, rhinorrhea and nasal obstruction. In patients with AR, the levels of neurotrophin nerve growth factor (NGF) and neuropeptides substance P (SP) increased. Therefore, they are considered important modulators in the development of AR. Olopatadine hydrochloride (olopatadine), is an anti-allergic agent with histamine H1 receptor antagonistic action. We reported that olopatadine inhibited the elevated levels of NGF and SP in the mouse model of chronic inflammatory dermatitis. Objective: To investigate whether olopatadine has an effect on the production of NGF and SP, we used TDI-sensitized rats as an animal model of nasal allergy. Methods: After the intranasal challenge of TDI, the numbers of sneezes were counted in a blinded way. To determine NGF and SP production in the nasal lavage fluids (NALF), rats were anesthetized and nasal lavages were done. Olopatadine was orally administered orally before the nasal instillation of TDI. Results: In TDI-challenged rats, nasal allergy-like behavior (sneezing, rhinorrhea and inflammation) was provoked after TDI challenge. The amounts of NGF and SP in the NALF were increased. Olopatadine reduced nasal allergy-like behavior. Moreover olopatadine inhibited the increases of NGF and SP production. Conclusion: Our findings suggest that the increase of NGF and SP production is one of the mechanisms responsible for nasal allergy-like behavior in TDIchallenged rats. These results suggest that the suppression of neurogenic inflammatory reaction might partially be involved in the improvement of allergy-like behavior by the treatment of olopatadine. Do topical steroids reduce subjective and objective measures of nasal congestion in persistent allergic rhinitis? Kivanc Gunhan, Halis Unlu, Ali Vefa Yuceturk, and Murat Songu. Celal Bayar University, Otorhinolaryngology and Head-Neck Surgery, Manisa, Turkey. Background: Nasal breathing is essential in maintaining the physiologic functions of the upper and lower airways. The predominant symptom of allergic rhinitis is nasal congestion, which also has a significant impact on quality of life and work productivity. In patients who suffer from persistent allergic rhinitis (PAR), a severe drug-resistant hypertrophy and increase in glandular structures of the inferior turbinates may develop, which leads to constant nasal obstruction. Objective methods are strongly recommended for use in the evaluation of pharmacologic agents that are expected to improve nasal airflow. Methods: This prospective, single-sited study randomized 50 patients with mild or moderate PAR who had substantial bilateral hypertrophy of the inferior turbinates to desloratadine (5 mg/day) or additional mometasone furoate monohydrate nasal spray (MFMNS), (2 sprays per nostril [total dose 200 6g] once daily) treatment groups. Patients with previous treatments, concomitant sinonasal disorders or systemic diseases were excluded. Both objective outcomes evaluated by total nasal resistance at anterior rhinomanometry and subjective outcomes assessed with endoscopic nasal examination and Quality of Life Questionnaire were analyzed before and at least 12 months after treatment. Results: The median total nasal resistance in patients treated with MFMNS decreased from 0,49T0,17 Pa/cm3/s to 0,39 Pa/cm3/s (p = 0,42), and at least 12 months later. Compared with pretreatment scores, the post treatment scores of these patients significantly improved in both 7 separate domain scores and overall Rhinoconjunctivitis Quality of Life Questionnaire scores (p = 0,004). Nasal symptomatology was reduced 2 months after MFMNS application. No adverse reactions including bleeding, infection, or hormonal disorders were encountered. The patients experienced a lasting benefit from this treatment. Conclusion: Nasal congestion affects most individuals with allergic rhinitis, and has a notable impact on quality of life, emotional function, productivity, and the ability to perform daily activities. These results suggest that topical mometasone reduces the volume of inferior turbinate at some point while significantly improving the quality of life in patients with PAR. Histopathologic and longer term studies with larger groups will enlighten the potential and mechanism of efficacy of topical steroid in management of inferior turbinate hypertrophy in patients with PAR. Topical treatment for perennial allergic rhinitis in children Felicia Manole. Faculty of Medicine Oradea, Otorhinolaryngology, Oradea, Romania. Background: Topical corticosteroid is now accepted as safe and most effective in controlling all symptoms of both allergic and nonallergic rhinitis. Mometasone furoate monohydrate nasal spray is a once daily topical corticosteroid preparation. Objective: To evaluate the efficacy and safety of mometasone furoate monohydrate nasal spray in children 6 to 12 years of age with perennial allergic rhinitis. Methods: A double-blind, placebo-controlled, parallel group of 112 recruited patients of whom 96 were evaluated. Treatment with once daily mometasone furoate monohydrate nasal spray 100 mcg once daily or placebo for 6 weeks followed by a 3-week follow-up period. Forty-eight patients of each group were treated with mometasone furoate monohydrate nasal spray or placebo by randomized assignment. Results: There was no statistical significance of the sex, mean age, weight, and height of the two groups. Patients treated with mometasone furoate monohydrate nasal spray showed a significant decrease in total symptom scores rated by physicians at 3 weeks and 6 weeks, respectively (P G .01, P G .05). The rhinitis symptom scores in treatment group rated by patients (nasal blockage, sneezing, watery rhinorrhea) were significantly decreased at 3 weeks (P G .05, P G .01). Nasal symptoms as assessed by doctors (turbinate swelling, color of nasal mucosa, secretion, and postnasal drip) also decreased at 3 and 6 weeks, but were not statistically significant, except for the secretion at 3 weeks and postnasal drip at 6 weeks (P G .05). There was no evidence of effects on adrenal function by morning plasma cortisol concentration between the two groups. Conclusion: Mometasone furoate monohydrate nasal spray was safe and effective, well tolerated in children aged 6 to 12 years with perennial allergic rhinitis with incidences of adverse events comparable to placebo. Effects of seasonal allergic rhinitis on fatigue levels and mood Ziad Adwan. Saha, Allergy, Reo De Janero, Brazil. Objective: Many allergy patients complain of fatigue, moodiness, and dysphoria during their allergy seasons. This study evaluated the effect of symptomatic allergic rhinitis on both fatigue level and mood. Methods: Symptomatic ragweed allergic rhinitis patients on no medications and healthy control subjects completed the Multi-Dimensional Fatigue Inventory and the Positive Affect-Negative Affect mood rating scales in an in-out-in ragweed season research design. Results: During ragweed seasons, allergic patients reported higher levels of general fatigue and mental fatigue, but not physical fatigue, as well as reduced motivation. Patients described experiencing feelings of greater sadness and reduced pleasurable engagement. Increased anxiety or emotional distress was not reported. Conclusion: These findings suggest that having allergic reactions to ragweed pollen causes significant fatigue and mood changes in at least a subgroup of patients. Psychoneuroimmunology and medical genetics research suggests that allergic reactions engender biochemical changes that directly affect the central nervous system. Sheen-Yie Fang. National Cheng Kung University Hospital, Dept. of Otolaryngology, Tainan, Taiwan. Background: Levocetirizine (Levo) is a potent latest-generation, non-sedating oral H1-antihistamine for which no data in Taiwanese population has yet been published. Objectives: Primary: patients`perception of Levo in the treatment of allergic rhinitis (AR). Secondary: adverse events (AEs) and incidence of intermittent (IAR) and persistent (PER) AR (as defined by ARIA) in Taiwan. Methods: A multicenter observational study (6 medical centers) conducted from May 2006 to March 2007 in Taiwan assessing the treatment perception of 236 AR patients on Levo. Runny nose, nasal and ocular pruritus, sneezing and nasal obstruction were measured (0 = absent, 3 = severe). Total 5 Symptom Score (T5SS, sum of the above symptoms (range 0Y15)) and asthma symptoms (per GINA): mild (91x/week, G1x/day); moderate (daily); severe (continuous) were determined. The observational period was 2Y4 weeks. The onset of action was rated very rapid (G30min), rapid (930min, G1hr), and moderate (91hr). A visual analogue scale (VAS) was used to evaluate global satisfaction of patients and physicians (range: 0Y10). Results: 236 patients were included and 217 completed the study. 19 patients (8%) were lost to follow-up due to AEs, none serious. 56 patients (24%) had concomitant asthma. PER was diagnosed in 191 (81%) and IAR in 45 (19%) patients. T5SS improved by 56%, from 10 (T3.3) at baseline to 4.4 (T2.3) at end of treatment. Overall efficacy and tolerability were assessed as Bgood/ excellent[ by 60% and 66% of patients, respectively. At least 61% of all patients and 75% of those with Bmoderate/severe^symptoms reported complete recovery or marked improvement of any individual symptom. 50% of subjects reported the onset of action as very rapid or rapid. Levo was reported as better than their previous therapy by 56% of patients. BGood/ excellent^improvement in quality of sleep and daily activities was reported by Results: Seasonal or intermittent allergic rhinitis. The treatment of the mild form of rhinitis with episodic symptoms the treatment should be begun with per oral or topic administration of antihistamine (non-sedative) drugs. Other variants of treatment are the topic decongestants (for no longer than 10 days) and per oral decongestants (which are not recommended for children). If eye symptoms are prevailed over the rhinitis symptoms or if they were not stopped with administration per orally of antihistamine preparations then the same preparations may be additionally used as eye drops. In cases of moderate severe and severe forms with episodic symptoms the treatment includes per oral antihistamine preparations with decongestants and topic glucocorticoids. The all-the-year-round or persistent allergic rhinitis. In cases of light clinical course when the symptoms of disease do not required special treatment and only the measures for elimination of allergen may be performed. Variants of medicamentous therapy include per oral or topic antihistamine preparations, per oral antihistamine preparations with decongestants and topic glucocorticosteroids. Efficacy of therapy performed must be evaluated in 2Y4 weeks. In moderate severe and severe forms of rhinitis the preparations of the first line are the topic glucocorticosteroids. In sharp disturbance of the nasal breathing this treatment may be added with short course of systemic steroid therapy. The effect of this therapy is evaluated in 2 weeks. Conclusion: The causes of insufficient efficacy of topic glucocorticosteroids may be: irregular dosing of preparation by physician or patients, insufficient administration of preparation into the nasal cavity because of sharp edema of mucous membrane, presence of concomitant pathology (deformation of nasal septum, chronic rhinosinusitis and others), power effect of unremoving allergen and irregularly established diagnosis. The impact of Flixonase on quality of life in paediatric patients with seasonal allergic rhinitis Maia Kherkheulidze, Nani Kavlashvili, Nino Adamia, and Eka Kandelaki. State Medical University, Pediatrics, Tbilisi, Georgia. The aim of our study was to investigate the efficacy of Flixonase in children with seasonal allergic rhinitis and to evaluate the impact of treatment on quality of life. 38 (4Y12 years old) outpatient children with seasonal allergic rhinitis were enrolled into the clinical study. We evaluated history, clinical symptom severity (nasal symptom severity Y such as sneezing, runny nose, itching, stuffiness; non-nasal symptom severity Y such as eye, ear, throat symptoms, chronic cough, and headache). Rhinoscopy (swelling and hyperemia of mucous) and evaluating allergic markers (eosinophyles) in nasal lavage detected diagnosis of rhinitis. We studied blood immunity data (IgE). For the assessment of the quality of life we used the Juniper Rinoconiuctivities quality of life (Pediatric and adolescent) scale. The baseline results showed that eosinophilia in peripheral blood was observed in 35%, patients, IgE hyper production in 72%. There was determined correlation link between IgE hyper production and severity of diseases and eosinophil count in nasal lavage. Flixonaze was administered 50 mcg once a day. The treatment with Flixonase (time of treatment 14 days) showed significant improvement of clinical symptoms and patients condition, reduction of nasal as well as nonnasal severity symptoms. Medication assessment average score was 6, 6 + 0.3. At the same time was observed decrease of eosinophil count in nasal lavage. After the two weeks trial period, the overall RQLQ scores of the treated patients improved by 68% + 7 from baseline. So, we conclude that using of Flixonazein paediatric patients with seasonal allergic rhinitis improved patient`s condition as well as their quality of life. Allergic rhinitis: especially of nasal's microflora in the children Background: Staphylococcus aureus is proved to play unduestionable role in development of atopic dermatitis. In this connection investigation of bacteria role in dermatitis pathogenesis is of considerable interest. The aim of the present study was to investigate microbial microflora of the nasal cavity in patients with persistent allergic rhinitis (PAR). Methods: A total of 89 PAR sufferers aged 3 to 17 years have been investigated, bacteriological study of nasal secretion being carried out. Results: Staphylococcus haemolyticus in diagnosticaly insignificant titer was isolated in every second person in the group of PAR patients (n = 38), Staphylococcus aureus in diagnosticaly insignificant titer being isolated in 23% of patients. High level of dissemination (more than 103Y104 KOE/ml) was noted in 10 out of 38 patients: Staphylococcus haemolyticus-15,8% (n = 6); Staphylococcus aureus-10,5% (n = 4). In PAR patients along with atopic bronchial asthma (n = 42) rising Staphylococcus haemolyticus titers-24% (n = 10), Staphylococcus aureus-33% (n = 14) was observed. In 6 out of 14 patients isolated Staphylococcus a ureus strain was combined with Klebsiella, Escherichia coli, Citrobacter spp. Isolation of permissible for normal microflora values was equal to 19% for Staphylococcus haemolyticus (n = 8); and 16,7% (n = 7) for Staphylococcus aureus. Streptococcus haemolyticus was isolated in 3 patients. In the group of PAR patients along with atopic dermatitis (n = 9), Staphylococcus aureus (in 7 patients under study) was the main representative of the nasal cavity microbiocenosis. Conclusion: Prevalence of staphylococcal found out in the course of study in nasal cavity of PAR patients may suqgest the ability of these microorganisms to support allergic inflammation. Efficacy topical desensitization in chronic rhinosinusitis and nasal polyposis with association nonsteroidal anti-inflammatory drugs hypersensitivity Background: Subject suffering from aspirin and over nonsteroidal antiinflammatory drugs (NSAID's) hypersensitivity frequently develop chronic rhinosinusitis (CRS) with nasal polyposis (NP). Previous studies showed that aspirin-lysine topical desensitization may be effective to CRS and NP treatment and prevent recurrent NP. The Aim of the Study: To evaluate the efficacy of topical desensitization by sodium diclofenac in patients (SD) with NSAID's hypersensitivity and chronic rhinosinusitis with nasal polyposis. Methods: Ten subjects (5 male and 5 female, mean age 52.5 T 2.1 years) after intranasal polypotomy with positive results of nasal challenge with SD underwent topical SD desensitization and daily treatment with SD 20 mg during 1 year. Additionally 5 patients were receiving intranasal corticosteroids in stable dose. Results: There were significant reductions of the total endoscope count (10.4 T 2.8 to 5.8 T 1.9, p G 0.05) and nasal symptom score (6.6 T 2.2 to 3.2 T 0.8, p G 0.05) in the subgroup with combination desensitization and topical corticosteroid therapy. In both subgroups there were improvement of nasal inspiratory peak flow (75,0 T 13,8 l/min to 86,5 T 9,2 l/min, p G 0.05) and pulmonary function. Using of inhalation corticosteroids were decreased. There were no recurrences of nasal polyposis in all patients after one year observation. Conclusion: Topical sodium diclofenac desensitization is an effective treatment in chronic rhinosinusitis and nasal polyposis with association NSAID's-hypersensitivity. Influences of allergic rhinitis to the outcome of functional, endoscopic sinus surgery Sylvi Meuret, Heidrun Mueller, Gero Strauss, and Andreas Dietz. University Clinic Leipzig, ENT Department, Leipzig, Germany. Background: The exact pathogenesis of nasal poyposis (NP) is still unknown. Empiric data show that patients who suffer from NP and allergic rhinitis (AR), do not benefit as much of functional, endoscopic sinus surgery (FESS) as patients without AR. Methods: In this study we want to present our expirience in FESS in patients with AR. 120 patients were included and devided into three groups: 1. 45 patients with AR without NP and without FESS 2. 43 patients with AR and NP and FESS a) 32 patients with SIT b) 11 patients without SIT 3. 32 patients with NP without AR. In our protocol we included a score of symptoms, the endoscopis findings and a CCT of the sinuses. Results: In general, patients in group 3 have a better outcome of FESS than patients in group 2. Group 2a benefits more of FESS than group 2b. Conclusion: In this investigation, we can show that there is a tendency that patients with NP and AR who are treated by SIT have a better outcome of FESS than patients without AR. Expressions of mammaglobins A and B are not different between nasal polyps with and without allergic rhinitis Supinda Saengpanich 1 , Chuntima Phannaso 2 , Siraprapa Tongkobpetch 3 , Songklot Aeumjaturapat 1 , Yong Poovorawan 3 , Kanya Suphapeetiporn 3 , and Vorasuk Shotelersuk 3 . 1 Chulalongkorn University, Department of Otolaryngology, Bangkok, Thailand; 2 King Chulalongkorn Memorial Hospital, Department of Otolaryngology, Bangkok, Thailand; 3 Chulalongkorn University, Department of Pediatrics, Bangkok, Thailand. Background: Nasal polyposis is a chronic disease of nose and sinuses. Its actual causes remain unclear. Mammaglobins have been implicated in the pathogenesis of nasal polyps. However, their association with the occurrence of nasal polyps in the presence of allergic rhinitis has not been explored. Objective: The aim of this study was to compare the expression levels of mammaglobins A and B between the nasal polyps with allergic rhinitis and without allergic rhinitis. Methods: 31 patients with bilateral nasal polyposis underwent skin prick test to specific aeroallergens. Nasal polyp tissues were obtained from all patients and divided into 2 groups as nasal polyps with allergic rhinitis and nasal polyps without allergic rhinitis depending on the skin prick tests`results. All polyp tissues were analyzed for the levels of mammaglobin A and mammaglobin B by using real-time quantitative polymerase chain reaction technique (RTQ-PCR). Results: Of the 16 samples from patients having nasal polyps with allergic rhinitis, only one expressed a detectable level of mammaglobin A (1/16). There was no detectable expression of mammaglobin A in tissues from the group of nasal polyps without allergic rhinitis (0/15). Expression of mammaglobin B was detected in all nasal polyp tissues from both groups. The mean expression of mammaglobin B was not significantly different between nasal polyps with allergic rhinitis (0.059; range 0.0002 to 0.343) and nasal polyps without allergic rhinitis (0.133; range 0.003Y0.628). Conclusion: Expressions of mammaglobins A and B are not different between nasal polyps with and without allergic rhinitis. Our findings suggest that mammaglobins`implication in the pathogenesis of nasal polyps is independent of an underlying allergic rhinitis. Characteristics and associated conditions of acute sinusitis in Thai children: a prospective evaluation of 140 patients Maleewan Kitcharoensakkul and Orapan Poachanukoon. Faculty of Medicine Thammasat University, Department of Pediatrics, Pathumthani, Thailand. Backgroud: Sinusitis is a common disease in children. Even though new research findings are continually being published on various aspects of this disease, reports of its clinical characteristics and associated conditions are still limited. Objective: To describe the characteristics and associated conditions of acute sinusitis in Thai children. Materials and Methods: Our study consisted of 140 acute sinusitis patients who presented to Thammasat University Hospital. Acute sinusitis is diagnosed on the basis of clinical presentation including persistent, severe or worsening nasal symptoms. An informed consent form was obtained from each patient before the study. Radiography has been used to aid in the diagnosis of acute sinusitis. Allergic skin prick test was also performed in this study. Results: Age range of the 140 patients were between 1.1 to 15 years with a mean (+/-SD) of 5.67 +/-2.85 years. History of allergic rhinitis, confirmed by positive skin prick test or nasal cytology, was found in 64%. The parental history of atopy was found in 50.4 %. Positive history of tobacco smoking was found in 25.9 %. Familial members with diagnosis of a cold or sinus infection during the same time was found in 43.2 %. 69.8% of the patients attended day care or school more than five days per week. 38.1% of the patients had a history of swimming or diving during the past 1 month. 51.8 % of the patients had a past history of sinus disease . The four most common symptoms were nasal blockage (88.5%), rhinorrhea (81.3%), nighttime cough (79.9%) and snoring (73.4%). The three most common signs were swelling of turbinates (75.5%), erythema of turbinates (74.8%) and tonsillar enlagement (48.2%). All paranasal sinuses X-rays were abnormal with maxillary sinus being the most commonly involved sinus (97.5%) followed by ethmoid sinus (58.8%). 58.8% of the patients had involvement of more than one sinus. Adenoid hypertrophy was positive in 36 % of all patients. The skin prick tests were positive in 62.6 % of all patients who the tests were performed. Conclusion: Nasal blockage, rhinorrhea, nighttime cough and snoring are very common in acute sinusitis. The present study demonstrates that allergic rhinitis and recurrent sinusitis is common in our patients. Alteration of paranasal sinus mucosa in children with allergic rhinitis Felicia Manole. Faculty of Medicine Oradea, Otorhinolaryngology, Oradea, Romania. The purpose of the study is to determine the alteration of paranasal sinuses in pediatric patients diagnosed as having allergic rhinitis. We have studied 80 patients aged between 8 and 16 diagnosed with allergic rhinitis in the period 2005Y2006. The associated diseases have been established: asthma, sinusitis, serous otitis media, hypertrophic chronic pharyngitis. Allergic rhinitis has been found to be perennial in 34 cases and seasonal in 46 cases. Associated sinusitis has been diagnosed in 42 patients, all of this have a high level of Ig E, from whom 28 patients had seasonal rhinitis. Main clinical symptoms: nasal discharge, nasal obstruction, sneeze, headache, eye itching, cough, postnasal drip. Clinical and radiological examination (X-ray) of paranasal sinuses has shown the alteration of maxillary, ethmoidal and frontal sinus mucosa. The mucosa is swollen or with serous effusion. The results of the study demonstrate that: maxillary and ethmoidal sinusitis is predominating affected in patients with seasonal allergic rhinitis. Radiological examination has shown predominant lesions of the type of sinusal swollen mucosa. Postnasal drip and cough associated with allergic rhinitis are indirect symptoms of the alteration of the paranasal sinuses. Allergic rhinitis in children must not be considered an isolated disorder, but must be considered in the context of a systemic allergic disorder. Sinus disorder in children is often misdiagnosed. Nasal mucosal edema at the level of the middle meatus favors associated sinus disorders. Presence of eosinophilic inflammation and airway remodelling signs in sinus tissues of patients with asthma and chronic rhinosinusitis Dilek Saka 1 , Cem Saka 2 , Filiz Cimen 1 , Sema Canbakan 1 , \stemihan Ak]n 2 , Funda Demirag 3 , and Nermin Capan 1 . 1 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Respiratory Medicine, Ankara, Turkey; 2 Diskapi Yildirim Beyazit Education and Research Hospital, Otorhinolaryngology-Head and Neck Surgery, Ankara, Turkey; 3 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Pathology, Ankara, Turkey. Background: Chronic rhinosinusitis exists in a high amount of asthma patients. It is still unknown whether a cause and effect relationship is present between the two diseases or they are the manifestations of the same underlying disease process affecting the whole respiratory tract. Objective: The aim of this study was to find out if sinus tissues in chronic rhinosinusitis have asthma related histological features such as eosinophilic inflamation and airway remodelling. Methods: The sinus tissues taken from 15 asthmatic patients with chronic rhinosinusitis and who had ungergone functional endoscopic sinus surgery were examined histologically. The patients did not have nasal polyposis. The 'Hematoxylin and eosin' staining was used to detect epithelial damage. Basement membrane thickening of the airway mucosa was evaluated with PAS staining. These two signs were accepted as signs of airway remodelling. Eosinophil semiquantitative scoring system (0Y4) was used to detect presence and amount of the eosinophils. Results: 15 patints with mean age of 37 T8.19 years were included in the study. 12 patients were female, 3 were male. 9 patients were found as atopic with skin prick test. 9 (60%) patients had epithelial damage with severe damage in 7 (46.6%). 11 (73.3%) patients had basement membrane thickening. 10 ( 66.6%) patients had eosinophils in the sinus tissue. 6 of these patients were allergic. The rate of allergy was the same in patients who had not eosinophils in sinus tissues. Conclusion: In patients with asthma and chronic rhinosinusitis without nasal polyposis, the histopathologic signs of asthma can also be seen in sinus tissues of the patients taken with functional endoscopic sinus surgery. This can be a clue of the same pathogenetic event of the whole respiratory tract. Alveolar concentration and bronchial output of exhaled nitric oxide in chronic rhinosinusitis Enrico Heffler, Iuliana Badiu, Giuseppe Guida, Luisa Bommarito, Pietro Marsico, and Giovanni Rolla. University of Torino -ASO Ordine Mauriziano, Allergy and Clinical Immunology, Torino, Italy. Background: Rhinitis and chronic rhinosinusitis (CRS), particularly with nasal polyps, are strong risk factors for asthma. Exhaled nitric oxide (FENO), a reliable non-invasive marker of airway inflammation, is known to be elevated in patients with asthma and rhinitis. In this study we investigated inflammatory and functional airway involvement in patients with CRS. Methods: We recruited 47 patients with CRS, in all of them respiratory questionnaire, spirometry and FENO measurement (at constant flow of 50, 100 and 200 ml/sec) were obtained. Alveolar concentration (FANO) and maximal bronchial output (QbrMaxNO) of nitric oxide were calculated for each patient using a nonlinear model described by Silkoff et al. All patients with asthmalike symptoms were further investigated by methacholine inhalation challenge and/or bronchodilating test. Results: 19 (40.4%) of our patients met the functional and clinical criteria of asthma, and 34 (72.3%) had nasal polyps. Among the 28 patients without asthma, 11 (39.3%) complained asthma-like symptoms. FENO was higher in patients with asthma (63.9, I.C. 95%: 47Y81 ppb) and asthma-like symptoms with normal lung function (64.7, I.C. 95%: 39Y91 ppb), compared with those without asthmatic symptoms (29.8, I.C. 95%: 21Y39 ppb), pG0.01. All the three groups of patients had FENO higher than healthy controls (13.4, I.C. 95%: 6Y19 ppb, pG0.01). QbrMaxNO was more elevated in patients with asthmatic symptoms, with and without lung function criteria for asthma (3.2 , I.C. 95%: 2.3Y4.1nL/s and 3.5, I.C. 95%: 1.5Y5.6 nL/s respectively) compared with patients without asthma-like symptoms (1.9, I.C.95%: 1Y2.8 nL/s), p G 0.05. FANO was higher in patients with asthma-like symptoms but normal lung function (4.5, I.C. 95%: 2.4Y6.6 ppb) compared with those with asthma (2.4, I.C. 95%: 1.6Y3.4 ppb) and those without asthmatic symptoms (2.8, I.C. 95%: 1.1Y4.5 ppb), p=0.03. Conclusion: Patients with CRS showed higher levels of FENO compared to normal controls, suggesting a diffuse airway inflammation. FENO values were particularly elevated, with increased maximal bronchial output of nitric oxide, in patients with asthmatic symptoms, irrespectively of lung function (asthmatics or not), while alveolar concentration of nitric oxide was significantly higher only in patients with asthmatic symptoms, but normal lung function, suggesting a more peripheral inflammation in these patients than in patients with asthma. Microbiology of patients with a clinical diagnosis of sinusitis The diagnosis of Acute Bacterial Rhinosinusitis (ABRS) involves clinical symptoms and signs, without the use of plain sinus radiographs. We wished to determine the microbiology of sinusitis using these criteria compared to endoscopic culture results, with and without plain sinus films. Methods: 103 patients diagnosed with acute sinusitis by clinical criteria were studied. Clinical criteria included biphasic illness, signs and symptoms of sinusitis and purulent sino-nasal drainage. After entry, patients had an endoscopic ostiomeatal culture and a Water_s view plain radiograph. Results: The pathogens identified in descending order were: Staphylococcus aureus (48.6%), Streptococcus pneumoniae (22.9%), Moraxella catarrhalis (11.4%), Pseudomonas (11.4%), Enterococcus (8.6%), Haemophilus influenzae (5.7%), Proteus mirabilis (5.7%), and other Strep species (5.7%). We also found 1 Methicillin Resistant Staphylococcus Aureus (MRSA) and 1 Aspergillus niger. The symptoms with the best combination of sensitivity, specificity, Positive Predictive Value (PPV) and Negative Predictive Value (NPV) were colored/purulent nasal drainage, congestion, and facial pain. Adding a positive x-ray produced a specificity of 68.7%, and NPV of 68.7%, but reduced sensitivity and PPV to 36.4% each (respectively). Utilizing the clinical criteria of colored nasal drainage, congestion and facial pain together (without x-ray) provides the best combination of sensitivity, specificity, PPV, and NPV. Mometasone furoate nasal spray in seasonal allergic rhinitis Ziad Adwan. Saha, Allergy, Swaida, Syrian Arab Republic. Background: Mometasone furoate nasal spray (MFNS) is effective for preventing and treating nasal symptoms in seasonal allergic rhinitis (SAR). Its effects on ocular symptoms have not been investigated. This retrospective analysis examined the effects of MFNS on ocular symptoms in subjects with SAR. Methods: Ocular symptom data were pooled and analyzed from four randomized, double-blind studies comparing MFNS 200 mcg once daily (n = 494) with placebo (n = 497). Subject-reported ocular itching, redness, and tearing were recorded at baseline and twice daily throughout treatment on a scale of 0 (none) to 3 (severe). Total ocular symptom score (TOSS) was defined as the combined 2-week average symptom scores. Results: MFNS produced a statistically greater reduction in TOSS from baseline as compared to placebo (j1.33 vs. j0.94, p G 0.05). Likewise, mean 2-week reductions in individual symptoms were significantly improved with MFNS (p G 0.05 for each symptom). In subjects with TOSS 4 at baseline, MFNS recipients (n = 298) reported a significantly greater reduction in TOSS as compared to placebo recipients (n = 304; j1.97 vs. j1.51, p G 0.05), with statistically significant benefits also observed in individual ocular symptoms (p G 0.05 for each symptom). Conclusion: MFNS has a beneficial effect on ocular symptoms, in addition to its established effects on nasal symptoms, in subjects with SAR. Background: Royal jelly is a health supplement widely consumed in the community and has perceived benefits ranging including boosting the immune system. However, royal jelly consumption has been linked with contact dermatitis, acute asthma, anaphylaxis and death. Case reports of allergy to royal jelly have been reported among atopics. Our previous study among local allergic patients demonstrated that royal jelly sensitivity was found to be present in this country. Several major allergens in the range of 39 to 71 kD have been detected. Thus, the aim of this study is to identify the major allergens of royal jelly using 2dimensional electrophoresis and mass-spectrometry analysis. Methods: Extract of royal jelly was fractionated by charge and molecular weight using 2-dimensional electrophoresis (2-D) . Immunoblotting of the 2-D profiles were performed to identify the allergenic spots of the major allergens using sera from royal jelly allergic patients. Spots were then excised from the 2-D gel, digested with trypsin and analyzed by MALDI-TOF MS and Q-TOF MSMS. For protein identification, the masses obtained were used to search databases to identify the proteins. Results: 2-D gel fractionated the royal jelly proteins to more than 50 different protein spots. Out of these, 31 spots demonstrated specific IgE to the sera tested. Digested peptides of most allergenic spots as compared to the amino acid sequence in databases identified the fragments of royal jelly homologus to major royal jelly protein 1 (MRJ1) and major royal jelly protein 2 (MRJ2). Our results confirmed that the major allergenic spots of royal jelly are MRJ1 and MRJ2. Identification of IgE-binding proteins of Thunnus tonggol (tongkol) by mass spectrometry analysis Background: Fish is one of the most frequent cause of food allergy. Parvalbumins represent the main major allergens and responsible for IgE cross-reactivity between various species of fish. Tongkol belongs to the genus Thunnus that includes several species. Tuna, which is widely consumed worldwide, also belongs to this genus. The aim of this study was to characterize IgE-binding components of Thunnus tonggol (tongkol/ Northern bluefin tuna). Methods: Uncooked and cooked extracts of the fish were prepared from the fish meat. Protein profile and IgE binding pattern was demonstrated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from subjects with fish allergy. The major allergens of the fish were then identified by two-dimensional (2-D) electrophoresis, followed by mass spectrometry analysis of the peptide digest. Results: The SDS-PAGE of raw extract revealed 26 protein fractions over a wide molecular weight range, while cooked extract demonstrated only nine protein fractions. The 1-D immunoblotting detected 17 IgE-binding proteins, ranging in molecular weight from 151 to~12 kD. Two protein fractions with molecular weight of~51 and 42 kD have been identified as the major allergens of this fish. Both proteins were sensitive to heat. The 2-D gel profile of the raw extract demonstrated about 9100 distinct proteins spots and immunoblotting detected at least 10 different major IgE reactive spots with molecular masses as expected and isoelectric point (pI) values ranging from 4.1 to 6.1. Comparison of the major allergenic spot sequences of the~51 and 42 kD proteins with known protein sequences in databases revealed extensive similarity with fish enolase and triose phosphate isomerase, respectively. In conclusion, this study demonstrated that enolase and triose phosphate isomerase are the major allergenic component of this species of fish. Interestingly, our study also detected a heat resistant protein at~12 kD as parvalbumin, which is similar as Sal s 1. However, this protein was only seen as a minor allergen for this fish. The analysis of mite and cockroach allergen in vitro test for allergic disease Baoqing Sun, Lili Wei, Hongyu Wang, Chunqing Zhang, Jing Li, and Nanshan Zhong. Guangzhou Institute of Respiratory Diseases, Allergy Dept, Guangzhou, China. Objective: To investigate the level of specific IgE (SIgE) of mite and cockroach in allergy patients' blood serum, and study their correlation. Methods: By ELISA test to examine the mite and cockroach allergen's sIgE from the blood serum of 199 outpatients who were recruited during 2004 Oct to 2005 Oct at Respiratory department and Allergy department. Results: In the mite (dermatophagoides pteronyssinus(d1), dermatophagoides farinae(d2), and Blomia Tropicalis(d201) allergic group, the children showed higher proportion of positive result than the adult group. Some strong positive results were all recorded from children. In the cockroach allergic patients, the positive results were all under the class IV level. For those patients who allergic to both of mite and cockroach, the mite allergen SIgE in blood was significantly higher then that of cockroach allergen (p G 0.01). Conclusion: Mite and cockroach are both important allergens of allergic diseases, from which mite played a dominant role. For those patients who are allergic to both mite and cockroach, they are proved to be more sensitive to mite allergen. Background: American cockroach (CR), Periplaneta americana is an important source of indoor allergens among Thais. Currently, screening and monitoring allergic status of allergic patients are performed by skin prick test using crude CR extract which varies in the allergenic composition from batch-to-batch. Thus, recombinant CR allergens may be a better standardized alternative. Thus, in this study, we produced recombinant tropomyosin (Per a7) which is one of the P. americana major allergens. The protein was tested for its ability to bind to IgE in sera of CR allergic Thai patients. Objectives: To produce recombinant tropomyosin of American cockroach, and to determine the IgE binding ability of the recombinant allergen. Materials and Methods: Sera of patients with CR allergy who gave positive skin prick test to CR extract and sera of non-allergic counterparts were collected. Total RNA was extracted from adult P. americana and first strand cDNA was produced by RT-PCR. Gene encoding tropomyosin was amplified using oligonucleotide primers and the cDNA as a template. The amplified sequences were cloned into a cloning vector and a protein expression vector. The latter was used to transfect an Escherichia coli host. E. coli transformant was grown and production of tropomyosin protein was induced by IPTG. The protein was extracted and purified from the bacterial lysate. Results: Gene encoding American cockroach tropomyosin (450 bp) was amplified and cloned into a cloning vector, i.e. pGEM T and a protein expression vector, i.e. pET 20 b+. Amplified sequence showed 100% homology to the tropomyosin gene sequence in the database. Recombinant tropomyosin was produced and purified from whole cell lysis of an E. coli transformant grown under IPTG induced-condition. The protein (~16 kDa) was found to bind IgE in sera of allergic Thai patients. Conclusion and Clinical Relevant: Recombinant tropomyosin of P. americana was successfully produced in pure form. The protein binds IgE in sera of CR allergic Thai patients. Thus, the protein may be used as a standard reagent for screening and monitoring of the allergic status of patients. Serum allergen profiles related to moth and house dust mite in cockroach-allergic patients in-vitro assay and correlation analysis Baoqing Sun, Nili Wei, Jing Li, Mei Jiang, and Nanshan Zhong. Guangzhou Institute of Respiratory Diseases, Dept of Medicine, Guangzhou, China. Objective: To look at the serum profiles of specific immunoglobin E (SIgE) related to moth and house dust mite allergens in cockroach-allergic patients and provide evidences for further research on insect hypersensitivity. Methods: Thirty-three patients who presented to our Respiratory or Allergy Clinics between October 2004 and May 2007, and tested positive of Blattella germanica allergen by both skin prick test and SIgE test, were further evaluated for allergens related to Periplaneta americana, moth and house dust mite using fluorescence enzyme-linked immunosorbent assay. Results: In patients who showed concurrent hypersensitivity to all of studied allergens, responses to the cockroaches and moth were weak-positive (below Grade IV), with significantly lower SIgE levels than as found for house dust mite (P G 0.001). Linear regression showed positive correlations in allergenicity between the two species of cockroaches and moth (P G 0.001), but not between house dust mite and moth or cockroaches. Conclusion: High response to house dust mite was demonstrated in patients with multiple allergies. Allergens from Blattella germanica, Periplaneta americana and moth appeared to share some IgE binding site in common. Production and identification of major allergens of two species of local crab: blue/ swimming crab (Portunus pelagicus) and red crab (Charybdis japonica) Rosmilah Misnan, Shahnaz Murad, Noormalin Abdullah, Siti Masitah Jamaluddin, Yuslina Yusoff, and Amir Shahmi Md Ali. Institute for Medical Research, Allergy and Immunology Research Centre, Kuala Lumpur, Malaysia. Background: Crab has been recognized as a source of potent allergens especially in countries where seafood forms a large portion of the diet of the community. Blue/ swimming crab (Portunus pelagicus) and red crab (Charybdis japonica) from Family Portunidae are among the most commonly consumed crab in Malaysia. The objective of this study is to characterize the IgE-binding proteins and major allergens of these species of crab. Methods: Cooked and uncooked extracts of both crab were prepared and fractionated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE). IgE-binding proteins were then demonstrated by immunoblotting tests using sera from 99 and 76 patients with positive skin prick test (SPT) to blue and red crab extracts, respectively. Results: Both species of crab had similar protein profiles. The uncooked extracts of both crab produced 19 protein bands in the range of 15 to 138 kD. Several protein bands between 17 to 25 kD and 50 to 71 kD which were present in the uncooked extracts appeared to be denatured in the cooked extracts. The immunoblotting of blue and red crab revealed 14 and 15 various IgE-binding proteins, in the range of 24 to 138 kD, respectively. The most common IgE-binding proteins were identified in the range of 34 to 46 kD. A 34 kD protein was identified as a major allergen for both crab. A 36 kD protein was also demonstrated as a major allergen in red crab and two proteins at 38 and 46 kD were also seen as major allergens in blue crab. Interestingly, these major allergens at 34, 36 and 38 kD were similar in size with tropomyosin, the main major allergen and responsible in the majority of allergic reactions to various seafood species including crab. Skin prick test reactivity to peanut and tree nuts in adult and children with atopy Noormalin Abdullah, Shahnaz Murad, and Rosmilah Misnan. Institute for Medical Research, Allergy and Immunology Research Centre, Kuala Lumpur, Malaysia. Background: Nut allergy (peanut and tree nut) is a common cause of allergy and can even cause severe anaphylactic reactions. Peanut allergy is more common than tree nut allergy, but many subjects develop hypersensitivity to both peanuts and tree nuts. Peanuts are known as ground nuts, earth nuts and monkey nuts while tree nuts refer to nuts such as Brazil nut, hazel nut, almond, walnuts and cashew nuts. The peanut plant, Arachis hypogaea, belongs to the botanical family Leguminose and do not related botanically to nuts such as Brazil nut, hazel nut or almond. There are no reports of prevalence of peanut and tree nuts allergy in Malaysia neither is there reports of fatal anaphylaxis due to nut allergy. Methods: A total of 621 children and adults with history of atopy (asthma, allergic rhinitis, hives and food allergy) were skin tested to a panel of peanuts and tree nuts (commercial and in-house extracts). Results: We found that children have higher skin test positivity to peanuts and tree nuts compared to adults. In children, skin prick test showed 8.5% positivity to raw peanut and walnut, 6% to big peanut and 4.3% to almond, and cashew nuts extracts whilst skin prick test positivity to both hazel nuts and walnut were lower at 2.6% and 1.7% respectively. In adults, the most common skin test reactivities are peanuts, 5% and 4% to cashew nuts. Conclusion: It appears that skin test reactivity to nuts exist in subjects with no clinical presentation of nut allergy. A large number of individuals have skin test reactivity to multiple nuts and this is probably due to crossreactivity. Allergen-specific IgE-values to inhalant and food allergens in China -comparison between two commercial immunoassays: Baoqing Sun, Jing Li, and Nanshan Zhong. Guangzhou Institute of Respiratory Diseases, Dept of Medicine, Guangzhou, China. Purpose of the Study: Comparison of two in vitro assays for the detection of allergen-specific IgE (sIgE) and Total IgE (TigE) (Dr. Fooke ALLERG-O-LIQ, Neuss, Germany, www.fooke-labs.de; Pharmacia CAP-System, Upsala, Sweden, www.pharmacia.com) re-analyzing sera with sIgE to common inhalatans and food allergens and TIgE. Materials and Methods: Allergens: Inhalants: housedust mite D. pter. d1, D.farinae d2, Blomia tropicalis d5, Cat (Epithelia) e1, Dog (Epithelia) e5, German Cockroach i6, Aspergillus fumigatus m3; food allergens (FA): hen's egg f1, cow's milk f2, Crab f23, shrimp f24. Patients Sera: Sera from allergic patients (bronchus asthma, allergic rhinitis and chronic cough) were collected from Oct. 2004 to May. 2007 at Guangzhou Institute of Respiratory disease out patient department. Clinical data were not evaluated. Performance: sIgE was measured in single runs according to the recommendations of the manufacturers. Assay features and methodological differences are listed in table 1. Calculation: Associations between quantitative sIgE-levels of different assays were calculated using non-parametric Spearman-Ranktest and depicted with log scales. Sensitivity, Specificity and Agreement per allergen was calculated, using following formula: 1) Sensitivity (%) = (# of LIQ+ results)/(# of CAP+ result per allergen) 2) Specificity (%) = (# of LIQ-results)/(# of CAP-result per allergen). Background: Autism is a neuropsychiatric condition that presents with abnormal, bizarre behaviour patterns and accompanied by learning difficulties. Peptides known as casamorphin (from milk proteins) and gliadinomorphin (from gluten protein) are absorbed from the gut into the blood stream leading to the theory of Bleaky gut syndrome[. Methods: Eighty-four autism children, divided into 2 groups: younger age (less than 5 years) and an older age group (5 to 19 years) were skin prick tested (SPT) to a panel of food allergens. Blood test for specific Immunoglobulin E (IgE) to cow's milk, wheat, soya and oat, specific Immunoglobulin G (IgG) and Immunoglobulin A (IgA) to alpha-lactalbumin, beta-lactoglobulin, casein and gliadin were carried out. Questionnaires on behavioural pattern of children were filled by parents. Results: The majority of autism children were Chinese (76.2%) and male (88%). In the younger group, we found that SPT to soya 8.3%, wheat 3.5%, and cow's milk 2.3%. Specific IgE to cow's milk 9.52%, wheat 2.3%, and soya 1.2%. 8.3% had detectable levels to IgG alpha-lactalbumin, 5.9% to IgG beta-lactoglobulin, 1.2% to IgA beta-lactoglobulin and 4.76% to IgG casein. No IgA alpha-lactalbumin, IgG casein, IgG gliadin, IgA casein and IgA gliadin were detected in this group of children. In the older children, 23% has positive SPT to wheat, 17.8% to soya, and 9.5% to cow's milk. Specific IgE to cow's milk and wheat was 9.52% whereas to soya 8.3%. 11.9% had detectable IgG alpha-lactalbumin, 7.1% to IgG beta-lactoglobulin, 3.5% to IgG gliadin while 2.3% had detectable levels of IgG casein, IgA alpha-lactalbumin, beta-lactoglobulin, gliadin and 1.2% to IgA casein. As to behavioural pattern, the study showed that 70.6% of younger group and 64.2% of older group presented with unawareness of the world, 88.4% in younger group has poor eye contact compared to 61.2% in older group. There is not much difference in general learning ability and antisocial behaviour observed in both groups. In conclusion this study showed children 5 years and above had a higher SPT positivity to wheat and cow's milk. Casein was found to be low in both age groups and no gliadin was detected in the younger group. Behavioural pattern seemed to be improving as they became older and has no correlation with cow's milk and gluten allergy. Background: In order to diagnose allergic patients with more relevant allergenic species which they are directly exposed to, indigenous species were collected and prepared commercially along with other allergens under a collaborative project with M/s Inmunotek, Madrid, Spain. These allergens were tested in allergic patients in the region to evaluate the efficacy and reactivity. Methods: A total of 30 allergenic extracts with 50% indigenous species were included in the Diagnostic Panel. These allergenic species were selected after comprehensive aerobiological studies in many parts of the country and their growth and availability in the region. Glycerinated extracts were prepared by Inmunotek under a brand name of Allergotek. Standard prick tests with histamine and saline controls were conducted on 541 allergic individuals in the Middle East and Africa. The method is considered to be convenient and economical providing nature of sensitizing allergen(s) and an opportunity for a possible successful immunotherapy. Maximum combined reactivity with pollen allergens for all sites were recorded with Cynodon dactylon (11.8%), Chenopodium murale (10.8%), Phoenix dactylifera (10%) Salsola imbricata (9.2%) Prosopis juliflora (8.9%) and Lolium perenne (8%). The maximum indoor allergens reactivity was recorded with D. farinae (15.7%), D. pteronyssinus (15.7%), Felis domesticus (12.5%) Periplaneta americana (10.4%), Blatella germanica (9.7%) and Blatta orietalis (8.4%). The results of this efficacy trial of indigenous allergens revealed that majority of these allergens were effective with moderate to severe reactions. Asthmatic and allergic individuals were found to be comparatively more sensitive to indoor allergenic species than the outdoor allergens. While the cultural habits and climate appear to have played a role, socioeconomic conditions did not influence the overall sensitization pattern. It is further suggested that, if possible, regional species and/ or allergenic material should be included in the diagnostic test panel. Prevalence and sensitization to weeds pollen in Saudi Arabia Methods: SPT on 500 individuals having asthma and other allergic manifestations attending allergy clinics in six different regions was conducted using commercial extracts. The selection of allergens was made after an extensive nation wide aerobiological survey using Burkard Volumetric Spore Traps. The major pollen components of the Kingdom_s environment were identified as Amaranthus viridis., Atriplex polycarpa, Chenopodium album, Cyperus rotundus, Rumex crispus and Plantago spp. Results: The SPT data revealed a comparatively higher degree of sensitization to weeds pollen. In the south, mountainous region (Abha), 21.8%, while in an agricultural setting (Gassim) 75.5% patients reacted to weeds pollen. In an another location in the Eastern region (Hofuf) 16.7% of the patients while close to Red Sea region (Gizan) 9% of the patients reacted positively to various weeds pollen, which included Atriplex polycarpa, Chenopodium album, Salsola tennifolia and Rumex crispus. Individual pollen releaved Chenopodium album with maximum reactivity (81.8%) in agriculture setting (Gassim) followed by Salsola tennifolia (75.5%), (25% Al-Hofuf), Rumex crispus 27.3% (Gassim) and 18.1% (Gizan). Apart from Cynodon dactylon, a grass pollen and Prosopis juliflora, a tree pollen, highest skin reactivities were recorded by members of the chenopodiace weeds in all regions. The study indicates that sensitization and exacerbation of symptoms in patients during pollination season may be caused by desert weeds growing in the Kingdom, and may possibly be a major contributor of respiratory allergy in Saudi Arabia. Specific IgG antibodies (total and subclasses) to saffron: a study of their correlation with specific IgE and immediate skin reactions Saffron (crocus sativus) cultivation is one of the most agricultural products in Iran. Saffron contains an aeroallergen that causes reactive respiratory allergic reactions in exposed subjects. To investigate the role of specific IgE and IgG in this type of allergic reaction, saffron specific IgE and IgG subclasses were determined in the sera of 38 exposed subjects (test group) and 20 normal subjects (control group) by ELISA. Immediate skin reactions were assessed in exposed subjects. The mean optical densities of saffronspecific total IgG and IgE antibodies were significantly higher in the test group than the control groups. Specific IgG4 antibodies were lower in the test group. Other IgG subclasses did not have any significant difference in the two groups. The immediate skin reactions were positive in 80% of the test group. The specific IgE antibody levels correlate with the specific total IgG antibody levels and the positive skin reactions (R = 0.67 and R = 0.42, respectively). A reverse correlation was found between the specific IgE antibody levels and the specific IgG4 antibody levels (R = 0.03). Can specific IgE testing in primary care be cost saving? Background: Allergy reduces quality of life and places a considerable burden on society. Specific immunoglobulin E (sIgE) testing can improve management and potentially reduce costs. The purpose of the study is to assess the cost-effectiveness of allergy testing for patients with respiratory and/or skin problems in primary care in Italy. Methods: A cost-minimization analysis was carried out in which the costs of the two treatment alternatives were compared and assessed using a decision model that includes costs for pharmaceuticals, physician visits in primary care, and the costs of allergy testing. In the no-test-strategy part, patients were classified as allergic, non-allergic or uncertain, on the basis of clinical judgment without the test. In the test-strategy part, classification of patients was also based on sIgE results. Results: Agreement between physicians_ classification and test results was substantially elevated in the test-strategy part (0.88) vs the no-test-strategy part (0.52). Costs per patient over 2 years decreased from 784 in the no-test strategy to 535 in the test strategy, owing mostly to reduced medication costs. Sensitivity to the prevalence of allergy and price of medications was demonstrated. Conclusion: Allergy testing in primary care is cost-saving compared with not testing. Decrease in false-positive or uncertain diagnoses is the main component of cost reduction and enables more appropriate patient management. Study of a clinical cohort of patients tested for allergy in a diagnositic immunolocgy clinic Heidemarie Saxarra, Glenn Reeves, and Karla Lemmert. John hunter Hospital Haps, Immunology Haps, Newcastle, Australia. A large number of patients (500) were studied and their allergy resuts collated. The specific allergen tests to dust mite, grass mix, food mix, animal epithelial mix, weed mix, latex and peanut were tabulated. This group of patients was then analysed for trends in any of the following parameters: (clinical notes and medical records were used ) age, sex, medication, general health, symptoms of allergy and severity of the symptoms. The study was limited in that all of the parameters were not available for the cohort studied. No significant trends noted. However useful guidelines established for future reviews of a large patient cohort in the diagnositic laboratory. How can the royal college of pathologists of australasia immunology qap help your laboratory? Robert Heddle 1 , Sue Jovanovich 2 , and Peter Roberts-Thomson 2 . 1 Flinders Medical Centre, Department of Respiratory Medicine, Adelaide, Australia; 2 Flinders Medical Centre, RCPA Immunology QAP, Adelaide, Australia. Why does in vitro allergy testing survive? despite generally being less sensitive, slower and more expensive than skin testing? 1. Applicable when: & skin disorders (e.g. eczema, dermatographism) & medications (e.g. H1 anti-histamines, tricyclic antidepressants) & risk of anaphylaxis (e.g. initial testing for drug allergy; extreme nut allergy) would prevent or inhibit skin testing 2. Does not require clinician to carry an expensive range of allergens 3. Lends itself to quality control, standardisation and regional collaboration Why quality control in vitro allergy testing? 1. Some errors are clinically crucial; e.g. false negative result in nut allergy 2. Credibility of laboratory reagents, equipment and performance depends on satisfactory performance in QC 3. Understanding discrepancies can enhance products and Appropriateness of reducing the number of pollen allergens to three Karuna Keat 1 , Karen Byth 2 , and Connie Katelaris 1 . 1 Westmead Hospital, Immunology Department, Westmead, Australia; 2 Westmead Hospital, Westmead Millenium Institute, Westmead, Australia. Background: Skin prick testing (SPT) is an essential tool in the diagnosis of allergic disorders. The optimal number and type of allergens used in different settings remains undefined. We aim to describe SPT in our clinical practice and propose the appropriateness of reducing the number of pollen allergens to three. The aim is to improve cost-effectiveness and reducing time spent on allergen testing, particularly in the community setting. Methods: Consecutive patients who attended the private rooms of 2 immunologists and the allergy/immunology clinic in a tertiary referral hospital who required skin prick testing were evaluated from June 2006 to November 2006. Statistical analysis was undertaken with the use of Pearson`s Chi-square and Fisher`s Exact test to assess for significance. Multivariate analysis was also performed. Results: There were a total of 273 skin prick test sets performed. There was no significant difference between the rates of SPT positivity with common pollen allergens between the two clinics. A positive SPT to Perennial Ryegass (Lolium perenne), Timothy (Phleum patense) or Bermuda grass (Cynodon dactylon) had a sensitivity of 100% to Bent or Orchard grass (Dactylis glomerate), with sensitivities of 97%, 96.3% and 94.8% to English Plantain (Plantago lanceolata), Bahia grass (Paspalum notatum)and Dock/Sorrel (Rumex sp) respectively. Use of Perennial Ryegrass, Timothy or Bermuda grass also detected tree pollen sensitivity with sensitivities in Birch mix (Betula sp) of 92.6%, Acacia (96.4%), Casuarina (89.1%), Platanus sp (92.3%) and Privet(Ligustrum sp) (88.6%). Conclusion: The use of 3 common grass pollen allergens in SPTs (Lolium perenne, Phleum pratense and Cynodon dactylon) detected 90% of atopic individuals with sensitivity to many pollen types. This information may be useful in defining the most appropriate allergens to determine pollen hypersensitivity in community settings. Clinical evaluation of a new allergy lateral flow assay for professional and home use Michael Mahler 1 , Lorenz Christine 2 , Ralf Lucassen 1 , Margrit Fooke 3 , and Jörg Kleine-Tebbe 2 . 1 Dr. Fooke Laboratorien, Development, Neuss, Germany; 2 Allergy and Asthma Center Westend, Allergy unit, Berlin, Germany; 3 Dr. Fooke Laboratorien, General manager, Neuss, Germany. Background: Specific immunoglobulin E (sIgE) is a hallmark in the diagnosis of type I allergic reactions and atopic diseases. A new allergy screening test (Allergy Lateral Flow Assay; ALFA TM ) for qualitative detection of sIgE in human whole blood, serum or plasma is based on a test device, allowing linkage to a variety of allergens. Objective of our study was the evaluation of ALFA TM for professional and home use. Aspergillus niger (m33)] with capillary blood. Each serum was tested for specific IgE to all single allergens contained in both ALFA TM tests by ALLERG-O-LIQ. Furthermore, skin prick tests (SPT, Allergopharma) were performed. Volunteers were defined allergic if patient`s history was concordant with SPT and sIgE in-vitro results. ALFA TM results and patient's diagnoses were analyzed by kappa agreement, Chi-square test, positive (PPV) and negative predictive value (NPV) and diagnostic efficiency (DE). Results: ALFA TM results were obtained from 91 (S) and 83 volunteers (P Sensitization to five common aero-allergens in children suffering from atopic eczema as examined by atopy patch tests, skin prick-tests and specific IgE Martin Liska, Petr Panzner, Vladimir Hrasko, and Vaclava Gutova. Faculty Hospital Pilsen, Institute of Immunology and Allergology, Pilsen, Czech Republic. Background: Although the role of allergy in atopic eczema (AE) is still controversial, some patients with atopic eczema suffer from exacerbation of skin lesions after contact with or inhalation of aeroallergens. From the histological examinations of the skin after contact with aeroallergenes is known that the delayed-type hypersensitivity reactions mediated by allergen-specific T cells can take a part in pathogenesis of atopic eczema. Atopy patch tests (APT) represent a useful tool for detection of such hypersensitivity. Methods: We examined hypersensitivity to common aero-allergens (birch pollen, grass pollen, cat dander, house-dust mites) using APT, skin prick-tests (SPT) and specific IgE in 27 children suffering from atopic eczema. Results of all methods were then compared. Results: Delayed-type hypersensitivity was found out (using APT) in 16 patients (59%), immediate type of hypersensitivity was found out (using SPT ) in 13 patients (48%), using specific IgE in 15 patients (55%). Only immediate type of hypersensitivity was proved in 5 patients (18%), only delayed-type hypersensitivity in 6 patients (22%). Both types of hypersensitivity occurred concomitantly in 11 patients (41%). In 32 cases the type of hypersensitivity differed in the same allergen. A significant (pG 0.0005) positive correlation was found between SPT and specific IgE. Correlation of clinical symptoms of AE and positivity of tests was in 7 patients (26%) in IgE mediated hypersensitivity and in 10 patients (37%) in delayed-type hypersensitivity. Conclusion: Various aero-allergens can influence substantially the course of atopic eczema not only via specific IgE, but as well by specific T cell-mediated reactions. Therefore testing for hypersensitivity to aero-allergens both using SPT and/or specific IgE, and atopy patch tests could be useful. Bogdan Petrunov 1 , Georgi Nikolov 1 , Antoaneta Michova 1 , Tzveti Ivanova 1 , Julia Radenkova-Saeva 2 , and Hristo Taskov 1 . 1 National Center of Infectious and Parasitic Diseases, Allergology and Immunology, Sofia, Bulgaria; 2 Emergency Hospital BPirogov[, Toxicology Department, Sofia, Bulgaria. Background: The aim of the study is to assess the diagnostic potential of two in vitro methods for IgE diagnosis and to compare them with the skin-prick test (SPT) as a gold standard. Methods: 131 patients with positive case history and SPT to grass pollen, house dust mite, moulds, bee and wasp venom suffering of bronchial asthma or allergic rhinitis /hay fever and 10 clinically health controls were studied. The in vitro quantity of serum allergen-specific IgE (UniCap, Pharmacia) and the percentage of allergen-specific basophil's degranulation (FasImmune, BD) were evaluated. The correlation and the percent of coincidence of the results from the three methods were analysed (Statistica 5.5). Results: Significant statistical correlation between the resuls from the three methods in patients sensitized to grass pollen and house dust mite were found. Strong positive correlation (Spearman, pG0.05) between the SPT and the quantity of specific IgE-R=0.67 and R=0.61, between the in vivo test and FasImmune -R=0.66 and R=0.62 and between the both in vitro methods -R=0.67 and R=0.53 were determined. Data from patients, allergic to insect's venom, showed a high percent of coincidence between the three methods -from 70% to 90%. Respectively a cincidence of 60% between the SPT and the quantity of specific IgE in the group sensitized to moulds was established. Conclusion: The results from the invitro methods represent positive correlation and coincidence with SPT, especially for the allergens of grass pollen, house dust mite, bee and wasp venom. Their application ensures more preceise diagnosis of patients and contributes to the complex assessment of IgE mediated allergy. Rawil Fakhrullin 1 , Rustem Fassakhov 2 , Victor Vinter 1 , Elena Sukmanskaya 2 , and Olga Konovalova 1 . 1 Kazan State University, Department of Biochemistry, Kazan, Russian Federation; 2 Kazan Reseach Institute of Epidemiology and Microbiology, Department of Allergology and Immunology, Kazan, Russian Federation. Background: Allergic disease have a significant impact on clinical practice due to their high prevalence. The total IgE quantification is one of the important steps in the classic atopic disease diagnostics.The most widely used methods for IgE detection are time-consuming and complex. Biosensors are interesting tools offering certain operational advantages over standard photometric methods, notably with respect to rapidity, ease-of-use, cost, simplicity, portability, and ease of mass manufacture. Methods: QCM work as sensors based on the relationship between frequency change and mass loading on the surface of the crystal according to Sauerbrey equation (Sauerbrey, J Phys, 1959) . When antigens react with coated antibodies on the surface, a frequency shift occurs and this change is proportional to the mass loading. Results: The monoclonal anti-IgE were successful immobilize in Nafion polymeric matrix on silver electrodes of piezoelectric quartz resonator. The optimal conditions for anti-IgE immobilization procedure and for piezoelectric immunoassay have been determined. Only 10 microlitres of serum and 45 minutes reaction time is required to measure total IgE. It was found that biosensor is capable to differentiate blood serums of patients with low, intermediate and high level of IgE. Conclusion: The quartz crystal microbalance immunosensor offers a number of significant advantages over the currentlu available in vitro techniques for the detection of total IgE. It is supposed that such biosensor can be used in laboratory practice for IgE determination. Background: The purpose of this study is to determine over 12 months, 2/1/ 06Y1/31/07, the rate of SRs to both P and ID ST, the symptoms reported, and the response to immediate treatment with epinephrine IM. Methods: A retrospective review over a one year period was conducted to evaluate SRs to P and ID ST to 20 to 50 allergens (trees, grasses, weeds, animals, molds, foods, medications, and Hymenoptera) in 1,456 subjects. A standard form was used to record symptoms, signs, and treatment. No vasovagal reactions were included. Nurses as instructed by the attending physicians administered epinephrine 1:1000 v/v, 0.2mL IM as soon as any signs or symptoms of anaphylaxis occurred. Results: 52 patients (3.5%) had SRs, 43 (83%) female and 9 (17%) male. The average age of the patients with SRs was 40.6 years (range 13Y70, median 35.5 years). 17/52 (33%) had asthma. Symptoms reported: pruritic eyes, nose, and/ or pharynx (40%), worsening cough (27%), sensation of difficulty swallowing (17%), worsening nasal congestion (15%), rhinorrhea (13%), chest tightness and/or shortness of breath (13%), generalized pruritus (12%), sneezing (12%), urticaria (4%) , and wheeze (4%) . No severe asthma, shock, hypotension, unconsciousness, or late phase responses occurred. Treatment: 52 (100%) patients received epinephrine (average dose, 0.2 cc, 1:1000 IM), 48 (92%) oral prednisone, 9 (17%) oral prednisone to take 6 to 8 hours after reaction, 50 (96%) oral antihistamine (H1) , and 6 (12%) nebulized beta agonist. Conclusion: SRs occurred in 3.5% of patients skin tested and readily responded to early intervention with epinephrine. This early administration of epinephrine by nurses appears to prevent more serious and late phase reactions. Clara cell protein in irritating and sensitizing effects of inhaled benzalkonium chloride in rats Background: Benzalkonium chloride (BAC) is a bacteriostatic agent used in the pharmaceutical industry as a preservative and is known to cause bronchoconstriction in asthmatic subjects. The aim of our study is qualification of results of inhalation exposure to BAC in rats, with particular reference to the effect on the remodelling of the respiratory system. Conditioned allergic reactions that impair lung function, such as allergic asthma, can be evaluated by specific lung biomarkers. It is known that the irritant fumes affect nonciliated epithelial Clara cells, which release antiinflammatory and immunosuppressive Clara cell protein (CC16) into the respiratory tract. Materials and Methods: Female Wistar rats were exposed to BAC aerosol at 30 mg/m3 for 5 days (6h/day) and on day 16 were re-exposed to BAC for 6 h. After the exposure, bronchoalveolar lavage fluid (BALF) was collected. BALF concentration of total protein, CC16, IgE, MMP-9, hyaluronic acid (HA), IL-6, TNF-", MIP-2 and activity of lactate dehydrogenase (LDH) were determined. CC16 as the marker of bronchiolar epithelium was assessed by latex immunoassay. In the lung, histological examinations were done and the activity of glutathione S-transferases (GST) was determined. Additionaly total and differential cell number (lymphocytes, neutrophils and macrophages) were measured. Results: Benzalkonium exposure after challenge induced statistically significant increases of BALF cytokines, LDH and IgE in BALF and serum. CC16 level in BALF was significantly reduced. Significant negative correlation of CC16 concentration in BALF with mediators (IL-6) of inflammatory processes was seen. Huge increase of LDH correlated with the level of total protein, MIP-2 and IgE in serum. Negative relationship was shown to occur between CC16 and LDH. IgE in serum and BALF correlated with MMP-9. In histopathology examination, focal agglomerations of alveolar macrophages were noted as well as proliferation of peribronchial lymphatic tissue. Conclusion: CC16 play a protective role in allergic inflammation and take part in remodelling effects of low molecular weight sensitizers. CC16 can be used as a diagnostic marker for early detection of impaired respiratory function. Pulmonary irritation after inhalation exposure to benzalkonium chloride in rats Radoslaw Swiercz 1 , Tadeusz Halatek 1 , Wojciech Wasowicz 1 , Barbara Kur 2 , Zofia Grzelinska 1 , and Wanda Majcherek 1 . 1 Nofer Institute of Occupational Medicine, Department of Toxicology and Carcinogenesis, Lodz, Poland; 2 Nofer Institute of Occupational Medicine, Department of Immunotoxicology, Lodz, Poland. Background: Benzalkonium chloride (BAC) is a quaternary ammonium compound in which the alkyl groups have a chain length from C8 to C18. BAC exerts toxic effects on microorganisms. This property has been utilized in the cosmetic industry and medicine, where it is used as an effective germicide and preservative agent. Various BAC-containing preparations used by people may produce a number of adverse effects on the human body. Bearing in mind that BAC is widely used in different branches of the national economy, its toxic effect may constitute a major health problem. Materials and Methods: Female Wistar rats IMP: WIST of body weight 165Y185 g were exposed to BAC aerosol at the target concentration of 30 mg/m3 in the dynamic inhalation chamber for 6 h and 3 days (6 h/ day). After the exposure and 18 h after termination of exposure to BAC aerosol, bronchoalveolar lavage fluid (BALF) was collected from each animal and BALF concentrations of total protein, Clara cell protein, matrix metalloproteinase-9 (MMP-9) hyalurnic acid (HA), immunoglobulin E (IgE) and cytokines (TNF-", IL-6 and MIP-20) and the activity of lactate dehydrogenase (LDH) and GSH-S-transferase (GST) were determined. Results: All the rats survived inhalation exposure to 30 mg/m3 BAC. A significant reduction of body weight was noted in the animals exposed repeatedly by inhalation to BAC. Lung weight, total protein, HA level and LDH activity in BALF were higher in rats after single and repeated exposure to BAC, compared to control. Decreased concentrations of CC16 in BALF of rats were observed after the single and repeated inhalation exposure. A significantly higher level of IL-6 and IgE were noted in the BALF from the animals exposed to the single and repeated dose. Concentrations of MMP-9, TNF-", and MIP-2 in BALF of rats exposed to BAC were similar to those found in the control animals. Conclusion: BAC showed a strong inflammatory and irritating activity in the lungs of the rats already after 6 hours of inhalation exposure. BAC stimulates the dynamic patterns of IL-6 and IgE production and infiltration of protein from blood circulation system to BALF. Continued exposure resulted in changes involving cellular destruction, statistically increase of LDH activity and a continuous reduction of CC16 concentration in BALF. Allergic bronchopulmonary aspergillosis in an asthma clinic using essential minimal criteria Soo-Keol Lee 1 , Doo-Kyung Yang 1 , Choon-Hee Son 1 , Ki-Nam Kim 2 , and Ki-Nam Lee 2 . 1 Dong-A University, Internal Medicine, Busan, Republic of Korea; 2 Dong-A University, Diagnostic Radiology, Busan, Republic of Korea. Objective: Allergic bronchopulmonary aspergillosis (ABPA) occurs in cases of atopic asthma and may result in important lung disease. Early diagnosis is essential as this disease is responsive to corticosteroids. However, there is still no consensus about the diagnostic criteria, because patients in different stages of ABPA may not fulfill the criteria. In this study, we evaluate the prevalence of ABPA or ABPA-like disease in an asthma clinic using essential minimal diagnostic criteria. Methods: A prospective evaluation of patients with bronchial asthma for ABPA from July 2006 onward. ABPA was diagnosed using essential minimal criteria ?asthma, skin prick testing (SPT) positivity to Aspergillus fumigatus (Af), elevated serum total IgE (CAP), elevated serum Af-specific IgE (CAP), and central bronchiectasis on CT scans. Results: Ninety consecutive patients with bronchial asthma were enrolled. Forty-four of 90 patients were atopic (49.0%), 7 of 44 (18.0%) were positive to SPT to Af. Five of 44 patients (11.0%) showed only elevated serum Af-specific IgE without positive response to Af on SPT. A secure diagnosis of ABPA, satisfying all essential minimal criteria, was evident in 4 of 12 patients (33.3%). Conclusion: There is high prevalence of ABPA in asthmatic patients presenting our hospital. Further evaluations are required to differentiate ABPA from asthma patients sensitizing to Af without ABPA. The role of serum Af-specific IgE as a screening tool in diagnosis of ABPA should be redefined. Clinical presentation in 12 patints with allergic bronchopulmonary aspergillosis Abdelmonem Sharara, Manaf Hijazi, Hythem El-Khushman, Jafer Momany, and Mohammed Enjada. King Hussein Medical Center, Department of Chest Disease, Amman, Jordan. Purpose: Allergic bronchopulmonary aspergillosis (ABPA) is an immunologically mediated lung disease charecterise by a Complex hypersensitivity reaction in patients with asthma which occurs when bronchi become colonized by Aspergillus. Repeated episodes of bronchial obstruction, inflammation, and mucoid impaction can lead to bronchiectasis, fibrosis, and chronic lung disease. Our aim of the study is to increase awareness of this disease. Methods: We described a study of 12 cases with Allergic bronchopulmonary aspergillosis Twelve patients (6 men and 6 women) were diagnosed in chest department at King Hussien Hospital between 1993Y2003. The main criteria for the diagnosis were A history of asthma, immediate skin test reactivity to Aspergillus antigens, Serum total IgE concentration greater than 1000 ng/mL, peripheral blood eosinophilia more than 500/mm3, Lung infiltrates and proximal bronchiectasis. Results: Demographic data for 12 patients with Allergic bronchopulmonary aspergillosis. Conclusion: ABPA is a rare disease, diagnosis is depending upon certain criteria. Clinical Implication: We have to think about the diagnosis of ABPA in any patient with a history of asthma, lung infiltrates and peripheral blood eosinophilia. The significance of the diagnostic profile of the ophthalmic allergies in excluding mimicking clinical conditions(MC)* which may pose therapeutic difficulties M Ishaq, I Khan Sameera, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Introduction: Patients with (AC) with/without concomitant allergies in some cases is a therapeutic dilemma. Methods: In the series of patients, ages 10Y35 years usually with intermittently red. eyes(shot redness), intractable itching of eyes, tearing (stringy discharge) with/without seasonal association. On laboratory investigations, is found raised tears & blood eosinophils counts, total eosinophils counts. Total serum IgE measurement in most of the cases had been higher than 200 to 300 kU/l. supported by, the. rise in titers of allergen-specific IgE, by the radioallergosorbent test (RAST) method, Skin prick test with a mixture of allergenic extracts had a conclusive evidence of an allergy cause for the red eye. On ophthalmoscope examination. Found pinkish papillae, with a central vessel & characteristics, serous, watery conjunctival secretion. Conjunctival scrapings, and tear cytology performed after topical ocular allergen challenge to sensitized subjects have shown significant increases in neutrophils and eosinophils, and their presence evidenced a positive diagnostic criterion. Results: Confirmation of a suspected allergic sensitization by skin prick test for the diagnosis of immediate hypersensitivity is the most sensitive, fastest and cheapest method to confirm an allergic sensitization. However, it carries a small but a significant risk of systemic anaphylaxis. Conclusion: Challenge tests are the only way to relate the specific allergen to the triggering of ocular symptoms. but with a variable degree of systemic anaphylaxis. *Bacterial, chlamydial and viral conjunctivitis, superior limbic, phlyctenular, conjunctivitis, keratoconjunctivitis, rosacea-associated conjunctivitis, erythema multiforme, eoiscleritis/scleritis, and ocular cicatricial pemphigoid. Electrodiagnostic study of phrenic nerve function in patients with systemic lupus erythematosus Nancy Mahmoud Abdelaty 1 , Mahmoud El Prince 1 , S.Maher Labib 2 , and Mohammed Hefny 3 . 1 Faculty of medicine, Suez Canal University, Chest Department, Ismailia, Egypt; 2 Faculty of medicine, Suez Canal University, Medicine Department, Ismailia, Egypt; 3 Faculty of medicine, Suez Canal University, Physical Medicine Department, Ismailia, Egypt. Objective: 21 patients with SLE were screened for the presence of Phrenic nerve neuropathy and to determine whether neurophysiologic findings correlate to clinical respiratory signs, spirometric abnormalities or serological examination in patients with Systemic lupus erythematosus. Methods: A total of 21 patients(18 female & 3 male)with systemic lupus erythematosus (SLE) (age range, 16Y36 yr) were included and studied by physical pulmonary examination, chest radiography, respiratory function tests, as well as serological examination and bilateral transcutaneous phrenic nerve conduction studies. Results: 14(66.6%) patient complained of dyspnea ,only one patient showed paradoxical abdominal movement .Pulmonary function tests showed proportional reduction of the forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1), suggesting a restrictive process which was severe in 23% of patients. All patients were on corticosteroids, only 10 (47.6%) patients were on immunosuppressive medication to include methotrexate or cyclophosphamide. Phrenic nerve evaluation using transcutaneous stimulation studies showed delayed latencies of RT, LT & both phrenic nerve in 17 (81%), 19(90%) and 17 (81%) patients respectively confirming a demyelinating neuropathy. Also Phrenic nerve stimulation evoked a low-amplitude response from the right, left and both in 17 (81%), 15 (71%) and 14 (66.6%) of patients respectively confirming axonal neuropathy. There was no significant correlation between electrical phrenic nerve stimulation and serum immune markers, except there was decreased action potential amplitude in SLE group with positive results for Anti DNA as14 (66.6%) of patients had Anti DNA + ve, all showed reduced amplitude of rt phrenic nerve & 13(93%) of them showed reduced amplitude of lt phrenic nerve. Fourteen (66.6%) patients presented with dyspnea and all of them showed abnormal phrenic nerve conduction studies. While 11 patients showed abnormal CXR with small but clear lung fields, no evidence of major parenchymal lung or pleural disease was found. There was no significant correlation between electrical phrenic nerve stimulation and CXR abnormalities. Conclusion: Diaphragmatic weakness in patients with SLE is both common and is very likely to be caused by a phrenic neuropathy with evidence of bilateral involvement. The search engine as a diagnostic tool in complex immunological and allergic case reports: is google useful? Objective: To evaluate the utility of searching clinical information with Google in order to obtain a correct diagnosis of complex immunological and allergic (CIA) case reports. Study Design: Comparative cross-sectional study. Methods: Firty-five CIA case reports were randomly selected by an independent investigator from peer-viewed medical journals. Clinical data was presented separately to three observers blinded to final diagnosis. Observer A is a Consultant in Internal medicine and Allergy with an expert knowledge of these fields and basic computing skills. Observer B is a Registrar in Internal medicine and Allergy. Observer C is a research nurse. Both observers B and C had a more familiar knowledge of the regular use of computer search engines. An internet-based search using Google was conducted. In order to perform this, the observers individually studied each text and independently selected five search terms, of their own choosing, from each case record to enter into the standard Google search engine. The observers then recorded for each case the single most prominent diagnosis that was evident from within the first three results pages of the conducted Google search. Since Google does not necessarily include diagnoses within the search results page itself, observers were permitted to select the diagnosis that best fitted the case record from information after opening each direct results link only. The independent investigator then compared the diagnoses obtained by each observer with the definitive diagnoses as published in the Journals. The main outcome measure of this study was the percentage of correctly obtained diagnoses achieved by each observer. Results: Observer A identified the definitive diagnosis in 30/45 cases (66%, 95%CI 52Y79). Observer B in 39/45 (86%, 95%CI 76Y95) and Observer C in 29/45 (64%, 95%CI 50Y77). Most diagnostic inaccuracies for both observers were those related to primary immunodeficiency or pediatric cases. Conclusion: This Google-based search was useful to achieve an appropriate diagnosis in CIA cases. Computer and Internet-based search skills could influence the results. The anti-allergic properties of potassium humate Johanna Meeding, Gandy Justin, Constance Medlen, and Jacques Snyman. University of Pretoria, Pharmacology, Pretoria, South Africa. Background: Although the anti-inflammatory properties of humate derived from peat, sapropeles and mumie have been described, no clinical studies has been done on the anti-inflammatory effects of humate derived from coal. Leonardite humate compared favourably with prednisolone in suppressing contact hypersensitivity in a rat model. According to a report by the European Agency for the Evaluation of Medicinal Products on toxicity studies (Feb 1999), humic acids extracted from brown coal has no toxic effects on rats in a chronic study at oral dosages as high as 1g/kg BW, whereas the LD50 in rats, after oral administration of humic acids, has been reported to be greater than 11g/kg BW. This report has recently been confirmed by a separate study. The objective of this study was to establish the safety and therapeutic efficacy of oral potassium humate in reducing the signs and symptoms of hay fever in atopic patients during the grass pollen season. Methods: In this parallel double-blind placebo controlled phase II study potassium humate was randomly assigned, at a dosage of 1.8g in divided doses/day, to atopic patients (n = 40) presenting with acute symptoms of hay fever. The blood and nasal samples were used to determine the safety and the effects of potassium humate on basophil activation, cytokine levels and eosinophil migration. A skin prick test was used to determine its anti-allergic effects. An in vitro neutrophil adhesion test was used to determine the effects of the product on the adhesion of human neutrophils to ICAM-1expressing baby hamster kidney cells. Results: A significant decrease in the skin prick test results (presented elsewhere) and eosinophil counts was observed. No significant differences were observed with regard to neutrophil adhesion nor were there any differences observed with regard to the stimulation of basophils. However decreases were observed in the expression of IL-4, IL-5, IL-8 and IL-1â after treatment, although not reaching statistical significance. The product had no effect on neutrophil adhesion to ICAM-1. Conclusion: This study confirmed, without doubt, that this product possesses anti-inflammatory as well as anti-allergic properties possibly due to a decreased recruitment of eosinopils to the site of inflammation. Anti-inflammatory effect of LipoPGE1 on therapeutic intervention for intractable skin ulcer Yorihisa Kotobuki, Hiroyuki Murota, and Ichiro Katayama. Course of Integrated Medicine Graduate School of Medicine, Osaka University, Dermatology, Osaka, Japan. Background: Intractable skin ulcer is common in our daily practice. Although patients' QOL is severely impaired by this skin lesion, until now effective treatment protocols have not been established. Wound healing process can be separated into inflammatory phase, proliferating phase, and remodeling phase. Recent reviews have described that prolonged inflammatory phase might partly affect the pathogenic mechanisms of intractable skin ulcer. During the inflammatory phase, various kinds of inflammatory cell infiltrates ware observed in the affected area, and recruited macrophages secrete some inflammatory cytokines including IL-6 and VEGF. Prostaglandin E1 (PGE1)treatment is believed to be one of the promising treatment for skin ulcers, and LipoPGE1 also demonstrated improvement in a drug delivery system. To note, it has been reported that infiltrated macrophages uptake LipoPGE1. Taken these results together, we hypothesized that LipoPGE1 might have an antiinflammatory effect and thus contribute to the improvement of intractable skin ulcer. Methods: Patients with various kinds of intractable skin ulcers were administered with intra-venous injection of 10 microgram/day LipoPGE1 (Palxus\) for two weeks, and the size of the ulcer area and serum concentration of CRP, IL-6, VEGF, and sICAM1 were measured before and after treatment. Results: LipoPGE1 effectively reduced the size of the ulcer area and the serum inflammatory markers after two weeks of LipoPGE1-treatment. Conclusion: These results indicated that anti-inflammatory effect of LipoPGE1 might contribute to the improvement of the intractable skin ulcers. Comparative efficacy of levocetirizine, desloratadine, clemastine, kvifenadine and sekvifenadine on histamine prick test induced weal reaction, blood perfusion evaluated by laser Doppler flowmetry. Randomized, double-blind, placebo-controlled, crossover design study Background: Evaluation of weal reaction and laser Doppler flowmetry are valuable methods for evaluation of efficacy of different pharmacological agents. The aim of our study was to compare the influence of different H1-antihistamines on histamine induced weal reaction, increase of skin blood perfusion and sedation. Methods: Histamine prick test induced weal area in mm2, percentage of blood perfusion change and area under curve during peak perfusion period (AUCmax) was measured with Periflux System 4000 (Perimed AB, Sweden) 2 hours after intake of 5 mg levocetirizine, 5 mg desloratadine, 1 mg clemastine, 50 mg kvifenadine, 50 mg sekvifenadine and placebo. Sedative effect was measured in mm by visual analogue scale (VAS). Results: Results were expressed as mean T 95%CI. Mean weal reaction area was 6.9 (j3.9;+10.7); 17.5 (j12.6;+23.1); 20.2 (j14.9;+26.2); 18.1 (j13.1;+23.9); 17.8 (j12.8;+23.5) and 29.0 (j22.6;+36.1) mm2 respectively. Statistically significant difference was observed between active treatment and placebo (pG0.05), and levocetirizine and other H1-antihistamines (pG0.001). Increase of blood perfusion was 393.1% (j221.3;+613.8); 626.2% (j403.0;+898.3); 756.5% (j508.8;+1053.2); 741.0% (j496.2; +1339.1); 1001.5% (j712.8;+1339.1) and 1033.2% (j739.6;+1375.8) respectively. Significant decrease of augmentation of blood perfusion was observed after pre-treatment with levocetirizine and desloratadine vs. placebo (pG0.05) and levocetirizine vs. kvifenadine, sekvifenadine and clemastine (pG0.05). AUCmax was 1298.7 (j781.2;+1947.0); 2197.3 (j1504.5; +3020.9); 2454.3 (j1718.3;+3321.0); 2633.2 (j1868.6;+3528.6); 2551.7 (j1800.0;+3434.2) and 3166.2 (j2321.4;+4141.7) U*s. AUCmax was significantly lower after pre-treatment with levocetirizine vs. placebo and other antihistamines (pG0.05). Sedative effect was 24.5 (j17.9;+32.1); 21.1 (j15.1;+28.1); 28.2 (j21.2;+36.2); 17.6 (j12.0;+24.1); 15.1 (j10.0;+21.0) and 19.9 (j14.1;+26.7) mm of VAS. Significant difference of sedation we observed between levocetirizine vs. sekvifenadine, clemastine vs. kvifenadine, sekvifenadine and placebo (pG0.05). Conclusion: Levocetirizine induced significant decrease of weal and flare reaction and skin blood perfusion compared to placebo and other H1antihistamines. Influence of kvifenadine and sekvifenadine on weal reaction area was similar to desloratadine and clemastine. Sedative effect of clemastine was more pronounced than kvifenadine and sekvifenadine. Characterising the pharmacological properties of fluticasone furoate, a novel enhanced-affinity glucocorticoid Methods: Binding affinity of FF for the human lung glucocorticoid receptor (GR) was determined by elucidation of association and dissociation rate constants. Molecular interactions of FF with the GR were identified by X-ray crystallography of the GR ligand-binding domain. Cellular onset of action was determined by measuring nuclear translocation in human lung epithelial cells. Measurement of human progesterone (PR), mineralocorticoid (MR), androgen (AR), and oestrogen receptor (ER) activities was used to assess the steroid hormone selectivity of FF. Cellular protection of FF to elastase or mechanical wounding was determined in 16HBe human lung epithelial cells and antiinflammatory effects of FF in the lung were determined using the Brown Norway Ovalbumin Rat model. Results: FF has very fast association with, and slow dissociation from, the GR, with a relative receptor affinity (RRA) of 2988T135 with reference to dexamethasone (RRA: 100T5), higher than all other currently available clinical glucocorticoids: mometasone furoate (MF) 2244T142, fluticasone propionate (FP) 1775T130, beclomethasone-17-monopropionate 1345T125, ciclesonide active principle 1212, and budesonide 855. FF has hydrogen bond interactions with the GR through the 3-keto (with Gln570 and Arg611) and the 11$-hydroxy (with Asn564) groups; the 17 $-fluoromethylthioester group also forms a favourable electrostatic interaction with Asn564. FF induces a rapid translocation of GR into the nucleus (G20 minutes to maximum effect) and has high selectivity for GR (30-, 790-, 9330,000-and 9330,000-fold) versus PR, MR, AR and ER, respectively. FF confers substantial protection against elastase-and mechanically induced damage, with more potent protection than budesonide, FP and MF. FF completely prevents lung eosinophilia, an effect greater than that with FP. Conclusion: FF has enhanced affinity for the GR compared with other available glucocorticoids, which translates into more potent protection against cellular damage and lung inflammation. Coupled with its fast cellular onset of action and high selectivity for the GR, these properties may contribute to a favourable clinical efficacy and safety profile for FF. Downmodulatory effects of cetirizine and levocetirizine on cytokine/chemokine production and CD54 expression in keratinocytes Miwa Kobayashi, Kenji Kabashima, and Yoshiki Tokura. University of Occupational and Environmental Health Japan, Dermatology, Kitakyushu, Japan. Cetirizine is an antihistamic drug of the second generation. Besides its anti-histamic activity various actions have been reported in this antihistamic. In epidermal keratinocytes, cetirizine inhibits the expression of costimulatory molecule ICAM-1 and the MHC class II molecule HLA-DR. Moreover, it exerts anti-inflammation actions by suppressing the production of cytokines and chemokines in various immunocompetent cells. Levocetirizine (L-cetirizine) is the optical isomer of cetirizine and widely used for the treatment of allergic disorders in European countries. In this study, we investigated whether there are differences between cetirizine and levocetirizine in the cytokine and chemokine production by normal human epidermal keratinocytes (NHEK).While NHEK were stimulated with interferon-,(IFN-,) and tumor necrosis factor-" (TNF-") cetirizine or levocetirizine was added to the experimental cultures. Three day-culture supernatants were measured for the concentrations of IL-1", IL-8, RANTES, Mig, I-TAC and MDC. The IFN-,/TNF-"-augmented levels of IL-1", IL-8 and I-TACK were significantly suppressed by the addition of either cetirizine or levocetirizine to the culture in dose-dependent manners (10 j9 Y 10 j7 M). RANTES, Mig or MDC was not suppressed by cetirizine. To examine the effects of these two reagents on the expression of CD54 (ICAM-1) molecules, NHEK were incubated with IFN-, with or without cetirizine or levocetirizine for 48 hrs. Cetirizine and levocetirizine at 10 j8 M downmodulated the expression of CD54 molecules at similar levels to each other.This study demonstrates that cetirizine and levocetirizine have comparable effects on the immunological function of keratinocytes. It is noted that levocetirizine has slightly but significantly stronger effects than cetirizine in the production of RANTES and Mig. Efficacy and safety of levocetirizine 5mg as continuous or on-demand treatment for persistent allergic rhinitis over 6 months Giorgio Walter Canonica, Enrico Compalati, Laura Guerra, Anthi Rogkakou, Cinzia Gamalero, Pierangela Massacane, Federica Fumagalli, Christian Zanella, and Ilaria Baiardini. University of Genoa, Allergy & Respiratory Disease Department, Genoa, Italy. Background: We aimed to document the efficacy and safety of levocetirizine 5mg as continuous (CT) or on-demand treatment (OnD) of persistent allergic rhinitis (PER), as defined by ARIA. Methods: This was a single-center, randomised, open-label study comparing CT vs. OnD treatment of PER patients with levocetirizine 5mg, once daily, over 6 months. Patients were allowed to have mild asthma treated with a shortacting beta-agonist. Sneezing, rhinorrhea, nasal and ocular pruritus (T4SS=sum of these 4 symptoms) and nasal congestion were measured daily on a 0 (absent) to 3 (severe) scale. Rhinasthma questionnaire (RQ) was used to assess subjects`quality of life (range 0Y100; 30 item questionnaire: 1=not bothered at all, 5=bothered very much). Quality of sleep was reported on a VAS scale (0=worst; 10=best). Results: 31 patients were enrolled per group; 18 in the OnD and 22 in the CT group completed the study. No patients discontinued for drug-related serious adverse events (AEs). Improvement from baseline in T4SS was significantly higher in favour of CT during months 5 (pG0.01) and 6 (pG0.03). The maximal T4SS improvement was 80% for OnD and 87% for CT. The maximal improvement in nasal congestion was 75% for OnD and 85% for CT. Quality of sleep considerably improved at end of study: from baseline VAS=5.77 (OnD) and VAS=5.63 (CT) to VAS=7.82 (OnD) and VAS=7.25 (CT). No serious (AEs) were observed. 3.3% of subjects in the CT and 9.4% in the OnD groups reported drug-related treatment-emergent AEs. Conclusion: Our study confirms previous data that, when taking a potent antihistamine, like levocetirizine, PER symptoms (including nasal congestion) are effectively controlled over a 6-month treatment period. In addition, most of the individual symptoms were controlled significantly better when treated continuously. Regardless of the regimen, levocetirizine improved the patientsq uality of life and sleep, and was very well tolerated with fewer patients reporting AEs in the CT group. Our results support the long-term continuous treatment of PER with a potent and well-tolerated antihistamine. Oxatomide-treated children with atopic dermatitis complicated by food allergy and prevention of asthma development Norifumi Ogawa, Toshiaki Saeki, Kawano Yutaka, and Takeshi Noma. Kitasato University School of Medicine, Pediatric Department, Sagamihara, Kanagawa, Japan. Background: Recent epidemiology suggests the increasing prevalence of allergic diseases in the industrialized countries including Japan, which necessitates the analysis of the mechanisms of allergic diseases and development of the effective treatment. Oxatomide (OXM), an antihistaminic drug, has been shown to be clinically effective for the treatment of hypersensitivity and childhood asthma. Its mode of action has been elucidated to increase IFN-g activity as well as anti-histaminic reaction. Objective and Methods: Peripheral blood mononuclear cells were obtained from 41 patients with atopic dermatitis allergic to hen-egg ranging from 2 months to 2 years 10 months in age. The patients had recurrent eczema, pruritus and positive skin reactions to egg white and/or cow`s milk. Patients also had positive responses to the oral provocation test to raw hen egg and or cow`s milk. Diagnostic criteria for atopic dermatitis was based on the criteria of Hanifin and Rajk. To clarify the mode of action whereby OXM ameliorates the conditions of the children with food allergy-complicated atopic dermatitis and whether development of bronchial asthma is prevented, OXM-related alterations of the clinical symptoms and examination, seen in patients during the course of 8 months`to 6 years and 10 months`treatment was evaluated. Results and Discussion: Scores for itching, sleep disturbance and skin lesions (inflammation, lichenification, cracking) was improved from 10 to 2.7 (mean) during the course of 8-16 weeks' treatment with 2mg/kg of OXM in addition to elimination diets, treatment of skin care (shower, Isodine[R], non-steroid ointment), administration of hydroxyzine and/or oral sodium cromoglicate. In further study, OXM efficiently suppressed incidence of asthma to approximately 9% of the patients (control:42.9%) and both of total IgE value and peripheral eosinophils count was not elevated after OXM-treatment and were lower than those in age-matched asthma patients un-treated with OXM. Conclusion: OXM, which is a significant candidate for one of the therapeutic modalities against children with food allergy-induced atopic dermatitis, and based on the clinical study, was also found to be effective prophylaxis for development of childhood asthma. A comparison between intramuscular dexamethasone and fluticasone propionate inhaler in treatment of croup Jamal Faghihinia. Isfahan Univercity, Pediatric, Isfahan, Islamic Republic of Iran. Introduction: Croup is a common viral disease in children under 6 years old with incidence rate of 2Y6%. The mainstay of treatment is airway management. Treatment focuses on respiratory distress, using cold mist, epinephrine, heliox and corticosteroids. In this study we tried compare the effectiveness of Fluticasone spray with intramuscular Dexamethasone. Materials and Methods: In this clinical trial, 107 children with croup randomly assigned into two groups. The study group was treated by Fluticasone Propionate and the control group was treated by intramuscular Dexamethasone. Croup scoring was performed at the 6th and 12th hours from initial administration according to Westley croup score. Results: Improvement was observed in 83% of the study group and 66% of the control group, 6 hours after initiation of treatment. In both groups 10% of the patients didn`t respond to treatment (p = 0.03). 12 hours after treatment the study group response was 85% and the control group response was 90% (p = 0.4 rhinitis and chronic idiopathic urticaria in pediatric and adult populations. The clinical and pharmacological profiles of DL have been extensively investigated and DL is available in a number of formulations, including tablets and syrup. The bioequivalence of DL tablets and syrup has not been studied to date in a Chinese population. This randomized, open-label, single dose crossover trial studied the pharmacokinetics of DL 5mg administered as a 5mg tablet or 10 ml of 0.5 mg/ ml syrup in 24 healthy adult male Chinese subjects. After providing written informed consent and undergoing screening, subjects were admitted to a clinic for baseline assessments. Subjects were randomized to receive one of the two DL formulations in a fasting state. Blood tests for pharmacokinetics were taken over 5 days (subjects remained in the clinic for the first 24 hours), and after a 14 day washout period the subjects were crossed over to the other DL formulation and underwent identical pharmacokinetic analyses. The main pharmacokinetic variables for the two formulations were the log-transformed AUC(I) and the Cmax for DL and 3-OH-DL. Biochemical and hematological tests, ECG data and vital signs were also assessed during the study and adverse event (AE) reports were collected. Results: DL was safe and well tolerated when administered in the tablet or syrup formulations; no AEs were reported. The Tmax, T1/2, Cmax, and AUC(I) values for DL and 3-OH-DL were similar for both formulations. There were no statistically significant differences between the tablet and syrup DL formulations on the basis of log-transformed Cmax and AUC(I) values for DL and 3-OH-DL (P90.05). The 90% CIs of AUC, and Cmax were 91.61Y103.97% and 86.04Y99.92% respectively for DL, and 94.22Y101.71% and 88.01Y101.35% respectively for 3-OH-DL. The relative bioavailbility of the DL syrup was 99.4% for DL and 98.79% for 3-OH-DL, which met the criteria for bioequivalence of the two formulations. Conclusion: Both syrup and tablet formulations of DL 5mg were safe and well tolerated. When administered as a syrup formulation DL was bioequivalent to the tablet form of DL in healthy Chinese subjects. The pharmacokinetic and safety profiles of desloratadine in healthy korean volunteers Background: Desloratadine (DL) is a non-sedating, selective and potent H1receptor antagonist that is effective and well tolerated in the treatment of subjects with allergic rhinitis and chronic idiopathic urticaria. The pharmacokinetics (PK) of DL have not been studied in a Korean population, to date. Methods: This was a double-blind, dose escalation study of the PK and tolerability of single doses of DL 5mg, 10mg and 20mg in 36 healthy male Korean subjects. In each dose group 10 subjects received DL and 2 received placebo. Safety was demonstrated at the 5mg DL dose before escalation to the next dose group. Subjects were screened for eligibility during a 3 week prebaseline period. Subjects were confined on Day -1 and baseline blood/urine tests and ECGs were performed. DL was administered fasting at 9 am on Day 1 and subjects remained confined until Day 3. Blood sampling was performed for biochemistry, hematology and PK from DL administration until Day 8 and the Tmax, terminal T1/2, Cmax and AUClast for desloratadine and it 3-OH-DL metabolite were calculated. Vital signs, physical examinations and ECGs were performed regularly and adverse event (AE) reports were collected. Background: In elderly patients with allergic rhinitis, the second-generation H1-antihistamines have not been adequately studies, although they are widely used and assumed to be safe. Objective: To evaluate cardiac safety of loratadine in the treatment of allergic rhinitis in elderly patients. Methods: A total of 40 patients with perennial allergic rhinitis were enrolled in the study. There were 25 males and 15 females, aged 50 to 88 years (mean, 64.4-years-old). 17 cases (42.5%) had a history of cardiovascular diseases and/ or presented abnormal ECG parameters, but had no prolonged QT-interval. The subjects received loratadine 10mg once-daily for 30 days. A series of baseline ECG recordings was obtained before treatment. ECG effects of the treatments were then compared with the baseline ECGs. Results: There were no changes in sinus rhythm in all patients after 30 days treatment by loratadine. No statistically significant difference was found between the heart rates, P durations, PR and QRS intervals at baseline and endpoint ECGs (P 90.05), with no significant prolongation of the QT as well as QTc corrected for heart rate using Bazett formula (P90.05). The results suggest no cardiotoxicity of loratadine, at the usual recommended dose, in long-term treatment of allergic rhinitis in the elderly. Propranolol cytotoxicity on human leukemic MOLT-4 cell line Background: Propranolol, a beta-adrenergic blocker has been used for treatment of a large number of cardiovascular diseases. This drug is also an inhibitor of phosphatidic acid (PA) phosphohydrolase and phosphatidic acid biosynthesis. Phosphatidic acid is a growth factor for tumor cells. In addition, the inhibitory effect of Propranolol on the development of a tobacco-induced pulmonary adenocarcinoma and also its cytotoxicity on rat and human lung macrophages and human lung tumor cell line has been reported. The widespread and long-term use of propranolol in lots of heart diseases as well as its cytotoxicity against some tumor cells, prompted us to investigate its cytotoxic effect on a human T leukemic cell line . Methods: The MOLT-4 cells were cultured in complete RPMI medium and then incubated with different concentrations of Propranolol ( 0.0004 Y0.4 mM) for 10 and 20 hours. The cytotoxicity was then assessed by 3-[4,5-dimethyl thiazolY2,5-diphenyltetrazoliumbromide (MTT) reduction and also trypan blue dye exclusion methods. Results: Propranolol induced a significant dose dependent cytotoxic effect on human MOLT-4 cell line in less than 10 hours compared to untreated control cells. The results showed that human T leukemic cell line was dose dependently sensitive to Propranolol. Further studies investigating the in vivo effect of Propranolol on leukemic patients and also other leukemic cells are warranted. The clinical features, diagnoses and laboratory investigations in these subjects were prospectively collected, and laboratory staff blinded to these details performed low-stringent reverse transcription-polymerase chain reaction (RT-PCR) assays using 12 pairs of primers that detect constant regions of HCoVs (i.e. pancoronavirus). Results: 1139 subjects (57% males) were recruited, with mean (SD) age being 5.1 (3.6) years. The main discharge diagnoses were pneumonia (n=239), upper respiratory infection (URI; n=227), asthma (n=191), seizure (n=107), bronchiolitis (n=105), roseola infantum (n=98), croup (n=31), and others (n=141). Twenty-eight (2.5%) of these NPA samples were positive for HCoVs. The clinical diagnoses associated with these HCoV isolates included asthma (n=7); seizure (n=6); URI (n=5); bronchiolitis, pneumonia, tonsillitis and roseola infantum (n=2 for each); and croup and otitis media (n=1 for each). HCoV infection was not related to age, highest respiratory rate and maximal temperature (P90.3). HCoV infection was not associated with wheezing illnesses as defined by Fasthma', Fbronchitis' or Fbronchiolitis' (2.7% versus 2.4%; P=0.870) or with lower respiratory infections (the above three plus Fpneumonia'; P= 0.341). HCoV cases were more likely to suffer from seizure (5.6% versus 2.1%, P=0.040). Complete blood count and C-reactive protein were not related to HCoV infections (P90.15). Conclusion: HCoVs are uncommon yet important pathogens causing seizure disorders in local hospitalised children. On the other hand, HCoV infections are not associated with wheezing illnesses in Hong Kong children. Are there predominant strains of Staphylococcus aureus in atopic dermatitis patients? : Genotypic characterization of staphylococcus aureus isolated in adolescent and adult patients with atopic dermatitis nization with S. aureus and clinical severity / skin barrier function has been demonstrated. Qualitative analysis, especially a genotypic characterization of S. aureus isolated from atopic patients, however, has rarely been reported. Methods: This study aimed to find the genotypic characterization of S. aureus from atopic dermatitis patients. We performed newly-developed typing methods -spa typing, multi-locus sequence typing (MLST) and toxin gene assay, by a multiplex polymerase chain reaction, with 165 isolates of Staphylococcus. The results showed that there was no predominant clone of S. aureus with a high heterogenicity of spa typing and MLST. A toxin gene assay showed very interesting results that all S. aureus strains had at least two kinds of toxin genes; sea and tsst-1 being the most prevalent. Role of primary and secondary low-grade rhinovirus infection in allergic airway inflammation in a murine model of allergic asthma Mykola Korzh. Kharkov National University, Fundamental Medicine, Kharkov, Ukraine. Background: Rhinovirus respiratory syncytial virus (RSV) infection is known to develop and exacerbate asthma in young children. In adult, RSV causes recurrent but asymptomatic infections. However, the impact of asymptomatic RSV infection on adult asthma is yet to be determined. The aim of this study was to determine the effects of primary and secondary low-grade rhinovirus infections on allergic airway inflammation in a murine model of allergic asthma. Methods: A low-grade rhinovirus (2 x 10 (3) plaque-forming units/mouse) was inoculated, and this caused neither pulmonary inflammation nor symptoms but induced significant IFN-gamma production in thoracic lymph nodes. To investigate interaction between low-grade virus and Dermatophagoides farinae (Df), airway hyperresponsiveness, lung inflammation and cytokine production from thoracic lymph nodes were compared after primary and secondary low-grade rhinovirus infections in four groups of mice; control, Df allergen-sensitized, rhinovirus-infected and Df-sensitized rhinovirusinfected mice. A direct comparison between low-and high-grade rhinovirus infections was also performed in primary infection. To investigate the role of IL-5 during secondary rhinovirus infection, anti-IL-5 monoclonal antibody (anti-IL-5 mAb) was injected in mice and similar parameters were compared in four groups of mice. Results: Primary high-grade rhinovirus infection increased allergen-induced airway inflammation, while primary low-grade rhinovirus infection attenuated allergen-induced airway inflammation concomitant with significant IFN-gamma production in lung-draining lymph nodes. In marked contrast, secondary lowgrade rhinovirus infection increased both IFN-gamma and IL-5 production, resulting in exacerbation of allergen-induced airway inflammation. Anti-IL-5 mAb treatment in secondary low-grade rhinovirus infection and Df allergensensitized mice attenuated virus and allergen-induced airway inflammation. Conclusion: Low-grade rhinovirus infection per se does not cause pulmonary inflammation, whereas it induces a significant immunological response in the allergen-sensitized host. These results indicate that subclinical and recurrent rhinovirus infection may play an important role in exacerbation and maintenance of asthma in adults, wherein IL-5 is critically involved. Hai Lee Chung, and Sang Mi Kwon. School of Medicine, Catholic University of Taegu, Pediatrics, Taegu, Republic of Korea. Background: Zinc is one of the dietary antioxidants. Previous studies have shown that zinc is crucial for normal development and function of cells mediating non-specific immunity. Recently, zinc supplementation was reported to reduce acute lower respiratory infections and prevent severe pneumonia in children. Our purpose was to examine zinc levels in the serum of the young children who had recurrent early wheeze and evaluate the clinical and laboratory findings in relation to zinc status. Methods: Seventy-three patients (aged from 8 months to 6 yrs) admitted with acute respiratory infection with wheezing were enrolled. All children had experienced more than 3 episodes of wheezing before admission. Zinc levels were measured in serum samples collected on admission using inductively coupled plasma-optical emission spectrometry (ICP-OES) and the value of G 64 mg/dl was defined as zinc deficiency. Clinical and laboratory findings in the children with zinc deficiency were examined and compared with in the children who had normal values. Zinc levels in sixteen age-matched controls were also studied. Results: Median value of zinc levels in the patients was significantly lower than in controls (PG0.001). 36 patients were found to have zinc deficiency (49.3%), which was significantly higher than in controls (12.5%). Zinc deficiency was observed in 56% of the patients = 2 yrs of age and 40.6% of 92 yrs of age. There was no significant difference in total WBC count, lymphocyte count and atopic status in relation to zinc status in the patients. CD4/CD8 ratio was significantly lower in the patients with zinc deficiency (PG0.05), however, other immune profiles were within normal limit. Conclusion: This study showed that median value of zinc level was significantly lower and zinc deficiency was more frequently found in the patients with recurrent early wheeze compared with in age-matched controls. Our results suggest that zinc deficiency may be associated with frequent respiratory viral infections, a likely trigger for recurrent early wheeze in the young children. Aurora Losada Pena, Ma Luz Díez Gómez, Aythamy Henríquez Santana, and Emma Gonzalez Seco. Hospital Ramon y Cajal, Allergy department, Madrid, Spain. Aim: Mycobacterium avium intracellulare (MAV) is the atypical Mycobacterium most commonly associated with human disease. The pulmonary disease is the most frequently clinical presentation and appears with higher prevalence in immunosuppressed patients. We present the case of a 46 years old woman, nurse as profession , with cough and dysnea for a period of nine years. No wheezing, fever nor constitutional syndrome were referred. Skin prick tests with common aero-allergens and latex, spirometry and bronchodilatation test were performed. Total IgE, complement study, proteins electrophoresis, immunoglobulins determination, cellular immunity study, HIV, X-ray study and thoracic CT-scan, mantoux, zielh and sputum culture were done. Results: Positive skin prick test for pollens and dog and cat epithelia were obtained. The patient had normal spirometry values and a negative bronchodilatation test. All the laboratory tests were in normal levels. Determination of total IgE was 483 KU/l. The chest X-ray showed cavities in both lungs with interstitial infiltrates. The CT-scan confirmed these findings. Mantoux, Zielh with 50 BAAR/field and MAV culture were positive. Mycobacterium tuberculosis was excluded by CRP. Cellular immunity, complement, proteins electrophoresis and immunoglobulins determination were in normal range. HIV test was negative. Conclusion: We present the case of a patient with rhinoconjuntivitis due to pollens hypersensitivity and persistent cough with pulmonary infection for Mycobacteium Avium associated. The Mycobacterium Avium was not described as human pathogen until 1950, when many series described pulmonary infections for MAV. This mycobacterium mainly attacks immunosuppressed patients. This infection is less frequent in patients with normal immunity. Our patient did not have immunosuppression nor risk factors. At the present time she is being treated with antibiotics (ethambutol and claritromicin) and she is in good general condition with no need of hospitalisations. Chronic Granulomatous Disease (CGD) is an inherited phagocytic disorder caused by mutations in NADPH oxidase subunits. Patients with CGD have life-threatening bacterial and fungal infections. Childrens Medical Center at Tehran University is the referral center for immunodeficiency in Iran. During two years of study forty five families with clinically diagnosed CGD were referred to this center. Neutrophil functional assays performed for affected children and their mothers; no activity or residual activity was detected in affected neutrophils. PMN (Poly Morpho Nuclear) oxidative burst revealed mosaic pattern in 12 mothers. Western blot analysis revealed gp91phenotype in all their sons. Mutation screening in CYBB gene using SSCP analysis followed by sequencing, showed 9 different mutations including one novel mutation. Western immunoblot subtyping of patients whose mother showed no mosaic pattern by DHR123 revealed 24 patients with p47 null expression, 7 and 2 patients with p22 and p67 defect, respectively. %GT screening in Ncf1 gene for p47-patients, revealed 8 patients with this mutation. Mutation analysis for the rest of Ncf1 gene for these patients is understudy. CYBA mutation analysis revealed 6 different mutations including three novel mutations in p22-patients. Overall, the number of autosomal recessive patients with CGD in Iran is high and it seems consanguineous marriages is one of its causative factors. All appropriate papers and information were presented to volunteers. Motivation, understanding of vaccine properties and trial procedure, risk behavior, ability to perform the trial protocol were estimated. Volunteers enrolled in trials after confidential medical investigation and informed consent undersigning. Results: 65 persons of 298 enquiries were screened as potential volunteers. The main reasons for participation: help to HIV-infected people (65%) including persons in nearest surround (wife, husband, friend, child) (41%); contribution in development of HIV/AIDS vaccine (35%); possibility of immune protection against HIV (18%), free insurance and/or medical care (4%) . 20% (13 of 65) of persons refused to participate in trials. Reasons of refusal: doubts in vaccine safety and fear to receive HIV-infection (61%), fear of side effects (30%), fear of vaccine-induced HIV seropositivity (15%), impossibility to perform the trial protocol (7%), low compensation (2%), unexplained (15%). 30 persons not refused to participate in clinical trials, but decided to do it in future (clinical trials phase II). Reasons: planned pregnancy or temporal impossibility to perform the trial protocol. 17 volunteers (14 men, 1 woman, age 22Y31) were enrolled in trials. 2 of them leaved trials after 1st vaccination (unexplained reason), 9 successfully passed the trials protocol and 6 are at final step of trials. Conclusion: The first cohort for the clinical trials of preventive HIV/AIDS vaccines was created in Russia and partially enrolled in clinical trials of VICHREPOL vaccine. Cohort is stable (88%), expandable and may be included into international multicenter trials. A novel mutation within exon 12 of the CYBB gene resulting in severe form of x-link chronic granulomatous disease Chronic granulomatous disease (CGD) is a primary phagocytic disorder with defective superoxide formation and intracellular killing. The most common form, X-linked CGD (X-CGD) resulted from the mutation of CYBB gene on chromosome Xp21.1. We evaluated a Thai boy who had multiple Salmonella septicemia, Aspergillus pneumonia and brain abscess. His nitroblue tetrazolium (NBT) test was reportedly abnormal. The dihydrorhodamine (DHR) flow cytometry assay was performed and the fluorescence pattern upon stimulation was compatible with typical X-CGD. CYBB analysis revealed a novel complex mutation atggacgY ttca in exon 12 (base pairs 1532Y1538). As a result, 3 amino acids Tyr 511, Gly 512 and Arg 513 were deleted and replaced by 2 amino acids, Phe and Gln. The DHR and mutation analysis of his mother showed normal DHR pattern and no mutations in exon 12 of CYBB gene. Over 300 CYBB mutations have been registered in an internationally maintained X-CGD database. Most mutations are distributed throughout the 13 exons or at exon/intron boundaries, and almost 200 of these mutations are unique. We reported a novel mutation within exon 12 of CYBB gene which was on the nicotinamide adenine dinucleotide phosphate (NADPH)-binding domain in a CGD patient. Functional defect was demonstrated by almost absence of fluorescence upon stimulation of granulocytes on DHR histogram. This defect leads to a severe form of X-CGD. Background: Nutrition is an important factor that influences immunity, and nutritional deficiency can impair resistance to infections.Malnutrition is the most common cause of immunodeficiency worldwide. Trace elements such as zinc, selenium, iron, and copper can influence several components of immunity. Primmary antibody deficiency disorders are a group of disorders characterized by an unusual susceptibility to infections and malnutrition. Impaired nutritional status has been reported in immunodeficient patients. The aim of this study was to determine anthropometric indices and trace elements status in these patients. Methods: Thirty-eight children (28 males, 10 females, aged 2Y18 years) with primary antibody deficiency referring to Children,s Medical Center of Tehran University of Medical Science were enrolled in this research. Primary immunodeficiency disorders consisting of CVID, XLA, IgA deficiency, IgG subclass deficiency, and hyper IgM were assessed. Anthropometric indices, comprised of height, weight that were measured and body mass index (BMI) was calculated. Height-for-age (HAZ), weight-for-height (WHZ) and weightfor-age (WAZ) were determined according to Z-score to study mild, moderate and severe malnutrition. Serum copper, zinc, selenium and iron levels were measured by an atomic absorption spectrometer. Results: The most common disorders were CVID 52.5% and X-linked agammaglobulinemia 27.5%. Based on BMI measuredment 21.1% of patients had malnutrition. According to HAZ, 13.2%, 13.2%, and 36.8% had severe, moderate and mild malnutrition, respectively. According to WAZ, 10.5%, 18.4 %, and 28.6% had severe, moderate and mild malnutrition, respectively. Regarding to WHZ, 14.3%, 28.6 %, had moderate and mild malnutrition, respectively.Low selenium levels and high copper levels were obsevd in 37.5% and 70.3%, respectively. Conclusion: Anthropometric data showed that the frequency of malnutrition in these patients was higher than the CDC standard. Low serum selenium levels and high serum copper levels were observed, suggesting further research is needed on these parameters. Most of the patients had serum zinc and iron levels within the normal range. It is recommended that clinical immunologists and nutritionists should make a collective effort to provide these patients with standard or specialized diet so as to decrease the risk of infection. A registry of primary immunodeficiencies in a University Hospital in Thailand: an 18 year-review Patchanee Benjasupattananan 1 , Nualanong Visitsunthorn 1 , Pakit Vichyanond 1 , Voravich Luangwedchakarn 2 , and Orathai Jirapongsananuruk 1 . 1 Mahidol University, Siriraj Hospital, pediatric, Bangkok, Thailand; 2 Mahidol University, Siriraj Hospital, Immunology, Bangkok, Thailand. Background: Primary immunodeficiencies are group of rare diseases which are difficult to diagnose and manage. Most presenting symptoms were infections from uncommon organisms leading to high morbidity and mortality. Objective: To evaluate clinical characteristics of pediatric patients with primary immunodeficiencies in Thailand. Methods: Medical records of all patients diagnosed and treated for primary immunodeficiency in the past 18 years, at Siriraj Hospital, Bangkok, Thailand, were reviewed. Patients with secondary immunodeficiencies and chromosomal abnormalities were excluded. Results: A total of 85 pediatric patients (58 males and 27 females) were registered. The earliest onset of symptom was in the newborn period. IgG subclasss deficiency was the most common primary immunodeficiency diseases (47%), followed by severe combined imuunodeficiencies (17.6%), specific antibody deficiencies (14%), agammaglobulinemia (7.5%), common variable immunodeficiencies (4.5%) and chronic granulomatous disease (4.5%). The most common presenting symptoms of antibody deficiencies were upper respiratory tract infections (49/56, 87.5%). In patients with T-cell immunodeficiencies, most common presentations were PCP pneumonia (9/15, 60%) with septicemia being the most of common presentations among patients with phagocytic defect (2/4, 50%) . There is an increasing trend of early detection of primary immunodeficiency and decreasing diagnostic lag month over the past 18 years. History of death in family is the important information for rapid diagnosis. Conclusion: Establishment of registry of primary immunodeficiencies may provide the information for early detection and proper treatment to improve the prognosis of these patients in Thailand. Paucity of adverse events associated with administration of ivig Constance Katelaris 1 , and Marjorie Bennett 2 . 1 University of Western Sydney, Immunology ans Allergy, Sydney, Australia; 2 Westmead Hospital, Immunology and Allergy, Sydney, Australia. Introduction: Intravenous immunoglobulin (IVIG) is a valuable treatment for many immune-mediated disorders and it is vital management for those with humoral immunodeficiencies. These individuals receive regular, longterm therapy with IVIG. Adverse events associated with administration of IVIG have been reported. In our specialist unit at Westmead, hospital, Sydney, approximately 850 courses of IVIG are administered annually. Aim: As we have a large experience of IVIG administration, we have performed a retrospective analysis of adverse events (AE) experienced by patients attending our unit. Methods: Retrospective chart review of all IVIG infusions delivered in the last four years was performed. Factors examined were patient demographics, diagnosis, previous administration of IVIG infusion and symptoms of AE. IVIG products utilized were Intragam-P (CSL Bioplasma,Melbourne,Australia.), Sandoglobulin (ZLB Bioplasma A G, Bern, Switzerland.), Octagam (Octapharma AG, Lachen, Switzerland.). Patients receiving IVIG were not routinely pre-medicated with either antihistamines or steroids. Results: In our centre 3,320 infusions of IVIG were given in a 4 year period to 112 patients (46 males -66 females). 49 of the 112 patients had PID. 63 had various autoimmune conditions or treatment induced hypogammaglobulinemia. 50 of the patients were long standing recipients of IVIG and 62 were naBve to any IVIG products. During the 4 years there were 7 AEs observed. 3 of these patients were naBve to any IVIG products. 4 were long-term recipients. 3 patients had PID and the remainder had autoimmune conditions. Discussion: Our review showed that the incidence of AEs associated with IVIG infusions is very low (0.4%) for 3,320 infusions, which are similar to figures reported elsewhere. All AEs may have been preventable as three of the patients were naBve to IVIG and may have benefited from pre-medication and three of the reactions occurred when recommended maximum infusion rates were exceeded. The other reaction occurred due to patient delay between infusions. All patients whom experienced AEs have continued on IVIG without any reoccurrence of AEs and only one of these is routinely premedicated. Therefore, in conclusion, with due regard to infusion rates, timing between infusions and use of pre-medication IVIG is a safe treatment for humeral immunodeficiences and autoimmune disorders. Lilian Varga 1 , Zsuzsanna Kelemen 1 , Gábor Széplaki 1 , É va; Németh 1 , Judit Gács 1 , George Füst 1 , Beáta Visy 2 , and Henriette Farkas 1 . 1 Semmelweis University, 3rd Department of Internal Medicine, Budapest, Hungary; 2 Heim Pál Children Hospital, Department of Internal Medicine, Budapest, Hungary. The diagnosis of hereditary angioedema (HAE) is based on complement tests, however the relationship between the clinical symptoms and the complement levels is poorly studied. In our study we compared complement values in 90 patients with HAE and 212 patients with angioedema of unknown origin. In addition the complement parameters (CH50, C1q, C3, C4, antigenic and functional C1-INH) tested at the time of diagnosis were correlated with age, sex, severity of the disease in 99 patients with HAE type I and 7 patients with type II. Unlikely to the previous findings we have found that out of the complement parameters tested the functional C1-INH had the highest specificity, but the lowest sensitivity in the diagnosis of HAE, while highest specificity was observed with the antigenic C1-INH assay. We did not find correlation between complement levels and age at the time of diagnosis. No association was found between the complement levels and sex or HAE type. We found significant association of baseline functional C1-INH (p=0.0144), and CH50 (p=0.054) levels with the severity of disease. As a conclusion we demonstrated that both antigenic and functional C1-INH is required for the correct diagnosis of HAE and testing of C4 highly validates the results. Determination of funcional C1-INH and C4 may have clinical significance. Regular evaluation of these parameters can be a useful tool in the strategy of long term prophylaxis, however further studies are required to confirm these associations. Henriette Farkas, Lilian Varga, Gabor Szeplaki, György Temesszentandrasi, Laszlo Jakab, Bela Fekete, George Fust, and Istvan Karadi. Semmelweis University, 3rd Department OF Internal Medicine, Budapest, Hungary. Background: HAE is characterized by recurrent edematous swellings of the subcutaneous or submucosal tissues. Bradykinin plays an essential role in the development of angioedema. This study was conducted to investigate the effects of the selective bradykinin-B2-receptor-antagonist Icatibant in the treatment of attacks in patients with HAE due to C1-inhibitor deficiency. During an open-label extension phase of a randomized, double blind, controlled phase III study (FAST-2) patients with cutaneous and/or abdominal attacks of hereditary angioedema were treated with Icatibant. Within 6 hours after the onset of a moderate/severe attack they received a single injection of 30 mg Icatibant subcutaneously. Symptoms were assessed by patient (VAS, symptom score) and by physician (symptom score global assessment, clinical global impression). Results: 6 angioedema attacks in 2 patients with HAE were treated with Icatibant. The affected sites were twice the extremities and once the genitalia in Patient #1. Patient #2 experienced two abdominal attacks, while another attack affected the gastrointestinal tract and one extremity. Clinical symptoms of all attacks improved quickly. Time to first improvement of symptom was 10Y23 minutes. The duration of complete resolution of symptoms was shorter in abdominal attacks compared to attacks involving the subcutaneous tissues (180Y240 min versus 480Y720 min). Drug related adverse events and relapses were not experienced. Local skin reactions at the injection site were tolerable and resolved within 4 hours. Conclusion: IcatibantYwhich differs from earlier drugs concerning both the mode of action and the method of administration as wellYhas been found effective and safe in the treatment of angioedema attacks in patients with HAE. The possibility of subcutaneous administration of Icatibant may improve the daily life of patients suffering from HAE. In the present cases repeated use of Icatibant did not result in reduced efficacy. The potential of Icatibant for the treatment of other bradykinin mediated angioedema has to be tested in further studies. Leukocyte adhesion deficiency-1 (LAD-1) is a genetic immunodeficiency disease characterized by life-threatening infection results from the mutations in the leukocyte integrin, CD18 molecule with severe and moderate phenotype. We report a case of severe LAD occurred in a 1 month old girl and a case of moderate LAD which was presented with aggressive periodontitis. Case 1: Clinical features include delayed separation of the umbilical cord, omphalitis, severe bullose and /ulcerative skin eruption which mimic picture of staphylocolal scaled skin syndrom in neonatal period and persistent leukocytosis. The immunological workup showed pathological values of CD11b, CD18 (2.3%, 0.3% respectively). The patient is waiting for BMT now. Case 2: A 4/5 year old boy with aggressive periodontitis in the primary dentition which pathologic examination showed actinomycosis ginigivits and normal separation of umbilical cord. He had CD11b __ 4/6% and CD18 __ 3/ 7% in peripheral blood flow-cytometery compatible with moderate phenotype of LAD1. He is receiving antibiotic prophylaxis. Conclusion: Although LDA is a rare form of congenital immuno -deficiency, severe LDA1 should be considered when delayed wound healing and recurrent bacterial skin infections are present in a newborn and moderate LAD1 in aggressive periodontitis with normal wound healing. Clinical and molecular characteristics of Thai families with X-linked chronic granulomatous disease Prapaporn Vilaiphan 1 , Pantipa Chatchatee 1 , Jarungchit Ngamphaiboon 1 , Siraprapa Tongkobpetch 2 , Kanya Suphapeetiporn 2 , and Vorasuk Shotelersuk 2 . 1 Chulalongkorn University, Department of Pediatrics, Division of Allergy and Immunology, Bangkok, Thailand; 2 Chulalongkorn University, Department of Pediatrics, Division of Medical Genetics and Metabolism, Bangkok, Thailand. Rationale: X-linked chronic granulomatous disease (X-CGD) is an immunodeficiency disorder characterized by defective intracellular killing of microorganisms due to neutrophils' inability to generate superoxide ions. Although it is caused by mutations in the same CYBB gene, clinical and molecular characteristics vary among different ethnic backgrounds. Materials and Methods: Two unrelated Thai boys presented with severe persistent pulmonary infections at the age of two months. Their abnormal DHR assays supported the diagnosis of X-CGD. Mutation analysis was performed by polymerase chain reaction (PCR) amplification and sequencing of the entire coding regions of CYBB. Mutations identified were confirmed by restriction enzyme analyses. Results: PCR-sequencing of the entire coding regions of CYBB identified nonsense mutations, 271C9T (R91X) in exon 4 and 456T9A (Y152X) in exon 5, in probands of each family. Both of the probands' mothers were found to be carriers. Conclusion: This observation supports that CYBB is the gene responsible for X-CGD across different populations and nonsense mutations are associated with severe phenotypes. Background: Inflammatory responses to prior infections due to improved immune responses to latent pathogens with paradoxical clinical worsening observed in HIV-infected patients initiating potent antiretroviral therapy. No case report of cytomegalovirus (CMV) associated CNS IRIS in children. Methods: We report a case with serious atypical adverse events after initiate ARV. Results: A 7 years old Thai girl, CDC C due to wasting syndrome, presented with history of fever and oral candidiasis with CD4% 6%, CD4 count 17 cells/ml, viral load 163,000 copies/ml. After 2 weeks cotrimoxazole for PCP prophylaxis, she started antiretroviral therapy (ARV), stavudine, lamivudune and nevirapine, at Chulalongkorn hospital. One week later, she developed right sided hemiparesis without any history of headache, vomiting and loss of consciousness. The neurological exam revealed right sided weakness with motor power grade IV at both upper and lower extremities, normal reflexes and clonus. The differential diagnoses were cerebral toxoplasmosis, brain abscess and tumor. The CT brain with contrast media revealed generalized brain atrophy without mass or ring enhancement. Lumbar puncture was performed. The cerebrospinal fluid (CSF) was colorless and clear with WBC 2 cell/mm3, no RBC, protein 35.9 mg/dl and CSF sugar/ plasma sugar 70/96 mg/dl respectively. CSF bacterial culture was no growth but CSF cytomegalovirus (CMV) was positive through PCR method. MRI brain was shown hypodensity lesion at the right cerebellum. Ophthalmologic examination was normal.After 2 weeks of ARV, her CD4%, CD4 count and viral load were 17%, 202 cells/ml and 440 copies/ml respectively. The diagnosis was Immune Reconstitution Inflammatory Syndrome (IRIS). However, we did not prescribe any steroids because of the bacteremia. During this period, she continued to take ARV as usual. One month after her hospitalization, she was reexamined to be fine with full motor power. After a total of 6 months of ARV, she was fine with CD4% 13%, CD4 count 356 cells/ml, viral load 50 copies/ml. Conclusion: We reported the CMV IRIS, right hemiparesis, after a week of initiating antiretroviral therapy in an HIV infected girl with severe immunosuppression. The diagnosis of atypical CNS symptoms after initiate ARV with rapidly increased CD4 and decreased viral load should consider as IRIS. Usefulness of quantification of C-reactive protein (CRP) for diagnosis of bacterial infection in the febrile neutropenic child Martin Penagos 1 , Fortino Solorzano 2 , Miguel A Villasis 2 , Andrea Tapia 2 , Hugo Rivera 2 , and Roberto Bernaldez 2 . 1 Pediatrics Hospital, CMN, Pediatrics Section, Mexico City, Mexico; 2 Pediatrics Hospital, CMN BSiglo XXI[, IMSS, Pediatrics Section, Mexico City, Mexico. Background: Diagnosis of bacterial infection in a neutropenic child with fever becomes difficult due to a poor inflammatory response. Currently, a method reliable and fast to identify patients with bacterial infection is not available. CRP has been evaluated in these patients with controversial results. Objective: To determine the sensitivity, specificity, predictive values and likelihood ratios of CRP for diagnosis of bacterial infection in neutropenic children with fever. Design: Cross-sectional study. Methods: We included patients less than 16 years of age with fever and severe neutropenia (G500 AN/mm3). Blood cultures and CRP were taken in the initial evaluation. All of them, had empirical antimicrobial treatment; their evolution was followed up until discharge. Blood cultures were processed by the automated system BACT-alert; CRP by nephelometry. Patients were classified in 4 groups: Group I: Clinical and bacteriological infection, Group II: Infection clinically defined, Group III: Fever due other causes than infection and Group IV: Patients with cancer, neutropenia without fever. Diagnostic test analysis, receiver operating curves (ROC) and likelihood ratios were performed. Results: One hundred twenty seven episodes were included from 113 subjects. Leukemia was the most frequent disease (61%). Twenty nine, 47, 20 and 31 episodes were included in groups I, II, III and IV respectively. We found microbiological isolation in 29 episodes, Staphylococcus aureus and Escherichia coli were the most common isolated germs (27.6% and 17.2%). Median for CRP levels (IQR) was of 282 mg/L (174Y385) in group I; 205 mg/L (119Y267) in group II; 27,3 mg/L (12.3Y55) in group III and 5,1 mg/L (2.4Y13) in group IV (pG0.001). By COR curves a CRP level higher than 60 mg/L showed a sensitivity (S) of 94%, specificity (E) 94%, positive PV (PPV) 96% and negative PV (VPN) 92% when we compared groups I and II vs. III; at this cut level, likelihood ratio for a positive result was 15,6 and 0,06 for negative result. Among those patients with diagnosis of solid tumor, S, E, PPV and NPV were of 96, 100, 100 and 95%. We found that none patient with neutropenia, fever with negative blood cultures and without focal infection, with a CRP level lower than 30 mg/L had not bacterial infection. All patients with CRP levels higher than 100 mg/L were infected. Conclusion: CRP is a useful, fast and economic test to identify bacterial infection in the neutropenic patient with cancer and fever. Methods: The patient was observed by traditional clinical and paraclinical methods of investigation. Results: Clinical signs: 1) Recurrent furunculosis and episodes of skin cold staphylococcal abscesses. An onset of streptostaphylodermia is 2 month age. It was the first clinical manifestation of disease. A frequency of exacerbation is 2Y4 times per year. 2) Recurrent bronchitis since 5 month. 3) Recurrent pneumonia complicated pleuritis and pneumatocele since 4 year old. The child had 1Y3 pneumonias per year. The child was operated at the age 4 yearlobectomy of upper right lung lobe; at the age 8 year -lobectomy of upper left lung lobe; and at the age 9 year -toracotomy and sewing up of the bronchopleura fistula of left lung. 4) Recurrent abscesses in abdominal cavity since 8 year. The child was operated many times -at 8 year due to acute appendicitis complicated peritonitis; at 10 year -a huge abscesses of big omentum; at 11 year -an abscess in a region of afteroperation scar; in 3 month -an abscess in left underdiaphragm region and an abscess of upper pole of spleen. 5) Presence atopic dermatitis since 3 month. The eczematous skin eruption does not have a seasonal variation and is present, to some degree, at all times. 6) Presence skeleton anomalies: scoliosis, generalized hyperextensibility of joints, facial abnormalities, retention of 4 primary teeth with unresorbed roots, absent 2 lower premolaries, 1 upper premolaries, 2 lower and 2 upper molaris. Lab Findings: high eosinophilia (7 Y 24 %), lymphocytosis (till 42 %), thanneutrophilia (till 86 %), shift to the left (band neutrophils 6 Y 11 %), eleveted ESR (25 Y 43 mm/hour), high level IgE (60498 IU ml). Imaging Studies: dental abnormalities -delay of primary teeth shedding owing to lack of root resorption, absent of some permanent teeth (X-ray); thin-walled cysts in both lungs (X-ray); cysts in the liver and in the lungs (Ultrasonic, Computer tomography). The patient was treated by prolonged course of antibacterial therapy, fluconazole for mucocutaneous candidiasis, onychomycosis and topical steroids for eczematous dermatitis. Conclusion: This clinical case gives us an experience in observation and treatment of children suffering from a rare primary immunodeficiency disorder. The evaluation of clinical and paraclinical in twenty CVID patients and detection Of IL-2 as a significant cellular function in these by invitro way Marzieh Heidarzadeh Arani 1 , Abolhassan Farhoudi 2 , Asghar Aghamohammadi 2 , and Gholamreza Ghasemiyeh 3 . 1 Shahid Behashti, Pediatric, Kashan, Islamic Republic of Iran; 2 Children Medical Center, Immunology & Allergy, Tehran, Islamic Republic of Iran; 3 Shahid Behashti, Farmacology, Kashan, Islamic Republic of Iran. Subjective: Common variable immunodeficiency is the primary immunodeficiency disease. CVID is a heterogeneous group of immunologic disorder of un known etiology, characterized by impaired antibody responses, associated with number and function cells defect including lymphopenia,anergy,impaired lymphocyte proliferation and deficit cytokine secretion that the most important of them is IL-2.most prevalent of the Objective: The goal of this study was evaluation of the clinical and paraclinical characterized 20 CVID patients,interlukine 2 production and compaired these to healthy control group. Methods and Materials: We done a randomized cross sectional study in 20 CVID patients from department of immunology and allergy in children medical center and 10 healthy control. The evaluation of the patients was done by questioner and peripheral blood mononuclear cells of the twp groups were cultured with PHA and supernatant were collected for quantitation of IL-2 by ELISA. Results: Mean of age the patients was 15.9+/j11.23 and distribution of gender was 9 female and 11 male. There were high frequency hospitalization in 80% of cases (over the two time).The most frequent of infection was respiratory tract infection. There were opportunistic infection in 5Y10% of them, autoimmunity in 40% of cases,un response DTH in 65% of cases and IL-2 production was zero in15 patients and the 5 patients had level of IL-2 lower than corresponding levels in the healthy control (P valueG0.005). Conclusion: In our the study, all of the patients showed that T cells of them exhibit deficient production of IL-2 and manifestations of this deficiency were high frequency of autoimmunity,granolumatose disease, bronchectasia and recurrent infection in spite of given monthly IVIg in patients. A 3 years old female with SCID T(j)B(j)Nk(+) treated with halogenic bone marrow transplantation. Case report Horacio Del Olmo Tellez, and Francisco Espinosa Rosales. National Institute of Pediatrics in Mexico, Allergy and Immunology, Mexico City, Mexico. A 3 years old female is the second of two children born to nonconsanguineous mexican parents after an uncomplicated, term pregnancy. Birth weight was 3900 g, length 53 cm. She had breast feed until 2 months of age. She didn't receive immunizations. She has history of a 9 years old brother who suffered totipotent cell transplantation because severe immunodeficiency. He cursed a neurologic infection and died. At age 45 days she had abundant, liquids, and blood stools. She received ceftriaxone treatment with partial improvement. After that a stool cultive test was positive for campylobacter yeyuni so she was treated with claritromicyn for 14 days. She also had oral and esophageal candidiasis treated with miconazol. Because the brother with immunodeficiency, when she arrived to our hospital the following studies were done: cd3 0.2%, cd 20/19 0%, cd4 0.1%, cd8 0.1% cd 53 83%, igm 4 mg/dl (20Y40) igg 415 mg/dl (310Y852), iga 6.6 mg/dl (3.5Y67). A severe combined immunodeficiency diagnosis was made with t (j) b (j) nk (+) features. We supposed deficiency in enzimes of receptor's recombination. We restituted totipotent cells with halogenic bone marrow transplantation. The hla compatibility was: patient: a68 (28) y a2, b39 (16) y b61 (40), dr4 y dr 11(5), dq8 (3), dq7 (3) patient's father: a68 (28) y a2, b61 (40) homozigous, dr4 y dr 11(5), dq8 (3) y dq7 (3). High resolution study for dr patient: drb1 0407 patient's father: drb1 0407. We decided to admistered only partial hidrolized formula because the posible risk to adquired infections by breast feeding. She was treated with immunosupresor as acondicionated treatment with busulfan 4 mgkgday 4 days, day j8 to j6, cyclophosphamide 60 mgkgday day j5 to j4. In september 19 2003, our patient received hallogenic bone marrow trasplantation with prior medication with h1 blockers and steroids. We adminitered 120 ml of aferesis products with a total count of 189,000 Â 10 3. 98.5% mononuclears. The patient presented host againts disease with papules, petechias in abdomen. She was treated with metotrexate, ciclosphorine and methilprednisolone. She had citomegalovirus blood culture test positive and was treated succesfully with ganciclovir. Nowadays she has normal blood citology, immunoglobulin levels, lymphocyte subpopulations and she has completed her immunization schedule two and a half years after the stem cell transfusion. 2 Berufsgenossenschaftliches Unfallkrankenhaus Boberg, Department of Dermatology, Hamberg, Germany; 3 University of Erlangen/ Nürnberg, Med. Informatics, Biometry and Epidemiology, Erlangen, Germany. Background: Staphylococcus aureus (S. aureus) is a well known trigger factor of atopic dermatitis (AD). Besides staphylococcal superantigens alphatoxin ("-t) which is produced by one third of skin-colonizing S. aureus strains in AD may influence the cutaneous inflammation. "-t can be detected both in the upper epidermis and in the dermis of AD patients. Sublytic concentrations of "-t have been shown to induce T-cell proliferation and secretion of T-cell cytokines. Objective: To explore the association of sensitization to inhalant allergens and "-t producing skin colonizing S. aureus in AD. Methods: We investigated 127 adult patients with AD according to their skin colonization after treatment with antiinflammatory and antiseptic substances, medical history, severity of AD and sensitization to different allergens. Results: 48 out of 127 patients where colonized with S. aureus. S. aureus colonized patients suffered from a more severe AD (SCORAD: S. aureus positive patients: 46, S. aureus negative patients: 33) and showed a higher sensitization level to different inhalant allergens. Surprisingly, they also suffered more frequently from allergic asthma (S. aureus positive patients: 69%, S. aureus negative patients: 44%). 30 of the 48 S. aureus skincolonizing strains produced "-t. The severity of AD was similar in patients being colonized with "-t negative S. aureus ("-t-pts) compared to patients with "-t producers ("-t+ pts). "-t+ pts had a significally higher specific IgE to birch pollen (median: "-t+ pts: 100kU/l, "-tpts: 32kU/l) and a trend of higher total IgE values (median: "-t+ pts: 6849kU/l, "-tpts: 2215kU/l) and specific IgE to timothy grass pollen (median: "-t+ pts: 80kU/l, "-tpts: 12kU/l). Conclusion: Following topical treatment the colonization rate was lower (38%) than expected. But there was a high rate of patients who were skin colonized with "-t producing S. aureus. Colonization with S. aureus was associated with a higher severity of atopic dermatitis, higher sensitization, and a higher frequency of allergic asthma. Cutaneous colonization with "-t producing S. aureus was associated with a higher sensitization to saisonal inhalant allergens in AD. If this is a consequence of direct chronic cutaneous T-cell stimulation by sublytic doses of "-toxin penetrating constantly into the skin needs further investigation. Trichophyton allergy: review of 98 cases A.G. Palma-Carlos, and M.L. Palma-Carlos. Clinical Allergy Immunology Center, CAIC, Allergology, Lisbon, Portugal. Background: The association of mycosis or onicomycosis by dermatophyte fungi and allergic diseases including urticaria/angioedema, rhinitis and asthma has been described for over 70 years but large series of confirmed cases have not been published. Methods: From 9087 patients seen in the last few years 98 (1,07%) 20 to 66, 61 males (62,2%), 37 females (37,8%) had a clinical history and previous or actual visible skin or nails infection and positive tests to Tricophyton. Skin tests have been done in all the patients by prick in a first step and by intradermal with 1/1000 and 1/100 dilution in a second step. Specific IgE has been looked by UNICAP for Tricophyton rubrum in 78 cases. Treatment with antifungal itraconazol has been tried as a first option in all the patients. When allergic symptoms subsisted specific immunotherapy by parenteric route has been started with a slow release vaccine during a minimal period of 3 years. Results: Fungal infection presentation was tinea pedis in 66 patients (67,3%), onicomycosis in 36 (36,7%), tinea cruris in 27 (27,6%) and remaining pruritus after clear infection in 3 (3,0%) . Dual infections were present in 33 patients (34,4%). Total of clinical mycosis 129. Allergic disease: urticaria/ angioedema in 64 patients (65,3%), rhinitis in 14 (14,3%), asthma plus rhinitis in 11 (11,2%), eczema in 8 (8,2%) and conjunctivitis in 1 (1,0%). Fifteen patients (15,3%) all rhinitic had also positive skin tests to house dust mites. Prick tests for Tricophyton were positive in 61 patients (62.2%). Intradermal tests were always positive. Specific IgE was positive in 52 (66,6%) cases with a range of 0,50 to more than 100 Ku/l. Treatment with antifungal cleared allergic disease only in 20 (20,4%). Specific immunotherapy has been tried in 60 patients with very good results (complete clearing) in 23 (38,3%), good in 36 (43,3%) and moderate or nil in 11 (9,4%) . Conclusion: Tricophyton IgE mediated allergy play a role in the pathogenesis of Urticaria/angioedena, asthma and rhinitis in most patients. Fungal skin and nails infection must be searched, skin tests and specific IgE used for diagnosis. Sensitivity and specificity of skin prick tests and specific IgE are roughly comparable. Specific immunotherapy must be tried if anti fungal therapy failed with good results in most cases. Profile of inflammatory cell in erythema nodosum leprosum patients, treated with minocycline-prednisone combination drugs and prednisone Background: A clinical trial study was conducted with B pre-post treatmentd esign, to know effectiveness minocycline-prednison combination drugs and prednisone only in the erythema nodosum leprosum (ENL) patients by profile of cells inflammation. Methods: Sixty ENL patients released from treatment (RFT) were included in this study. They were devided into 2 groups, 30 patients were treated with minocycline-prednison combination drugs (group A) and the other 30 patients were treated with prednisone-placebo (group B). To evaluate result this study, we performed histopathology examination by skin biopsy, haematology test by peripheral blood sample that obtained from cubital venous puncture, and the changes of clinical feature. Results: The results, that the ENL patients group A were shown more improve histopathologically (pG0.05) haematology finding not showed difference between 2 groups (p90.05). There was more improve change clinical feature in the patients get combination therapy. Conclusion: Combination therapy, prednisone-minocycline, better than just prednisone only in ENL patients, so that it may be alternative drug to reduce steroid dependency in the ENL patients. Multiple sclerosis is one of demyelinating diseases based pathophysiologically on changes in vasculo-myelin system. Immunological basis of these diseases is failure of immunological tolerance for brain antigens with development of autoimmune cellular reactions. Important role in this failure and development of demyelinating process is attributed to bacteria possessing antigenic determinants common with myelin basic protein (MBP) molecule. Therefore we performed in multiple sclerosis patients study of IgG and IgM antibodies against Clebsiella pneumonia, E. coli, St. aureus, H. influenzae, and pertussis anatoxin. The study was performed with serum from 78 patients with different types of multiple sclerosis. Titer of antibodies against Clebsiella pneumonia, E. coli, St. aureus, H. influenzae, and pertussis anatoxin was detected by enzyme immunoassay. Results were expressed as 1Y5 classes of reaction. The study showed that in recurrent type of multiple sclerosis exacerbation is accompanied by increase of IgM antibodies level (to 4Y5 class) for all abovementioned bacteria. Remission is marked by lowering of IgM level compared to exacerbation one, to 2Y3 class, and by increase of IgG level to 3Y4 class. In progressive type of multiple sclerosis level of IgG as well as IgM antibodies increased progressively (to 3Y5 class). If course of multiple sclerosis lasted more than 5 years, levels of IgG and IgM antibodies were the highest (mostly 4Y5 class). Based on data obtained, we concluded, that degree of manifestation of antibacterial immunity reflects the severity of multiple sclerosis and can be considered as negative prognostic factor. Local cytokine status and clinical morphological parameters of chronic viral c hepatitis The current research is aimed at investigating the levels of local cytokines in liver histology examination samples of the CCH patients taking into account clinical morphological parameters as well as biochemistry and virology investigations. Cytokine investigation in homogenizers of liver histology examination samples was performed in the period of CCH clinical manifestation accompanied with cytolysis. As a tool of control the samples of the hepatic tissue belonging to 5 donors who did not have any chronic diseases and markers of being infected with viruses of parenteral hepatitis were used to investigate cytokine levels in homogenizers of liver histology examination samples. In the process of investigation of liver histology examination sample homogenizers in CCH patients it was established that the they have the authentic increase of IL-1!, IL-4, TNF-! proportion and decrease of IL-2, IFN-+ proportion. The considerable fluctuation in proportion levels of each of them from the minimal to the maximal indices was noted and the total distribution differed from the normal one, which hampered the adequate assessment of the investigated indices. The patients were divided into two groups that differed from each other by the median of the low (V0,25) and upper (V0,75) percentiles of the quantitative contents of the local cytokin levels. The first group whose levels of practically all investigated cytokines in liver histology examination samples were authentically different from those of the second group. IL-4 proportion in the second group nearly twice exceeded its index in the first group. The increase of TNF-! level in liver histology examination samples of CCH patients of the second group six times as high as that of the first group was noted. The considerable decrease of the local proportion of IL-2 and IFN-ã in the second group (0,6 T 0,02 pg/ml versus 16,0 T 0,6 pg/ml, p G 0,001 and 4,2 T 0,65 pg/ml versus 18,9 T 2,7 pg/ml, p G 0,0001 correspondingly) was noted. No difference in IL-1á levels in investigated groups was noted (p 9 0,05). In accordance with the local cytokine levels along the median of the extreme percentiles (V0,25 and V0,75) two CVH patient groups were singled out, the groups having the authentic difference in clinical, biochemical and morphological parameters. HFRS is one of the clinical forms of Hantavirus infection that is widely spread on Eurasian continent. In Europe HFRS is predominantly associated with Puumala, Dobrava -Belgrad Hantaviruses (HV), in Asiawith Hantaan, Amur, Seoul. Another clinical form, which is common for the American continent is Hantavirus Pulmonary Syndrome (HPS), associated with Sin -Nombre, Andes serotypes. On the south of Far East region, where three serotypes of HV are collected, the similarity of the main pathogenical and clinical aspects of HFRS and HPS were revealed. It is established that in the pathogenesis the important role plays either HV, as the initiator of the process, or the reactions of cellular immunity. The more virulent HV (Sin -Nombre, Andes, Hantaan, Amur) and lass virulent (Puumala, Seoul) are known. The studying of immunopathogenesis in HFRS patients revealed the dependence of characteristics of immune response from the serotype of HV. On the onset of the disease the level of misbalance of serum cytokines (high levels of IFN -!, IL -1!, IL12p70, IL12p40, IL-8, IL-10 and low levels of IFN-+) and also the number of cytotoxic lymphocytes CD8 correlated with the severity of clinical symptoms and the serotype of HV. In HFRS, associated with Hantaan, Amur serotypes severe and complicated manifestations with hemorrhagic syndrome, hyperhydration of kidneys, respiratory tract, liver tissue and multiorganic insufficiency. Severe forms of Seoul -infection were less common and the characteristics of immune and cytokine status were not changed a lot. The studying of the levels of cytokines and metabolites of nitric oxide in urine and exhaled air condensates showed the absence of correlation of their production locally and systemically. That fact reveals the independent synthesis of the immune response mediators and allows supposing the respiratory tract, as the kidneys to be the target -organ of HV. Viremia begins after the respiratory infection of alveolar macrophages and then the central cells of pathogenesis -endothelial cells of the lungs, kidneys and other organs. So, the results of the study of cytokines in different biologic substrates (serum, urine, exhaled air condensates) in the onset of the disease showed the prevalence of immune reactions in the pathogenesis of HFRS without any dependence with the Hantavirus serotype. The comparison of allergenicity between L3ESP and L4ESP and the cytokine production profile in experimental infection of rats with Anisakis simplex Ju Hyeon Lee 1 , Haneul Nari Lee 1 , Sungae Cho 2 , Han-Gyum Kim 3 , Kyung-Whan Joo 4 , Joon-Sang Lee 4 , Guan Gyu Song 5 , and Sung-Weon Cho 4 Background: Anisakis simplex is the marine parasite which belongs to ascaroidea. This worm normally live within marine mammals, however frequently induces allergy in the infection of mammals on land by third stage larvae (L3) carried by marine fishes. When L3 infect mammals, these worms develop into fourth stage larvae (L4). Each state larva produces different excretory-secretory products (ESP) in main protein constituents. This investigation compared ESP from L3 (L3ESP) with L4ESP in antigenicity and allergenicity to develop on efficient method to diagnose the allergy by L3, and analyzed cytokine profile in reinfection of L3, which is critical period in allergy development. Methods: The kinetics of specific antibody production of sera harvested from rats infected by L3 was analyzed by indirect ELISA using either L3ESP or L4ESP as antigen immobilized on ELISA plate. The cytokine kinetics of IL-4 and IFN-r was analyzed with rat sera harvested for 5weeks from L3 reinfection. Results: The kinetics of antibody production showed that specific antibody level against L4ESP was higher in antigenicity than the level against L3ESP in IgG1 IgG2b IgG2c and IgM, and very similar in IgG2a. However, the latter had evidently higher allergenicity than the former in specific IgE level. In L3 reinfection sera, the level of IL-4 was relatively constantly maintained. In contrast, that of IFN-Y j +; was decreased continuously. Conclusion: These results indicated that L3ESP be better than L4ESP for allergic state analysis and Th2 cytokine be comparatively dominant in Anisakis simplex larva reinfection. Miki Fukuda 1 , Kengo Kobayashi 1 , Yuriko Hirono 1 , Hisae Ishikawa 1 , Emenike C Ejiogu 2 , Masaharu Sawai 3 , and Minoru Takeuchi 1 . 1 Kyoto Sangyo University, Biotechnology, Kyoto, Japan; 2 Origins Japan Co. Ltd, CEO, kyoto, Japan; 3 TAKARA SHUZO Co. Ltd, Development, Kyoto, Japan. Introduction: Jungle honey (JH) is collected from timber and blossom by wild honey bees that live in the tropical forest of Nigeria. This is used on traditional medicine for cold, skin inflammation and burn wound as well as health care. However, the effect of Jungle honey on immunomodulatory activity is not yet clearly. We have investigated the effect of Jungle honey on immune system and anti-tumor activity in mice. Methods: JH was provided by Nihon origins. JH was dissolved with distilled water, and freezed dry, and then adjusted with PBS at each concentration. JH was fractionized to Fraction (Fr.) 1, 2, 3, 4 and 5 by HPLC size chromatography. Female C57BL/6 mice were injected with JH at dose of 1mg/mouse/day, 7 times intra-peritoneal. After 7 times injections, peritoneal cells (PC) were obtained by peritoneal lavage with PBS. Total numbers of PC were counted with homocytometor. Cell differentials were observed by microscope after Giemsa stain. Expressions of surface antigens (CD3, CD11b, CD19, NK, and Gr-1) on PC were analyzed by FACS. H 2 O 2 production of PC was assayed by FACS using DCFH-DA. Chemotactic assay was analyzed by TAXIScanTM using neutrophils from peripheral blood of human or guinea pig. IL-1$ mRNA expression of PC were analyzed by RT-PCR. Anti-tumor activity was assayed by using Lewis Lung Carcinoma/2. Results: Total numbers of PC were increased in JH-injected mice compared with control mice. In Dot Plot analysis by FACS, neutrophils were increased in JH-injected mice. Percent of Gr-1 surface antigen positive cells and intensity of Gr-1 antigen expression were also increased and H 2 O 2 production of PC was increased by JH. Chemotactic activity for Neutrophil from human or guinea pig was expressed at concentration of 1, 5mg/ml or 1, 5, 10mg/ml of JH. IL-1$ mRNA expression of PC was increased by JH or Fr.2. Molecular weight of Fr.2 was approximately 261. Inhibitor of tumor growth was showed by JH. Conclusion: Chemotactic activity for neutrophils, increase of neutrophil counts and H 2 O 2 production of PC by JH may prevent bacterial and viral infections. A case report of 11-month-old boy infant with Toxocara canis eosinophilic meningitis Nuntiya Pitaksit, Pantipa Chatchatee, and Jarungchit Ngamphaiboon. King Chulalongkorn Memorial Hospital, Pediatrics, Bangkok, Thailand. An 11-month-old boy infant was referred due to prolonged fever for 4 weeks, drowsiness and bulging at the anterior fontanelle. The clinical was no response with high dose cefotaxime for 2 weeks that treated as bacterial meningitis. Neurological examination revealed meningeal irritation and bulging at the anterior fontanelle. Marked increase of absolute eosinophils count (1,893/mm 3 ) was presented. A lumbar puncture showed 270 leukocytes/microliters with 60% of eosinophils; protein 50 mg/dL; CSF culture was no growth and negative PCR for TB. The immunoblotting assay was positive for Toxocara canis in both serum and in CSF. The CT scan of brain showed communicating hydrocephalus. The patient was close contact with cats with poor sanitation. A diagnosis of eosinophilic meningitis due to Toxocara canis was made and started treatment with albendazole 200mg oral twice a day for 4 weeks, prednisolone 2 mg/kg/day and tape off within 8 weeks. The clinical was improved and discharged at 5 days after start both medications. Follow up monthly for clinical symptom and monitored for absolute eosinophil count. No abnormal neurological symptoms were detected. The parents had got rid of the cats out of the household area. Serum absolute eosinophil count and lumbar puncture results turn to normal within 11 months after treatment. The serological follow up 8 weeks after treatment for Toxocara canis was positive in both serum and in CSF by the use of immunoblotting assay but decrease in intensity. He had normal growth and development. Then long term follow up was done and aware for reinfection in this patient. Reinvestigation for immunoblotting assay and follow titer for Toxocara canis will be done. Activity of allium ascalonicum (shallot) and myrtus communis extract as two novel antibacterial agents against acne vulgaris Ahmad Farajzadeh Sheikh, Mohammad Ali Mashhdizade, and Mohammad Radmanesh. School of Medicine, Jondishapor University of Medical Sciences, Ahwaz, Iran, Microbiology, Ahwaz, Islamic Republic of Iran. Background: Acne vulgaris is a common skin disease. Its etiology and pathogenesis is not well known, and treatment is not satisfactory, therefore study for novel drugs to replace to common drugs will be necessary. Methods: From 136 patients who suffered from acne lesions, swabs were obtained. Each swab was cultured on 2 blood agar plates and stored aerobically and anaerobically condition. Sensitivity test from 2 herbal extracts against bacterial isolations done by disk diffusion method, and were compared with tetracycline disks. Results: From 176 bacterial strains isolations, 113 (64.2%) Coagulase Negative Staphylococci (CONS), 52 strains (29.5%) Propionibacteria spp., 7 strains(4%) Diphtheroids, 2 strains (1.13%) E. coli and 2 strains (1.13%) Streptococci spp. were isolated. 53% of CONS and 10% of Propionibacterium isolates were resistant to tetracycline, whereas only 8% of CONS and14% of them was resistance to Shallot and Myrtus communis water extract respectively. No Propionibacterium strains and only 4% of Propionibacterium were resistant to shallot and Myrtus communis water extract respectively. Also 11% of CONS Isolates were resistant to shallot alcohol extract and 12% of them were resistant to the Myrtus communis alcohol extract. Two percent of Propionibacterium strains were resistant to shallot alcohol extract and 3% of them were resistant to the Mmyrtus communis alcohol extract. Conclusion: Our finding revealed that bacterial agents of acne vulgaris are highly resistant to tetracycline and this antibiotic is not efficient for treatment of the disease. It is suggest that these herbal extracts may have potential for acne treatment. Galactomannan antigen detection in the diagnosis of invasive pulmonary aspergillus Objective: The invasive aspergillus occurs almost exclusively in immunocompromised host. Aspergillus fumigatus is now the leading cause of infectious mortality in many hematology bone marrow and allograft transplantation. Galactomannan is hetropolysaccharid present in the cell wall of aspergillus species. A rapid and important method for invasive aspergillus diagnostic could be the circulating galactomannan aspergillus (GM) measurement in the serum and BAL (Bronchoalveolar lavage) samples of the patients. The aim of this study was determined the value of galactomannan detection in serum to diagnosis of invasive aspergillus. Methods and Patients: Fifty three patients (with recurrent pulmonary infection suspected to aspergillus) were included in our study group. None of the selected patients had received any anti fungal therapy prior to the study. Direct microscopy and fungal culture of BAL (Bronchoalveolar lavage) was done and galactomannan estimation in serum and BAL were measured. Results: 32 patients (960 %) were considered galactomannan test positive from a total 53 cases (30 male and 23 female) and 28 of these patients were isolated aspergillus in microscopy and culture. Conclusion: The GM EIA assay had greater sensitivity than culture and microcopy in detection of aspergillus spp in BAL fluid in experimentally induced invasive pulmonary aspergillus. GM positivity also allowed the anticipation of invasive aspergillus diagnosis (from 3 to30 day before mycological culture). Revisiting the hygiene hypothesis: Cross-reactivity studies between house dust mites and Ascaris lumbricoides Background: The causal relationship between allergy and ascariasis remains a matter of controversy. Despite growing evidence for protective immunity exerted by helminth infections against allergic diseases, the molecular and immunological mechanisms involved needs to be elucidated. In this study, cross-reactivity of allergens from house dust mites (HDM) Blomia tropicalis (Blo t), Dermatophagoides pteronyssinus (Der p), and D. farinae (Der f) with antigens from Ascaris lumbricoides (Asc l) was determined. Methods: Enzyme-linked Immunosorbent Assay was performed to determine the allergenicity of HDM extracts and antigenicity of Asc l extracts to allergic (n=100), ascariasis (n=60), and healthy control (n=100) patients. Inhibition assays and Western blot analyses were done using positive sera from allergic (n=15) and ascariasis (n=15). Specific IgE levels of allergic (n=50) and ascariasis patients (n=50) to a recombinant paramyosin peptide (Blo t 11-fD) was determined. Results: The allergenicity of HDM extracts was significantly higher among allergic patients (Blo t=81, Der p=72, and Der p=80% positive reactions) than ascariasis patients (20, 20, and 28.3%, respectively) while the antigenicity of Asc l extracts among allergic patients (70%) was significantly lower than ascariasis patients (87%). Cross-inhibition assay showed that Asc l antigens can inhibit up to 92% of the IgE reactivity of allergic patients to HDM allergens while up to 54% of the IgE reactivity of ascariasis patients to Asc l antigens was inhibited by HDM allergens. Western blot analysis showed multiple sensitizations of allergic patients to HDM allergens with molecular weights ranging from 14Y240 kDa and of ascariasis patients to Asc l antigens ranging from 15Y250 kDa. Positive reactions to rBlo t 11-fD was observed among allergic (80%) and ascariasis (46%) patients. Conclusion: Multiple cross-reactive antigens are present in HDM and Al extracts and may play a role in the complex immunological relationship between allergy and ascariasis. These antigens might share homology or similarity in epitope recognition sites and must be identified in future studies. The potential role of paramyosin as a specific cross-reactive allergen present in HDMs and A. lumbricoides was described. These findings provide support for the Hygiene Hypothesis and may serve as basis for novel forms of treatment and diagnosis of allergic diseases and helminthic infections. . This model allows us to define basic principles of pathogenesis of infection on cellular level. We are able to define precausions of effective macrophage activation leading to subsequent infection elimination. Methods: We followed up the expression of specific macrophages surface markers: CD 86, CD 54 (ICAM-1), CD 16/32 (FcgIII/IIR) and CD 25, including specific isotypic controls. The phenotypic changes expressed as mean fluorescence intensity (MFI) or percentage of positivity, respectively, were evaluated by flow cytometry. Morphologic changes were documented by immunofluorescence microscopy. Murine macrophage-like cells (J774.2) were incubated in cultivation flasks (2x106 cells/10ml of medium Dulbecco's MEM with Glutamax-1 with 10% BSA). The cells were activated with 5, 10,50 ng of LPS per 1ml of medium or with 10, 100, 1 000, 10 000 I.U. of IFN, per 1ml of medium respectively and infected by F. tularensis LVS with multiplication of infection 1:100 in particular time schemes. Results: Infection of F. tularensis does not result in any activation of host cells, as seen from stable phenotype profile of infected cells in time. In contrary, stimulation of J774.2 cells by IFN, or LPS results in predictable, time and dose dependent phenotypic changes. The phenotype profile of IFN, activated cells is characteristic. These cells display a harmonic increase in CD86 and CD16/32 surface expression in time. In contrary, activation by LPS results in isolated CD16/32 expression elevation without increase in CD86 expression. Moreover, prior F. tularensis infection of macrophages prevents subsequent activation, especially by LPS. We demonstrate an essential role of IFN, in infection control. Minimal concentration of IFN, resulting in active infection elimination is 1000 IU/ml. Conclusion: We assume, that F. tularensis infection is able to interfere with intracellular signaling triggered especially by LPS, less significantly by IFN,. This interference leads to immunosupression and seems to be the efficient escape mechanism of F. tularensis. Effects of cigarette smoke exposure on immune functions in alveolar macrophage reported the inhibition of antibody production by alveolar macrophages (AM) from cigarette smoke (CS) exposed mice. However, the mechanism of immune suppression by CS on AM functions is not clearly understood. Therefore, we investigated effects of CS on phagocytosis and antibody production, expression of surface antigens, IL-1$mRNA in AM associated with immune functions. Methods: Female C57BL/6 mice were exposed to 20 cigarettes /day during 10 days. After 10 days, AM were obtained by bronchoalveolar lavage (BAL). Phagocytosis activity was analyzed by FACS using FITC labeled seep red blood cell (SRBC). Expression of surface antigens (Class II, B7.1, CD11b, CD16/32, CD14, TLR-2) on AM were analyzed by FACS. IL-1$mRNA expression of AM was analyzed by RT-PCR. Antibody production was analyzed by plaque forming cell (PFC) assay using SRBC antigen. Results: Phagocytosis of AM was significantly decreased in smoked mice (SM) compared with non-smoked mice (NSM). Surface antigens positive cells in AM were decreased in SM compared with NSM. IL-1$mRNA expression of LPS non-stimulated AM was increased, while in case of LPS stimulated AM was decreased in SM compared with NSM. Antibody production was significantly decreased by AM at induction phase, but not expression phase in SM compared with NSM. Conclusion: These results suggest that the inhibition of antibody production is caused by the inhibition of phagocytosis and expression of surface antigens in AM. Such inhibition of AM functions may be increased the risk of bacterial and virus infections. Katsuhiko Matsui, and Akemi Nishikawa. Meiji Pharmaceutical University, Department of Immunobiology, Tokyo, Japan. Background: Atopic dermatitis (AD) is a chronic inflammatory skin disease with immunopathologic features that vary depending on the duration of the lesion. The lesional skin of AD patients shows an increased number of eosinophils in the dermis and superficial Staphylococcus aureus colonization. Our previous study showed that percutaneous application of peptidoglycan (PEG) from S. aureus induced eosinophil infiltration in murine skin through RANTES production by epidermal Langerhans cells (LCs). Although it is well known that PEG is an agonist of Toll-like receptor (TLR)-2, it is unclear whether other TLR agonists are able to induce RANTES production by LCs. In this study, therefore, RANTES production by murine LCs in response to different TLR stimuli, and the signaling pathways involved, were investigated. Methods: PEG was applied to barrier-disrupted abdominal skin of mice every 5 days. Twenty days later, eosinophils in the abdominal skin were detected. LCs were purified from epidermal cells by the panning method using anti-IA d monoclonal antibody. RANTES production in the skin and by the cultured LCs was investigated by RT-PCR, immunohistologic analysis and ELISA. Analysis of the signaling pathways responsible for RANTES production by LCs was performed by ELISA using N-acetyl-L-cysteine, SP600125, PD98059 and SB203580, which are specific inhibitors of NF-e?B activation, JNK, ERK and p38 MAPK, respectively, and was finally confirmed by Western blot analysis. Results: The results of RT-PCR and ELISA showed that PEG, poly(I:C), LPS and CpG DNA, whose signals are transduced through TLR-2, TLR-3, TLR-4 and TLR-9, respectively, strongly induced the production of RANTES. Although treatment with an inhibitor of NF-e?B activation inhibited PEG-, poly(I:C)-, LPS-and CpG DNA-induced RANTES production, treatment with a JNK inhibitor did not inhibit CpG DNA-induced RANTES production. Furthermore, treatment with a p38 MAPK inhibitor affected only PEG-and LPS-induced RANTES production, and the inhibition of RANTES production was correlated with that of p38 MAPK phosphorylation. Conclusion: These results suggest that the signaling pathways involved in RANTES production by murine epidermal LCs in response to different TLR stimuli are not necessarily the same, and that inhibition of p38 MAPK may be a more specific therapeutic strategy for eosinophilic inflammation in AD patients with S. aureus colonization. Mansour Amin 1 , and Balo P. Kapadnis 2 . 1 *Dept. of Microbiology, School of Medicine, Ahwaz Jondishapor University of Medical Sciences, Ahwaz, Iran, Microbiology, Ahwaz, Islamic Republic of Iran; 2 Pune university, Microbiology, Pune, India. Background: Studies in past decade confirm that the growth of both gram positive and gram negative bacteria, yeasts and mold can be inhibited by garlic, onion, cinnamon, cloves, thyme, sage, and other spices. The Latin name for shallot is Allium ascalonicum. Shallots belong to the lily family. The name Allium is known to be derived from the Celtic word Allium (pungent), whereas the name ascalonicum could either be derived from its original site of cultivation, Ascalon, an old Palestinian town, or from the French name Echalogne. Methods: In this study the extract of shallot was extracted by organic solvents and purified with help of column chromatography and TLC. The pure compound was named shallomin. Antimicrobial activity of this compound was tested against 23 strains including pathogen and non pathogen microbes using modified E test and broth dilution technique. : All fungi and bacteria tested were sensitive to shallomin. The MIC values of shallomin for microorganisms tested ranged from 2Y10 6g/ml. fungi were more sensitive than bacteria to this novel antimicrobial compound. MLC values were slightly greater than MIC indicate cidal nature of antimicrobial at low concentration. Conclusion: Shallot is commonly used as a folklore medicine, and used to cure earache, fever, antidote for snake venom and also as an aphrodisiac. In this study antimicrobial properties of shallot were investigated for discovery of a new antibiotic. Based on this shallomin can be an effective medicine for treatment of dermatomycosis and other infectious diseases. Hydatid disease of the lung Nemeth Agnes 1 , Banfi Andrea 2 , Peterffy Erzsebet 2 , Baktai György 2 , Kosa Lajos 2 , Goschler Adam 1 , Kurti Sandor 3 , and Fekete György 1 . 1 Semmelweis University, Faculty of Medicine, 2nd Department of Pediatrics, Budapest, Hungary; 2 Pediatric Institute of "Svabhegy", Bronchological Department, Budapest, Hungary; 3 Hetényi Géza County Hospital, Pediatric Department, Szolnok, Hungary. : Human hydatid disease is caused by metacestode of Echinococcus granulosis. Hydatid disease of the lung appears more frequently in childhood. The infection may result in asymptomatic to severe disease which may be fatal. The conventional treatment of hydatid disease is the surgical intervention. Several reports proved that the patients who have widespread disease and no surgical resection is possible, the medical treatment remains the only oportunity. Percutaneous aspiration and treatment of hydatid cysts was believed to be contraindicated due to anaphylaxis and spillage of scolices, however several investigators have reported neither anaphylaxis nor dissemination. The authors present a successful treatment in pulmonary hydatid disease with bronchoscopic investigations combined with mabendazole/albendazole therapy. A nine year old boy was admitted to a county hospital because of intense black vomit without any severe disease in his previous medical history. He lost 8 kgs within a month and once he had bloody split earlier. Chest X-ray showed decreased left side basal transparency. CT scan suggested a tumor and he was referred to the 2nd Department of Pediatrics of Semmelweis University. Laboratory data indicated the possibility of lung infection and we started antibiotic therapy. The patient improved after few days and bloody vomit was stopped. Repeated CT scan did not prove any improvement and bronchoscopy was required. Cysts were found in the samples removed in the course of bronchoscopy. Histological and serological investigations confirmed the diagnosis of pulmonary hydatid disease. Repeated bronchoscopy, lavage and 50 mg/ bwkg/day mabendazole then 10 mg/bwkg/day albendazole therapy were continued. After 10 months of combined therapy the patient is in good medical condition (he gained weight, he regulary performs physical activities). Control bronchoscopy and CT scan showed mild bronchiectasis on left side in the S6 segment. After one year from the beginning of the hydatid disease the patient recovered nearly completely. Conclusion: The authors demonstrated the safety and efficacy of combined medical and bronchoscopic treatment in pulmonary hydatid disease. Prevalence and clinical characteristics of recurrent wheezing infants in the south of Brazil Herberto Jose Chong Neto 1 , Nelson Rosario 1 , Dirceu Solé 2 , and Javier Mallol 3 . 1 University of Parana, Pediatrics, Curitiba, Brazil; 2 University of Sao Paulo, Pediatrics, Sao Paulo, Brazil; 3 University of Santiago, Pediatrics, Santiago, Chile. Background: There are a few data regarding recurrent wheezing in infants around the world. The aim of study is to verify the prevalence of recurrent wheezing in infants from south of Brazil. Methods: This is a cross-sectional study. A standardized questionnaire was applied to parents of infants with 12 to 15 months-old attending Health Centers for regular immunization between August/2005 and December/2006. This instrument was previously validated and had questions about clinical characteristics, wheeze, respiratory infections and risk factors. Among 107 Immunization Centers 35 were randomly selected in order to maintain a homogeneous selection of population sample. Results: Three thousand and three parents answered the surveys. Fourty five percent have had at least one wheezing episode in the first twelve months of life, starting with 5.5 T 3.1 months of age, and 678 (22.6%) have had 3 or more wheezing episodes. In this group, 84.6%, 18.5%, 24.3% and 5.4% have used $ 2 -agonists, inhaled steroids, oral steriods and antagonists of leukotriene receptors, respectively. Night-time symptoms, breathless and emergency room visits were as frequent as 58.9%, 46.2% and 57.6% in all of wheezing infants, 12.7% had hospitalization for asthma and in 10.9% asthma was diagnosed. Night-time symptoms, emergency room visits, severe symptoms, asthma hospitalization and diagnosis of asthma were more frequently among wheezers with equal or more than 3 episodes (p G 0.001). Conclusion: The prevalence of recurrent wheezing in infants in the south of Brazil is high, it start early and carriers on great morbidity. Infants possibly represent a high number of persistent asthmatics. Pediatric asthma severity score BPASS^and pulmonary score BPS^as clinical tools compared to others instrumental measures in acute asthmatic children Khaled Taman, Malak Shaheen, and Lammia Mokhtar. Ain Shams, Pediatrics, Cairo, Egypt. Background: International guidelines for treatment of acute asthma call for measurement of peak expiratory flow rate (PEFR) or other objective tests of pulmonary functions. Such measures, however, are frequently difficult to obtain in young children and those unfamiliar with the technique of PEFR. Therefore a variety of clinical scoring systems have been developed for evaluating the severity of acute exacerbations of asthma in children. Aim: To cmopare between both pediatric asthma severity score BPASS[ and pulmonary score BPS[ as clinical tools to others instrumental measures as peak flow meter, pulse oximetry and arterial blood gases for assessing the severity of acute asthmatic attacks in children. Also, our aim to find if clinical assessment tools are of value for rapid intervention in treatment of acute asthma, compared to laboratory measurements. Patients and Methods: These clinical study was done on 100 ashtmatic children attending ER with acute attacks of asthma. Their age ranged from 5Y16 years during the period from January 2005 to January 2006. Clinical assessment of severity was done by using both BPASS[ and BPS[. The results of assessment were compared to BPEFR[, oxygen saturation and arterial blood gases. Results: There was significant association between BPASS[ and BPEFR[ before treatment, 20 minutes and 24 hours after treatment.There was also significant association between BPS[ and BPEFR[ before treatment, 20 minutes and 24 hours after treatment. There was significant association between both BPASS[ and BPS[ scores and oxygen saturation, after 24 houres of treatment. Conclusion: BPASS^and BPS^are of clinical utility in assessing the severity of asthmatic attacks especialy if the patiant is unable to use the peak flow meter or when pulse oximetry or the peak flow meter are not available. Key words: Acute asthma, assessment, pediatric asthma severity score, pulmonary score, peak flow meter, pulse oximetry, arterial blood gases 375 Some clinical and immunologices characteristics at children with the pathology of the biliary systems proceeding on the background of allergic diseases Shahla Rustamova, Sanubar Rustamova, and Aslan Qasanov. Azerbaijan State Advanced Training Institute for Doctors, Department of Pediatrics, Baku, Azerbaijan. Background: To study features of immune infringements at children with a pathology biliar systems proceeding on a background of allergic diseases. Methods: Under supervision there were 36 children with bronchial asthma (BA). To all children determined a level of general IgE, absolute quantity of basofils in a peripheral blood. Results: At all surveyed children deformations of a bilious bubble (Sfigurative, prezense of excesses in the field of a body, a bottom and cervix ) have been revealed. Thus similar features of a structure of a bilious bubble frequently were found out in parents of surveyed children. Clinical displays were characterized by an abundance and polymorphism of complaints. The incidental nausea and vomiting was observed at 21 % surveyed with BA. Dispepsia frustration were observed at 7 %, locks at 28 %, meteorism at 10 %, abdominal a painful syndrome at 18 % of surveyed children. At the majority of patients complaints of vegetative character were marked. The increase of local or general sweating was established in all the patients. Patients complaints of undue fatigability, emotional lability, sleeplessness, headaches, dizzy. Disturbance of thermoregulation manifested itself by chilling and fall in temperature of distal parts of extremities. Significant meteotropism was discovered in children with BA (46,4 %), that caused the aggravation of general condition in change of weather. At children with the expressed clinical displays of a pathology biliar systems the tendency to higher parameters of the general IgE (514 T 303 IU/ml), than at children with asymptomatic current of illness (320 T 200 IU/ml) was marked. At 15 % of children IgE was within the limits of norm. Correlation analyses between level of general IgE and quantity of basofile showed the positive correlation r=+0.63. One of in children with the level of general IgE in limit norms observed increasing quantity of basofiles peripheral blood. May to suppose that in these children happen IgEindependent degranulation fat cell, connected, possible, with breach of vegetative innervation. Conclusion: Thus, the lead researches dictate necessity of more profound inspection immunologices parameters at the given contingent of children. Prognostic importance of IgE for the evolution of recurrent wheezing into the bronchial asthma in children : The recurrent wheezing associated with childhood can evaluate in the bronchial asthma in 30Y70%. Aim: The study was performed to determinate the potential forecast of IgE for the development of the recurrent wheezing into the bronchial asthma. Methods: 37 children ages 2Y4 years old with the recurrent wheezing were included in the study. The study has been done as a part of the detailed medical cheek up in the follow-up period 3Y5 years. We used the stepwise selection Student (t) criterion of statistical analysis, coefficient of contingent (Ä2), U-Fisher (F) criterion for the selection of forecast parameters in the development of asthma in childhood. Results: The statistical discriminative BStepwise[ analysis identificated the level of IgE a parameter with potential informative prognostic for selection the children with risk of asthma. The prognosis of asthma_s risk in children with hiperimmunoglobulinemia E (582,03 T 83,27 IU/ml) which have specificity 56,16%, sensitivity 62,96% and prognosis index 59,46%. The favorable evolution with complete recovery of children with wheezing is determinated by IgE (pG0,01) more increased (391,75 T 54,47 IU/ml) and potential prognosis 77,78%. Knowledge of such bronchial asthma risk_s needs elaboration of a program for effective measuring to prevent asthma in children with recurrent wheezing. Conclusion: The level of hiperimmunoglobulinemia E in child with recurrent wheezing permits to prognosis the risk of the development of bronchial asthma. Thalassotherapy (seaside-treatment) in allergic children Jaroslava Simonickova, Gabriela Polakova, and Vit Petru. Na Homolce Hospital, Ctr Allergol Clin Immunol, Prague, Czech Republic. Background: It is generally held that a comprehensive therapy for allergic children includes thalassotherapy, i.e. seaside treatment. We have attempted to objectify the effect of this therapy in two groups of children: one with the diagnosis of bronchial asthma, the other with atopic eczema. Methods: The effect of a three weeks`seaside stay was evaluated in a group of 31 asthmatic children, compared to 23 asthmatic children in a control group. The evaluation was based on the following criteria: change of activity score, pulmonary functions, price of an anti-asthmatic pharmacotherapy, number of school absences and parent questionnaires. In the second group, which included 25 children with atopic eczema, the effect was evaluated on the basis of the extent of skin affliction, intensity of the symptoms, SCORAD index and parent questionnaires. Results: In the group of asthmatic children, the score fell from 3.8 to 1.9 (with no change in the control group). The difficulties became less frequent, dyspnea on exertion was alleviated, drug consumption fell, school absences and the price of anti-asthmatic therapy were reduced. Pulmonary functions remained unchanged. In the group of eczematous children the extent of skin affliction fell from an average of 8.68 % to 1.88 %. The score reflecting the intensity of difficulties fell on average from 2.44 to 0.56 points with each child. The average value of the SCORAD index in the whole group before treatment was 10.26, after treatment 2.32. The changes of all evaluated parameters are of statistical significance. Conclusion: A three week therapeutic seaside treatment reduces the activity of bronchial asthma and atopic eczema and markedly improves an allergic patient`s quality of life. 378 Abstract withdrawn 379 Ready-to-use HDM atopy patch test (APT) in the diagnosis of sensitization to HDM in children Christophe Dupont 1 , Pascale Soulaines 1 , Nathalie Donne 2 , Pierre Henri Benhamou 1 , and Nicolas Kalach 1 . 1 Hopital Saint Vincent de Paul, Pediatrics, Paris, France; 2 Dbv Technologies, Dbv Technologies, Boulogne Billancourt, France. Background: Sensitization to allergens derived from house dust mites (HDM) is strongly associated with asthma, perennial rhinitis, and atopic dermatitis. Its diagnosis is considered crucial in the allergic work up of these conditions. An increasing number of reports demonstrate the accuracy of atopy patch test (APT) in the diagnosis of HDM allergy. Aim: The aim of this study was to assess the accuracy of a new ready-to-use HDM atopy patch test (APT) (Diallertest\) in the diagnosis of sensitization to HDM in children. Methods: A prospective study was carried out in 47 children, age 57.4 T 42 (mean T SD, ranges 7-176 mo), 18 girls. Patients exhibited isolated or combined atopic dermatitis (AD) (n=28, 59.57%). Children were tested for specific HDM-IgE [against D. pteronyssinus (DPT) and D. farinae (DF)], and skin testing based on HDM (DPT and DF) skin prick test (SPT) and ATP, using HDM-Diallertest\) Results: Among the 47 enrolled children, 15 cases (31.9 %) exhibited sensitization with positive specific HMD-IgE titers against both DPT and DF and 16 (34.04) and 17 (36.1 %) positive SPT against both DPT and DF respectively. The HDM-Diallertest\ was positive in 16 (34.04 %). Among these 16 positive HDM-Diallertest\), 9 exhibited an eczematous reaction and showed an excellent concordance with DPT and DF-SPT and specific IgE against DPT and DF, respectively 93.3%, 97.77%, 90.47% and 90.47%. Conclusion: The 3 diagnostic techniques exhibited a comparable level of accuracy for the diagnosis of HDM allergens sensitization. The excellent concordance with other techniques of HDM-Diallertest\) with eczematous reactions strongly supports its use as a reliable non invasive diagnostic tool of HDM sensitization. Prevalence of asthma, allergic rinoconjunctivitis and atopic dermatitis in the southwest of Mexico City, ISAAC model study Alvaro Pedroza, Jose Huerta Lopez, Francisco Espinosa, Gabriela Trevino, Francisco Rivas, Cintia Ramos, Nadia Ramirez, and Horacio del Olmo. Instituto Nacional de Pediatria, Pediatric Allergy and Immunology, Mexico City, Mexico. Introduction: The allergic diseases are the most frequent chronic entities in the childhood, through the time their prevalence has been increased in the last three decades, that is why it was deasigned a task force called International Study of Asthma and Allergy in Childhood (ISAAC) that promotes the epidemiological researches in asthma and allergic diseases. Objective: To approach the prevalence and severity of asthma, allergic rhinoconjunctivitis and atopic dermatitis in children between 6 and 7 year and 13 and 14 year old in the southwest of Mexico City. Methods and Materials: It is an observational, transversal, descriptive, prospective study, it includes students of first and second grade of primary school whit age of 6 and 7 year old and junior high school students from 13 to 14 year old. The study was authorized by the Public Education Institute, school principal and parents. The statistical analysis was made by Epi info version 6 whit a significative value of p e 0.05. Results: There were 58 schools enrolled, with 6184 students from wouthwest of Mexico city, 32 primary schools with 3093, 1585 girls (51.24%) and 1508 boys (48.75); junior high schools included 3034 students and 26 schools with 1538 girls (50.69%) and 1496 boys (49.30%). In the scholastic population the antecedent of only one wheezing episode was present in 20.43% (632) and the proportion of patients with severe asthma was 6.7% (207), from this data only 4.3% (134) had asthma diagnosded, in the teenagers only 172 (5.66%) had diagnostic. The severe rhinoconjunctivitis was present in 399 kids (25.99%) and only 103 teenagers (11.26%). The major part of students, kids and adolescents did not give correctly the information, about atopic dermatitis, only 9 (0.29%) joined with pets (cats and dogs). Discussion: Similar studies made around the world reveals that asthma and allergic diseases are a public health problem, with implications in quality of life, it may lead to temporal o permanent incapacity, if they are not diagnoses and treated in early approach. The variation in the prevalence are expected because various factors, the most important is the diversity of each zone about risk factors, in fact the results can not be compared with a different population, they can be used for the epidemiological knowledge of this zone. Bronchial asthma and child physical activity in Lithuania Egle Vaitkaitiene. Kaunas Medical University Institute for Biomedical Research, Social pediatrics, Kaunas, Lithuania. : The background of the study. Prevalence of child asthma is 2-3 % in Lithuania. Asthma is not only clinical, but social problem also. The aim of this study was to evaluate physical activity and its limitation in asthmatic children. The method used. The study group included children suffering from asthma between the ages of 7 and 17. The severity of asthma was assessed by grading the asthma as mild, moderate or severe. We used standardized and diseasespecific PAQL questionnaire (national version). Responses of 186 asthmatic children were analyzed. The results obtaines. Only one third of our respondents were involved in extra activity: sport, dancing and singing. 62.9 % of parents noticed that their children have no extra activity. 17,7% of children with asthma were not attending sports at their school. We asked childrens to name three activities, in which they had been bothered by asthma. 16,6 % of respondents indicated more than three daily activities, restricted by asthma. Asthma for boys was the most disturbing while running (85.2%), playing with pets (23.8%), playing ball (25%) and sleeping (26%). Girls marked different activities, such as running (64%), housekeeping (46.9%) and school sports (25%). Asthma bothered boys on the run and playing ball more often than girls. Girls were bothered by asthma more often than boys when doing chores and singing. Statistically significant differences were found comparing all the three main activities for boys and girls: the first activity -Â 2 = 44.9, df = 15, p G 0.001; the second activity -Â 2 = 42.8, df = 26, p = 0.02; the third activity -Â 2 = 47.9, df = 23, p = 0.002. The conclusion reached. Asthma is a limiting factor in the childrenFs level of physical activity. It prevents children from doing various activities. Due to asthma, children were not able to run, play ball, do their chores, play with their pets. Boys more often complained thet asthma had limited their physical activity (running, playing games), whereas girls indicated that the disease had limited both, their physical activity (such as running) as well as passive activities (such as doing chores, in addition to dancing and singing). Analysis of allergen-specific serum IgE in children with asthma or cough variant asthma Xin Song, and Jing Zhao. Capital Institute of Pediatrics, asthma center, Beijing, China. Objective: To investigate the positive rates of food allergen (Fx5), mixed molds (mx1), phadiatop and house dust mites (d1) allergen-specific serum IgE (sIgE) in children aging from 6 months to 16 years diagnosed of asthma or cough variant asthma (CVA). Their age distribution characteristics were also compared and analyzed. Methods: Blood samples of 597 asthmatic children were applied to fx5, mx1, Phadiatop and d1 tests using fluoroenzyme-immunometricassay, UniCAP100. The positive rate of fx5,mx1,Phadiatop and d1 were compared. Age distribution characteristics of children with positive SIgE were analyzed. Results: 1, The positive rate of fx5, mx1, Phadiatop and d1 were 34.6%, 30.%, 35.2% and 23.5% respectively; 2, The highest positive rate of Phadiatop and d1 were highest in children of 8-16 age group (73.6% and 49.1%); The positive rate of fx5,mx1 peaked in children of 2 years and 5 years separately(44.1% and 53.6%); 3, Among 181 children with mx1 positivity 90 cases manifested phadiatop negativity. 4, 65% of children with clinically diagnosed asthma or CVA presented positivity in test in fx5,mx1 or phadiatop at least. Conclusion: Two-thirds asthma and CVA patients existing sIgE; Asthmatic infants mainly manifested food allergen sensitization. The positive of Phadiatop increased with age. Positivity of mx1 increased with age in children e 5 years , and it decreased in older children (Q 6 years). Mx1 was one of the main allergens of asthmatic children. An observational study on the effectiveness of montelukast in children with chronic rhinitis Background: Impacts of chronic rhinitis in children especially nasal congestion included learning disorder, impaired school performance and quality of life. Intranasal corticosteroid is the most effective medication for chronic rhinitis treatment. Recently, leukotriene receptor antagonist has been added to the modes of therapy approved by the US Food and Drug Administration of allergic rhinitis. Objective: To assess the effectiveness of add-on montelukast with intranasal corticosteroid in children with chronic and persistent rhinitis. Methods: We enrolled children attending the allergy clinic, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, with poorly controlled rhinitis during the period of April, 2005 to December, 2006. Diagnosis of allergic rhinitis was made by positive prick skin test to at least one aeroallergen. Intranasal corticosteroid was the first-line medication for treatment of persistent symptoms. Montelukast was added in cases that no response to intranasal corticosteroid, mostly due to nasal congestion. Efficacy was assessed by symptom improvement (nasal congestion, rhinorrhea, itching, sneezing) reported by patients and physical examination by allergist. Results: There were 10 boys and 16 girls (average age 13). The mean duration of symptom before diagnosis was 4.7 years. Twelve children were diagnosed to be allergic rhinitis and 14 children were non-allergic rhinitis by prick skin testing. The mean duration of intranasal steroid use was 4.4 year in children with allergic rhinitis and 2.7 years in children with non-allergic rhinitis. Symptom improvement, especially nasal congestion, was reported in 8 of 12 (66.6%) allergic rhinitis subjects and 11 of 14 (78.5%) non-allergic rhinitis subjects after add-on 10 mg of montelukast daily. Montelukast was overall well tolerated. Mean duration of montelukast therapy was 1.7 years in both groups. Intranasal corticosteroid was able to stop in 8 of 14 (57%) children with nonallergic rhinitis and unable to stop in all children with allergic rhinitis, but all of the patients were satisfied with these treatment. Conclusion: Montelukast have an advantage in additional treatment of either allergic or non-allergic rhinitis children, but particularly more benefit on children with non-allergic rhinitis. Sputum eosinophils in childhood asthma: Correlation with PEFR in controlled asthmatics and with exacerbation Maria Cristina Edquilag, and Agnes Andaya. University of Santo Tomas Hospital, Department of Pediatrics, Section of Allergy, Manila, Philippines. Background: Sputum eosinophilia is a non-invasive airway inflammatory marker. Airway inflammation which is the characteristic functional abnormality peculiar of asthma, leads to variable airflow limitation. Significant correlation between PEFR and absolute eosinophil counts have been observed among patients with asthma. However, these studies are limited and others show conflicting results. Objective: To determine the correlation between sputum eosinophilia and PEFR among patients with asthma. Methodology: Following clinical assessment and peak flow determination, sputum eosinophil count was determined among patients with stable asthma (29) and acute exacerbation (36). Patients were classified as to intermittent and persistent asthmatics. Age, sex, atopic status and asthma duration were obtained. Sputum eosinophil count of = 3% was considered significant. Results: Patients with sputum eosinophilia had lower PEFR, compared to those without sputum eosinophilia (p = 0.004). Patients with sputum eosinophilia were more likely to have abnormal PEFR compared to those with normal PEFR (OR = 6.64, 95% CI = 2.02-21.90). Patients with persistent asthma were more likely to have sputum eosinophilia compared to those with intermittent asthma (OR = 3.06, 95% CI = 1.02Y9.18). Similarly, patients in acute exacerbation were more likely to have sputum eosinophilia compared to those with stable asthma (OR = 3.42, 95% CI = 1.07Y11.04). Conclusion: This study demonstrates that there is a direct correlation between sputum eosinophilia and PEFR among patients with acute exacerbation. However, no correlation could be made among patients with stable asthma since all patients had normal PEFR. Serum levels of VEGF, TGF$2, IL-4, IL-2R and ECP in children with acute astma bronchiolitis Kadir Kocak 1 , Bilal Yildiz 2 , Yesim Kural 2 , and Omer Colak 3 . 1 Eskisehir Osmangazi University, Faculty of Medicine, Pediatric Allergy, Eskisehir, Turkey; 2 Eskisehir Osmangazi University, Faculty of Medicine, Pediatrics, Eskisehir, Turkey; 3 Eskisehir Osmangazi University, Faculty of Medicine, Biochemistry, Eskisehir, Turkey. Background: Asthma is leading cause of chronic illness with increased vascularity, vascular permeability, airway and lung tissue remodeling. Although, various growth factors and cytokines have been implicated in modulating in these processes, the role of factors and cytokines are not known very well in asthmatic children. Aim of this study is to determine the serum levels of vascular endothelial growth factor (VEGF), transforming growth factor beta-2 (TGF$2), interleukin-4 (IL-4), interleukin-2-receptor (IL-2R), and eosinophilic cationic protein (ECP) in acute astmatic children (AAC) and compare with acute bronchiolitis and healthy subjects and to assess the changes with treatment. Methods: These parameters were measured by ELISA techniques in AAC and bronchiolitis patients before and after inhaled or systemic steroid (SS). Results: Serum VEGF, TGF$2, IL-4 and IL-2R levels were increased in patients with AAC and bronchiolitis than in controls before therapy. These parameters were higher in AAC than bronchiolitis except IL-2R (Table-I Serum VEGF, IL-2R and ECP levels were higher in AAC than in controls after treatment. After steroid therapy, VEGF, IL-4 and ECP levels were higher in AAC than in bronchiolitis but IL-2R levels were higher in patients with bronchiolitis. Only serum IL-4 levels in bronchiolitis were similar with controls after treatment. By the severity of disease, only VEGF levels were increased in modarate AAC and bronchiolitis higher than in mild patients (p G 0,0001). Only VEGF levels were lower in AAC who were treated with prophylactic drugs than in AAC without prophylactic treatment (p G 0,05). According to modality of steroid treatment, only VEGF levels were lower in AAC who were treated with inhaled steroids compared with SS(p G 0,001). In AAC; VEGF, IL-4, IL-2R and ECP levels were higher than in controls after treatment with SS (p G 0,0001). But in AAC who were treated with inhaled steroids, VEGF and IL-4 levels were similar with controls after treatment (p 9 0,05). Conclusion: VEGF, TGF$2, IL-4 and IL-2R may play role in pathogenesis of asthma and bronchiolitis. These parameters aren't useful for distinguishing asthma and bronchiolitis. VEGF could be useful parameter for distinguishing mild and moderate disease in asthma and bronchiolitis. According to our findings, inhaled and SS could be combined for appropriate treatment in moderate AAC. Also, anti-VEGF therapy may be useful in AAC. Effect of self-management education on asthma complication and pulmonary function in Iranian children with asthma Background: Despite global efforts to manage asthma, the adverse outcome of this disease is increasing. Effective self-management and treatment compliance is important in achieving good symptom control in asthma. The aim of this study was to determine whether asthma nurse intervention during clinical period could increase knowledge and improve self-management and whether this would influence the frequency and severity of symptoms, and pulmonary function test. Setting: A university hospital pediatric outpatient clinic. Methods: Randomized controlled trial of 60 children with intermittent and mild to moderate persistent asthma were assessed to indicate whether the educational self-management intervention would improve lung function and the use of inhaler with spacer score, increase feeling of control over asthma, and decrease the use of health care services. 60 children with asthma, who were categorized based on disease severity, duration of disease, age were randomly assigned to an experimental group (n=30) which received an educational program or to a comparison group (n=30) which received common education. The education program consisted of 4 group sessions which focused on improving the patients and parents' self-management skills. Background: Chronic cough lasting 8 weeks or more often seems an intractable problem in childhood. Toxocara infestation is associated with an increased prevalence of airway symptoms and may be the possible etiologic agent of chronic cough. Methods: Spirometry, bronchial reversibility tests and nonspecific bronchus provocation tests were carried out in patients who could cooperate in these tests. Measurements were made of the peripheral blood eosinophil count, the serum total IgE level, the serum total magnesium (Mg) concentration. The serological examination for Toxocara canis, based on the most widely used enzyme-linked immunosorbent assay of IgG (Toxocara canis IgG ELISA, Novatec, Germany) and more sensitive Western blotting (Toxocara Western Blot IgG, LDBIO, France) methods, was performed. All patients with positive serology for T. canis index of positivity 9 9 were treated with albendazole in a dose of 10 mg/kg/day, patients weighing 9 40 kg received 400 mg albendazole a day for 7 days. All patients with bronchial asthma and CVA were treated with inhaled corticosteroid (ICS). Further, short-term beta2-mimetics could be administered in cases of dyspnoea Bas needed^. Results: One-hundred and thirty-six of 425 children (32%) aged 2Y17 years were seropositive for T. canis antigens. Ninety-three children were adequately assessed, diagnosed and followed up during 1 year. Bronchial asthma was diagnosed in 40%, cough variant asthma (CVA) in 27% and nonasthmatic eosinophilic bronchitis (NAEB) in 33% of children with chronic cough. The eosinophil cell count, the Toxocara canis IgG, the total IgE levels and the total Mg concentrations in the serum are predictors of the improvement or the decline of the patients`condition. We could significantly decrease the dose of ICS in 23 of 37 (62%) children with bronchial asthma. Anthelminthics and avoidance were sufficient for children with NAEB; none needed ICS. ICS therapy could be stopped 2-3 months later in 17 (68%) of 25 patients with CVA. We found that 8 of 25 patients with CVA (32%) presented asthmatic symptoms at the end of the 1-year period. Conclusion: In Hungary, T. canis may be the potential sensitizer for chronic cough in seropositive children. Deworming therapy will then alleviate the airway symptoms without exacerbation in patients with bronchial asthma, and definitively treat children with NAEB and the majority of children with CVA. It is supposed that prenatal Th1/Th2 polarization of immune response is possible, but the markers of such polarization, trustworthy associated with atopy forming are not known. Aim: To evaluate the connection between IgE level, CD26, CD30 expression and intracellular interferon (IFN)-, production by cord blood mononuclear cells (CBMC) and the risk of infant atopic dermatitis (AD) forming. Methods: We defined per cent content of CD26+and CD30+in CBMC by using flow cytometry as well as the concentration of total IgE in 117 full-term newborns. Simultaneously, intracellular IFN-, expression as well as early activation marker CD69 (separate for total mononuclear cells population and CD4+lymphocytes) were examined (95 newborns). We estimated AD presence in 41 enrolled newborns in the age of one year (medically diagnosed AD was marked in 25 of them). Data are shown as median (25-75% quartiles) of absolute cells count (10(-)6/l). Statistical analysis was performed using the Mann-Whitney test. Relative Risk (RR) was present with 95% CI. Results: Only for nonactivated CBMC, producing IFN-, (subtype CD69-/ IFN-,+), the association with AD forming has been found. The number of CD69-/IFN-,+CBMC in AD infants was 3.3 (0-6.4), as opposed to infants without AD -9.4 (5.3-16.5), p = 0.006. Relative risk for AD forming in newborns with low CD69-/IFN-,+( G 5.0 -median for our cohort) was 3.4 (1.19-9.69 ). Significant differences in the number of CD69-/IFN-,+CBMC was kept after adjusting to such factors as family atopic history, place of living (city/village), birth order (first/second or later), the number of family members living together, bedsharing smoking, pets owing and the newborn gender.We revealed the negative concentration of total IgE in cord blood with the ratio CD26+/CD30+CBMC (r = j0.37, p = 0.0005). We did not find any association of CD26, CD30 expression, the CD26/CD30 ratio, as well as the level of total IgE with AD forming in infants (medians were chosen as cut off points). RRs were 1.5 (0.67-3.33), 0.67 (0.37-1.19), 0.78 (0.42-1.45) and 1.24 (0.59-2.61), respectively. Conclusion: Thus, the decreasing of intracellular IFN-, production by nonactivated CBMC, proving the prenatal displacement in Th1/Th2 balance, is the strong predictor of infant AD forming. However, the decreasing of CD26/ CD30 ratio as well as the level of total IgE in cord blood are not the predictors of atopic phenotype forming in infants. Sputum eosinophil level in children with mild, moderate to severe exacerbation and stable asthma Maria Cristina Edquilag, and Agnes Andaya. University of Santo Tomas Hospital, Department of Pediatrics, Section of Allergy, Manila, Philippines. Background: Sputum eosinophilia is a non-invasive marker in the evaluation of airway inflammation as well as asthma control. Increased proportion of sputum eosinophils have been observed among subjects with asthma, especially among those with exacerbation. However, there are also studies that show airway eosinophilia among patients with controlled asthma. Objective: To determine sputum eosinophil level among asthmatic patients in exacerbation and those with stable asthma and determine its association with severity of disease. Methodology: Following clinical assessment and peak flow determination, sputum eosinophil level was determined among patients with stable asthma (n = 29), mild (n = 18) and moderate to severe (n = 18) asthma exacerbation. Age, sex, atopic status, asthma duration and medications were obtained. Sputum eosinophil count of Q 3% was considered significant. Results: The mean eosinophil count in children with stable asthma, mild exacerbation and moderate to severe exacerbation were 4.65% T 12.53%, 1.94% T 4.30%, and 8.83% T 11.12% respectively. Sputum eosinophils were significantly higher in children with moderate to severe exacerbation than in mild exacerbation and stable asthma (p = 0.0086). The percentage of patients with significant sputum eosinophilia was higher among those with moderate to severe exacerbation (17%) than mild exacerbation (6%) and stable asthma (8%). Statistically significant difference was seen among the three population (p = 0.0234). Logistic regressions showed that wheezing (p = G 0.0001), peak expiratory flow rate (p = 0.0010), sex (p = 0.0110) and asthma duration (0.0232) were significantly associated with sputum eosinophilia (p = 0.432). No statistically significant difference was seen with respect to long term asthma severity classification (p = 0.432). Conclusion: This study demonstrates that significant sputum eosinophilia is seen among patients with moderate to severe asthma exacerbation compared to those with mild exacerbation and stable asthma. Sputum eosinophilia was also significantly associated with wheezing, lower peak expiratory flow rate, male sex and longer asthma duration. Long-term asthma severity is not a defining factor in the evaluation of sputum eosinophilia among asthmatics but the severity of exacerbation. Allergy in children: managing patients at risk of anaphylaxis Srinivas Bandi 1 , and Colin MacDougall 2 . 1 Birmingham Heartlands Hospital, Paediatrics, Birmingham, United Kingdom; 2 University Hospital, Paediatrics, Coventry, United Kingdom. Aim: To report the management of children with allergy in a district general hospital, in particular to review prescribing practices of epinephrine auto injectors (Epipen) and to compare the management of children seen in allergy clinic with those seen in general paediatric clinic. The clinical records of all children who were prescribed Epipen or Epipen junior between November 2003 and October 2005 were retrospectively reviewed. 39 patients were included of which 23 were seen in allergy clinic and 16 were seen in general paediatric clinics. There ages ranged between 6 months to 17 years (median of 7 years). Results: The common allergens were Nuts/seeds (79%) followed by egg (23%), milk (18%) and fish (8%). 15 children had positive skin prick tests and 18 had positive RAST (total 33/39 had positive tests). Information on treatment with Epipen was provided to 22/23(95%) children seen in allergy clinic compared to 10/16 (62%) children seen in general clinics. Practical instruction of Epipen administration was provided to 21/23(91%) compared to General paediatric clinic Metabolic responses to treatment with salbutamol and theophylline in asthmatic children Mohammed Ragab. Alexandria University, Faculty of medicine, Alexandria, Egypt. : Drug therapy and its abuse have been incriminated as a main contributive factor in asthma mortality. The effect of treating asthmatic with salbutamol inhalation (0.15 mg/kg body weight up to 5 mg/dose, group A), theophylline (5mg/kg body weight, group C) or both drugs (group B) was investigated in 30 asthmatic children. Ten healthy children of matching age and sex were also included to serve as control. Serum glucose increased significantly in group A and B, but did not change significantly in group C as a compared to pretreatment level. Serum inorganic phosphate decreased significantly in group B and C, with no significant change in group A. serum sodium level was significantly decreased an all asthmatic children before treatment. Group B and c showed significant reduction in serum sodium level as compared to pretreatment values. Serum sodium level did not change significantly in group A. Serum potassium decreased significantly in the three studied groups as compared to pretreatment values. These metabolic changes could possibly help in explaining the observed increase in cases of fatal asthma, when using a combination of inhaled beta-agonist and oral theophylline preparation. Multymeasuring phenotyping and automated prevention system of Bronchial asthma in children Tbilisi State University, Stomatology Department, Tbilisi, Georgia. Background: Prevention in early age period significantly decreases morbidity of allergic diseases and improve quality of life. Sometimes doctors are unable to manage well this pathology, because of neglected early diagnostic. According to the most actual and perspective direction of nowadays allergology it is working out new scientific approach of resolution of bronchial asthma (BA) problems in young population. Hereditary predisposition plays the most important role in the developing of disease, but it is not everything recognizable, it may be not demonstrated clinical symptoms in parents and other close relatives of affected individual. Aim: To elaborate the new automated assessment cost-effective system which would allow early diagnostic and prevention BA by using Bformalized cards^by providing multymesure phenotyping in Georgian children population. This system is based on the primary and secondary prognosis of BA. Methods: Choosing of markers: HLA-markers (Terrasaki micro lymphocytetoxic tests) typing was performed with 2nd class (DR, DQ) and 1st class (A, B, C) antigens locus`s; Erythrocyte markers: ABO, Rhesus and MNS systems, Clinical-genealogic method (Falconer D., 1965) : study of genealogical tree of investigated families; Genetic-statistic analysis Y Informative Criteria of different signs: ontogeny, phenotypic, environmental factors (by Kulback, Goubler E.B, 1990) . Method of multi-dimensional phenotyping: agglomerative-hierarchical cluster analyse of chosen systems of signs. Analyse was performed in the interactive order. Results: Multi-dimensional phenotyping shoves phenotype classes: Classes of basic predisposition, including up to 80% of individuals with manifested asthma; Classes of relative predisposition, including less then 80% of individuals with manifested asthma; Phenotype classes including a few patients; Phenotype classes not predisposed to BA, basically including healthy persons. Conclusion: Elaborated computing table is enough simple and opportune for comprehensive use. It is proposed, for primary prognostic of BA, to practical doctors working in outpatient clinics and dispensaries, as well for military doctors working with enlistment committee or in military units. This method gives the possibility for prognosis predisposition to bronchial asthma in young population. TBXA2R gene polymorphism and its pharmacogenetic effect to leukotriene receptor antagonist in children with asthma Background: Thromboxane A2 receptor(TBXA2R) gene polymorphism has been associated with atopy and asthma. However its role in children has not been defined. We investigateed associations between asthma-related phenotypes and TBXA2R polymorphism, and also to analyze whether TBXA2R polymorphism has an effect on the efficacy of the leukotriene receptor antagonist(LTRA), montelukast, in asthmatic children with exercise-induced bronchoconstriction (EIB). Methods: Asthmatic children(n = 695) and control children(n = 159) were evaluated for asthma-related phenotypes including total IgE, pulmonary function test, and BHR to methacholine or exercise. Genotypes were detected by PCR-RFLP. In the montelukast study, exercise challenge was performed before and after 8-week montelukast treatment. Results: The TBXA2R polymorphism was not associated with asthma susceptibility and clinical parameters of asthma. However, asthmatic children with combinations of the TBXA2R + 795T 9 C and + 924T 9 C risk alleles had significantly higher total IgE levels (P = .01), total eosinophil counts (P G .01) and lower FEV1 (P = .02) and FEF 25Y75% (P = .02) than those carrying the common alleles. When compared with individuals with the common alleles, in patients with the TBXA2R + 924T 9 C TT homozygote and TBXA2R+795T 9 C hetero-or homozygote (CT or CC) had a 3.67-fold poor response to 8-week montelukast treatment regarding to maximum percent fall in FEV1 after exercise (odd ratio, 3.67; 95% CI, 1.15-11.15). Conclusion: A combined effect of TBXA2R + 795T 9 C and + 924T 9 C risk alleles may be linked to IgE production, eosinophilic inflammation, and severity of asthma. In addition, the combined genotype of TBXA2R may be a predictive marker of clinical response to the LTRA in Korean asthmatic children with EIB. A randomized prospective double blind controlled trial on effects of long-term consumption of fermented milk containing Lactobacillus casei in pre-school children with allergic asthma and/or rhinitis Institute, Università Cattolica del Sacro Cuore, Piacenza, Italy; 3 University of Milan-San Paolo Hospital, Paediatric Department, Milan, Italy. Background and Aim: Probiotics may have immunomodulatory functions, exerting beneficial effects in allergic disease. We investigated whether prolonged daily consumption of Lactobacillus casei DN-114 001 fermented milk improve health status of children aged 3-6 years allergic to inhalants. Methods: In a randomised prospective double blind controlled trial patients were randomly assigned to receive for one year 100 ml per day of L. casei DN-114 001 fermented milk (109 cells/ml), or no probiotics (controls). Clinical evaluations occurred every 3 months. Faecal flora composition was assessed every 6 months (30 treated, 15 controls). Outcome measures were time free from and the number of episodes of asthma/rhinitis, number of fever or diarrhoea episodes, IgE-G-A serological levels (12 months vs baseline). Results: Participants (187; 92 treated) were similar regarding gestational age, breastfeeding, family smokers, pets, siblings, day-care admission. No statistical difference between intervention and control group occurred in asthmatic children. In children with rhinitis, the annual number of rhinitis episodes was lower in the intervention group, mean difference (95%CI), -1.6 ( -3.15 to -0.05); the mean duration of an episode of diarrhoea was lower in the intervention group, mean difference -0.81 (-1.52 to -0.10) days. Faecal analysis showed Lactobacillus casei DN-114 001 in the gut flora of 9 78% of supplemented children through the study period. Conclusion: Long-term daily consumption of Lactobacillus Casei DN-114 001 fermented milk may positively influence on the clinical and immunological status of allergic children. Prevalence of pediatric allergic diseases in the first three years of life Chizuko Sugizaki, and Motohiro Ebisawa. Sagamihara National Hospital, Clinical Research Center, Department of Allergy, Sagamihara-City, Kanagawa, Japan. Background: We have maintained birth cohort study in Sagamihara-City since 2002 to clarify the prevalence of atopic dermatitis (AD), food allergy (FA), bronchial asthma (BA), and Japanese cedar pollinosis (JCP) from infancy to childhood. Methods: Using the mass medical examination system at the age of 4 months (mo) old, we first had obtained information of eczema, nutrition method, family history of allergic diseases, and results of allergy examination by questionnaire. We followed up the subjects, whose parents agreed to participate in this study, at the age of 8 mo, 12 mo and 3 years (y) old using questionnaire by mail. Furthermore the risk factors for the subjects to develop BA by the age of 3 y were analyzed from the information obtained in the study. Results: Informed consent was obtained from 5247 parents of 4 mo infants out of 5932 parents from 1/1/02 to 12/31/02. We could follow up 4214 infants at the age of 8 mo, 4068 infants at the age of 12 mo, and 2888 children at the age of 3 y. The incidence of chronic eczema to suspect AD at 8 mo, 12 mo and 3 y was 18.8%, 13.3% and 14.8%, respectively. The incidence of food elimination due to FA reported by parents at 8 mo, 12 mo and 3 y was 17.0%, 12.8% and 5.4% respectively. Interestingly, the incidence of doctor-diagnosed FA at 8 mo, 12 mo and 3 y was 2.4%, 2.8% and 5.1% respectively. Confusion related to FA and AD during infancy seems to exist between parents and doctors. The prevalence of BA at 12 mo and 3 y was 2.9% and 8.7% respectively. The risk factors for infants to develop BA by 3y were related to the possession of AD or FA at 8 mo, 12 mo and 3 y, family history of allergic diseases and history of indirect cigarette exposure. The incidence of JCP at 3 y was even 3.0%. Conclusion: Since the population of children in Sagamihara-city represents about 5% of that of Japan, we can now estimate the prevalence of allergic diseases from infancy to childhood by the data obtained in this study. However, it was very difficult to estimate the prevalence of FA especially during infancy due to the under-evaluation by doctors and over-reaction by parents. It is important for both doctors and parents to fix the confusion in our society. The data can make us possible to clarify the prevalence of pediatric allergic diseases and to reveal the transition of the diseases. Update of the guideline on allergy prevention Torsten Schäfer, Cathleen Borowski-Muche, and ABAP, Consensus group. Medical University Luebeck, Institute of Social Medicine, Luebeck, Germany. Background: Prevention is crucial to counteract the rising trend of allergies. In 2004, we published an evidence-based and consented guideline on primary and secondary prevention of asthma, allergic rhinitis and atopic eczema. In order to update this guideline, we repeated a systematic literature search in 2007 and present the results here. Methods: A literature search was performed in Medline for the years 2003 through April 2007. The used mesh-term groups were as follows. Endpoints: asthma, allergy, allergic, atopic, hay fever, dermatitis, eczema, rhinitis. Interventions: prevention, risk factor, epidemiology. Study types: randomized controlled trial, clinical trial, control study, systematic review, meta analysis, case control study, cohort study. Terms were combined with Band[ within groups and Bor[ between groups. We included studies on humans published in English or German language and excluded therapeutic and drug trials. A first selection process was performed on the basis of titles and abstracts. Results: We obtained 1551 hits of which 139 potentially relevant studies were identified. The later consists of 6 reviews, 9 RCTs, 91 cohort-and 33 casecontrol-studies. After an initial review, the studies could be allocated to the areas breast feeding, hypoallergenic formula, introduction of solid food, diet of the mother and the child, smoking, mould and dampness and vaccination. The recommendations of the guideline to these areas were supported by the actual evidence. There is new evidence which allows a specification of the current recommendations for the topics body mass index and exhaust. There is indication that the recommendations on house dust mite, pet keeping and unspecific immune modulation need to be revised. Conclusion: By this literature search it is possible to update the current evidence-based recommendations on allergy prevention and to adapt the recommendations according to the current literature. T Schäfer, C Borowski, TL Diepgen, M Hellermann, I Piechotowski, I Reese, T Roos, S Schmidt, H Sitter, T Werfel, U Gieler und die Konsensusgruppe des Aktionsbündnisses Allergieprävention. Evidenz-basierte und konsentierte Leitlinie BAllergieprävention [. Allergo J 2004; 13:252-60. 398 A specific mixture of short chain galacto-oligosaccharides and long chain fructo-oligosaccharides induces an anti-allergic immunoglobulin profile in infants at risk for allergy Background: In a prospective study in infants with a family history of atopy a specific prebiotic oligosaccharide mixture (90% short chain galactooligosaccharides and 10% long chain fructo-oligosaccharides (GOS/FOS) (IMMU-NOFORTIS) reduced the cumulative incidence of atopic dermatitis at six months of age. In a subgroup of these infants it was possible to obtain a blood sample at six months of age to analyse the potential effect of these dietary oligosaccharides on the immunoglobulin profile. Methods: In this prospective double-blind randomized, placebo controlled, study the infants received a hypoallergenic formula with either 8g/l GOS/FOS or 8 g/l maltodextrine (placebo) for six months. At three months of age, children were vaccinated against diphteria, tetanus ad polio (DTP). At six months of age total plasma levels of IgE, IgG1, IgG2, IgG3, and IgG4 as well as cow's milk protein (CMP) and DTP specific immunoglobulins were measured. Results: Supplementation of GOS/FOS has lead to a significant reduction in the plasma level of total IgE (p = 0.007), IgG1 (p = 0.0054), IgG2 (p = 0.029) and IgG3 (p = 0.0343) immunoglobulins whereas no significant effect on IgG4 was observed. The plasma levels of CMP specific IgG1 was significantly decreased (p = 0.015) in the GOS/FOS group. The levels of CMP specific IgE were very low and no effect of GOS/FOS supplementation could be observed. CMP specific IgG4 was not detectable in the samples. No influence of GOS/ FOS supplementation was found on any vaccine specific antibody isotype levels. Conclusion: Evidently GOS/FOS supplementation induced an anti-allergic immunoglobulin profile in infants at high risk for allergic diseases whereas desired specific immune responses were not affected indicating the potential role of oral GOS/FOS exposure for primary prevention of allergies. Background: An altered microbial exposure may underlie the increase of allergic disease in affluent societies. Early colonisation with Bifidobacterium (B.) and Lactobacilli (L.) have been postulated to prevent children from developing allergy, while Clostridium (C.) difficile colonisation has been associated with allergic disease. Previous studies have mainly been performed with culture dependent techniques. However, lately the sequencing of bacterial genomes has made it possible to study the gut flora with molecular techniques. The Real-time PCR technique uses primers targeting conserved genes of bacteria, leading to efficient qualitative and quantitative analysis of bacterial DNA from faecal samples. Methods: Presence and amounts of bacterial DNA in infant faecal samples, collected at 1 week, 1 month and 2 months, were measured with Real-time PCR and related to allergy development in Swedish children, followed prospectively at 6, 12, 24 and 60 months of age. Children regarded as allergic (n=14) had developed allergic symptoms and sensitisation to food and/or inhalant allergens during their first five years of life while non-allergic children (n=23) were non-sensitised without symptoms. Primers binding to C. difficile, B. bifidum, B. longum/infantis, B. adolescentis, B. breve, Bacteroides fragilis, Lactobacilli group I (L. rhamnosus, L. paracasei, L. casei) and Lactobacilli group II (L. gasseri, L. johnsonii group) were used. Results: The most abundant bacteria were B. longum/infantis while few children were colonised with C. difficile. At one week of age none of the allergic children was colonised with Lactobacilli group I compared to 56% of the children who remained non-allergic (p=0.004). Also, at 1 month of age B. adolescentis was more common in non-allergic than allergic children (p=0.008). Furthermore, the persistent colonisation with these bacteria were more common among non-allergic children, p=0.018 and p=0.060 respectively. Conclusion: A more diverse gut flora might prevent allergy development and the early colonisation might be of major importance. Aim: To evaluate the efficacity of educational activity from School of Asthma in children with asthma and/or their family. Methods and Materials: In the school of asthma children the specialists have performed educational activities in 172 children with different forms of asthma and their parents. Educational seminars consisted of 5 sessions which approached the following subjects: triggers, asthma symptoms, prophylaxis and asthma medicines. Each session also included individual interrogation of the trained to estimate their knowledge of asthma before and after the education session according to the theme of the working day. The quality of knowledge of some children with asthma and/or their parents was also tested after 3Y12 months and afterwards visits. Results: It was found that the knowledge-of children and parents before the training at the School of Asthma Children was incomplete and incorrect. Children with severe asthma had wrong knowledge about triggers (47,73%), about prophylaxis (52,76%) comparatively with those who suffer of mild asthma (58,06% and 60,76% respectively) and moderate (51,81% and 55,94% respectively). Knowledge of children with mild asthma referring in triggers has grown to 90,77%, of children with moderate asthma to 83,72% and those with severe asthma to 80%. Also was established a growth in knowledge about prophylaxis in all groups of children: mild asthma-83,82%, moderate asthma -85,72%, severe asthma -79,26%. Knowledge of children regarding clinic manifestations constituted: mild asthma -5,5%, moderate asthma -74,79%, severe asthma -78,28%. Knowledge level under the subject "treatment" is the most incomplete in all groups of the questioned persons: mild asthma -37,93%, moderate asthma -36,96%, severe asthma -38,78%. Towards the end of the course they were appreciated as follows: mild asthma -63,17%, moderate asthma -72,67%, severe asthma -73,0%. The average of knowledge in all groups of children after 3Y12 month referring triggers was 79,72%, about prophylaxis 79,01%, regarding clinic manifestations -69,75% and referring treatment -52,44%. For compare, the average of knowledge before educational activity was: referring triggers -52,43%, prophylactic measures -56,32%, clinic manifestations -45,95% and treatment -37,89%. Conclusion: The knowledge of those questioned has little decreased during 3Y12 months, but it remained net superior to the level before the seminars. The usage of paracetamole \ \ The subsequent development of allergy and asthma -Study of matched patients-siblings Background: Many studies on this field have come to the suggestion that, the intake of paracetamole during pregnancy and during the first Months of life is risky as it increase the risk of childhood asthma. Our aim is to deterring the asocial between paracetamole usage during pregnancy and the first ten moths of life and childhood allergy, allergy asthma and asthma. A matched patients-sibling study comparing patients with allergic asthma with their healthy siblings without any symptoms of allergic diseases. Allergic in patients and their siblings was determined by skin prick tests. We assumed that children having up to one positive skin prick test were considered to be allergic. Intake of paracetamole was assessed by standardized interviewer Yadministered, questionnaire. 22 pairs of allergy asthma patients`vs. non-allergic siblings were compared to determine the risk factors for allergic and asthma. While 12 pairs of allergic asthma patients vs. allergic siblings were compared to determine the risk factors for asthma. However, 35 pair of allergic asthma patients vs. non-asthmatic sibling ( with and without allergy) were compared to determine the risk factors for asthma, also 15 allergic siblings were compared with 19 non-allergic siblings (with asthma ) to determine the risk factors of Allergy. Results: Intake of paracetamole during pregnancy was associated with allergic asthma (P = 0,04). Intake of paracetamole between birth and ten months of age and between four to seven moths of age, was also found to be associated with non-allergic asthma (p = 0.004 and p = 0.03) respectively. Usage of paracetamole during pregnancy and during early months of life may play a role in the development of allergic and non Yallergic in children. Conclusion: Intake of paracetamole during pregnancy and during first month of life is associated with an increase risk of childhood asthma. Cow's milk allergy in premature infants: a ready-to-use cow's milk atopy patch test before starting an amino-acid formula Christophe Dupont 1 , A. Lapillone 1 , Pascale Soulainesv, Pierre Henri Benhamouv, and Nathalie Donne 2 . 1 Hopital Saint Vincent de Paul, Pediatrics, Paris, France; 2 Dbv Technologies, Dbv Technologies, Boulogne Bllancourt, France. Background: Cow's milk (CM) allergy (CMA) is an infantile disease, usually appearing in the first months of life, with very few cases up to now reported in the premature infants. The study was designed to detect CMA in premature infants with digestive symptoms, based on a ready-to-use Atopy Patch Test (APT), Diallertest\. Methods: During the year 2006, 13 premature infants (31 wT2 weeks, 4 girls), aged 42T18d , receiving formula for prematures with CM and presenting with digestive symptoms (rectal bleeding, 5, vomiting,1, diarrhea, 3, severe reflux, 2, others, 2) were tested for CMA by Diallertest\. Whatever the results, all children received an amino-acid formula (Neocate\ and the outcome of symptoms was evaluated one month later. Results: Among the 13 premature infants tested, the digestive symptoms disappeared under amino-acid formula in 10. Diallertest\ was positive in 7 cases. All infants with a positive Diallertest\ improved with the amino-acid formula (no false positive) and 1 child with a negative Diallertest\ improved with the diet (1 false negative). Conclusion: CMA is a frequent and mostly underestimated cause of digestive symptoms in premature infants, mostly fed with CM based formula. APT seems an appropriate method to diagnose CMA in this age range and Diallertest\ might thus be a useful tool in the neonatal ward. Which is more important regarding psychological consequences: level of asthma control or overall disease severity in paediatric asthma? Alexandra Szabó, Györgyi Mezei, and Endre Cserháti. Semmelweis University, First Department of Pediatrics, Budapest, Hungary. Aim: The study's objective was to examine the consequences of overall asthma severity and current asthma control regarding psychological symptoms in paediatric asthma. Methods: 108 patients, age: 11.75T3.10 (meanTSD) years; (boys 11.6T2.8 years and girls 12.1T3.7 years) completed the Child Depression Inventory (CDI), the State Trait Anxiety Inventory for Children (H.STAIC), the Pediatric Asthma Quality of Life Questionnaire (PAQLQ), and a symptom score. FEV1 was also measured. Results: Mean FEV1% was 97.4T12.8. Twenty-three patients (21%) had intermittent asthma, 40 patients (37%) had mild persistent, 43 patients (40%) had moderate persistent, 2 patients (2%) had severe persistent asthma. The only factor associated with depression is: FEV1% less than 100. Factors associated with anxiety are: night symptoms, asthma control. Factors associated with quality of life are: the age at the beginning of asthma, asthma duration, current symptoms, night symptoms, day symptoms. Factors that have no association with the above are: disease severity, other allergies, skin test positivity. Conclusion: Not the overall disease severity, other allergic co-morbidities, but more the current asthma status and level of control alter the psychological status of the asthmatic child. They can distinguish between very small changes in the level of control. Thus professional asthma care should focus on achieving the best possible asthma control at all times. Changes of induced sputum cytology by leukotriene antagonists Tetsuya Takamasu, Yukinori Enomoto, Naoka Ito, Chisato Inuo, and Kazuyuki Kurihara. Kanagawa Children's Medical Center, Department of Allergy, Yokohama, Japan. Inhaled corticosteroids $B!J (BICS $B!K (B and leukotriene receptor antagonists $B!J (BLTRA $B!K (Bare major controllers for bronchial asthma. In our previous observation, ICS decreased eosionophils, and increased neutrophils in induced sputum. In this study, we examined changes of induced sputum cytology after starting LTRA. Methods: Subjects were 8 patients (mean age was 11T3 years, 6 boys and 2 girls) who were prescribed LTRA for asthma control. Before and 1 month after starting LTRA (pranlukast for 6 cases, montelukast for 2 cases), we obtained sputa by inhalation of 4.5% saline. Samples were treated for cell counts and differential cells by Eosin stain. Results: Cell numbers before and after starting LTRA were 31T30Â10 4 cells/ ml, and 21T17Â10 4 cells/ml, respectively (n.s.). The percentage of eosinophils were 13T11%, and 3T2% (pG0.05), and that of neutrophils were 79T17%, and 76T27% (n.s.). Conclusion: LTRA decreased sputum eosinophils as well as ICS, however did not increase neutrophils. Abnormal lung function and bronchial hyperreactivity in early infancy Y Y Y a predictor for severe respiratory syncytial virus infection? Porntiva Poorisrisak, Lotte Loland, and Hans Bisgaard. Gentofte Hospital, Danish Pediatric Asthma Centre, Hellerup, Copenhagen, Denmark. Background: Two-three percent of infants react with severe bronchiolitis to infection with Respiratory Syncytial Virus (RSV). The cause for such exaggerated response to a common infection is unknown, but it has been suggested that it may be associated with pre-existing abnormal pulmonary function. Aim: To compare lung function and bronchial hyperresponsiveness in newborns who later develop severe RSV bronchiolitis and infants who do not develop such severe infection. Methods: A prospective birth cohort study (Copenhagen Studies on Asthma in Childhood) was conducted in 411 infants of asthmatic mothers. Lung function was scheduled at 1 month of age by means of the raised volume rapid thoraco-abdominal compression technique during sedation measuring forced expiratory volume at 0.5 s (FEV0.5). Bronchial responsiveness was determined by metacholine challenge using transcutaneous oxygen as endpoint (PD15-TcO2). We defined severe RSV bronchiolitis as a child admitted to hospital or requiring medication with high-dose inhaled budesonide (9800mcg) or oral prednisolone for RSV-verified bronchiolitis. Results: In this prospective cohort study 23 children developed severe RSV infection before age 2 (mean age, 8 months). All completed baseline lung function measurements before subsequent RSV infection. Children with severe RSV infection and controls never presenting severe RSV bronchiolitis did not differ significantly at baseline lung function (FEV0.5) or metacholine challenge (log PD15-TcO2); p-value90.1 for all comparisons. Conclusion: Severe RSV infection in infancy was not associated with preexisting abnormal lung function and bronchial hyperreactivity. Background: Several studies have explored the role of probiotics in the treatment or prevention of atopic diseases. The findings were however inconclusive. Objective: To assess the effect of Bifidobacterium longum and Lactobacillus rhamnosus GG supplementation in the first 6 months of life on the incidence of eczema and allergen sensitization in the first year in high risk Asian infants. Materials and Methods: Infants (n = 253) with a first degree relative with a family history of allergic disease and sensitization to dust mites were randomized in a double blind, placebo-controlled manner to receive a cow`s milk based infant formula with or without probiotics (daily dose of 1x10 9 colony-forming units (CFUs) of Bifidobacterium longum and Lactobacillus rhamnosus GG) for the first 6 months of life. Subjects were assessed at 1, 3, 6 and 12 months for atopic diseases and skin prick test was conducted at the 12 month visit. Results: Out of 253 subjects, 235 were followed-up to 12 months (drop out rate 7%). The subjects in the two treatment groups were comparable in terms of demographic and birth characteristics, except for gender and birth order. Incidence of eczema in the probiotic (n = 33/122; 27%) group was found to be similar to placebo (n = 33/113; 28%) (OR, 1.07; 95% CI, 0.60 to 1.89). There was no difference in the rate of sensitization to common allergens (probiotic = 25% vs placebo = 21%, OR, 0.78; 95% CI, 0.42 to 1.45). Sensitization to dust mite allergens (Dermatophagoides pteronyssinus, Blomia tropicalis) was the most common (Probiotic = 20% vs Control = 18%), followed by eggs (Probiotic = 6% vs control = 5%). None of the subjects were sensitized to cow`s milk or soy. Potential confounding factors including birth order, household size, smoking exposure, presence of pets and breastfeeding did not affect these outcome measures. Adjustment for imbalance of gender and birth order between treatment groups did not affect the findings significantly. Conclusion: This study did not show a protective effect of probiotic supplementation for the first 6 months of life on eczema or allergen sensitization in high risk Asian infants at 1 year of age. The prevalence of eczema in our cohort is also lower than in reported studies. Detection of fecal Bifidobacterium infantis in the first year of life in infants at risk of atopy supplemented with Lactobacillus rhamnosus GG and Bifidobacterium longum from birth till 6 months old G.C. Yap 1 , K.W. Mah 1 , C. Lay 2 , L.P.C. Shek 1 , M. Aw 1 , K.Y. Chua 1 , G.W. Tannock 3 , and B.W. Lee 1 . 1 National University of Singapore, Department of Paediatrics, Faculty of Medicine, Singapore, Singapore; 2 A*STAR, Institute of Microelectronics, Singapore, Singapore; 3 University of Otago, Department of Microbiology and Immunology, Dunedin, New Zealand. Background: Studies have suggested that oral administration of probiotic bacteria in early life may positively modulate the immune system of infants at risk of atopy towards a non-allergic state. Aim: This study aims to monitor the intestinal transit of a probiotic supplementation (Lactobacillus rhamnosus GG and Bifidobacterium longum BB536) in the first 6 months of life in at risk infants (first degree relative with allergic disease) participating in an ongoing randomized double-blind placebo controlled trial. Effect of this supplementation on Bifidobacterium infantis prevalence was also investigated. This particular bifidobacterial species has been implicated as a potential signature of infants at low prevalence of atopy. Methods: Newborns at risk of allergies received either probiotic preparation (n = 55) or placebo (n = 48) daily for the first 6 months of life. Analysis of fecal bacteria and clinical examinations were conducted serially at birth, 1, 3, and 12 months. Bacterial DNA extracted from fecal samples was evaluated with nucleic acid amplification approach using specific PCR primers targeting Lactobacillus rhamnosus GG and Bifidobacterium infantis. Results: All extracted DNA samples were amplified with universal primers targeting a conserved region of 16S rRNA to ensure extraction of intact DNA. Lactobacillus rhamnosus GG was more commonly detected in the probiotic group compared to the placebo group [at 3 days (94.1% vs 6.8%), 1-month (92.7% vs 14.9 %), 3-month (86.5% vs 21.3%, respectively) (pG0.0001)] but not significantly at 1-year old (28.8% vs 14.9%, p90.05). Infants supplemented with probiotics were also more likely to harbour Bifidobacterium infantis at 3 months (n = 19/52; 36.5%) compared to placebo (n = 7/47; 14.9%) groups (p = 0.0267). Conclusion: Our data suggests that probiotic supplementation from birth may modulate the intestinal bifidobacterial species composition. Its effect on allergy outcomes remains to be examined. Objective: The main objectives of this study was to establish allergen sensitization in a large proportion of children who required hospitalisation for a severe exacerbation of their asthma. Material and Methods: Patients hospitalised at the Victor Fouche Hospital were monitored from 2002 to 2005. Patients were contacted by phone after leaving the hospital. Afterwards, they were skin tested with 16 common aeroallergens, including 4 mite species, 3 cockroaches, cat and dog, grasses and 4 foods. A detailed questionnaire concerning environmental and social risk factors was answered by the parents. Results: A total of 632 hospitalisations were counted in this period of time. A total of 492 children (294 boys and 198 girls; mean age 5.93 years) were hospitalised. The mean number of hospitalisations per child was 1.3 (1 to 8 times). The mean days of admission per patient was 2.81 (1 to 27). The number of hospitalisations in 2002 were 156, in 2003, 191; in 2004, 148 and in 2005 137 . A total of 269 patients (54.67%) (159 boys and 110 girls) agreed to undergo skin testing. From this group, 11 had been skin tested prior to the entry in the study and 3 had started, and abandoned, immunotherapy. One hundred ninety six (72.86%) children had at least 1 positive skin test; 158 (58.74%) were sensitised to at least 3 allergens. 183 (68.03%) were positive to at least 1 mite species; 99 (36.8%) to at least 1 cockroach species; 63 (23.42%) to at least one food, among these, 61 were sensitised to shrimps; 18 (6.69%) were sensitised to cat and 14 (5.2%) to dog and just 5 (1.86%) to grass pollen. Conclusion: We have identified a high rate of sensitization to aeroallergens in a large cohort of young paediatric patients with severe asthma that required hospitalisation in Martinique. Mites, cockroaches and shrimps accounted for the largest number of sensitisations. These results are similar to those obtained in a control group of allergic children being evaluated for allergic respiratory diseases during the same period of time. Background: In tropical regions, there is a large proportion of the population which has been in contact with intestinal parasites, including the roundworm Ascaris spp. There are conflicting reports on the protective effect or harmful influence of sensitization to Ascaris spp. in patients with allergic respiratory diseases, such as allergic asthma. Objectives: to investigate specific IgE levels in children hospitalised with severe asthma in the tropical island of Martinique. Materials and Methods: From 2002 to 2005 we investigated specific IgE levels in the serum of children hospitalised for severe asthma in Fort de France, Martinique. As controls we used a group of 484 children (mean age 7.42) evaluated for allergic respiratory diseases at an outpatient clinic. Specific IgE to Ascaris spp. was measured by the CAP system (Phadia). Results: A total of 117 children (68 boys and 49 girls; mean age 6.45) were investigated. Mean number of hospitalisations and of days stayed in the hospital were 1.35 and 3.02 days, respectively. A total of 60 children (51.28%) had a positive determination (mean value 6.69 kU/L). The mean number of hospitalisation in the positive group was 1.28 and the mean days stayed at the hospital, 3.05 days. 47 children stayed 3 or more days in the hospital; in this group there 22 Ascaris spp. negative and 25 Ascaris spp. positive children (NS). In the negative group, the mean number of hospitalisation was 1.42 times and the mean days stayed at the hospital, 2.98 days. In the control group, 215 (44.42%) had a positive specific IgE determination to Ascaris spp. Conclusion: In contrast to what has been proposed for other tropical countries, sensitization to Ascaris spp. seems not to influence the severity of asthma in Martinique. Similar rates of sensitization to Ascaris spp. are detected in allergic children who do not require hospitalisation for their allergic asthma. Demographic of monitoring and treatment of childhood asthma in Surakarta, Central Java, Indonesia Johannes Ridwan T. Sugiarto 1 , and Prof. Hugo Van Bever, PhD 2 . 1 Dr. Oen Hospital, Pediatrics, Surakarta, Indonesia; 2 National University Hospital, Pediatrics, Singapore, Singapore. Background: In Surakarta, one of the districts in Central Java, Indonesia, with a population of around one million, many areas house industrial companies, leading to high pollution, that can trigger childhood asthma. As the most common chronic illness in children, childhood asthma causes school absences and limitations of children`s activities, making asthma especially serious for them. Methods: Standardized questionnaires were distributed randomly to 200 doctors in Surakarta, who are general physicians, general pediatricians and pediatric pulmonologist. Results: Of the 200 questionnaires, 78 were returned: 74 from general physicians and 3 from pediatricians. Most of the doctors never use score cards to monitor childhood asthma, never used spirometry and never used peak flow meters. For the treatment of acute asthma methylprednisolone was first choice in 83%, while 17 % used dexamethasone. For maintenance treatment of childhood asthma no physician used montelukast because this drug is not available in Surakarta. However, as a first choice maintenance treatment, most physicians used a long-acting beta-agonist (LABA) in combination with an inhaled corticosteroid in infants, preschoolers and older children. Conclusion: Striking results were the low usage of score cards, lung function testing in monitoring childhood asthma, and the high usage of a LABA in combination with inhaled corticosteroids as first choice maintenance treatment, suggesting that guidelines on asthma monitoring and treatment are not strictly followed in Surakarta. Frequency of infections, atopic eczema and asthma outcome in infancy John Warner 1 , Augustin Huret 2 , and Marie-Etienne Pinelli 3 . 1 Imperial College, Paediatrics, London, United Kingdom; 2 Business EffiScience, Science strategy, Paris, France; 3 UCB Pharma, Global Medical Affairs-Allergy, Brussels, Belgium. Studies of association between frequent early infection and allergic disease outcomes have produced diametrically opposed results. Some have associated reduced frequency and others increased frequency of infection with more allergic disease. We have employed a unique algorithm (Business EffiScience) to the analysis of a large database from early prevention of atopic asthma in children (EPAAC) trial. 510 infants between 1Y2 years of age with atopic eczema (SCORAD9 10) entered the study and 434 completed 18 months follow up. As there was no difference between active and placebo intervention all subjects were merged for the purposes of this analysis which were to establish which factors at recruitment either singly or in combination had the greatest impact on the subsequent development of asthma defined as greater than or equal to three episodes of wheezing and/or paroxysmal cough disturbing sleep for at least three consecutive nights. To analyze the impact of early infection on asthma we analysed the effect of the number of infections per year of infants from birth to asthma onset or end of the study when asthma was not observed. A bell shaped curve was identified with 27% infants less often developing asthma if they had 1.5 or less infections per year, 25% more often to have asthma with 1.5 to 4 infections per year and 11% less often to have asthma with more than 4 infections per year. The only other interaction was with proximity to a factory as a surrogate for pollution exposure which only increased the risk of asthma if there was a frequency of infections between two and five per year. Thus the combination of atopic eczema with raised IgE and proximity to a factory was associated with a 77% uplift in frequency of developing asthma if the frequency of infections was between 1.5 and 4 per year. The bell shaped curve of frequency of infection in relation to asthma risk may well explain the discrepant results in various studies. Very low frequency of infection occurs in infants with an effective TH1 response and therefore reduced probability of developing allergic disease while those with a very high frequency of infection have a greater probability of inducing TH1 responsiveness while normal rates of infection in an atopically predisposed individual will not modify the outcome. Challenges of Fpeanut challengeS rinivas Bandi, and Scott Hackett. Birmingham Heartlands Hospital, Paediatrics, Birmingham, United Kingdom. Background: The diagnosis of peanut allergy has important consequences for patients and their families. An accurate diagnosis of peanut allergy is essential as it is usually life long and can potentially be fatal. The cornerstones of diagnosis are a detailed history, skin prick tests (SPT), IgE estimation with or without an oral peanut challenge. Objective: 1. To demonstrate the importance of peanut challenges in diagnosing children with peanut allergy 2.To see whether we can predict children who are likely to outgrow peanut allergy 3.To assess the safety of peanut challenges. Methods: A retrospective study of peanut challenges performed at a tertiary care paediatric hospital allergy clinic between January 1997 and December 2006. Results: Of the 79 peanut challenges performed, (data available for 77), 14 (18%) were positive. All children had a history suggestive of peanut allergy and 72 had a skin prick test (SPT) or IgE RAST performed prior to the challenge (SPT-63, IgE-6, SPT and IgE-3) The results (72) of SPTTIgE RAST: &23 were negative (SPT-20, IgE-2, SPT+IgE-1) &49 were either positive for RAST or SPT or both . Positive challenge n=14 Negative challenge n=52 Q10 mm 3 2 6 Y 9 mm 7 7 e5 mm 3 21 negative 1 22 Only 1 child who had a negative SPT developed a positive challenge (4%), whereas 36/49(73%) children with positive SPT or RAST had a negative challenge. Further analysis of these children who had a negative challenge shows that majority had a SPT of e 5 mm (21/36), where as 7 had SPT of 6Y9mm, 2 had SPT of Q 10mm. 3 had positive IgE RAST and 2 had both SPT and IgE RAST positive. SPTs e 5 mm was strongly associated with a negative peanut challenge; Fishers Exact test p = 0.02 Adverse clinical effects of positive peanut challenges included urticaria, lip swelling, vomiting and in one anaphylaxis; 6 required medical treatment (5-antihistamines, 1-adrenaline). Conclusion: The majority of children with a diagnosis of peanut allergy, based on history and skin prick / antibody tests had a negative challenge. Peanut challenges are not warranted in patients with a definite history of peanut anaphylaxis with positive SPT. We recommend that where the history is suggestive but SPT is negative or borderline positive, i.e. e 5 mm, a peanut challenge is necessary to confirm the diagnosis. We plan to SPT and RAST test all patients before they have peanut challenge to see if this improves sensitivity and specificity of our challenges. Effect of lysed Enterococcus faecalis FK-23 (LFK) on intestinal microflora in antibiotic treated mice Takashi Shimada 1 , Tadao Enomoto 2 , and Lei Cheng 3 . 1 Nichinichi Pharmaceutical Co., Ltd., Central Research Laboratories, Iga-city Mie, Japan; 2 Japanese Red Cross Society Wakayama Medical Center, Department of Otolaryngology, Wakayama, Japan; 3 Nanjing Medical University, International Centre for Allergu Research, Nanjing, China. Background: Antibiotic use in infancy may be associated with an increased risk of developing allergic diseases. Lysed Enterococcus faecalis FK-23 (LFK) could suppress the allergic responses; however, the mechanism responsible for this phenomenon remains unclear. Objective: To evaluate the effect of LFK on intestinal microflora in antibiotic treated weaning mice. Methods: Three-week-old BALB/c mice were sensitized with cedar pollen allergen to establish an experimental model. Orally administered erythromycin, one kind of macrolide antibiotic, was used for the experiments. The intestinal microbiota, and the allergen-induced accumulation of eosinophils and IgE level in sera were determined in the control, antibiotic, LFK and antibiotic-LFK groups (n = 7, respectively). Results: The total aerobes, total anaerobes and Enterococcus of intestinal microflora were not significantly different among all groups. Lactobacillus was distinctly eliminated in the mice exposed to erythromycin on day 7 and totally recovered in erythromycin-treated mice with LFK intervention on day 28, but could not recovered in the erythromycin-treated mice without LFK intervention. Conclusion: LFK improved the intestinal ecosystem disturbed by antibiotic use, and may prevent subsequent development of allergy. Background: Mycobacterial tuberculosis is a major cause of mortality and morbidity worldwide.Infection with this bacteria is known to induce the development of autoantibodies and a few of these antibodies are also known to be diagnostic markers for some other diseases. ANCA`S are one of autoantibodies used in clinical setting for diagnosing systemic vasculitic syndrome . More than 20 studies investigating ANCA positivity in diseases other than small vessel vasculitis . This study was undertaken to determine the prevalence of ANCA in pulmonary tuberculosis (TB) which could lead to false diagnosis of wegner`s granulomatosis or vice versa. Materials and Methods: In a caseYcontrol study 32 consecutive smear positive pulmonary TB patients and 32 normal individuals were studied . All cases and controls screened for ANCA by indirect immunoflurecent (IIF) and also myeloperoxidase antibodies (anti-MPO) and proteinase 3 antibodies (anti-PR3) were tested by ELISA. Results: Perinuclear pattern (P-ANCA) was detected in 25% of cases and 6.25% of controls and cytoplasmic pattern(C-ANCA) in 3.1% of both cases and controls by IIF assay. ANCA specificities by ELISA in cases revealed that 75% had anti-myleperoxidase and 12.5% had anti-Proteinase 3. In controls 3.12% had anti-MPO and no person had anti Y PR3 . The positive ANCA significantly correlated with tuberculosis (p valueG0.018) and also positive anti-MPO significantly correlated with TB (p valueG0.01). Conclusion: Autoantibodies especially ANCA`S may present in both disease, Wegner Granulomatosis and Tuberculosis .The presence of autoantibodies in TB patient could have a multifactorial etiology. The presence of human T lymphocytes reactive to heat stress proteins may be an important target of immune response against certain intracellular autoantigens such as anti-MPO from PMN added to the mechanism of molecular mimicry would explain the association of ANCA and TB. The cells in psoriatic skin Rationale: The psoriasis is an autoimmune disease with significant role of the immune system. Methods: We investigated the group of 16 patients with active form of chronic psoriasis vulgaris before and after treatment with cyclosporine A. The results was compared with the control group of the patients without any skin disease. In the samples of the skin tissue we detected the cells with surface markers CD14, CD45, CD3, CD19, CD46+56, CD4, CD8, HLA-DR, CD25 (we have used the monoclonal antibodies Immunotech, Becton-Dickinson). Results: The lymphocytes represent 7,3% in the samples of the skin in patients in the active state of psoriasis, 2,1% in control group (pG0,0001) and 3,1% in patients after treatment (pG0,001).Cytotoxic T-cells (CD3+CD8+) represent 37,8% in the samples of the skin tissue in psoriatic patients, 32,4% in control group and 28,4% after treatment, The cytotoxic T-cells expressed HLA-DR in 25,6% in psoriatic patients before treatment, in 10,7% after treatment and in 11,4% in the control group. We found 44,3% of the lymphocytes with CD3+CD4-CD8-phenotype in the skin of patients before treatment, 29,6% (pG0,05) after treatment and 10,3% in the control group. The expression of the surface marker CD25 was lower in the active stage in comparison with control group ( 9,6% vs 13,1%), its expression increased after treatment at 38,4%. Introduction: Rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) are autoimmune diseases with respectively T cell mediated and B cell mediated mechanisms. Endotoxin (ET) from gram negative flora can rapidly induce or reactivate experimental arthritis, probably by stimulation of secretion of Th1 inflammatory cytokines. ET is a potent polyclonal Blymphocyte activator. This study investigates anti-endotoxin humoral immunity in patients with RA and SLE. Methods: 27 patients were studied including: Group I-18 patients with RA; and, Group II-9 patients with SLE. The age range was 30Y50 years. Disease duration was from 6 months to 10 years. Disease status based on clinical, including joint involvement, and laboratory data were less than or equal to class II degree of activity. Peripheral blood was obtained and serum analyzed. Quantification of IgA, IgM, and IgG anti-ET antibodies was done by ELISA. ET from Esherichia coli K30 (09:K; O:H12) was employed as the antigen. The control group included 32 healthy donors. Results: Patients with RA had levels of IgA anti-ET and IgM anti-ET which were 2.3 fold less (pG0.001) than the average in healthy controls. The levels of IgG anti-ET antibodies were not different from the healthy control group. Patients with SLE had levels of IgG anti-ET antibodies which were 2.2 fold greater (pG0.01) than normal controls. These same SLE patients had levels of IgA anti-ET and IgM anti-ET which were respectively 2.3 (pG0.001) and 2.1 (pG0.01) fold less than in the healthy control group. Reduced IgA anti-ET levels occurred in 56.7% (pG0.001) and reduced IgM anti-ET levels occurred in 53.6% (pG0.01) of SLE patients. Reduced IgA anti-ET levels occurred in 56.7% (pG0.001) and reduced IgM anti-ET levels occurred in 56% (pG0.001) of RA patients. Conclusion: Anti-ET IgG levels in RA patients demonstrate a normal level of response while in SLE patients greater than normal responses to ET occur Dysfunctional anti-ET immune responses occurred with respect to reduced IgM and IgA anti-ET antibodies in both SLE and RA patients. There may be a role for gut flora and immune responses to ET as factors modulating immune responses in RA and SLE patients. Activity of systemic lupus erythematosus in Mexican children. Correlation between medical clinical evaluation and evaluation based on the use of five indicators of activity Rosa Maria Cortes Grimaldo, Francisco Espinosa Rosales, Marcia del Carmen Perez Ruiz, Martin J. Penagos Paniagua, and Daniel Garcia Imperial. Instituto Nacional de Pediatria, Allergy and Immunology, Mexico, Mexico. A computer program has been developed to evaluate five of the most important SLE AI: SLEDAI, SLAM, BILAG, LAI, ECLAM. These indicators have a close correlation with each other in the case of adult patients and also, when they are compared with the expert opinion. On the contrary there is sparse information of the usefulness of these indicators in children with SLE. Objective: To determine whether is there a correlation between 5 different SLE AI in children and to assess the correlation between the clinical evaluation of activity by physicians of the Department of Immunology and that provided by the use of the five AI of SLE. Materials and Methods: A transverse analytical study was undertaken in patients under 18 years of age diagnosed with SLE at the National Institute of Pediatrics in Mexico City. A questionnaire was used containing every variable included in the five indices of SLE activity. Statistical analysis. The median and interquartil limits were used as descriptive ststistics because the variables did not show a normal distribution with the Kolmogorov-Smirnov test;. To ascertain the degree of association among quantitative variables, correlation test of Spearman was used. Sensibility, specificity and predictive values were estimated at different cut-off levels selecting the best one using Receptor Operator Curves (ROC). Results. We included 30 patients with SLE; 28 females (94%) and two males. Median age was 15 years (liq 13-17). The median time of evolution from the time of diagnosis was 41 months (liq 16-5). According to the expert 14 patients had active SLE (47%); according to the SLEDAI 13 patients had active SLE (43%) (p = 0.79). Conclusion: The indices with the greater correlation with the SLEDAI were the opinion of the expert and the SIS; those that best correlated with the appraisal of the clinician were the SLEDAI and the SIS. Sensibility and specificity for each one of them were 92% and 89% for the expert; 85% and 88% for the SIS and 85% and 89% for the SLEDAI respectively. The value of the *corticosteroids(CSP) in countering the residua of the protracted tuberculous infection/inflammation(TI) I. Khan Sameera, Munir Imran Khan, Ali Iftikhar, and M. Ishaq. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Purpose: (CS) BThe double edge sword^in function the favorable therapeutic advantage against disadvantage should be weighed when the indications arise. Methods: (TI) being a lingering process is associated with an enhanced degree of exudation,caseation fibrogenesis etc.there may be binding ,obstruction.constrictoion,compression of the surrounding inflamed surfaces,also the constitutional symptoms as a consequence of the release of the inflammatory products.Exudative changes in the cerebrospinal fluid pathways were associated with features of space occupying lesions. (TI) as pleural, pericardial and peritoneal; effusions were associated with fibrous bands formation, also exudative changes in and around the inflammed joint spaces may lead to interference with the function of these structures. In all these cases corticosteroids with in the therapeutic dose had been associated with a remarkable degree of improvement without any fibrous residua and prodromal manifestations were relieved favourably. In relatively seriously ill patients also patients with in the terminal stages of illness (at the verge of death) from advanced tuberculosis and inanition,corticosteroids may be indicated purely on humanitarian grounds as these(CS) detoxicate the patients and allow the chemotherapeutic agent to take their principal effects. Dosage regimen for adults 40mg/day in divided dose for the first day then 20mg/day in divided dose for 3Y6weeks to be tapered by 5mg at 3Y5 days intervals.For children 2mg/kg/day for those less 2 years, 1.5mg/kg/day for those more than 2years but less than 10 years age. Results: Patients with a reasonably certain degree of diagnosis of (TI), (CS) in concomitant anti-tuberculosis drugs had a remarkable therapeutic response evidenced as fall of temperature ,improvement in appetite and vigor. Conclusion: With un-certain diagnosis of (TI), (CS) may mask the untowards effect of chemotherapeutic agents,creating a false sense of improvement and may precipitate steroid dependence. Clinical Implication: The safety of (CS) is only out of question,if these had been indicated for the shortest possible time and then tailed off in a step ladder pattern.*Prednisolon sulphate. The prophylactic value of the low dose intravenous immunoglobulin(IVIG) in reducing the incidence of the infection in chronic lymphatic leukemia(CLL) and secondary hypogammaglobulinaemia M. Ishaq, I. Khan Sameera, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Objective: In the study concerened low dose of IVIG was as effective as high dose in reducing the risk of infection in patients with Chronic Lymphatic Leukemia(CLL) and secondary Hypogammaglobulinaemia without the inclusion of the control group, behind the idea to assess the IVIG superiority over the empirical therapy. Materials and Methods: 5 Patients with CLL with the background of hypogammaglobulinaemia(IgGG600mg/dl ) with a history of 2 infection episodes in almost 6 months were included, all had been randomly allocated to receive regularly an infusion of 300mg/kg IVIG every 4 weeks for 6 months then for financial reasons 4 could not afford the expense while 1 was continued with the ongoing IVIG schedule. Results: In response to IVIG there were immediate and accumulative increase in the serum IgG levels and an associated decrease in total and serious infections. In the 12 months study only 4 incidence of infection (1 with severe and 3mild) with 1 neutropenia observed. Four patients having completed 6months IVIG had an overall 50% reduction in the incidence of infection, while 1 patient with 12 months IVIG had 65%reduction in the incidence of the infection. Conclusion: Infection as the main cause of morbidity and mortality consequent upon the hypogammaglobulinaemia with the background of cell mediated immunity was associate with protracted cytotoxic drugs therapy. In the trials concerened, the protective values of IVIG had been time related than dose related (longer the period of IVIG therapy les was the incidence of infection). Bearing in mind the cost, IVIG is only advisable in pateints with life threatening infection and then too with lower dose to reduce the expense incur upon the treatment. khan et al aljh nsr nwfp pk. A six year old girl with recurrent varicella infections associated with a mutation in CD16 (FCGR3A, FcyRIIIA) Suthida Kankirawatana, Yuling Dai, and T. Prescott Atkinson. University of Alabama at Birmingham, Division of Allergy and Immunology, Department of, Birmingham, United States. Background: Natural killer (NK) cells are a subset of lymphocytes that play a pivotal role in innate immunity. NK cells recognize antibody-coated target cells using CD16 (FcyRIIIA), and kill the target cells by the process called antibody-dependent cell-mediated cytotoxicity. Quantitative and qualitative defects in NK cell function result in susceptibility to intracellular microbes. Methods: Lymphocyte subpopulations in peripheral blood were determined using flow cytometry. NK cell surface expression of CD56 and CD16 and the intracellular expression levels of perforin and granzyme B were analyzed by flow cytometry. Patient cDNA for CD16 was sequenced to screen for mutations within the coding region. Results: The patient is a 6 year-old girl who suffered from recurrent upper and lower respiratory tract infections. She had experienced two episodes of varicella despite receiving the vaccine, and, despite this, varicella antibody was negative. She also has numerous warts on her fingers. Immunologic studies revealed low serum immunoglobulins (IgG 413, IgA 38, and IgM 37 mg/dL at 42 years), but she had exuberant antibody production in response to pneumococcal vaccine. Peripheral blood lymphocyte phenotyping revealed slightly low T cell numbers for age with preservation of the CD4/CD8 ratio, B cells were normal in percentage and number. CD56+ NK cells were 6 % of the lymphocyte gate, but the expression of CD16+ was less than 1 % using Leu11c/B73.1. However, analysis of NK cells using a different antibody to CD16, NKP46/9E2, revealed normal CD16 surface expression. Sequencing of peripheral blood cDNA demonstrated a homozygous TA transversion at base pair 489 in the CD16 mRNA sequence producing a Leu to His substitution at residue 102 in the peptide sequence, a mutation previously described in a patient with recurrent orolabial HSV exacerbations and respiratory infections (Jawahar et al 1996, Clin. Exp. Immunol. 103:408 Background: Cicatricial pemphigoid (CP) is a rare autoimmune subepithelial blistering disorder, characterized by predominant involvement of the mucous membrane and occasionally the skin, chronic course, and tendency towards scarring of the affected areas. Most common sites of involvement are oral mucosa and conjunctivae. In our hospital, this is the second case of CP, and the first case with skin involvement for the last five years. Case: A 72 years old male had undergone dysphagia and blistering disorder in the skin for six month. Clinical manifestation were erythematous and erosions in oral mucous, tongue, and crust in lips. Skin lesions were tense hemorrhagic blisters with negative Nikolsky sign, erosions and crust in chest, axilae, abdomen and left lower arm. Few slight atrophic scars were also found. Pharyngoscopy showed erythematous and brittle of the pharynx, soft palate and tongue. The histopathology examination showed subepidermal bullae, epidermal atrophy, sparse perivascular and periappendageal inflammatory infiltrates consisted of mostly lymphocytes with few polymorphonuclear leukocytes and eosinophils and fibrosis in the dermis. The patient favorably responded to oral steroid, prednisone 45 mg daily in combination with dapsone 100 mg daily, and improved after seven weeks. Discussion: CP is a rare autoimmune subephitelial blistering disorder, characterized by predominant involvement of the mucous membrane. According to the literature, skin involvement in CP occurs in about onefourth of cases. Diagnosis in this case was suggested by the presence of scarring lesions in pharyngeal mucous membrane with dysphagia, slight atrophic scar in the skin and confirm by histopathology. Prednisone and dapsone are used due to severe disease and to prevent potential severe complication. Conclusion: CP with skin involvement is uncommon. Diagnosis is based on clinical manifestations and, confirmed by histopathology. Therapy with prednisone and dapsone shows good result in CP. Introduction: IgA deficiency (IgAD) is the most common immunoglobulin deficiency and one of the most frequent primary immunodeficiencies. The prevalence of IgAD in Icelandic blood donors has recently been estimated to be 1:570. Correlations between having IgAD and various autoimmune diseases, including autoimmune thyroid disease (AITD), have been rapported. Materials and Methods: IgA was measured in serum from 319 patients with thyrotoxicosis and 199 patients with DM TI. In comparison IgA levels from 609 blood donors was evaluated. IgA was measured with Beckman Array 360 System Nephelometry. IgA levels were also studied in relation to titers of TSH Receptor Antibodies (TRAb) and Thyroid Peroxidase Antibodies (anti TPO). TRAb was measured with ELISA and anti TPO was measured using FEIA. Results: None of the 319 thyrotoxicosis patients had IgAD (IgA in serum G 0,05 g/L). Women with thyrotoxicosis had significantly lower levels of serum IgA compared to men with thyrotoxicosis ([IgA] male = 2,67 T 1,16 g/L vs. [IgA] female = 2,11 T 1,04 g/L; PG0,001). However, similar gender difference was also found in the blood donors. IgA levels increased significantly with age in patients with thyrotoxicosis (PG0,01) and a similar correlation was found in the blood donor group. A negative correlation was found between levels of IgA in serum and TRAb (correlation coefficient=j0.322, P = 0,0455) and also between levels of IgA and antiTPO (correlation coefficient=j0,376, P = 0,0204). One individual with DM TI had hypogammaglobulinemia, but none had selective IgAD in that cohort. Interestingly, men with DM TI had lower serum IgA than women. Conclusion: These results are a strong indication that the prevalence of IgAD is not increased amont patients with thyrotoxicosis or DM TI in Iceland. The negative correlation between IgA levels and thyroid autoantibodies suggests a possible role of IgA in the pathogenesis of autoimmune thyroid diseases. IgA might have a protective role in the formation of autoantibodies against thyroid structures or a mutual autoimmune mechanism might cause brake-down of IgA and the formation of thyroid autoantibodies. ALLERGIC & IMMUNOLOGIC MECHANISMS Background: Asthma is a chronic inflammatory disorder of the airways associated with reversible airway obstruction and airway hyperresponsiveness. It has been thought that Th2 response play a critical role in the pathogenesis of asthma. However, recently, there have been reported that Th1 response also play a important role in pathogenesis of asthma. In Th1 immune response, IL-12 and STAT4 are key molecules. IL-12 play a important role in Th1 development, and promotion of Th1 responses and the production of interferon-gamma. And STAT4 are believed that it also play a important role in Th1 immune rsponse. But it still remains unclear whether these molecules play key roles in asthma development in vivo. Methods: To evaluate the roles of IL-12 and STAT-4 in murine model of asthma, 6-week-old IL-12 receptor $2 (j/j), STAT4 (j/j) null mutant, and their littermate wild type control mice were used, respectively. The mice were sensitized four times with 756g of OVA at days 0, 1, 2, and 7 in the presence or absence of 106g of LPS, and then challenged 4 times intranasally with 506g of OVA at days 14, 15, 21, and 22. STAT4 (j/j) and wild type control mice were sensitized and the challenged in the same manner. After challenging, we tested the airway hyper-responsiveness (AHR) against methacholine challenge. The mice were sacrificed 48 hours after last allergen challeng, and various immunological parameters were evaluated such as BAL fluids cellularity. Results: Total cell number of BAL fluids was increased in the mice sensitized with LPS and OVA compared with mice sensitized with OVA only. BAL fluids flammatory cells were similar between IL-12 receptor l$2 knock-out mice and wild type mice, but significantly decreased in STAT4 null mutant mice. Interestingly, IP-10 level in BAL fluids was partially decreased in IL-12 (j/j) mice, but markedly decreased in STAT4 (j/j) mice. Conclusion: In Th1 experimental asthma, airway inflammatory responses are partially dependent on IL-12, but dependent on STAT4 signaling pathway. The role of the sensitized alveolar macrophage in T cell proliferation Bo Ram Bang, Eunyoung Chun, Heung-Woo Park, Yoon-Seok Chang, Sun-Sin Kim, Sang-Heon Cho, Kyung-Up Min, and You-Young Kim. Seoul National University, College of Medicine, Institute of Allergy and Clinical Immunology, Seoul, Republic of Korea. Background: It is well known that alveolar macrophage (AM), which composes more than 90% of the bronchial alveolar cells, is the first barrier protecting lung from the harmful antigens. However, few different studies suggest that sensitized alveolar macrophage could exacerbate the asthma phenotype such as airwayhyperresponsiveness (AHR). The exact roles of alveolar macrophages in asthma have been unknown. Thus we undertook to determine whether sensitized alveolar macrophage could induce T cell proliferation in asthma animal model. : BALB/C mice were intraperitoneally sensitized by ovalbumin with alum on day1 and 7, followed by challenge with 1% ovalbumin on day 21, 22, and 23. The lung cells were isolated and analysed by FACS analysis. T cells isolated from the bronchial lymph node and bronchial alveolar lavage (BAL) cells were cocultured with or without ovalbumin in vitro in order to determine ability of sensitized alveolar macrophages to induce T cell proliferation. Results: After challenge, the percent of CD11chigh/CD11blow lung cells (alveolar macrophage) were not changed, but the MHC II expression of CD11chigh/CD11blow lung cells increased compared to PBS group. The proliferation of T cells increased 3 times in IP group than other groups without stimulation of ovalbumin. Furthermore, the BAL cells of IP group with stimulation of ovalbumin induced 8 times increase of T cell proliferation compared to that of other groups. Conclusion: It is suggested that the sensitized alveolar macrophages in BAL could facilitate the proliferation of T cells in asthma animal model. Toll-like receptor 2, 4, and 6 expression and function in peripheral blood mononuclear cells from asthma patients Background: Asthma is a chronic inflammatory lung disease that is caused by impairment of adaptive immune system like many immunologic diseases. However, the pathogenesis of this unwanted response has not well determined. Toll-like receptors have been shown to play a pivotal role in both innate and adaptive immune responses. Recently, many research groups have hypothesized that these TLRs are likely to play important roles in asthma pathogenesis, but, most studies are focused on the effect of TLR polymorphism. Therefore, we investigated whether the expressions of several TLRs are different in asthma patients compared to normal subjects and if so, whether the functionality of these receptors could be related to this change. Methods: The expressions of TLR2, 4, and TLR6 on peripheral blood mononuclear cells (PBMCs) from asthma patients and normal subjects were analyzed by flow cytometry. To study the functional responses of these receptors, PBMCs were stimulated with PGN and Pam3Cys as TLR2 ligands, LPS as a TLR4 ligand or FSL-1 as a TLR2/6 ligand for 24 hours and the amounts of TNF-" and IL-1$ were determined by ELISA. Results: The expression of TLR2 was up-regulated on PBMCs from asthma patient as compared to normal subject. Upon stimulation with PGN or Pam3Cys, TNF-" and IL-1$ production significantly increased in asthma patients. In contrast, the response to LPS (TLR4 ligand) stimulation on PBMCs was higher in normal subject than asthma patients, although the expression of TLR4 was not significantly different between asthma patients and normal subject. In case of TLR6, surface expression was significantly reduced in asthma patients. However, no difference was observed in the amounts of TNF-" and IL-1$secreted from PBMCs treated with FSL-1. Conclusion: Our data suggest that the difference of the phonotypical expression and the functional responsiveness of TLRs might be related with pathogenesis of asthma. Influences of anti-IgE antibody omalizumab on allergen-induced airway inflammation and bronchial hyperresponsiveness in murine models of asthma Background: The aim of this study was to examine the effects of treatment with omalizumab, an anti-immunoglobulin E antibody, on allergic-airway responses in mice after inhalation of the naturally occurring aeroallergen Aspergillus fumigatus (Af) and to examine the effects of omalizumab on specific immune responses to a defined protein antigen with the use of an ovalbumin (OVA) model of asthma. Methods: Mice were subjected either to repeated, intranasal application of Af extract or to intraperitoneal immunization with OVA, followed by inhalation challenge. Omalizumab or a control fluid was given daily throughout the sensitization process. Immunoglobulin E (IgE) levels, bronchoalveolar lavage-fluid cytokines and cytology, lung histology, and physiologic responses to methacholine were assessed in the allergen-treated mice. Anti-OVA IgE responses and OVA-driven T-cell cytokine production were examined. Results: Treatment with omalizumab did inhibit bronchial inflammation and bronchial hyperresponsiveness in both Af-and OVA-treated mice. This inhibition required that omalizumab be administered concurrently with allergen sensitization, indicating that the attenuation of bronchial hyperresponsiveness and inflammation was not caused by anticholinergic receptor effects. OVA-responsive T cells from omalizumab-treated mice exhibited depressed production of IL-4, IL-5, and IL-13 and normal amounts of interferon-gamma. The amounts of IL-5 and IL-13 were also diminished in the bronchoalveolar lavage fluid. Conclusion: Omalizumab, given at the time of exposure to the allergen, inhibits the induction of allergic pulmonary inflammation, and bronchial hyperresponsiveness. These results suggest that omalizumab or similar agents given during times of antigen exposure might alter disease progression in patients with respiratory allergy. The non-proteolytic major house dust mite allergen Der p 2 induce proasthmatic responses in bronchial epithelial cells partly through NF-0B and MAPK pathways Camilla Osterlund 1 , Hans Gronlund 2 , Sofia Sundstrom 2 , Guro Gafvelin 2 , and Anders Bucht 1 . 1 FOI CBRN Defence and Security, Resp. Medicine and Allergy, University Hospital, Umeå, Sweden; 2 Karolinska Institute, Dep. of Medicine, Clinical Immunology and Allergy, Stockholm, Sweden. Background: House dust mites (HDM) belong to the most common sources of airborne allergens worldwide and sensitization to them is strongly associated with development of allergic airway diseases. Many proteolytic HDM allergens can activate respiratory epithelial cells to produce proinflammatory mediators. In contrast there is limited knowledge regarding potential similar effects of non-proteolytic allergens, although they include many major allergens. One of them is Der p 2 of Dermatophagoides pteronyssinus. To investigate whether Der p 2 activate respiratory epithelial cells to produce mediators involved in pathogenesis of allergic asthma and to elucidate the mechanism of such activation, we exposed the human bronchial epithelial cell line BEAS-2B to this allergen. Methods: Bronchial BEAS-2B cells were exposed to recombinant Der p 2 (1-80 6g/ml). After 2-24 hours the mRNA levels and the secreted amount of soluble mediators as well as the expression of cell adhesion receptors involved in recruitment, survival and binding of inflammatory cells, for instance GM-CSF, IL-6, IL-8, MCP-1, MIP-3" and ICAM-1, were analyzed. After exposure the adhesion of leukocyte cells U937 to the epithelial cells was also studied. In order to study if the activation was dependent on signalling through NF-0B and the MAP kinases ERK1/2, p38 or JNK specific inhibitors were used. Results: Der p 2 induced dose-dependent up-regulation in gene expression and protein secretion of granulocyte-macrophage colony-stimulating factor GM-CSF, IL-6, IL-8, MCP-1 and MIP-3". Expression of ICAM-1 was also up-regulated, which was associated with a subsequent increased adhesion of leukocytes to the epithelial cells. These responses were partly dependent on NF-0B and MAPK activation, since the specific inhibitors reduced the activation. Conclusion: Taken together these results imply that Der p 2 may potentiate asthmatic responses in airways by direct activation of lung epithelial cells in a protease-independent manner. Human eosinophils play important roles in the pathogenesis of allergic diseases, specifically asthma. Interleukin 6, IL-8 (CXCL8), and Monocyte chemotactic protein 1 (MCP-1/CCL2) play a pivotal role in mediating the infiltration and activation of immune cells into pathogenic lesions, including the lung and skin. The aim of this study was to examine whether the house dust mite Dermatophagoides pteronissinus extract (DpE) affects the mRNA and protein expression of IL-6, IL-8 and MCP-1 in the human eosinophil cell line, EoL-1 by performing RT-PCR and ELISA. DpE increased the mRNA and protein expression of IL-6, IL-8 and MCP-1 in EoL-1 cells. The increased expression of MCP-1 and IL-8 was inhibited by PP2, an inhibitor of Src, rottlerin, an inhibitor of protein kinase C & (PKC&), and PD98059, an upstream inhibitor of extracellular-signal-regulated kinase (ERK). It indicates that DpE increases MCP-1 and IL-8 expression through Src, PKC & and ERK. We also found that IL-6 expression due to DpE was related to Src, PKC &, and p38 mitogen-activated protein kinase (MAPK). In early signal pathway, the expressions of IL-6, IL-8 and MCP-1 are regulated by Src family tyrosine kinase and PKC & pathway activated by DpE. This finding may contribute to the elucidation of the pathogenic mechanism triggered by DpE. Perturbations of NK cell regulatory functions in respiratory allergic diseases Background: Allergic diseases are characterized by abnormal responses to allergens favored by an inappropriate regulation of the Th1-Th2 polarization. NK cells give rise to a complex NK/DC cross-talk that would help Th1 response. By analyzing Peripheral Blood NK cells from 12 patients suffering from either allergic rhinitis or rhinitis and intermittent asthma, we evaluated whether, these cells were impaired in their ability to interact with DC. Methods: Different circulating NK cell subsets were analyzed by flow cytofluorimetry. Mixed NK/DC cultures were performed to assess the reciprocal functional interactions. NK cells were analyzed for their ability to induce DC maturation and cytokine production, and to kill immature DC. In addition, DC were assessed for their ability to induce cytokine production by NK cells. Results: We first analyzed the CD56+CD16+/jcells: a subset of circulating NK cells that is able to respond to DC by proliferating and producing IFN-,. Our analysis revealed that this NK cell subpopulation was sharply reduced in most patients. This was reflected by reduced NK cell-mediated IFN-, production in response to DC. Also the capability of promoting DC maturation and/or to kill immature DC, a function sustained by CD56+ CD16+NK cells, was reduced in most patients. Conclusion: We suggest that allergic diseases are accompanied by a partial impairment of the NK cell capability of promoting and maintaining appropriate Th1 responses. Humoral and cellular immune responses to blomia tropicalis and its concanavalin a-binding fractions in atopic patients Ernesto Taketomi 1 , Ronaldo Alves 1 , Deise Silva 1 , Jorge Fernandes 1 , Karine Almeida 1 , Leandro Ynoue 1 , Cristiane Bernardes 1 , Priscila Moreira 1 , Mônica Sopelete 1 , Margareth Gennari-Cardoso 2 , and Sun-sang Sung 3 . 1 Federal University of Uberlândia, Laboratory of Allergy and Clinical Immunology, Uberlândia, Brazil; 2 State University of Santa Cruz, Immunology, Ilhéus, Brazil; 3 University of Virginia, Rheumatology and Immunology, Charlottesville, United States. Background: Blomia tropicalis (Bt), Dermatophagoides pteronyssinus (Dp) and D. farinae (Df) are the prevalent house dust mites (HDMs). Concanavalin A-binding components derived from B. tropicalis (Bt-ConA extract) have shown to be highly immunogenic in allergic diseases. This study aimed to evaluate the humoral and cellular immune responses to B. tropicalis in sensitized patients. Methods: A total of 137 patients with allergic rhinitis with/without asthma and 109 non-atopic subjects were selected and analyzed for skin reactivity (SPT), total serum IgE and specific IgE levels to both Bt-total and Bt-ConA extracts, proliferative response and cytokine (IFN-+ and IL-5) production by peripheral blood mononuclear cells stimulated with both extracts. Results: SPT showed that 70% of patients were sensitized to Bt (Bt+) and no patient was monosensitized to Bt. Similar levels of specific IgE to Bt-total and Bt-ConA extracts were found in Bt+patients, while higher levels of total serum IgE were found in atopic than non-atopic subjects. Significant PBMC proliferation was observed in response to Bt-total extract in Bt-sensitized, but not in Bt-non-sensitized patients and non-atopic subjects, while Bt-ConA extract was able to induce increased proliferative responses in all patient groups. Significant IFN-+ production was observed only after Bt-ConA stimulation in Bt+patients, while Bt-total extract showed no changes. IL-5 production was consistently seen in Bt+patients after allergen-specific stimulation or even with no stimulus. Conclusion: We can conclude that Bt-ConA extract may contain relevant antigens that are involved in both humoral and cellular immune responses, with potential use in diagnostic procedures. Ken-ichi Kanai 1 , So Watanabe 1 , Atsuko Furuta 1 , Kazuhito Asano 2 , and Harumi Suzaki 1 . 1 Showa University, Otorhinolaryngology, Tokyo, Japan; 2 Showa University, School of NRS, Division of Physiology, Tokyo, Japan. Background and Purpose: Eosinophils are well accepted to be the most important cells in the development and maintenance of the clinical conditions of allergic diseases such as allergic rhinitis and asthma. It is also accepted that nasal mucosal epithelial cells secrete several types of cytokines and chemokines, which affect eosinophil accumulation into the site of the disease and cell survival. However, the influence of second-generation antihistamines, which are used for the treatment and prevention of allergic diseases are not well understood. The present study, therefore, was undertaken to evaluate the influence of epinastine hydrochloride (EP), the most famous secondgeneration of antihistamine in Japan, on cytokine secretion and eosinophil survival. Materials and Methods: Epithelial cells from nasal polyps were stimulated with 25 ng/ml TNF-! with or without EP for 24 h. Cytokine levels in culture supernatants were examined by ELISA. Human peripheral blood eosinophils were cultured with epithelial cell conditioned medium (CM) and EP. Eosinophil survival was assessed by Trypan blue dye exclusion test. Results are expressed as the mean SE of cytokine concentrations (pg/ml) and eosinophil survival index (%). Results: Addition of EP into cell cultures dose-dependently inhibited the ability of cells to produce granulocyte-macrophage colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) and IL-8, which are increased by TNF-! stimulation. EP also could inhibit eosinophil survival induced by CM and the minimum concentration of EP that caused significant suppression of the survival was 20.0 ng/ml. Conclusion: The inhibitory effect of EP on inflammatory cytokine production from epithelial cells and on eosinophil survival contributed to its therapeutic effect on allergic airway diseases, including allergic rhinitis. Inhibitory action of epinastine hydrochrolide on the production of anti-angiogenesis factors from mouse peritoneal mast cells in vitro Atsuko Furuta 1 , Ken-ichi Kanai 1 , Kazuhito Asano 2 , and Harumi Suzaki 1 . 1 Showa University, Otorhinolaryngology, Tokyo, Japan; 2 Showa University, Division of Physiology, School of NRS, Tokyo, Japan. Background: H1 receptor antagonists, so called antihistamine, are accepted to have anti-angiogenesis properties in addition to being H1 antagonists. Epinastine hydrochloride (EP) is the most famous H1 receptor antagonists in Japan and used for treatment of allergic upper airway inflammatory diseases such as pollinosis with remarkably success. However, the influence of EP on pro-angiogenesis factor production is not well understood. In the present study, therefore, we investigate the influence of EP on the production of angiogenesis factors, vascular endothelial growth factor (VEGF), keratinocyte-derived chemokine (KC) from murine mast cells in vitro. Materials and Methods: Murine mast cells were stimulated with ovalbumin in the presence of various doses of EP for 24 h. The levels of vascular endothelial growth factor (VEGF), keratinocyte-derived chemokine (KC) and tumor necrosis factor (TNF)-! levels in culture supernatants were examined by ELISA. mRNA expressions of these factors were also examined by semiquantitative RT-PCR in 4 h-cultured mast cells. Results: EP could suppress the production of VEGF, KC and TNF-! induced by an IgE dependent-mechanisms in dose-dependent manner. The minimum concentration of the agent that caused significant suppression was 45ng/ml. EP also suppressed mRNA expression examined when the agent was added to cell cultures at a dose of 45 ng/ml. Conclusion: These findings strongly suggest that anti-angiogenesis activity of EP may confer the attenuating effect of the agent on allergic diseases, including allergic pollinosis. Reduced levels of total serum IgE and FcRI expression in releaser and non-releaser basophils FcRI mediated signal pathway in basophils and mast cells leads to release of histamine and other mediators in-vitro and in-vivo systems. Interestingly, basophils from 18-20% of the population do not release histamine and other mediators on activation of the IgE signal transduction pathway and this has been attributed to the absence of tyrosine kinases Lyn and Syk. The present work deals with the histamine releasibility in Indian subjects to assess relationship between releasers and non releaser basophils with expression of FcRI and total serum IgE levels.Basophils from peripheral blood of healthy adults was purified by density gradient centrifugation and negative immuno-selection technique. Histamine release assay was performed flourometrically. Assessment of IgE receptor expression was carried out by flowcytometry and total serum IgE was estimated by ELISA method.Histamine release after ConA challenged varied from 0Y100% in Indian subjects. Eighteen percent subjects showed less than 5% histamine release and were considered non-releasers while those with more than 20 % were considered as releasers. Flowcytometric analysis revealed a significantly reduced expression of FcRI in non releaser basophils (p G 0.05 ). Interestingly, total serum IgE levels were also significantly (p G 0.05) reduced in non-releasers suggesting a common regulator of the phenotype. An in depth evaluation could lead to identification of a potential target for the development of therapeutics for allergic patients. An Investigation into the interaction of IgE with truncated recombinant CD23 (FcåRII) fragments Naser Nazari. Kermanshah University of Medical Sciences, Department of parasitology, Medical School,, Kermanshah, Islamic Republic of Iran. CD23, low-affinity receptor for IgE, has been widely implicated in the synthesis of IgE as well as in IgE-mediated immune and inflammatory functions. There are two forms of CD23 in humans-CD23a and CD23b, their cell expression and functional activities are different. Structurally, CD23 presents a single membrane-spanning domain followed by an extracellular domain that consists of three regions: the á-helical coiled coil stalk region, which mediates the formation of trimers, followed by the lectin head, which binds IgE, and at the C terminus, a short tail containing an inverse RGD (Arg-Gly-Asp) sequence. Aims of the project is to express defined truncated fragments of CD23 and to assess the physical and biological properties of these truncated fragments with regard to: IgE independent mast cell /basophile secretayogue activity. After obtaining all the necessary information concerning the different generation of CD23 fragments as well as the vector PET-14 and pIRES-EGFP, the appropriate primers were designed. The primers are essential for PCR amplification of the desired sequences and for subsequent cloning into the PET-14 and pIRES vectors. CD23 a and b YRGD sequence and also the whole things of CD23 a and b have been cloned into pIRES vector for transfection and subsequent evaluation of IgE in functional assays. Overlap PCR was used to produce the different truncated fragments of CD23 molecule, consisting of CD23 a and b minus the RGD sequence ( adhesion part). In the meantime, the plasmid vectors PET-14 and pIRES were introduced to XL1-E.coli cells in order to be obtained using the Mini and Midi preps protocol (gene transfection). Their DNA and protein was purified and obtained the characterisation by electrophoresis. CD23 a and b-RGD sequence and their whole things was transfected into the J558L (mouse myeloma cells) in order to study of expression of EGFP and biological activity by FACS (Fluorescence activated cell sorting) and FACS analysis for investigating the interaction of the IgE with different expression of CD23. Binding of IgE-Fc fragments to cell surface and expressed human CD23 were assessed using flow cytometry to detect the binding of IgE to cell surface receptors, using a biotinylated anti-IgE, followed by a streptavidin phycoerythrin conjugate. The results show that there is no expression of CD23 a-RGD sequence on J558L cells and these cells can express CD23 b-RGD sequence but it is not stable. Allergenic and hypoallergenic isoforms of the major birch pollen allergen Bet v 1 are differentially uptaken by DCs of allergic and healthy individuals Background: The major birch pollen allergen Bet v 1 is present in structurally slightly different isoforms that vary in their allergenic properties. Whereas Bet v 1a represents a potent allergen, the naturally occurring isoforms Bet v 1d and the engineered Bet v 1mut display strongly reduced allergenic activity. Although the biology of T cells in allergy is well understood, little is known about the mechanisms that control the initial T cell polarization by DCs in response to allergens. Objective: The present study investigated the different properties of Bet v 1 isoforms to be uptaken by human monocyte-derived DCs (MoDCs) of birch pollen allergic and healthy individuals. Methods: Eight birch pollen allergic and 7 healthy donors were included in this study. MoDCs generated from peripheral blood and taken at day 7 of differentiation were incubated with various concentrations of fluorescein isothiocyanate (FITC)-labeled recombinant Bet v 1a, Bet v 1d, and Bet v 1mut for 10 to 120 minutes at 37-C and studied by means of flow cytometry. Mechanisms of antigen internalization were investigated by the same experimental setup in the presence of inhibitors of pinocytosis, endocytosis, and receptor internalization. Results: FITC-labeled proteins were taken up by MoDCs in a dose-and time dependent manner. The percentage of FITC positive cells increased from 1 to 5 2g/ml of labeled proteins whereas the cells internalized most proteins between 10 and 30 minutes. At the lowest concentration of 1 2g/ml, the uptake of Bet v 1a was lower compared to the uptakes of Bet v 1d and Bet v 1mut in DCs of both study groups. This fact did not change for healthy individuals when higher concentrations of proteins were used whereas in allergic individuals Bet v 1a uptake at higher concentrations was similar to that of the two other isoforms. Inhibition of the uptake was generally lower using DCs from allergic compared to those of healthy individuals. Conclusion: All different agents showed inhibitions indicating that the uptake of Bet v 1 isoforms is not only mediated via one pathway but an interplay of mechanisms like receptor-mediated internalization, endocytic processing and macropinocytosis. Our observations implicate that different mechanisms are involved in Bet v 1a uptake in allergic individuals possibly leading to the induction of signals for Th2 polarization. This study was supported by the SFB F1802 of the Austria Science Fund and the Austrian Academy of Sciences. HLA-DRB1 polymorphism in atopic dermatitis with egg white allergy in korean children Purpose: It is known that polymorphism of the human leukocyte antigen (HLA) class II can restrict specific IgE responses. We investigated whether particular HLA-DRB1 polymorphisms contribute to the development of AD and EW sensitization in Korean children with AD. Methods: A total of 185 patients with AD and 109 normal controls (NC) with no personal and family history of allergy were included. HLA-DRB1 typing was done using PCR-SSO (sequence specific oligonucleotide) and PCR-SSCP (single strand conformation polymorphism) methods. Phenotype frequencies of the HLA-DRB1 alleles of AD patients were compared with those of NC. AD patients with allergy to EW was defined as group A (96 patients) whose EW specific IgE was over 2 kU/L in less than 2 year old age and over 7 kU/L in greater than 2 year old age. Group B (89 patients) was defined that EW specific IgE was negative among AD patients. Phenotype frequencies of the HLA-DRB1 alleles in group A and group B were compared with those of NC. HLA-DRB1 alleles were classified into functional groups (A, De, Dr, E, Q, R, a) and frequencies of HLA-DRB1 functional groups in group A and group B were compared with those of NC. Results: HLA-DRB1*1101 was present at significantly higher frequency in AD patients compared with NC (12.4% vs 1.8%, P=0.002, OR=7.796, CI 1.775Y32.883) and was regarded as a factor associated with AD susceptibility. The result was significant after Bonferroni correction (Pc=0.048). The frequency of HLA-DRB1*0803 (10.8% vs 19.3%, P=0.043) was decreased in AD compared with NC, showing a weak protective effect against the development of AD. HLA-DRB1*0802 was decreased in group A compared with group B (2.1% vs 10.1%, P=0.021) and was regarded as a weak protective factor against the development of egg allergy in AD. HLA-DRB1*1501 was increased in group A compared with group B (22.9% vs 11.2%, P=0.036) and was regarded as a weak susceptibility factor associated with the development of egg allergy in AD. HLA-DRB1 functional group "a", in which DRB1*1501 is included, was also weakly associated with the development of egg allergy in AD. However, none of theses results remained significant after Bonferroni correction. Conclusion: There was a significant association between HLA-DRB1*1101 and AD. Weak association between HLA-DRB1*1501 with susceptibility to and HLA-DRB1*0802 with protection against the development of EW allergy in AD were observed. Maria Background: Food allergy is an abnormal response of immunological system, especially of mucosa immunological system on antigens supplied per os. There are very complicated and still unexplained to the end immunological mechanisms, which lead to hypersensitivity reaction. Most often food hypersensitivity is identified as the effect of atopy, which is connected with humoral response (specific IgE antibody). On the contrary cell immunological response are less investigated, however they can be very important, especially as a significant factor to initiate pathological allergic processes. Aim: The study concern the usefulness of flow cytometry to estimate specific sensitization of subpopulation of lymphocytes to food allergens in the allergy diagnosis. Methods: The investigations were performed on 60 children since 6 month to 5 years old: 20 children with CMA IgE dependent, 20 with CMA IgE independent and 20 healthy children. IgE total, sIgE, IgG, IgA, IgM, basic immunological panel, CD 23, CD25, CD26, CD30, CD69, PCNA were measured. Results: We noticed decrease of expression of CD4+CD30+between I and II examine (p=0,029), between I and III (p=0,009); decrease of expression of CD8+CD26+between I and III test (p=0,038); decrease of expression of CD19+CD23+between I and II examine (p=0,012) in I type of hypersensitivity. We observed decrease of expression of CD4+CD25+between I and III examine (p=0,026) and decrease of expression of CD4+CD26+between I and III examine (p=0,036) in IV type of hypersensitivity. Expression of CD69 was decreased after diet in allergy IgE dependent. Values of expression of PCNA are similar in I and IV type of hypersensitivity in children with CMA. Decrease of expression of PCNA in II examine was observed in both cases. Allergen reintroduced caused increase of expression of PCNA in both types of allergy (p=0,048 and p=0,041). Conclusion: Our recent research confirm changes of the expression of T lymphocytes activation markers. It is connected with in vivo stimulation to allergen or with allergen elimination. The study of expression of activation markers using flow cytometry in food allergy in children can be helpful in observation of the dynamic progress process, but it cannot be used as a single diagnosis test. Background: Probiotics are defined as live non-pathogenic microorganisms that beneficially affect the host. It has been suggested that administration of probiotics may have therapeutic and/or preventive benefits by improving the Th1 responses in atopic dermatitis (AD). Aim: We investigated the possibilities that skin can be used as a target for the prevention of AD and Lactobacillus rhamnosus, one of the most commonly used in the therapeutic intestinal probiotics, can induce the Th1 responses through keratinocytes. Methods: Keratinocytes were treated by L. rhamnosus for 2 hours and then cultured with new media for 24 hours. The supernatant (KCM) of L. rhamnosus-treated keratinocytes were collected and used for the maturation of immature dendritic cells from 5 normal and 5 AD individuals. CD14+ cells and naive T cells were isolated from peripheral blood of normal and AD individuals using isolation kits. CD14+cells were differentiated to immature dendritic cells (iDCs) with GM-CSF (200 ng/ml) and IL-4 (20 ng/ ml) for 6Y7 day. The iDCs were activated by KCM or LPS lipopolysaccharide (LPS) (positive control) for 48 hours and cocultured with naive T cells for 7 days. Results: We could confirm the maturation of iDCs by KCM and LPS through morphological changes and the expression of mature DC specific markers (CD80, CD83, and CD86) in normal and AD individuals. ELISA analysis showed that the mature DCs activated by KCM could induce the Th1 reponses. So, the IFN-+ was significantly increased in the co-cultured supernatants of mature DC and naive T cells, but IL-4 level was almost based. Furthemore, we found that IL-8 and HBD-3 were highly increased in keratinocytes after L. rhamnosus treatment. Conclusion: Probiotics, L. rhamnosus, can improve the Th1 responses through keratinocytes. The effects of probiotics will be mediated by IL-8 and/ or HBD-2 secreted by keratinocytes. However, there is no difference for the effects of probiotics between normal and AD individuals. Interaction of aeroallergens with the respiratory interphase: degradation, processing and transmission via the epithelial barrier Research Center Borstel, Biological Chemistry, Borstel, Germany. Background: Antigen presenting cells are considered as the most important immune cells giving rise to allergy or tolerance development. But APCs are already embedded in a cytokine milieu, which in part predicts the immune response. Epithelial cells form a barrier to the environment and they are activated upon contact with molecules e.g. allergens. Therefore, they might play a pivotal role in the sensitization process. We studied the fate of the major grass pollen allergen Phl p 1 and its molecular modifications after contact with the respiratory interface. Methods: The influence of mucosal secretions (nasal secretion, bronchial lavage fluid) was investigated by immunoblotting and zymography and the uptake and transmission of the allergens by epithelial cells (cell lines and nasal and lung biopsies) were studied by FACS analysis, determination of cytokine release and immunohistology. Results: Nasal secretions of individuals suffering from bronchial inflammations or allergy to aeroallergens showed proteolytic activity. Experiments using supernatants from mast cells or neutrophils, similar to acute inflammation, resulted in a partial cleavage of the allergens. To investigate the allergen uptake two epithelial cell lines were used mimicing different sections of the lung, A549 (derived from alveolar pneumocytes) and Calu-3 (from the upper respiratory epithelium). Both cell lines were activated by Phl p 1 as demonstrated by the release of IL-8 and IL-6. Calu-3 cells in contrast to A549 cells expressed MHCII, a prerequisite for antigen processing and presentation. A549 cells, on the other hand, take up allergens by macropinocytosis and probably perform transcytosis. First results of human lung slices incubated with Phl p 1 showed only a faint allergen uptake, while macrophages in the alveolar space showed a considerable uptake. Whether macrophages after allergen uptake enter the interstitium again and/or whether the epithelial cells transfer allergens or fragments to professional APCs in the interstitium is still under investigation. Conclusion: The secretions of the respiratory tract cause a more or less incomplete fragmentation of allergens which may facilitate the allergen uptake by epithelial cells. Our results reveal differences in the uptake and transmission of allergens in the upper and lower airways. While the epithelial cells of the upper part probably degrade and process the allergens, cells of the lower part seem to transmit the unprocessed allergens. The estimation of influence of various programs of therapy on system and local level IL-6 and SR-IL-6 was made at development atopic march of teenager patients. The research allowed to reveal, that therapy with the help of topical steroids practically did not influence SR-IL-6 level in serum of blood (22,72T0,25ng/ml), but led to decrease in IL-6 level at patients with BA (0,17T0,001 pg/ml) and combination BÀ and À R (3,59T0,37 pg/ml). At combination of BÀ and À R the simultaneous decrease of IL-6 and SR-IL-6 was established while using topical steroids. On the contrary, having clinical BÀ and AD evidence steroid therapy led to one-stage increase cytokine (6,45T0,41pg/ml), and level of receptor to it (31,76T0,15ng/ml). Thus, research of nasal lavage can be offered as nonivasive method for the control of efficiency of therapy of patients having combination BÀ and À R while using topical steroids. Therapy with topical steroids of patients with BÀ and with combination BÀ with À R and À R with AD provided decrease of IL-6 level and did not lead to change of SR-IL-6 level in serum of blood. Therapy with topical steroids did not lead to decrease in content of IL-6 both in serum and in nasal secret only in group of patients with combination BÀ and AD. Previous work in type-I pollen allergies has focused on lymphocytes and immune responses. Here we begin to analyse with a systems biology top-down view the differences in nasal epithelium obtained from healthy and allergic subjects. Light and immunoelectron microscopic analysis showed that birch pollen Bet v 1 allergen bound to epithelial cell surfaces within minutes even during non-symptomatic winter seasons only in allergic, but not healthy individuals. Bet v 1 also travelled through epithelium together with lipid rafts/caveolae and reached mast cells only in allergic, but not healthy individuals within minutes. A putative viral entry protein E3 ubiquitin-protein ligase and three enzymes involved in lipid rafts/caveolae metabolism were among the few Bet v 1 binding proteins found in allergic subjects with affinity chromatography and LC-MS2. Nasal epithelial cell transcriptomics during non-symptomatic winter season identified a list of putative receptors by which Bet v 1 might be recognized. It also suggested that defence mechanisms (such as expression of histatins) could be impaired in allergic patients. Comparing nasal epithelial transcriptomics taken in winter and during symptomatic summer seasons provided hints to the cellular perturbations enabling the Bet v1 traffic through nasal epithelial cells and tissues. Thus application of discovery and hypothesis driven methodologies on human nasal epithelial tissue could provide new hypothesis worth further analysis of the underlying molecular mechanisms. The most significant aspect of this and other top-down explorative studies using whole genome or other large-scale analysis is that they can provide truly new hypothesis. Before this work few would have argued that the nasal epithelium is so markedly different in healthy compared to allergic subjects already during non-symptomatic winter session. Now based on these validated results the mechanisms of Bet v 1 pollen allergen binding to and traffic through the epithelium can be further explored and analysed. Furthermore it must be noted that no single data domain, whether is would haven microscopy, proteomics, transcriptomics etc. would have alone pointed out the above-described phenomenon. Thus systems level understanding of complex pathophysiological phenomena will need all possible wet lab techniques combined with computer sciences as the biology will be transformed into computer-readable format. Maternal tolerance achieved before pregnancy is transferred to the offspring and prevents asthma development in the next generation Tobias Polte 1 , Christian Hennig 2 , and Gesine Hansen 2 . 1 Martin Luther University Halle-Wittenberg, Department of Pediatrics, Halle, Germany; 2 Hanover Medical School, Department of Pediatrics, Hanover, Germany. In this study we hypothesized that immunologic tolerance acquired before conception can be transferred from the dam to the pup. In a murine model, we induced tolerance before conception by oral application of antigen. We than immunized the offspring of tolerized dams with the same antigen. While the offspring of naBve dams developed an asthma-like phenotype with airway hyperreactivity, inflammation, Th2-cytokine production after immunization, the offspring of tolerized dams was protected, even when immunized as late as 8 months after birth. Critically involved in tolerance transfer is allergenspecific IgG that was increased during pregnancy in the tolerized mouse, fetus and newborn. FcRn j/j mice, that cannot transport IgG via the placenta, transferred tolerance to the offspring only when the missing diaplacental IgG transfer was compensated by IgG transfer via breast milk from tolerant dams but not when the offspring was crossfostered by naBve mice. Inhibition of IFN-+, produced by memory B cells in the offspring, abrogated the protective effect of maternal tolerance demonstrating its crucial memory role in materno-fetal tolerance transfer. Our data show that maternal immunologic memory has significant and persistent impact on the immune response of the offspring indicating that e.g. allergy prevention strategies might be effective for more than one generation. Meditation as a suggestion for allergies and autoimmune diseases Parisa Karimi 1 , and Alireza Salekmoghaddam 2 . 1 Immunology, Asthma and Allergy Research Institute, Medicine, Tehran, Islamic Republic of Iran; 2 Iran University of Medical Sciences, Medicine, Tehran, Islamic Republic of Iran. Objective: The aim of the present study was to analyze the effects of a Meditation program on Complete Blood Cell count(CBC), IgG and C3 as a marker of specific and nonspecific humoral immunity. Methods: 78 subjects, aged 18Y56 years, of whom 62 were male and 20 female undergoing a period of one month Meditation, were excluded. They were all experiencing meditation for the first time. Before starting the meditation training, all of these volunteers have been visited by a physician and only healthy subjects(78 subjects), not taking any type of drug and with regular life habits were chosen to be at this clinical trial. Blood samples were taken from all subjects, the day before the study commenced, and again one month later, at the end of the study. The blood parameters investigated included the number of Leukocytes(total Leukocytes, Monocytes, Neutrophils, Eosinophils, Basophils, Lymphocytes),Red cell count(RBC), Hemoglobin(Hb), Hematocrit(Hct), Mean cell volume (MCV), Mean cell hemoglobin (MCH), Mean cell hemoglobin concentration(MCHC), Red cell distribution width (RDW),Platelet count, as well as the concentrations of immunoglobulins(IgG) and complement(C3). 4 of these volunteers has been excluded from research plan because of their sickness during the period. Results: Statistically significant differences were found between first and second blood samples of volunteers showing lower numbers of total Leukocytes(P=0.02) and Eosinophils(P=0.01),number and percentage of Monocytes(P=0.01),as well as complement C3 concentration(P=0.004). Conclusion: These findings demonstrate that one month of practicing Meditation, can decrease some immunological parameters. According to these results Meditation can be suggested for Hypersensitivities(Allergies and Autoimmune Diseases) although further studies seem to be needed. Key words: Meditation, Complete Blood Cell count(CBC), IgG, C3. A randomized, double-blind, placebo controlled trial on the effect of zinc supplementation on bronchial asthma as measured by sputum eosinophil count and asthma control test (ACT) in children Felix Bernard Cepeda, and Agnes Andaya. University of Santo Tomas, Section of Allergy, Asthma Immunology, Manila, Philippines. Backround: Zinc being a major dietary anti-oxidant has a protective role in the airway epithelium. It may therefore have important implications for asthma where the physical barrier is vulnerable and compromised. Objective: We investigated the effects of zinc supplementation on bronchial asthma as measured by sputum eosinophil levels and asthma control test (ACT) in children. Methodology: There were sixty-six asthmatic subjects age range 7 to 18 years old were randomized to receive zinc 20mg/day(n=29) or placebo (n=37). Sputum eosinophil count were checked before supplementation and 12 weeks later. Likewise asthma control test (ACT) scores were obtained before supplementations and at 4 weeks, 8 weeks and 12 weeks thereafter. Results: After 12 weeks of supplementation, the sputum eosinophil count decreased in both groups but the zinc group has the more significant change in eosinophils as compared to placebo at p=0.029 and p=0.059 respectively. However, ACT score from week 1 to week 12 showed no difference between the zinc and placebo group. (p=0.069). Conclusion: A dietary zinc supplementation significantly reduced sputum eosinophil count as compared to placebo. However, the decline in sputum eosinophil count was not associated with improvement in asthma control. Zinc should be considered in decreasing airway inflammation but not asthma control. Documentation of the relative efficacy and safety profile of the salbutamol delivery by slow infusion(SI) VS inhalation(IN) to patients with acute severe bronchial asthma I. Khan Sameera, M. Ishaq, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Purpose: Patients with acute severe bronchial asthma are in a moribund position. The art of devising an effective delivery system which may be followed by an optimal relief in the degree of bronchospam is the goal of resolving the imminent urgency of an acute severe bronchial Asthma. Methods: Patient's both sex age16Y50 years, with history of acute severe bronchial asthma have been included in the study. They had been Group A, medicated with Prednisolon 15Y20 mg immediately followed by 10 mg 6hourly for 2 days along with Salbutamol 4mcg/kg body weight in slow infusion. Group B, having being medicated with Prednisolon as in Group A, along with inhalational delivery of Salbutamol in dilution of 1.5ml+2.5ml and 1.25ml+2ml (for adult and childhood bronchial asthma respectively). Therapeutic response as observed had been as under. Results: Group B, had been followed with optimal bronchodilation in about 5Y10 minute while with group A, it was 15Y20 minutes. Conclusion: Group B had therapeutic advantages vs A (e.g.) optimal bronchodilation in a relatively shorter period, no hazard of parental administration and a simple technique of delivery that can be managed even by any one. The prevalence of asthma has increased worldwide since the 1960s. The incidence of asthma is high among children; however, a relative high annual incidence is also estimated in adults. Studies for risk factors have mainly analyzed cross-sectional design data based on prevalent cases of asthma. Such studies may determine either cause, consequence or both. Incidence studies have mostly focused on occupational asthma or smoking habits. Since the late 1990s, obesity has been reported to be associated with asthma, and an increase in the prevalence of obesity has been reported along with a parallel increase in asthma prevalence. However, a convincing relationship between asthma and obesity has not been established. Accordingly, this study was carried out aiming at demonstrating the effect of weight reduction on clinical, functional and serological parameters in obese asthmatics. In this study, obese asthmatic patients showed marked improvement clinically, functionally, with marked decrease in number of exacerbations and medications used. Feasibility and applicability of secondary prevention of asthma in allergy practice Mahesh Padukudru Anand 1 , Amrutha Holla Devidas 2 , A.K. Prabhakar 2 , Biligere S. Jayaraj 1 , C.P. Chathura 2 , Rao B. Chaitra 2 , and Pudupakkam K. Vedanthan 3 . 1 JSS Medical College, Department of Pulmonary Medicine, Mysore, India; 2 Allergy Asthma and Chest Center, Department of Allergy, Mysore, India; 3 University of Colorado Health Sciences, Department of Allergy, Colorado, United States. Background: The possibility for secondary prevention of asthma was highlighted by the prevention of asthma study, which demonstrated that immunotherapy in patients with allergic rhinitis, could prevent asthma in some patients. The feasibility of secondary prevention of asthma and the proportion of patients likely to be benefited need to be demonstrated as immunotherapy requires time to exert its beneficial effects. Methodology: All the patients who underwent a detailed evaluation including allergy testing during the period 2003 to 2006 were included in the study. A structured questionnaire was applied to collect data on demography, a detailed clinical history, the duration of asthma and allergic rhinitis and the proportion of patients whose rhinitis preceded asthma and the time interval between the development of rhinitis and asthma. Results: A total of 934 cases were included in the study. 701 (75%) patients had allergic rhinitis for varying intervals before they developed asthma. 143(15.5%) patients continued to have rhinitis for more than 5 years without developing asthma. 57(6%) patients had only asthma without rhinitis. 33(3.5%) patients developed asthma before they developed rhinitis. 327 (46.6%) developed asthma within 1 year of developing rhinitis. Of these, 164 developed asthma within 6 months and 169 between 6Y12 months. 374 patients developed asthma more than a year after the onset of rhinitis and the frequency gradually decreased with increase in duration of rhinitis. Conclusion: Majority of patients had rhinitis before they developed asthma. Nearly 50% of these patients with preceding rhinitis developed asthma within one year, who may not be amenable for secondary prevention with immunotherapy due to the delays in referral to the allergist and the time required for obtaining the necessary benefit from immunotherapy. Patients who develop asthma alone or develop rhinitis after the onset of asthma are not candidates for secondary prevention. Of the total cases studied, less than 40% can undergo interventions targeted towards secondary prevention of asthma. Intense education of the Family physicians and ENT specialists is needed to ensure prompt referral to the allergist to help these patients. Aim: to assess the relationship between ACT and different markers of airway inflammation in asthmatic subjects Methods: In a crossover design, we studied 106 patient suffering from mild or moderate asthma (median age 31years IQR 16Y45) ). Fifty five percent were only treated with short beta agonist on demand. On the same visit patients fullfiled the ACT questionnaire, exhaled nitric oxide (eNO) (NiOx Mini) was measured and bronchial challenge with hypertonic saline and sputum induction were performed. Results: On the whole sample ACT was modestly correlated with eNO (r= j0,40, pG 0,001), eosinophils in induced sputum (r=j0,30, pG 0,05) and the slope of hypertonic saline bronchial challenge (r= 0,43, pG 0,001). Patients with very good control (ACT 24 0r 25) had significantly lower levels of eNO (41 vs 72 ppb, pG0,001) and eosinophils (10 vs 6%, pG 0,05). Correlations of all these variables improved, when only the 55 patients treated with beta agonist on demand were included: eNO (r=j0,55, pG 0,0001), eosinophils in induced sputum, (r=j0,50, p= 0,002), and slope of hypertonic saline bronchial challenge (r= 0,52, pG 0,0001. ACT was not correlated to FEV1 of FEV1/FVC. Conclusion: asthma control assessed by the Asthma Control Test was correlated to direct and indirect markers of airway inflammation. ACT and pulmonary function variables were not correlated. Salbutamol metered-dose inhalation is as effective as nebulisation in managing acute asthma exacerbations in hospitalized children in accordance with an asthma pathway Anne Goh 1 , Oh Moh Chay 1 , Jenny Tang 1 , Ling Ho 1 , Oon Hoe Teoh 1 , and Kee Chong Ng 2 . 1 KK Women's and Children's Hospital, Department of Paediatric Medicine, Singapore, Singapore; 2 KK Women's and Children's Hospital, Children's Emergency, Singapore, Singapore. Background: Acute asthma exacerbations have traditionally been managed with nebulised bronchodilators in the emergency rooms as well as for inpatients. In 2003, Singapore was hit by SARS and the fear that the use of nebulisation could cause the spread of the virus prompted the development of an asthma pathway using MDI. Aim: To compare the length of stay in children admitted for asthma exacerbations before and after the introduction of an asthma pathway for managing acute exacerbations with salbutamol metered-dose inhalers(MDI) instead of nebulisation. Methods: Children presenting to the Children_s Emergency and admitted to the wards for asthma exacerbations were managed according to an asthma pathway. Results: There was no increase in admissions from the emergency department or to high dependency or to the intensive care after implementation of the asthma pathway with the use of Salbutamol MDI instead of nebulisation. The average length of stay decreased from a high of 2.78 days pre-implementation of pathway to a low of 2.19 days post-implementation. The average cost per patient decreased from $1,136.85 in 2001 to between $974.00 Y$1,011.05 post pathway implementation. Conclusion: The asthma pathway has shown that the use of salbutamol MDI is as effective as nebulisers in the management of acute asthma exacerbations and has led to a decrease in the average length of hospitalisation as well as decreased cost per patient. Pierangela Massacane, Anthi Rogkakou, Laura Guerra, Enrico Compalati, Antonio Scordamaglia, and Giorgio Walter Canonica. University of Genoa, Allergy and Respiratory Diseases, Genoa, Italy. Case Report: A 41-year-old asiatic man was admitted to the emergency room of our hospital with fever, wheezing, dyspnea, arthralgias. His past history was remarkable for allergis rhinitis and recurrent episodes of asthma since his arrival in Italy at the age of 34. The patient was treated with a course of systemic corticosteroids (80mg daily prednisolone), theophylline, inhaled budesonide/ formoterol 160/ 4.5 mcg (twice a day), montelukast 10 mg . His chest radiograph and full blood count including eosinophil count were normal. His spirometric test showed FEV1 1.57 ( 42%), FVC 2.52 (57%), PEF 240 l/min. One month later he was seen in the outpatients department and noted to be generally unwell, very wheezy and 2 days before the visit he has noticed the comparison of hemoptysis. His chest radiograph and full blood count, including eosinophil count, were normal. Antinuclear antibody and antineuthrophil cytoplasmic antibody (ANCA) were both negative. Sputum culture and urine microscopy were unremarkable. Aspergillus precipitins and radioallergosorbent test (RAST) against Aspergillus were negative. Cultures for mycobacteria were negative. The electrocardiogram and echocardiogram were both normal. Computed tomography of the paranasal sinuses dimostrated a neartotal opacity with the material of the soft parts of the normal sinus and right sphenoidal sinus, as well as of ethmoidal cell, the right maxillary sinus and nasal cavities. Thorax computed tomografy revealed generalized enlargement of the mediastinic lymph nodes. Treatment with montelukast was stopped and high dose oral prednisolone was commenced. The patient elected to discontinue his inhaled treatment .Three months later his symptoms of asthma returned. Chest-X-ray showed patchy bilateral infiltrates in the lungs. He had marked hypereosinophilia (15%), elevated serum IgE levels (465 IU/ml), and antineuthrophil cytoplasmic antibody (ANCA) were positive. His spirometric test showed FEV1 1.60 (45 %), FVC 2.35 (55%), PEF 240 l/min. He required a further short course of oral coricosteroids. His symptoms remained troublesome and his treatment was changed to cyclosporine 3 mg/ kg and his condition showed marked clinical as well as radiological improvement. Discussion: We must carefully diagnose and treat patients with middle-age onset of severe asthma, because the symptom may be a lung manifestation of CSS, in which various organs are involved as a result of systemic necrotizing vasculitis. Educational courses improve asthma management and treatment Angelko Gjorcev, Zlatica Goseva, Marija Zdraveska, Deska Dimitrievska, and Dejan Todevski. Clinic of Pulmology and Allergy, Asthma Department, Skopje, Macedonia, Fyrom. Background: Education of the patients is one of the first goals of asthma management in all the international treatment protocols. We made this study in order to estimate the need and the effect of an educational program prepared by the Macedonian Asthma Center. Methods: We studied 300 asthmatics. We prepared a pilot educational course, covering the most important and ''hot'' subjects concerning asthma, following the guidelines from the Global strategy for asthma management and prevention. (May 1996) , and a questionnaire with 10 questions treating the problems concerning the treatment problems in asthma management. The questions were grouped in 2 parts : I -general knowledge of asthma medications and the necessity of adequate and continuous anti-inflammatory treatment; and II -accent on the emergency interventions and actions during acute exacerbations. The questionnaire was completed by all of the patients, before and after attending the educational course. Results: Before visiting the course, only 46.3% answered correctly the questionnaire (39.9% correct answers on part I and 52.8% on part II). These results imply that more than 50%of the tested asthmatic population do not realize the need for preventive medication or would in fact take the wrong medication during worsening of their disease. After visiting the educational course the results are as follows: 54.8% overall correct answers (nonsignificant-NS). The results from part I showed that only 52.4% (NS) of the patients gave correct answers and 73.6% (significant) answered the II part correctly. Conclusion: We conclude that patients more willingly accept advice on emergency self management of acute asthma exacerbations, but it is the most difficult to overcome the ''fear of preventive treatment'' and to change the dogmatic opinion on the therapeutic issues of asthma management. The study shows the necessity of organizing further and continuous asthma educational programs on all levels of the health care. Clinical evaluation of severe asthma attack requiring tracheal intubation and respirator management Kiyoshi Sekiya 1 , Masami Taniguchi 1 , Keishi Sugino 2 , Hidenori Tanimoto 1 , Yuuma Fukutomi 1 , Emiko Ono 1 , Chiyako Oshikata 1 , Takako Hojyo 2 , Kazuo Akiyama 1 , and Sakae Honma 2 . 1 National Hospital Organization, Sagamihara National Hospital, Clinical Research Center for Allergy, Kanagawa, Japan; 2 Toho University Medical Center, Department of Respiratory Medicine, Tokyo, Japan. Background: Recently, the number of patients requiring hospitalization because of an asthma attack has decreased; however, there are still patients who require hospitalization for tracheal intubation and respirator management for a severe asthma attack. Therefore, we evaluated the background features of 20 asthmatic patients who required tracheal intubation and respirator management in our hospital. Methods: We evaluated 20 asthmatic patients who visited our hospital from January 2001 to December 2005 and required tracheal intubation and respirator management. All the patients had severely exacerbated asthma, as determined on the basis of the guideline of the Global Initiative for Asthma (GINA) 2006, and they required tracheal intubation and respirator management. We evaluated their history of smoking, the days from asthma attack onset to their visit to our hospital, the level of asthma control as determined from the GINA 2006 guideline, treatments taken before the patients visited our hospital, their frequency of visiting a hospital, the reason for tracheal intubation and respirator management based on the Asthma Prevention and Management Japanese Guideline 2006, and their prognosis. Results: The patients who required tracheal intubation and respirator management were the following: 1. smokers, 2. patients not taking or irregularly taking treatment, 3. patients who used inhaled short-acting "2 agonist only at the time of an asthma attack, and 4. patients not using inhaled glucocorticosteroids. The reasons for tracheal intubation and respirator management were as follows: PaO2 of less than 50mmHg despite maximum oxygen administration in 30% of the patients; sudden increase in PaCO2 leading to unconsciousness in 20%; both PaO2 of less than 50mmHg despite maximum oxygen administration and sudden increase in PaCO2 leading to unconsciousness in 25%; and severe ventilatory or cardiorespiratory disturbance in 25%. Conclusion: We observed that a thorough education of patients and treatment mainly using inhaled glucocorticosteroids are important for preventing a severe asthma attack that requires tracheal intubation and respirator management. Assessment of quality of life in caregivers of asthmatic children Shima Sayanjali, and Fariborz Zandieh. Tehran University of Medical Sciences, Asthma, Allergy and Immunology, Tehran, Islamic Republic of Iran. Background: Interest in the impact of illness on day to day function is leading investigators to include both disease specific and generic health related quality of life (HR QOL) questionnaires in a broad range of clinical studies and to gain a full picture of the impact of asthma on the lives of caregivers of asthmatic children, it is necessary to make direct measurement of health related quality of life. Methods: In response to this need, we used Juniper_s Pediatric Asthma Caregiver_s Quality of Life Questionnaire (PACQLQ) that has been developed based on guidelines for construction of over a dozen validated disease specific quality of life instruments.The PACQLQ that contains 13 items in two domains of emotional and activities disturbances. The study design consisted of an 18 month single cohort study. Patients participating in the study were 113 children, 7Y17 years of age, with a wide range of asthma severity and their caregivers. For each caregiver a PACQLQ was completed. One week before visit patients recorded morning peak flow rates, medication use and symptoms in a diary. After complete physical examination, for determining of asthma severity, spirometry was performed. Results: The questionnaires after statically analysis showed good levels of both longitudinal and cross sectional correlations with the conventional asthma indices and with general quality of life. Conclusion: We found a good relevancy between severity of asthma and QOL scores and more disturbances of QOL in caregivers of male asthmatic patients than caregivers of female asthmatic patients. Bronchial asthma as a chronic and devastating disease can affect not only life style of the patients but also their caregivers. Increment of our knowledge about these disturbances can help the physicians for better understanding of burdens of their patients. Unified treatment in school children with allergic rhinitis and asthma Jelena Moskovljevic. Emergency Center, Immunology, Nis, Serbia and Montenegro. Aim: To verify the efficacy of beclomethasone dipropionate (BDP), administered through nasal inhalation, in the simultaneous treatment of asthma and allergic rhinitis (AR). Methods: 30 patients with asthma and AR, aged 7Y17 years were randomly allocated. During eight weeks, 20 of them received BDP-CFC (at least 500 mcg/day) inhaled exclusively by the nose (mouth closed) using a large-volume (650 ml) spacer attached to a facemask. Control group (10 subjects) received conventional treatment, i.e., BDP by dual administration (aqueous intranasal and oral inhalation through the mouthpiece of the same spacer device). Every two weeks a clinical score for AR and peak expiratory flow (PEF) was assessed by independent observers. Spirometry was performed at admission and at the end of the follow up. A minimum decrease in 66% in clinical scoring was considered as therapeutic success. Results: Therapeutic success rate was of 50.0% for the experimental group and of 70.0% for the control group (p=0.11). PEF and FEV1weren_t statistically different in the two groups neither at admission nor by the end of the treatment (p 9 0.10). Conclusion: Results suggest that this alternative treatment could be recommended for the simultaneous treatment of AR and asthma, specially in developing world. Other advantages are higher compliance, lower costs and fewer side effects. Influence of topical intranasal steroids on improvement of ventilating capability of lungs in patients with bronchial asthma Tair Nurpeissov, Rimma Akpeissova, and Bibigul Ilyasova. Institute of Internal Diseases, Republican Allergological Center, Almaty, Kazakhstan. Background: Treatment of a bronchial asthma (BA) is often conducted without an allowance of a concomitant allergic rhinitis (AR), which can be a reason of unsuccessful results in an achievement of a maximized therapeutic effect. Research about an interaction between BA and AR is conducted in the republican allergic center of scientific research institute of cardiology and internal diseases under Ministry of Public Health of the Republic of Kazakhstan. Aim: To determine the degree of influence of allergic inflammation of the upper respiratory tracts on ventilating capability of lungs in patients with bronchial asthma. Materials and Methods: The thirty patients with severe BA and concomitant the AR were prescribed to intranasal mometazon spray in a dose of 100 microgram two times a day for a one month. This treatment followed after two weeks course of standard hospital therapy that includes steroids (intravenous, inhalation), b2-àgonists, etc. The age of the patients was ranged from 25 to 50 years old. Therapy effectiveness was controlled both by clinical and by laboratorial methods: spirography and daily pickflowmetry, determination of the resistance of a bronchial tree, rhinomanometry and identification of ECP. Results: All patients at the end of the first month of treatment mentioned the stable improvement of nasal breathing and sleep recovery. Also, a valid reduction of the resistance of nasal apertures was discovered (pG0.005). The majority of the patients (93.3%) have the improvement of clinical picture of the BA, which confined in less amount of asthma attacks and also improvement of the functional and laboratorial measures: PEF growth, decrease of its variability (pG0.05), reduction in the resistance of the bronhial tree (pG0.05) and normalization of ECP. Conclusion: Conducted research proved a valid increase in the effectiveness of supporting therapy of the BA with the combination of active treatment of the AR, which becomes apparent besides clinical improvement in an additional growth of measures of ventilating capability of lungs, level of the resistance of the bronchial tree and nasal apertures and normalization of ECP content. Cardiff, United Kingdom; 2 University of Wales Institute Cardiff, Statistics, Cardiff, United Kingdom. We audited the secondary care offered to acute asthmatic patients admitted to all 17 hospitals in Wales, UK during February 2006. The audit analysed referral source, care delivered by the ambulance service, emergency units, medical admission units and inpatient units in all cases of asthma aged 18 years or greater. All data was extracted by qualified medical personnel. The results show that 248 patients were admitted during that month (36 patients were excluded as not having asthma). 44 were acute severe/severe, 91 moderate, 75 mild and in 38 severity was not documented. Ambulance transfer was used in 99 cases, 63 patients calling the ambulance directly. Overall, ambulance staff measured vital signs well but peak flow (PFR) was measured only 8 cases. Most patients received nebulised bronchodilators/high flow oxygen in the transfer process but steroids were used rarely and observations were repeated in only 27 cases. Emergency units were the primary site of acute care. PFR was measured in only 67% of cases compared to 90% having oxygen saturations measured. Only 162 of 248 cases had a record of steroid administration and only 67% were reassessed. Of those admitted (99) only 51% were under specialist care and only 32% were treated in a specialist respiratory ward. Overall discharge planning both from emergency and inpatient units was disappointing with only 13% of inpatients having written management plans and only 4.5% of patients discharged directly from emergency units offered further review. Secondary care of acute asthma in Wales does not confirm to national guidelines for acceptable levels of care. Chunmei Zhu, and Yuzhi Chen. The Capital Institute of Pediatrics, Asthma Clinic and Education Center, Beijing, China. Objective: To investigate the approach (monitoring and treatment) of pediatricians and general physicians to childhood asthma in China. Methods: A standardized questionnaire survey, which was construced by APAPARI on monitoring and treating childhood asthma was send to physicians in 25 provinces and cities of China. Results: 1863 questionnaires were received, of which 912 were completed (48.95%) 57% of the doctors were general pediatricians, 20% were pediatric pulmonologists or pediatric allergists, 9% were adult pulmonologists or allergists who also treatment children, and 14% were others (non-specified). Working places were: 71% in a tertiary hospital, 23% in a secondary hospital, and 4% work in a first grade hospital.Most of the doctors (66%) used peak flow meter or spirometry to monitor childhood asthma, while 31% of them used diary cards. For treating acute asthma exacerbations, the first choice treatment was nebulized salbutamol/terbutaline every 20 mins (29%), the second choice was salbutamol inhalation with a spacer (22%) and the third choice was systemic corticosteroids (12%). Pediatric pulmonologists and pulmonologists tended to use ICS for acute asthma. The most frequently used systemic corticosteroids for acute asthma treatment were dexamethasone (86%), methylprednisolone (83%) and hydrocortisone (81%). The criteria applied by doctors to start a maintenance therapy in asthmatic children were the frequency of symptoms (36%), severity of symptoms (32%) and whether or not there was a severe disturbance in lung function testing (27%). For the selection of drugs, doctors tended to choose ICS. Generally, the first choice of drugs used in maintenance treatment of asthma in young children (0Y3y) were ICS (85%), the second was montelukast, and the third was ICS+ oral LABA(slow release sulbutamol) (67%), while in preschool and school children (4Y16y) the first choice was a LABA + ICS (84Y87%), the second was ICS (81Y83%) and the third montelukast (73%). For moderate to severe asthma , the first choice was ICS + LABA. The awareness and recognition of childhood asthma, the use of peak flow meter, the implementation of GINA guidelines and the use of ICS has improved among the doctors of the cities in China. Nevertheless, dissemination of the GINA program among non-specialists and doctors in countries and communities must be continued and re-enforced. Impact of patient education on knowledge, attitude, practice and self-efficacy in patients with asthma in India Mahesh Padukudru Anand 1 , Sabin Thomas 2 , and Parthasarathi Gurumurthy 2 . 1 JSS Medical College, Department of Pulmonary Medicine, Mysore, India; 2 JSS College of Pharmacy, Department of Clinical Pharmacy, Mysore, India. Background: Patient related factors like poor knowledge, individual beliefs and attitudes of patients influence control of asthma leading to nonadherence. This study was conducted to develop a validated KAPSE questionnaire for the Indian population and to assess the knowledge, attitude, practices and selfefficacy (KAPSE) of patients in Mysore regarding asthma and the impact of patient education on KAPSE in these patients. Methodology: A total of 225 adult patients participated in the KAPSE study and were part of the study that compared the efficacy and quality of life in patients receiving beclomethasone, budesonide or fluticasone. A questionnaire with 22 items was selected from Knowledge, Attitude and Self-Efficacy (KASE), a validated instrument for asthma and suitably modified to suit the cultural and educational level of the population. Two medical experts performed content validation and only those items with content validation index of above 0.75 were retained. Cognitive debriefing in 5 patients confirmed the suitability and acceptability of the questionnaire. Patient counseling individually and in groups and information leaflets about the pathology of asthma, drugs and delivery systems were conducted at every visit on 7 occasions, until 6 months. KAPSE was assessed at baseline before education and at the end of the study. Results: Patient education significantly improved most of the questions assessing Knowledge, Attitude, Practice and Self-efficacy compared to the baseline (pG0.05). Subgroup analysis revealed that KAPSE at baseline were not influenced by Age, Gender and Duration of disease (p90.05), but was related to educational levels and area of residence (pG0.05). Following patient education KAPSE of all the patients improved significantly (p G0.05) and there was no difference between patients of different areas of residence. Conclusion: This study demonstrated that regular patient education significantly improves different aspects of Knowledge, Attitude Practice and Self-efficacy of patients regardless of their age, gender, education or place of residence, which could positively influence patient medication adherence. Asthma -a curable disease Sivanandhan Kasimalliah. Sri Ragavendra Clinic, Paediatrics, Pollachi, India. Background: In normal individuals, the Serum Histamine Binding Capacity (SHBC) is 20 to 30% whereas it is only 0 to 5% in allergic patients. The SHBC can be raised with Histaglobulin (HG) if it is given after preparing the patient properly and drugs like mast cell stabilisers, bronchodilators, antihistamines and anti-inflammatory drugs are given along with HG therapy. In allergic patients serum IgE level is high which comes down after HG therapy and medicines. Patient becomes symptom free and cured. Methods: It was an observational study for 14 years. 161 patients were registered at different periods from out-patients consisting of all age-group, both sexes and from urban and rural areas. Immunoglobulin E (IgE) levels were estimated and other routine tests were done before starting treatment. Patients were prepared before HG therapy; HG was given in two phases: Phase I Y 6 primary doses, Phase II Y3 booster doses. Along with HG therapy, drugs like mast-cell stabilisers, and H1 receptor-specific Histamine antagonists (Loratadine, Rupatadine, Cromolyn etc.) bronchodilators (Theophylline), antihistamines (Cetirizine, Levocetirizine) and anti-inflammatory drugs (Monteleukast) were given. 2 to 3 months after 3rd booster dose IgE levels were estimated for 61 patients. Patients were examined periodically to assess clinical improvement. Results: Out of 161 patients, 152 patients (94.6%) are free from asthma and allergic rhinitis symptoms. Among 152, 96 patients are free for more than 4 years. 9 patients did not respond satisfactorily. Out of 61 patients, 55 patients (90.2%, pG0.0001) showed reduction in IgE level and are clinically free from asthma. 6 patients showed raised IgE level and correspondingly no clinical improvement. It is also found out that allergic manifestations are lowest up to 2 years of age; between 2Y5 yrs, it increased and more males are affected; the occurrence rate came down in the age group of 6Y15 yrs in both sexes. Above 15yrs, again occurrence rate raised and more females were affected. Conclusion: Thus in our study comprising of 161 patients having the complaints of allergic asthma with or without allergic rhinitis, the HG therapy given after preparing the patient and continuing the above-mentioned drugs, during HG therapy was found to be effective in curing the allergic asthma. HG is effective in raising the SHBC thereby improving the immunity of the patient and also in reducing the serum IgE level. Flavonoids and related compounds as anti-allergic substances Toshio Tanaka 1 , Toru Hirano 1 , Mari Kawai 1 , Shinji Higa 2 , Junsuke Arimitsu 1 , Yusuke Kuwahara 1 , Tomoharu Ohkawara 1 , Kaori Nakanishi 1 , Keisuke Hagihara 1 , Yoshihito Shima 1 , Atsushi Ogata 1 , and Ichiro Kawase 1 . 1 Osaka University Medical School, Respiratory Med. Allergy and Rheumatic Diseases, Suita City, Osaka, Japan; 2 Tondabayashi Hospital, Internal Medicine, Tondabayashi City, Osaka, Japan. : The prevalence of allergic diseases has increased all over the world during the last two decades. Dietary change is considered to be one of environmental factors that cause this increase and worsen allergic symptoms. If this is the case, an appropriate intake of foods or beverages with anti-allergic activities is expected to prevent the onset of allergic diseases and ameliorate allergic symptoms. Flavonoids, ubiquitously present in vegetables, fruits or teas possess anti-allergic activities. Flavonoids inhibit histamine release, synthesis of IL-4 and IL-13 and CD40 ligand expression by basophils. Analyses of structure-activity relationships of 45 flavones, flavonols and their related compounds showed that luteolin, ayanin, apigenin and fisetin were the strongest inhibitors of IL-4 production with an IC50 value of 2-5 mM and determined a fundamental structure for the inhibitory activity. The inhibitory activity of flavonoids on IL-4 and CD40 ligand expression was possibly mediated through their inhibitory action on activation of nuclear factor of activated T cells and AP-1. Administration of flavonoids into atopic dermatitis-prone mice showed a preventative and ameliorative effect. Recent epidemiological studies reported that a low incidence of asthma was significantly observed in a population with a high intake of flavonoids. Thus, this evidence will be helpful for the development of low molecular compounds for allergic diseases and it is expected that a dietary menu including an appropriate intake of flavonoids may provide a form of complementary and alternative medicine and a preventative strategy for allergic diseases. Clinical studies to verify these points are now in progress and we will present the result in the congress. Sublingual progesterone dilutions as bronchodilator in asthmatic females Background: Asthma is associated with hormones. Increases in asthma symptoms have been associated with menstrual variances. Pre-menstrual asthma has been noted in the literature and studies from as early as 1921 have suggested an allergic reaction to hormones. Bronchial asthma in female patients could be due to allergy to self-hormones particularly progesterone. Interestingly, progesterone is also a potent respiratory stimulant. The present study was aimed to evaluate efficacy of sublingual progesterone dilutions as bronchodilator. Methods: This study was approved by the ethics committee of Austin, Texas. Sixteen females who had a previous diagnosis of severe asthma and who were nebulization dependent were selected for the study. After obtaining written informed consent from each patient spirometric studies were performed on these subjects Using sublingual progesterone as a bronchodilator. We analyzed changes over time of the forced expiratory volume in one second (FEV 1 ), the forced vital capacity (FVC), and the peak expiratory flow (PEF). For each patient we measured lung function three times: (1) before treatment, (2) after sublingual normal saline treatment (3) after sublingual progesterone treatment. Results: After treatment with sublingual progesterone, twelve of the sixteen patients (75%) experienced a significant bronchodilator effect (greater than 12% increase) in either FEV 1 or FVC. Eight (50%) experienced significant increase in both FEV 1 and FVC. Eight (50%), had an increase of 27% or greater in PEF. Conclusion: Progesterone dilutions are candidate drugs for the treatment of bronchial asthma. The possibility that dilutions of progesterone can act as a progesterone antagonist and that bronchial smooth muscle may be sensitive to such dilutions of progesterone due to activation of the L-arginine-nitric oxide (NO) pathway is interesting because of the close relationships between the immune and neuroendocrine systems. The study might have important implications for the development of a novel safe non-invasive treatment strategy against bronchial asthma due to hormone allergy. A randomized, double-blind, placebo controlled trial on the effect of zinc supplementation on bronchial asthma as measured by sputum eosinophil levels and asthma control test (ACT) in children ages 12Y18 y/o Kristine Marie Gutierrez. World Citi Medical Center, Pediatrics, Quezon City, Philippines. : Zinc may have its role in bronchial asthma by regulating airway inflammation and subsequently preventing asthma exacerbation. Objective: To determine the effects of zinc supplementation on bronchial asthma as measured by sputum eosinophil levels and asthma control test (ACT) in children ages 12Y18 y/o. Methods: After consent was obtained, 30 children at a local community ages 12Y18 years old diagnosed with intermittent, mild-moderate persistent asthma were randomly allocated to receive zinc supplementation 20 mg/day or placebo for thirty days. A change in the sputum eosinophil level and ACT score was obtained before and after intervention. Results: Fifteen subjects were included in the zinc group and 15 subjects in the placebo group. There were no differences in the demographic profile between the two groups in terms of age, severity of asthma, atopy, use of maintenance medications and infections during the study period. Intragroup statistics showed a statistical significance in the zinc group for sputum eosinophil (p=0.045) and ACT score (p=0.004) while the placebo group showed no statistical significance for sputum eosinophil (p=0.954) but was statistically significant for ACT score (p=0.045). The mean difference in sputum eosinophils between zinc and placebo showed no significant difference between the two groups. (p=0.270). The mean change in the ACT score of zinc and placebo group showed a significant difference between the 2 groups (p=0.067). Conclusion: Supplementation with zinc sulfate improves bronchial asthma in terms of ACT score but not in sputum eosinophil count in children ages 12Y18 years old. The application of the small doses of the human recombinant alpha-2 interferon oromucosal for the patients with seasonal atopy (bronchial asthma and allergic rhinoconjunctivitis) Veronica Osipova 1 , Seda S. Grigorian 2 , and Alexandr G. Chuchalin 3 . The immuneYenzyme method in commercial test system Immuno CAP 100, Pharmacy, Sweden was used to determine ECP. Results: The analisis of the journals of the patients showed that in comparison with three previous years the intensity of the stuffiness in nose, rhinorrea owing to the preventive therapy HRá2-IFN were authentically lower in comparison with the group of control and were 0.85 T 0.75 and 1.00 T 0.97; in the group of control j1.64 T 1.03 and 1.75 T 0.79. The necessity in B2agonists was authentically two times lower in the analyzing group of patients than in the group of control so that it was in the analyzing group 3.25 T 4.12 days while in the control group it was 7.82 T 7.78 days. The level of ECP before the medical treatment was higher in the analyzing group 252.89 T 141.15 mkg/l (the norm is till 11.5 mkg/l), than in the control group 61.36 T 55.47 mkg/l. After the medical treatment the level of ECP authentically became lower in the analyzing group 100.57 T 124.68 mkg/l, however after the season of the blossoming of trees the unauthentic increase of it was registrated till 161.16 T 143.32 mkg/l. The level of ECP in the control group unauthentically increased after the medical treatment till 84.81 T 111.60 mkg/l, however the level of ECP in this group authentically increased after the season of the blossoming of trees till 151.02 T 183.98 mkg/l. Conclusion: we can assert that the prevention usage of small doses of HRá2-IFN guarantees positive clinical effect on the pollinosis (bronchial asthma and allergic rhinoconjunctivitis), provides for the lowering of the levels of ECP and therefore the decrease of inflammation process in the windpipe. So it can be recommended as the modern medicine for the prevention theraphy for the patients with the seasonal atopy. Polyprenol as a possible supplement for treatment of steroid resistance in patients asthma Galina Kuznecova, Inese Joksta, and Klara Jegina. Preventive Medicine Research Laboratory, Allergy Unit, Riga, Latvia. Background: Plant Polyprenol (PP) is approved as a substitute of Dolichyl Phosphate (DP) which decreases P-glycoprotein, enhances IL-10 synthesis and alpha GP isoforms expression in vitro. In a proof of our previous study, the effect of oral administration of PP was investigated in steroid resistant asthma (SRA) patients. Methods: The samples obtained from 58 patients with asthma: 27 patients with SRA, 31 patients with steroid sensitive astma SSA and 20 donors. The patient's forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) was measured with a microspirometer, following recommended procedures for standardization of pulmonary function measurements. Patients with forced expiratory volume in 1 s (FEV1) G 70% predicted were studied. SSA patients were with FEV1 increased 9 30% after a 1-wk course of oral prednisone 20 mg twice daily and SRA if they failed to increase 9 15%. Dolichol (Dol) in blood and urine were assayed by HPLC method. Dolichyl phosphate, alpha-and beta-GR isoforms expression were measured in CD4+ T-cells. Results: Blood Dol concentration in persons SSA was 255.6 + 25.9 ng/mL and urinary Dol concentration was 18.9 + 4.5 mg/mmol. Blood Dol in patients with SRA was increased up to six times making up 689.2 + 47.9 ng/mL. Urinary Dol concentration was increased up to 590.9%, making up to 48.8 + 9.7 mg/mmol. The normal levels of Dol in donors are 8,0 + 2,9 mg/mmol in urine and 110,4 + 12,6 ng/mL in blood. The synthesis of DolP was 8.8Y10.5-fold decreased in T-lymphocytes in patients with SRA. 10 days course of PP 5 mg supplementation in SRA patients returned DolP concentration in blood, urine and T-cells to the normal level. 8 of the 10 SRA patients demonstrated a significantly increase FEV up to 30% after 2-wk course of oral prednisone 20 mg twice daily and 5 mg of PP. Conclusion: Dolichyl Posphate Cycle (DPC) disorder is a rate limiting mechanism of steroid resistance in asthma and associated with a marked defect of glucocorticoid receptors (GR) N-glycosylation in CD4+ T-cells. Presented findings provide evidence that PP supplementation in patients with SRA enhanced the expression of alpha GP isoforms, restore the possibility to induce IL-10 synthesis and made CD4+ T-cells more responsive to steroids. Plant Polyprenol and prednisone increase FEV and FVC in SRA patients. These preliminary results may have important implications for the design of alternative treatment approaches for steroid resistant asthma. Uncontrolled severe asthmatic patients clearly improve under anti-IgE (Omalizumab) treatment. 14 cases reported Beatriz Huertas 1 , Miguel Hinojosa 1 , Jesus Gonzalez-Cervera 1 , Belen De la Hoz 1 , M Luz Diez-Gomez 1 , Moises Sanchez-Cano 1 , and Adalberto Pacheco 2 . 1 Ramón y Cajal Hospital, Allergology, Madrid, Spain; 2 Ramon y Cajal Hospital, Pneumology, Madrid, Spain. Background: Anti-IgE Monoclonal antibodies (Omalizumab) had been recently introduced in the treatment of bronchial asthma. Last October 2005, the EMEA authorized this drug in the European Union for allergic severe asthmatic patients as an add-on therapy. Experience with a 14 patienttreatment is reported. Methods: 14 patients, 11 females and 3 males, were studied, age average 57 years, all of them suffering from severe allergic asthma (step V present GINA guideline) from the last 5Y37 yr, average 19 yr. A positive skin prick test and/or specific IgE antibodies to a variety of common aeroallergens was observed in all patients. IgE sera levels were between a range of 40Y1000 IU/ml (overage 316 IU/ml). FEV1 were under 80% predicted in all patients, overage (63% predicted). All patients were under treatment with high doses of inhaled corticosteroids, long acting inhaled beta-agonists and leukotriene modifiers and eleven of them also were on treatment with oral corticosteroids, prednisone, in a range from 5Y20 mg. Omalizumab 150 to 600 mg per month were subcutaneously administrated to each patient once or twice monthly depending of IgE levels and body weight. Every month after onset of treatment the following parameters were evaluated: Asthma quality of life questionnaire (AQLQ, E. Juniper) Asthma control questionnaire (ACQ, E Juniper), FEV1, physician overall assessment and possibility of side effects. Patients were following up for two to ten months. Results: A marked improvement of AQLQ and ACQ could be observed in 11 out of 14 patients. Physician overall assessment also improved in the same patients. This improvement was moderate in the remaining 3 patients. Response to treatment was appreciated into the first two months in all patients. FEV1 also improved in all patients in a range of 8 to 35% predicted. One patient developed marked edema in both legs after 12 wk and another acute otalgia and erythema nodosum in legs after 20 wk of treatment. Both patients withdrew omalizumab treatment. Oral corticosteroids were withdrawn in two patients and lowed in other four. Conclusion: Omalizumab treatment has proved to be effective in patients with uncontrolled severe allergic asthma. A total control was shown in more than half of patients and an enough control was got in the remaining patients. No exacerbations were observed in any patient all over the omalizumab treatment. Adverse reactions possibly but not sure related to omalizumab were shown in two patients. Efficacy of budesonide/formoterol in the treatment of children with moderate-severe asthma Lina Chen 1 , Yuzhi Chen 2 , Chunmei Zhu 2 , and Shuo Li 2 . 1 West China Women and Children's Hospital, Sichuan University, Pediatrics, Chengdu, China; 2 Capital Institute of Pediatrics, Asthma Clinic and Education Center, Beijing, China. Background: Our previous survey indicated that Chinese pediatricians tended to choose iv corticosteroids when treating moderate-severe childhood asthma,while GINA recommends the combination of inhaled corticosteroids (ICS) and long-acting " 2 agonists (LABA) for the treatment of persistent asthma. In order to evaluate one combination product budesonide/formoterol in the treatment of moderate-severe childhood asthma, we investigated its efficacy and safety. Methods: In a 12-week study, 34 children with moderate-severe asthma (28 males, 6 females,mean age 9.7T2.5years; receiving no systemic corticosteroids 4 weeks prior to study and having no LABA or SABA 1 week before study) were given nebulized budesonide suspension and salbutamol plus ipratropium bromide to relieve acute exacerbations at the clinic, then were given budesonide/formoterol (160ug/4.5ug) one inhalation in the morning and two in the evening for 2 weeks and one inhalation twice daily afterwards till the end of 12 weeks. Oral prednisone for 1Y3 days and slow release salbutamol for 3 days were prescribed to severe asthmatic children. Lung function, symptom improvements and adverse effects were mornitored after inhalation therapy. Results: Lung function variables such as FEV1, FEV1%, FVC, FVC%, PEF, PEF%,MMEF and MMEF% were statistically improved 2 weeks and 1 month post treatment (PG0.01).The mean time for relief were 3Y4 days after inhalation therapy. 91.2% of the children gained complete asthma control in 2 weeks and all children gained complete control at 4 weeks, 8 weeks and 12 weeks post treatment. No child had emergency treatment and short-acting " 2 agonist inhalation during the period. Such side effects as hoarseness and pharyngeal discomfort were observed. Conclusion: Budesonide/formoterol is an effective and safe treatment option for children with moderate-severe asthma, such treatment regimen may reduce intravenous infusion, simplify therapy and improve patients' adherence to treatment. 467 Abstract withdrawn 468 Therapy of chronic airway diseases with nucleic acids Stefanie Thiele, and Andreas Pahl. University of Erlangen, Department of Pharmacology, Erlangen, Germany. Background: RNA interference (RNAi) holds considerable promise as a therapeutic approach to silence genes involved in inflammatory pathways. It has been reported that intranasal administration of small interfering RNA (siRNA) is able to down-regulate protein expression. Aim of the project is the development of a nucleic acid based therapy for asthma and other chronic airway diseases. Methods: Different knock-down strategies were evaluated in vitro. Various cell lines were transfected with siRNA or short hairpin RNA (shRNA) using different transfection reagents. Furthermore, the optimal route of application in vivo was investigated. The intranasal administration and the intratracheal administration of RNAi agents were investigated. For these studies constitutively GFP-expressing transgenic mice were used, which allowed the monitoring of cell specific knock-down effects. Finally genes relevant for chronic airway diseases will be knocked down in vivo. Results: In comparison to other transfection reagents, a cationic lipid showed highest transfection efficiency along with lowest toxicity in vitro as well as in vivo. Using this reagent in vivo an uptake of shRNA in 30% of lung cells could be detected. In comparison down-regulation of GFP in cultured lung cells from transgenic GFP-mice was analysed. siRNA directed against GFP silenced GFP expression by about 40%. Conclusion: The cationic lipid transfection reagent is suitable for RNAi in vitro and in vivo. Lung cells are amenable for RNAi agents as shown by intranasal application of a shRNA vector. Effect of immunotherapy with ISS-ODN and allergen in animal model of mugwort allergy Junwoo Kim, and Bin Zhou. University of Tennessee, Department of Medicine, Memphis, United States. Background: Despite a number of effective pharmacological options for the prevention and treatment of the pathophysiologic responses that occur in sensitized patients on allergen exposure, the termination of allergic hypersensitivities remains an elusive therapeutic goal. By specific immunotherapy (SIT) with allergen extracts, allergy can be cured, however it has a limited scope of efficacy. Immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) was known to provide potent allergen-independent immune redirection from Th2 into Th1. Objective: We investigated that with the animal model of mugwort allergy, the allergic inflammation is controlled by ISS-ODN and what is the mechanism of the immune redirection. Also we investigated that ISS-ODN with allergen as a model of potentiated SIT could have better anti-allergy effects on allergic inflammation induced by specific allergen over ISS-OND alone. Methods: Experiments were performed in four groups of mice which designed to develop mugwort allergy with sensitization and aerosol challenge with mugwort extract. Group 1 received 100 mg of ISS-ODN with 110 mg of mugwort allergen as a therapeutic group, Group 2, ISS-ODN with bovine serum albumin, Group 3, C-ODN (control ODN) with mugwort allergen, Group 4, ISS-ODN alone. Cytokine profiles of IL-4, IL-5, IL-10, IL-13, and IFN-+, degree of inflammation in lung histology, and differential counts of bronchoalveolar lavage cells were measured. Results: Significant decreases of allergic inflammation in lung were observed in ISS-ODN with allergn group and ISS-ODN alone group comparing to negative control groups. IFN-+ was significantly increased in ISS-ODN with allergen group and ISS-ODN alone group comparing to control group. Conclusion: Allergic inflammation is controlled by ISS-ODN with allergen, which could be a new treatment of human allergic diseases. Estrogen exerts protective effects in a murine model of asthma Methods: A mouse model of asthma was established by sensitizing and challenging female BALB/c mice with ovalbumin (OVA). In addition, some mice were given intranasal (IN) IL-13, which induces an asthma-like disease, including airway inflammation and airway hyperresponsiveness. For in vivo studies, the effect of IN E2 treatments on methacholine (MCh)-induced bronchoconstriction was analyzed by using a Buxco system, and airway responsiveness was recorded as PenH values. An increase in bronchoconstriction is reflected by an increase in PenH values. PenH was expressed as the mean T SEM. For in vitro studies, lungs of OVA-sensitized female BALB/c mice were sectioned and mounted in a perfusion chamber, and observed under a microscope. Images of selected airways were captured pre-and posttreatment with acetylcholine (ACh) or E2. Results: In vivo, the MCh-induced PenH of OVA-sensitized mice before treatment with E2 was 8.30T0.58. When this group of mice was treated IN with 1 nM E2 4-hr prior to MCh challenge, their PenH was lower (5.54T0.84). OVA-sensitized mice pre-treated with IN PBS, on the other hand, did not display any change in PenH. In preliminary studies, E2 also suppressed airway responsiveness in IL-13-treated mice (PenH of 5.35T2.39 versus 7.34T1.95 for E2 and control mice, respectively). In vitro, ACh-induced airway contraction (lumen area measured 99.7 2m 2 after 1 2M ACh compared to 116.8 2m 2 at baseline) was reversed when treated with 1 2M E2 (109.1 2m 2 ). Pre-treatment of the airways with 12M E2 pre-treatment blocked the ACh-induced contraction (lumen area measured 125.1 2m 2 after 12M ACh compared to 126.8 2m 2 at baseline and 125.8 2m 2 after E2). Conclusion: These data suggest that estradiol can inhibit and prevent AHR associated with antigen and IL-13 in a murine model of asthma. Thus, estrogen analogues might play a therapeutic role in the management of asthma. Allergens and rhinovirus infections are among the most common elicitors of asthma, a severe disabling disease affecting more than 300 million people worldwide. We report the construction of a recombinant combination vaccine for allergen-and rhinovirus-induced asthma. Using a peptide from one of the most frequent respiratory allergens, the major timothy grass pollen allergen Phl p 1, and the human rhinovirus-derived coat protein VP1 required for infection of respiratory cells a recombinant fusion protein was produced. Immunization with this fusion protein induced in mice and rabbits protective IgG antibodies which recognized the allergen and neutralized the infection of cells expressing the receptor for human rhinoviruses. The vaccine exhibited neither IgE nor T cell-mediated allergenic activities. The described principle may be used for the combined vaccination against allergic and infectious asthma. An Internet-based questionnaire study was conducted of patients with asthma and chronic obstructive pulmonary disease (COPD) and parents of children with asthma to evaluate adherence to regimens of treatment and convenience of use of inhaled and transdermal preparations. Valid responses were obtained from 1,470 patients. Among asthmatic patients, the percentage of those who selected Btaking as prescribed[ was 52.7% for inhalant users and 83.2% for transdermal users. Among patients with COPD, the corresponding values were 54.7% and 86.6%. There was a significant difference (pG0.01) in percentage between inhaled and transdermal preparations for both airway diseases. The most common reason for poor adherence was Bfrequency of administration[, and a high proportion of patients, 83.2%, preferred once-daily administration. In addition, patients who had used both preparations preferred transdermal to inhalant drugs, to a significant extent. In conclusion, health care professionals should further educate their patients about the importance of treatment with inhalants, since poor adherence to treatment with inhaled agents significantly hinders achievement of optimal efficacy. In addition, transdermal tulobuterol patch, which is administered once daily as a long-acting beta2agonist, appears to be useful for long-term control of both asthma and COPD. Aerosol characteristics of admixture of budesonide inhalation solution with beta2-agonist, procaterol Toshiko Itazawa 1 , Yuichi Adachi 1 , Hiroyuki Mochizuki 2 , Naoki Shimojo 3 , Toshishige Inoue 4 , Toshiyuki Nishimuta 5 , Akihiro Morikawa 2 , and Sankei Nishima 6 . 1 University of Toyama, Pediatrics, Toyama, Japan; 2 Gunma University Graduate School of Medicine, Pediatrics, Maebashi, Japan; 3 Chiba University, Pediatrics, Chiba, Japan; 4 Sumitomo Hospital, Pediatrics, Osaka, Japan; 5 Shimoshizu National Hospital, Pediatrics, Shimoshizu, Japan; 6 Fukuoka National Hospital, Pediatrics, Fukuoka, Japan. Background: Nebulizer solutions of asthma medications are often mixed together in order to simplify inhalation regimens, although not recommended. We therefore evaluated the effect of admixture on aerosol characteristics. Methods: An 8-stage cascade impactor was used to measure the particle size distribution of admixture of Pulmicort \ Respules \ (budesonide, 0.5mg/2mL) with Meptin \ Inhalation Solution Unit (procaterol hydrochloride, 302g/ 0.3mL) from a jet nebulizer, PARI LC Plus \ . Concentrations of each drug were assayed by HPLC. Physico-chemical compatibility was also assessed up to 48 hours after mixing. Results: Impaction analysis revealed no differences in aerodynamic size and output of each drug between admixture and single-drug solutions. The mass median aerodynamic diameter (MMAD) of budesonide from the admixture with procaterol was 2.92 T 0.03 2m, and 2.99 T 0.14 from single-drug solution. The respirable mass of budesonide from the admixture was comparable with that from single solution (146.8 T 2.9, 147.6 T 8.2 2g, respectively). There was no significant change in pH or visual identification of a precipitate in the admixture. Recovery rates of each drug kept more than 96% of the initial values during the observation period. Conclusion: Our study demonstrated compatibility of co-administration of budesonide with procaterol in the aspect of aerodynamic characteristics and physico-chemical stability. In vivo data will be needed for the clinical implications of our findings. Efficacy of sublingual immunotherapy in asthma control Sylvia Novakova 1 , and Rositsa Dimitrova 2 . 1 The IVth Outpatient Clinic, Allergology, Plovdiv, Bulgaria; 2 Regional Hospital of Pulmonary Diseases, Allergology, Veliko Tarnovo, Bulgaria. Background: It has been of great interest to evaluate the clinical efficacy of sublingual immunotherapy (SLIT) in achieving asthma control. This interest is justified as the most recent GINA guidelines put an emphasis on control of asthma as a baseline for asthma management. Furthermore, this is the first study on SLIT in Bulgaria since this rout of administration of immunotherapy has been available for two years in the country. Copyright @ 2007 World Allergy Organization. Unauthorized reproduction of this article is prohibited. The aim of the study was to investigate the potential effect of one year SLIT with dust-mites on asthma control. Methods: 50 patients with asthma and allergy to dust mites (Dermatophagoides pternyssinus and/or Dermatophagoides farinae or both) were included in the study. 21males and 29 females, aged from 12 to 35 years (mean 23,5). SLIT has been prescribed in 25 of them (12 men) for one year along with other antiasthma therapy. A control group of 25 patients (10 men) had been maintained on medical therapy alone. Asthma Control Test /ACT/ was performed to asses the level of control for the last four weeks. On the basis of number score, asthma control was estimated as: out of control, partially controlled and completely controlled. Assessments of FVC and FEO1 were performed too. Results: From the group of patients, subjected to SLIT + medical treatment, 12 (48%) were completely controlled, 9 (36%) Y partially controlled and 4 (16%) out of control for the last four weeks of the study. From those on medical treatment alone, 5 (20%) were completely controlled, 10 (40%) partially controlled and 10 (40%) were out of control for the past four weeks. FVC and FEO1 did not differ in both groups. Patients on SLIT were better controlled (pG0.05) after the first year of treatment. Conclusion: The results of the study suggest that SLIT affords additional benefit to medical treatment in patients with asthma and allergy to dust mites. The results suggest that SLIT, added to medical treatment is of advantage in achieving asthma control. Background: Depigmented and polymerised allergen vaccines have shown to be efficacious using objective and subjective outcomes. It is now recognized that treatments under evaluation should enable patients to feel better in their day-to-day life activities. The objective was to evaluate the impact in the quality of life of a group of 64 asthmatic patients sensitised to house dust mites. Materials and Methods: The study was double-blind and placebo-controlled. Sixty-four patients were randomly allocated to either receive active treatment (n=32) or placebo (n=32). The active treatment was a therapeutic vaccine containing a mixture of depigmented and glutaraldehyde polymerized mixture of allergen extracts of D. pteronyssinus and D. farinae. The Asthma Quality of Life Questionnaire developed by Marks was used. It was conducted at baseline and after each administration of allergen vaccine (19 times). This is a 20-item self administered questionnaire, developed to measure quality of life in adult subjects with asthma by measuring a total scale score together with subscale scores for breathlessness, mood disturbance, social disruption and concerns for health. The result was expressed as the mean of all 19 measurements, Mann-Whitney's test was used to evaluate the statistical differences between both groups, and Hodges-Lehmann for the effect size of these differences. Results: The AQLQ overall score was the sum of all individual scores. The values of the mean for these scores were 7.44 (95% CI: 5.78Y9.11) for the active group and 11.44 (95% CI: 9.67Y13.22) for the placebo. The differences between both groups were significant (p=0.043, Mann-Whitney's test) and relevant (Hodges-Lehmann test: j4.36, 95% CL: j8.69,j0.10). The active group showed an improvement of 34.98% over placebo. Conclusion: The results of this clinical trial show that the immunotherapytreated patients, in contrast with untreated, experience a significant improvement in the overall quality of life related to asthma. The statistical significance as well as the improvement on the active group compared with the placebo occurred also in the domains of breathlessness, social disruption and concerns for health, with the exception for mood disturbance. The leukotriene receptor antagonist pranlukast improves quality of life in poorly controlled asthmatic patients Hiroko Arioka. International Medical Center of Japan, Pulmonology, Tokyo, Japan. Background: Asthma is a chronic inflammatory disease, and inhaled corticosteroids have been shown to be very effective in controlling symptoms. However, it is important to minimize the dosage of corticosteroids used because of local and systemic adverse effects. The importance of leukotrienes in the pathogenesis of asthma is established and many studies have shown that leukotriene receptor antagonists can improve asthma control in patients with poorly controlled asthma despite the use of inhaled corticosteroids. Objective: The aim of this study was to assess the effects of 8 weeks' pranlukast treatment on quality of life and pulmonary function in patients with inadequately controlled asthma. Methods: This study was conducted at 312 hospitals and clinics in Japan from October 2004 to March 2005. We recruited 1546 outpatients (mean age 51.4T18.5 years) with mild-to-severe asthma (as defined by the Japanese Society of Allergology) who were not well controlled with daily antiasthmatic treatment. During the 8-week treatment period, patients received pranlukast (225mg, twice a day) in addition to their previous treatment. Patients were required to complete symptom diaries and record peak expiratory flow (PEF) on a daily basis. Quality of life (QOL) was assessed using the Asthma Health Questionnaire (AHQ). Results: A total of 839 asthmatic patients were enrolled. Significant improvements were observed in the AHQ total score and all subscale scores. When the patients were classified according to baseline disease severity (mild, moderate and severe), the improvement from baseline remained significant for all AHQ subscale scores other than the economics subscale in all patient groups. Patients also demonstrated significant improvements from baseline in PEF (+34.9 L/min; p G 0.001), forced vital capacity (+0.17L) and forced expiratory volume in 1 second (+0.14L; both p G 0.005). Conclusion: The leukotriene receptor antagonist pranlukast improves QOL and pulmonary function in asthmatic patients inadequately controlled with antiasthmatic therapy. Effectiveness of short-term oral corticosteroid for preventing relapse following the emergency treatment of acute asthma Tomoyuki Serizawa, Shoji Yoshida, and Itsuo Iwamoto. Asahi General Hospital, Research Center for Allergy and Clinical Immunolog, Asahi City, Japan. Background: Acute asthma is responsible for many emergency department (ED) visits. It has been shown that high-dose intravenous methylprednisolone in addition to standard emergency treatment for acute asthma induces the early termination of the asthmatic attacks and reduces the number of hospitalizations. In addition, it has been reported that about 15 to 30% of the asthmatic patients treated in the ED will relapse to require additional interventions within 2 weeks of ED discharge. However, little is known about the efficacy of oral corticosteroid in reducing the relapse in the asthmatic patients discharged from the ED after treatment of acute asthma. The purpose of this study was to determine the effectiveness of oral corticosteroid in reducing relapse following the emergency treatment of acute asthma. Methods: Patients with acute asthma were treated with nebulized salbutamol and intravenous 125 mg methylprednisolone and 200 mg theophylline and were then discharged from the ED. Upon discharge, some patients were given oral 20 to 30 mg/day prednisolone for 5 days. Results: The asthmatic patients who received oral prednisolone had a significant decrease in the need for repeated emergency care (5.4%, 4 out of 74 patients, pG0.0001) within 2 weeks as compared with those who did not (33.3%, 73 out of 219 patients). The mean dose of prednisolone was 24.4 mg/ day and the mean duration was for 4.4 days. Conclusion: It is concluded that a 5-day course of medium-dose oral prednisolone (0.5 mg/kg/day) in outpatients prevents the relapse following the emergency treatment of acute asthma. Management of childhood asthma in east-java indonesia Sulistiowati Santosa 1 , Handoko Tanuwidjaja 2 , and Hugo van Bever 3 . 1 Adi Husada Hospital, Paediatrics, Surabaya, Indonesia; 2 Dr. Ramelan Navy Hospital, Pulmonary and Allergy, Surabaya, Indonesia; 3 National University Hospital, Paediatrics, Singapore, Singapore. Background: Treatment guidelines for childhood asthma is available, but it is important to know whether this guideline is being used in East Java. Methods: Using a standardized questionnaire on monitoring and treatment of childhood asthma, distributed to doctors treating childhood asthma in East Java. Results: Only 70 (20%) out of 350 doctors participated in this questionnaire study. General practitioners were 75.7%, 5.7% paediatricians and 2.9% pulmonologists. 88.6% worked in policlinic or private hospitals, and only 5.7% worked in university hospitals. Symptom score, peak flow meter or spirometry never or seldom used to monitor childhood asthma (94%). Nebulised salbutamol/terbutaline was the first drug of choice (63.8%) for treating acute asthma attack, followed by iv or im corticosteroids (23.7%), and iv aminophylline (30.6%). The three main criteria for admitting acute attacks were based on personal clinical judgement (80.0%), no improvement after 2 or 3 doses of salbutamol inhaler (44.4%) and other factors such as home distance and travel time (37.0%). IV aminophylline was seldom used (60.0%). Systemic (57.1%) and inhaled corticosteroids (50.7%) were seldom used in outpatient setting. Corticosteroids were used in all admitted patients. The corticosteroid of choice were dexamethasone (50.0%), methylprednisolone(45.7%), and were used same the dose for 5 days. Oxygen therapy, based on pulse oximetry reading were used only for severe asthma (45.6%). Antibiotics were used only in case of pneumonia, otitis media or sinusitis (40.0%). Drug of choice in the maintenance treatment of asthma in all ages were the same: LABA was the first choice, followed by LABA + inhaled corticosteroid and inhaled corticosteroids. Maintenance treatment was seldom used in all ages. The main criterium to start a maintenance treatment was the severity of the symptoms (47.1%). Specific immunotherapy never been used (72.1%). Conclusion: Although treatment guidelines for asthma in children is available, in daily practice we still find many modifications, including the usage of a LABA as first choice maintenance treatment. More efforts in socialization of the guidelines and providing tools for monitoring and treatment of asthma in children are needed. The use of yoga as an adjuvant therapy in the management of bronchial asthma: based on a randomized controlled trail Ramaprabhu Vempati, and Deepak KK. All India Institute of Medical Sciences, Physiology, New Delhi, India. Background: Complementary and alternative medicine (CAM) therapies such as herbal therapy, acupuncture, yoga, chiropractic, relaxation techniques, nutrition and dietary supplements, continue to gain popularity as modalities for the management of asthma. However there is a dearth of qualitative research with strong methodology to understand plausible mechanism of action of these therapies on asthma. The present randomized controlled trial (RCT) examines the use of yoga as an adjuvant therapy in the management of bronchial asthma. Methods: The study was conducted on 57 adult patients having mild or moderate bronchial asthma who were allocated randomly to either the yoga group (n = 29) or the wait-listed control group (n = 28). The control group received conventional treatment whereas the yoga group received, in addition to conventional treatment, also an intervention based on certain yogic practices. The outcome measures were assessed in both groups at 0 wk (baseline), 2 wk, 4 wk and 8 wk. Results: In the yoga group, there was a steady and progressive improvement in pulmonary function. There was also a trend towards a reduction in exerciseinduced fall in timed vital capacity (FEV1) in the exercise-sensitive (exercise induced asthma) subgroup of subjects in the yoga group. However, there was no corresponding reduction in the excretion of urinary prostaglandin D2 metabolite (11"-PGF2!) in response to the exercise challenge, which is an indicator of mast cell activation. There was also no significant chan there was a significant reduction in serum soluble interleukin-2 receptor (sIL-2R) levels after 2 wk in yoga group alone. NNT for quality of life indicators worked out to be 2.41 in QOL symptoms, 1.66 in QOL activity limitation, 1.91 in QOL emotional function, 1.70 in QOL environmental stimuli and 1.82 in total quality of life. The results indicate that of yoga in addition to conventional treatment as an adjuvant therapy in asthmatic patients results in improvement in pulmonary function and quality of life; reduction of T-cell activation and response to exercise stimuli. These findings provide yet another substantial evidence towards the use of yoga as an adjuvant therapy in the management of bronchial asthma. Background: The effect of inhaled steroids on oxidative stress in asthmatics is unclear. In this study the levels of lipid peroxides in the serum of asthmatic patients, whose symptoms were controlled with inhaled corticosteroids and long-acting beta(2)-agonists, were measured. Methods: Twenty-four asthmatic patients and 15 matched, healthy controls were recruited. Oxidative stress levels were quantified by measuring thiobarbituric acid reactive substances. Results: After 3 months of treatment, the mean lipid peroxide concentrations were significantly higher in asthmatic patients than in the healthy controls (4.3 +/j 0.15 micromol/mL vs. 3.4 +/j 0.03 micromol/mL, respectively). Conclusion: The parameters of oxidative stress was higher in patients with asthma than in healthy controls, even when the asthma is well controlled after 3 months of treatment. Effect of thalidomide in murine model of asthma Background: Tumor necrosis factor-alpha (TNF-alpha) has been identified as a proinflammatory cytokine that might be important in airway hyperresponsiveness (AHR). Thalidomide exhibits diverse actions of anti-inflammation and immunomodulation including attenuation of TNFalpha production. Objective: To evaluate the role of thalidomide in murine model of asthma. Methods: Six-week-old female C57BL/6 mice were sensitized with LPS 0.1 mg/ml and ovalbumin (OVA) via intranasal route and were exposed to OVA via intranasal route for 3 days. Thalidomide 50mg/kg was given via gavage twice a day from a day before the challenge. Airway responsiveness, inflammatory cells, cytokines in bronchoalveolar lavage fluids (BALF), serum OVA-specific IgE, IgG1, IgG2a, and histological examination were carried out. Results: Airway hyperresponsiveness decreased significantly in mice treated with thalidomide. There were no differences in IL-4, IL-5, IL-12, IFN-gamma, and eotaxin levels in BALF. Conclusion: Thalidomide decreases AHR in murine model of asthma. Therapeutic comparison between low-dose sustained-release theophylline dry syrup and capsule in children with mild persistent asthma Theophylline has been recently reported to have concurrent antiinflammatory effects at low therapeutic plasma concentration which is below the dose at which significant clinically useful bronchodilatation is evident. Sustained-release formulation as capsule and dry syrup forms was developed to reduce its adverse effects and improve its clinical effects. We compared between therapeutic effects of theophylline dry syrup and capsules in children with mild asthma. Methods: Ninety children with mild asthma were randomized to receive either theophylline dry syrup (n = 44) or theophylline capsules (n = 46), 4 mg per kilogram of body weight twice a day for 12 weeks. Baseline and serial measurements of daytime and nighttime asthma symptom score were performed. Compliance score, drug swallowing score, and drug usability score were measured every 4 weeks. Each scoring rated on a scale of 0Y4. Serum theophylline concentration were mesured at 4 and 12 weeks. To examine anti-inflammatory effect of theophylline on asthma, Serum eosinophilic cationic protein as a marker of airway inflammation caused by eosinophil was measured 12 weeks pre-and post-administration. Results: Daytime and nighttime asthma symptom score of two groups after 4 weeks significantly improved than baseline score. Daytime and nighttime asthma symptom score in dry syrup group were statistically lower at all time points except for nighttime symptom score at 4 weeks. Compliance score, drug swallowing score, and drug usability score in dry syrup group were significantly higher at end time point. Only in dry syrup group, serum ECP at end time point was statistically lower than baseline. Conclusion: Low-dose sustained-release theophylline may be safe and effective in bronchial asthma and this effect may be mediated by its antiinflammatory action mechanisms. Especially, when used in children with asthma, dry syrup formulation is recommended because of its higher compliance than that of capsule formulation. Steroid-dependent bronchial asthma: how to reduce the dose of oral glucocorticosteroids and improve asthma control? Rustem Fassakhov, and Irina Reshetnikova. Kazan Scientific Research Institute of Epidemiology and Microbiology attached to Rospotrebnadzor, Allergology and Immunology Department, Kazan, Russian Federation. Background: Patients with chronic severe asthma are often dependent on the long term prescription of oral glucocorticosteroids. The use of steroids is associated with serious side effects. Physicians treating such patients continue to search for alternative therapies that reduce the need for chronic dosing oral steroids. Aim: To assess efficacy and safety of 24-months treatment with Salmeterol/ Fluticasone propionate (100/ 1000 mcg daily) and Fluticasone propionate (1000 mcg daily) in 105 steroid-dependent bronchial asthma patients aged 18 to 60 years, who had received oral glucocorticosteroids (predisolone equivalent) less than 10 mg daily (75,2%) and more than 10 mg daily (23,8%) for at least 6 months but not more than 10 years. Methods: This was comparative, randomized, multicentre, open study in parallel groups (15 visits at 2 weeks intervals). Oral glucocorticosteroids dose reduction phase started at 3-rd visit. Results: Inspite of continual treatment with oral glucocorticosteroids patients did not reach asthma control. After 6-months treatment maintenance dose of prednisolone was 6 times lower in comparison with baseline. Moreover, oral glucocorticosteroids were abolished in each second patient receving Salmeterol/ fluticasone propionate and each third patient receiving Fluticasone propionate. At the same time asthma control and quality of life significantly increased. Administration high doses of fluticasone propionate and decreasing dose of oral glucocorticosteroids lead to reliable increase the morning level of serum cortisol by the sixth month of treatment in both groups of patients. Conclusion: The treatment with Fluticasone propionate and Salmeterol/ fluticasone propionate allows reduce the dose of oral glucocorticosteroids, moreover in the most causes patients can abolish oral glucocorticosteroids maintaining good asthma control. Tatiana Ospelnikova. The Gamaleya Institute of Epiremiology&Microbiology RAMS, The Gamaleya Institute of Epiremiology&Microbiolog, Moscow, Russian Federation. : Respiratory infections (viral and bacterial) may be trigger of atopic bronchial asthma (ABA) exacerbations. Was revealed II-III degree Interferon (IFN)-a and IFN-g production deficiency of leukocytes by all 19 patients with moderate and mild ABA in unstable remission phase and also acute mixed (viral and bacterial) respiratory infections. The purpose of this study was to use low molecular IFN inducer cycloferon for to improve IFN system indices and to reduce ABA exacerbations rate. It is known, that low molecular IFN inducer cycloferon have both antiviral and immunomodulating activities, can be successfully used in acute respiratory viral diseases. Cycloferon have affinity to alveolar macrophage receptors and induce of IFN in lungs. After the administration of cycloferon IFN-a and IFN-g producing capacity of leukocytes was 2-fold higher in comparison before treatment (pG0,05). 82% of patients (n=9) Aim: The aim of the study was to compare the two different topical corticosteroids in the treatment of asthmatic patients. Material and Methods: 100 naive asthmatic patients with the baseline values of FEV1 from 60-80% were divided in two groups. The first group received BDP and second received B aerosol in total of 800 2g/day, for 4 weeks. Short acting "2 agonist (salbutamol) was used as needed. Clinical symptoms, FEV1, FVC, PERF and the usage of "2 agonist were recorded. Local adverse effects were also documented. Results: Indices of dynamics show an increase of the values of FEV1 and FVC during the four weeks_ therapy with B and BDP, were in average of 22% and 15,6%, respectively. In patients who took B, FEV1 was increased in average 25%, and in patients who took BDP 19%; the difference in proportions was not statistically significant, it was accidental in our sample (pG0,5). In patients who took B, FVC was increased in average 16,3%, and in patients, who took BDP 15%, the difference in proportions was not statistically significant, it was accidental in our sample (pG0,88). With B, minimal increase of FEV1 was 9,3%, and maximal was up to 51% of the baseline value. Minimal increase of FVC was 3,7%, and maximal increase of the value was 36% after therapy with B. With BDP minimal increase of FEV1 was 5,8% and maximal was up to 40% of the baseline value. Minimal increase of FVC was of 5%, and maximal increase of the value was 31% after therapy with BDP. The difference between morning and evening values of PEFR under the influence of B became reduced in 86% and under the influence of BDP in 92%, the difference in proportions was not statistically significant (pG0,08). 25 patients complied on sore throat 19 on hoarseness and in 6 Candida albicans was isolated. The therapy with topical corticosteroids improved the clinical symptoms and values of FVC, FEV1 and PERF in both study groups, but that difference between them was not statistically significant. Fluticasone propionate/salmeterol combination compared to Fluticasone propionate alone in asthma (our experience) Gorica Breskovska, Biserka Kaeva, Gencijana Stevcevska, and Jagoda Stojkovic. Pulmology and Allergology Clinic Medical Faculty, Pulmology and Allergology, Skopje, Macedonia, Fyrom. Background: The rates of morbidity and mortality associated with asthma are increasing.Asthma is chronic inflammatory disease characterized by recurrent attacks of dyspnea associated with wheezing. Inhaled corticosteroids are the effective controller medications currently available.ICS may not provide optimal control of asthma when taken alone and additional therapy is necessary. Objective: The objective of the research work is to identify the efficiency of Fluticasone/salmeterol (discus inhaler) a 250 2g tvice dauly compared to Fluticasone a 2502g twice dauly in patients with asthma. We have analyzed a group of 35 asthmatics (aged 21Y32) being sensitive to allergen dermatophagoides pt and grass pollen. We split them up into two groups:group I consisted of patients who have undergone FP/Sal. (no=19); group II were the patients who have been treated by Fluticasone alone (no=21). All of them have been given a skin sensitivity test to Dermatophagoides pt. from 2+ to 3+,symptom score (0Y30,FEV1,morning and evening expiratory flow. Results: The results of the nvestigation showed statistically significant improvement of thr score symptoms with the first group:the reduction of the number of asthmatic approaches,the increase of FEV1 from (78,1T3,2) to (95,3T2,1) and a cough prevention (pG0,01).Following a 12 weeks tratmwnt,93% of the patients from the first group and 73% of the patients from the second group have not shown any side effects during the treatment period.We have notice significantly greater improvements in morning and evening peak expiratory flow of the patients from the first group.Patients from the second group needed additional therapy of short acting beta2agonist. Conclusion: Patients on FP/Sal. had significantly greater improvement in asthma control than patients on Fluticasone allone. Study of inhalation therapy with special reference to inhalation technique and patient's perception Suresh Koolwal. Sawai Mansingh Medical College, Jaipur, Respiratory Diseases, Jaipur, India. : Drug delivery by aerosol rout is often regarded as a twentieth century innovation,but in fact records of inhalation therapy can be found in the writtings of ancient cultures, notably those in China, India, Greece, Rome and the Middle East. The modern era of inhalation dates back from 70 years ago. One of the major problem in the management of Bronchial Asthma and COPD lies not in the airways but on the prescription. Technique of inhalation is crucial for the management of such diseases. With this background the present study was designed to find out, How the patients are using their inhalation devices? How does patient education affect their inhalation technique? What are their views and apprehensions regarding inhalation therapy and problems they face with this mode of therapy. The study was undertaken in 284 patients, 196 using Dry Powder Inhalation and 88 using MDI. The results were evaluated and we found that about half of the patients were not using their devices correctly. Among incorrect users majority were those with MDI. Among correct users majority of patients were educated the technique by their physicians or paramedics. One fourth of patients were wrong while preparing for inhalation, more than 80% incorrect users were not breathing out before inhalation, half of the patients were inhaling drug with inadequate efforts and more than 85% were not holding breath after inhalation. Cough, sudden spasm and dysphonia were the pr5oblems faced by 20%, and fear of addiction was the most common apprehension of using inhalation. Details of the study will be presented. A psychosocial perspective: Asthma in the third world, a portrait of a society malady Arnaldo Capriles Hulett. Hospital San Juan de Dios, Unidad de Alergologia, Caracas, Venezuela. : Asthma in Latin America is a pressing and growing health problem affecting the young and mostly impoverished urban populations across the continent. Venezuela, an oil rich country with 80 % or more of the population living under variable conditions of poverty, has an acute asthma morbidity that ranks ahead of diarrheas and seconds the Bviral syndrome[ and thus representing more than one million visits per year at the ambulatory health services of the Ministry of Health (caring for the majority of the population), a tendency constantly on the rise for roughly 27 million inhabitants. Efforts for over 25 years by an existing National Asthma Program and the dissemination of GINA and similar guidelines have all been unable to reverse this trend; the prevailing asthma approach centers around a perverse paradigm of vicious cycles consisting of repeated acute asthma care; this boldly denotes a morbidly persistent lack of awareness in the minds of physicians (Health System), patients and lay public at large. A unique trait, valid alike for the upper rich as well as Venezuelas' poor and marginal classes, is acknowledged from many years of psychotherapeutic practice: it is a tendency of sorts to live in a Bstate of emergency[, a lacking in future provisions of any kind and an attitude of strong accent on the immediacy of life along with no risk prevention. Poverty, understood as a Bprecarious condition of daily life[, lies beneath these set of highly complex issues. Our asthma abounds in these attributes and have been in part considered a psychosomatic expression of this malady i.e.: Bif....when a next asthma attack or crisis comes, we'll then see about it[. The above represents an overwhelming historical challenge that heavily pounds on individual minds and society, undermining a much needed reflection and making the surge of a proper response no easy task. The implementation process of asthma guidelines in primary health care Eeva Ahonen and Risto Makinen. Espoo Health Care Centre, Finland, Leppävaara, Espoo, Finland. Background and Aim: To improve the clinical practices is known to be a challenging process. Here we describe the implementation of asthma guidelines and the development of the house rule and patient self management protocol. The setting is Leppävaara health care centre with six GP-nurse teams each providing primary care for 10 000 inhabitants. National Asthma Programme 1994Y2004 delegated the responsibility in preventing and treating asthma to the primary health care. The evidence based guidelines published 2000, updated 2006, gives the general outline in asthma treatment. Materials and Methods: Participatory observation and document analysis using qualitative methods Results: Two GPs and two nurses were nominated to asthma co-ordinators to act as internal consultants. They organized many workshops during 2000Y2007 to introduce the guidelines and to develop asthma management. The governmental Centre for Pharmacotherapy Development joined the implementation activities by providing training of trainers, and material and tutorial support. Therefore, the workshops after the year 2003 were problem-based and multi-professional, and used EBM, feed-back of prescription practises, and case reports as learning methods. The Leppävaara health centre team produced a written agreement of inter-professional collaboration and asthma management process, the Leppävaara house rule of asthma care, in a workshop in 2004. They arranged a new workshop in 2005 to assess the changes in asthma care. A multi-professional group developed a written self management protocol for the patients in 2006. The team renewed the house rule in a workshop in 2007 according to updated national asthma guidelines. Discussion: The evidence based asthma guideline was a valuable Bgolden reference[. To implement it the interactive multi-professional workshops were used to develop the house rule. This method seemed to support the professionals to commit to true team work and to jointly agreed practices. The implementation has been a necessary process, which has to be continued by supporting, giving feedback and evaluating. We will evaluate the professionals' attitudes towards the usefulness of the house rule, the rate of admissions to the secondary care, and changes in prescription patterns. Producing guidelines is not enough: The improvement of clinical practices demand the interesting and useful implementation process to every day work. Exercise-induced bronchospasm (EIB) is widely prevalent in asthmatic individuals particularly in children varying from 40% to 90% in various reports. There are, however, few studies addressing the effects of asthma severity on airway responsiveness to exercise from India. Aim: In examining the mechanisms of exercise-induced bronchospasm (EIB), it is important to determine which factors mainly affect the severity of EIB. We report such factors in patients with asthma by stepwise multiple-regression analysis. Methods: Sixty five asthmatic children (age range 10Y14 years), 200 adults between 20 to 44 years old were selected randomly as representative of persons living in the city of Mumbai. All subjects filled out a questionnaire on respiratory symptoms. Bronchodilator therapy was withheld three days before the study. The exposure to house dust mite allergens was estimated from dust samples obtained in the subjects' homes. Eosinophil count was done from peripheral blood. Serum Il-4 was estimated in adult asthmatics by ELISA method. Atopic status was labeled on the basis of Total serum IgE and Modified Skin Prick Test and subjected to exercise challenge. assessments of atopy. An exercise challenge test was done (8 min) bout of cycle-ergometer exercise): decrease in PEFR of 15% was considered positive for EIB. Results: Atopic adults showed significantly higher sensitization to house dust mite compared to children (p G 0.05). Significant rise in incidence of EIB was observed in adults (62.6%) against 40% in children ( p G 0.05) showing direct relationship of house dust mite sensitization in both the groups ( adults correlation coefficients: r = 0.955 p G 0.001, children r = 03135 p T 0.05). Relative risk of developing EIB associated with HDM sensitivity was higher in adults (2.85 at 95% CI ). Regression analysis showed direct relationship between the severity of EIB and serum IL-4 levels (r2 = 0.03 p G0.0001). Raised eosinophils were well correlated with serum IL-4 levels. Conclusion: The findings suggest that though EIB is common in children, adults were more significantly affected. Allergen specific IgE to house dust mite, serum IL-4 and eosinophilia were the major associated contributing factors in the development of EIB in asthmatic patients. Clinical Implications: Patients with exercise induced bronchospasm may benefit from novel tharapies specially designed to target the specfic mechanisms underlying airway inflammation. Valeria Ghiglione, Sara Balestracci, Fulvio Braido, Sara Cauglia, Ilaria Baiardini, and Giorgio Walter Canonica. University of Genoa, Department of Internal Medicine, Genoa, Italy. Background: As indicated in Asthma guidelines, treatments`goal should be the total control of asthma and consequently a minimal impact of asthma on patient`s life. Aim: The aim of our study was to investigate the relationship between FEV1, control of asthma and quality of life in real life. Methods: In a 1 month period, asthmatic patients were enrolled consecutively during their routine control visit. A spirometry was performed for every patient. Patients were asked to complete 2 different questionnaires: the Asthma Control Test (ACT), a brief instrument developed to assess asthma control of patients in a clinical setting, and the Rhinasthma, a 30 items tool aimed at evaluating the impact of respiratory allergy on quality of life. It provides 4 different scores: Upper Airways (UA), Lower Airways (LA), Respiratory Allergy Impact (RAI) and Global Summary (GS). Results: Seventy-four asthmatic outpatients (30 men and 44 women; mean age 44.11 + 14.83) were enrolled. 53 had concomitant rhinitis. For 39 patients (52.7%) the control of asthma was achieved; 35 (47.3%) didn`t obtain asthma control. FEV1 value (mean 88.45 + 16.13) correlates with ACT and with Rhinasthma, LA and RAI. ACT significantly correlated with the four Rhinasthma scores. Subdividing patients according to the score (optimal and good control vs non control) and comparing the level of control measured by means of ACT we obtained a significant correlation between ACT and LA (p. 0.0001), RAI (p. 0.01) and GS (0.0001) and no correlation with Rhinasthma UA score (p 0.053). The analysis of the patients' population showed that a well or total control was achieved in 53% of patients with asthma and 51% of patients with both asthma and rhintis. The subanalysis of 53 patients with concomitant rhinitis showed worse Rhinasthma scores for all the domains compared with patients with asthma alone (p. G 0.05). Discussion: Our results show that asthma control is related to an improvement in quality of life; nevertheless, asthmatic patients often refer also symptoms due to rhinitis that have an important impact on their Quality of Life. So, in order to improve Quality of Life of asthmatic patients, the treatment of patients with rhinitis and asthma includes both lower and upper airways treatment. Relationship between quality of life related to chronic cough and asthma level of control; a pilot study Sara Balestracci, Fulvio Braido, Valeria Ghiglione, Silvia Brandi, Ilaria Baiardini, and Giorgio Walter Canonica. University of Genoa, Department of Internal Medicine, Genoa, Italy. Background: The antiasthmatic treatments nowadays available can induce a good or total disease control. Recent evidences have also shown that the level of control is strongly related to health related quality of life (HRQL). Aim: The aim of our real life observational study was to evaluate the relationship between HRQL related to chronic cough and asthma level of control defined according to Asthma Control Test score (ACT). Methods: In a 1-month period, asthmatic patients were consecutively enrolled during their scheduled visits. Patients were asked to complete 2 different questionnaires: the ACT, a brief instrument developed to assess asthma control of patients in a clinical setting, and the Chronic Cough Impact Questionnaire (CCIQ), a 16 items validated questionnaire divided into 4 areas (sleep/ concentration, social relationship, mood and daily life impact). Results: Seventy-four asthmatic outpatients (30 men and 44 women; mean age 44.11 SD: +/<14.83) were enrolled. 53 had concomitant rhinitis. 39 patients (52.7%) had total or well controlled asthma (ACT score 9 20); 35 (47.3%) had non controlled asthma (ACT score G 20) . ACT didn`t correlate significantly with any of the CCIQ domains: sleep/concentration (p. 0.689), social relationship (p. 0.741), mood (p. 0.704), and daily life impact (p. 0.458). Taking into consideration that concomitant rhinitis could induce a post nasal drip related cough, the subanalysis of patients with both asthma and rhinitis didn`t show any correlation between CCIQ and ACT. Discussion: Previous studies showed a significant correlation between asthma level of control and HRQL. Despite these evidences, analysing the HRQL related to a specific symptom (cough) occurring in asthma, the correlation is lacking. Our pilot observations seem to demonstrate that ACT is an useful tool, but a specific instrument should ameliorate patients`evaluation. Moreover, the presence of other causes of cough should be investigated in well controlled asthmatic patients with persistent cough. Clinical effect of clean air administered directly to the breathing zone (Airsonett Airshower\) on perennial allergic asthma Background: Studies previously made on air cleaning has shown little or no effect on patients with perennial allergic asthma. We examined a novel treatment (Airsonett Airshower\) using a laminar airflow directed to the breathing zone of a patient during the night, on teenagers and young adults with moderately severe allergic asthma. We hypothesised that this directed treatment would have effect on the bronchial inflammation and consequently, the quality of life. Methods: 22 patients 12Y33 year of age (mean 18.8 yr) was randomized double-blind to 10 weeks active treatment respective 10 weeks placebo treatment. All patients received both active and placebo treatment (cross-over) with a 2 week wash-out period in between treatments. Maintenance treatment with inhaled corticosteroids was unaltered (400 6g budesonid/day or equivalent) during the trial period. Health related quality of life, miniAQLQ, was the primary effectiveness measure. Exhaled nitric oxide (FENO) and lung function was also investigated. Results: Active treatment resulted in an improved quality of life compared to placebo (mean score 0.54, p G 0.05, n = 20). Also an effect on the bronchial inflammation was detected with significantly lower FENO values during the active treatment period (mean <6.95 ppb, p G 0.05, n = 22 Anxiety symptoms in allergic patients: identification and risk factors Ziad Adwan. Saha, Allergy, Rio de Janeiro, Brazil. Objective: Multiple relationships between anxiety, allergic symptoms, and treatment difficulties have been observed. The aim of the present study was to estimate the prevalence of anxiety disorders in outpatients with various allergic diseases, to identify diagnostic cues or possible risk factors, and to test the usefulness of self-administered questionnaire screening at the allergy clinic. Methods: Six hundred forty-six (646) consecutive patients with rhinoconjunctivitis (59.3%), asthma (26.8%), or Bother[ allergy (13.9%), aged 16 to 65 years, completed self-administered questionnaires in six outpatient allergy clinics; 60 of the respondents also participated in structured psychiatric interviews. Anxiety was measured with the Spielberger State-Trait Anxiety. Results: According to the interviews, STAI-T 9 52 predicted with 86% accuracy a current psychiatric diagnosis, without differentiating between anxiety and depression. Using this threshold, the rate of anxiety and/or depressive disorders is estimated as 19% (95% CI: 15.9Y22.1) in our unselected allergic outpatient sample; 46% of these patients never received any psychopharmacological treatment, indicating that anxiety related disorders are underdiagnosed and undertreated. Risk indicators were female gender; asthma; perennial symptoms; sleep problems; nonspecific allergy triggers like strong emotions; stressful situations; and considerable limitation in everyday activities attributed to the allergic symptoms. Conclusion: Our findings confirm a high rate of anxiety and/or depressive disorders in patients visiting the allergy clinic. Self-administered questionnaires such as STAI-T provide reliable help for the identification of these frequent psychiatric problems. Key Words: anxiety, allergy, rhinitis, asthma, Spielberger State-Trait Anxiety Inventory, depression. Issues: Health care assessment system in Nepal is still take long time and high cost. Collection of data about prioritization of community health problems by the community is essential for planning and monitoring of programs and interventions for improving community health status. Scientific Rapid Community Health Assessment Methodology needs to be validated in rural community developing country like Nepal which is economic and less time consuming. Objective: To validate the Rapid Community Health Assessment Methodology (RCHA) for prioritization of community health problems in a rural community. Study Area: 14 Villages of rural areas. STUDY POPULATION: 34 Primary School Teachers from 10 randomly selected Primary Schools in above villages. (One Teacher each from Class I to V, in each School). VALIDATION: Heads of households from 500 Households (50 per village, systematically sampled). DATA Collection technique: Self-Administered Questionnaires for Teachers, Interview Schedules for Heads of Households. DATA ANALYSIS: with the help of EPI info program. Results: There was significant correlation between the responses of the school teachers and heads of households on community health problems viz., (a) the prioritization of ten village problems(r = +0.77, p G 0.02), (b) prioritization of utilization of services of various health functionaries for treatment of and advice for children`s illnesses (r = + 0.75, p G 0.05), and (c) prioritization of households using water from different sources(r = + 0.975, p G 0.02). The method was also found to be more rapid (3.3 times) and less costly (6.3 times) compared to the traditional household survey method. Conclusion: Rapid Community Health Assessment Methodology (RCHA) for prioritization of community health problems in a rural community is validated. The information thus obtained can be utilized for purposes of health policy and program planning, monitoring and evaluation. This is especially relevant for micro planning of child health services in developing countries. Repeated use of questionnaires for monitoring disease control programs must be carefully considered. Further studies to confirm and reconfirm the results of this study may be done before wider application of above methodology. Aim: To assess the temporal changes of medicines consumption in relation to risk factors for respiratory symptoms/diseases in a subsample of a general population of Italian adults. Methods: Within the Po Delta and Pisa epidemiological surveys, subjects who habitually used medicines at baseline were selected as cases. Controls were non-drug consuming subjects randomly drawn and matched for gender and age groups. 1663 subjects (89% of those invited) participated in a telephone interview (follow-up = 11 years) with questions on current drug consumption, respiratory health status, life habits, and comorbidity, beside socio-demographic characteristics. Main characteristics were: 37.2% males, mean age 63yrs, cases = 46.2%. We used logistic regression models accounting for smoking habit, gender, age, comorbidities, respiratory symptoms, occupational/passive smoking/environmental exposure. Results: There was a non significant increase between baseline and follow-up for prevalence of asthma (5.8 vs 7.1%) or rhinitis (20.6 vs 23%). Prevalence rate of habitual use of any medicine at follow-up was 70.1 vs 46.2% at baseline (p G 0.001). Among the users (N=1166), we found higher prevalences of broncho-pulmonary (7.5 vs 4.8%) (significantly) and antiallergic drugs (2.4 vs 1.3%) (non significantly) at follow-up. New regular use of any medicines at follow-up was 60.4%, while persistence and cessation were 37.6 and 18.6%, respectively. Only asthma resulted significantly related to the use of medicines at baseline (OR 1.69, 95%CI 1.04Y2.73), while at follow-up the relation was significant for both asthma (2.92, 1.51Y5.66) and rhinitis (1.55, 1.03Y2.33).). The association of the use of specific medicines (brochopulmonary/ antiallergic/cardiovascular/diuretic vs other medicine/no use) was significant with asthma at baseline ( Roya Sherkat. Alzahra University Hospital, Infectiouse Disease, Isfahan, Islamic Republic of Iran. Introduction: Chronic urticaria/angio-oedema has traditionally been defined as daily or almost daily symptoms lasting for more than 6 weeks. Chronic urticaria affects 0.5Y1% of individuals (life-time prevalence) and significantly reduces quality of life. Optimal management of chronic and acute intermittent urticaria depends on a thorough understanding of clinical presentation, etiologies, triggers and aggravating factors. An underlying extraneous cause for chronic urticaria cannot be identified in most patients, but infections may play a causative role in a few cases, and when present, chronic infections such as dental sepsis, sinusitis, urinary tract infections and cutaneous fungal infections should be treated. However exhaustive investigations searching for underlying infections are not indicated. Infection with Helicobacter pylori (HP) has been proposed as a possible cause, but the association is unlikely to be causal (particularly in otherwise asymptomatic children where the background prevalence of HP infection is high). Also complement activation can mediate or augment histamine release from mast cells via the anaphylatoxin C5a. This inflammatory pathway is triggered by antibody and antigen interacting to form immune complex-associated urticaria, e.g. in hepatitis C and hepatitis B, EBV, other viral and possibly parasitic infections. However common triggers for episodes of chronic urticaria are intercurrent viral infections and possibly stress. Human brucellosis is an infectious disease produced by Brucella species: small, coccoid or rod-like, aerobic, Gram-negative bacteria. Every organ and system of the human body can be affected in brucellosis a fact that underscores the importance of including brucellosis in the differential diagnosis in areas of endemic disease, even if clinical features are not entirely compatible. The infection can also affect the skin. We present a case of chronic urticaria who also had fever and chills at presentation for a week. His urticaria-like papules and plaques and a red, raised, itchy rashes especially in his face continue after his fever has stopped. In a vast array of laboratory investigation brucella serology, using both eliza and serum agglutination method, was positive. Only treatment of underlying infection has leaded to resolution of symptoms. Bronchial hyperresponsiveness in patients with chronic urticaria Zikica Jovicic, Mirjana Bogic, Sanvila Raskovic, Aleksandra Peric Popadic, and Ljiljana Stefanovic. Institute of Allergology and Immunology, CCS, Department of Allergology, Belgrade, Serbia and Montenegro. Background: Involvement respiratory system in patients with chronic urticaria is still a controversial subject. Large number of patients report dyspnea. The aim of this study was to investigate the present of bronchial hyper responsiveness in patients with chronic urticaria. Methods: A total of 23 patients (17 women, 6 men, age range 19Y56 years) who had urticaria for at least 6 weeks, without other known causes of urticaria, were enrolled in the study. Diagnostic programme of urticaria include: eosinophil count, skin prick test with inhalation and nutritive allergens, test for NSAID intolerance, pulmonary function tests and methacholine provocation. Patients are investigating during a phase of remission. Results: Three patients had (13%) abnormal pulmonary function tests, ten patients (43,4%) showed significant bronchial hyperresponsiveness on methacholine provocation, three (13%) had elevated eosinophil count, four (17,3%) had positive skin prick test with inhalation and/or nutritive allergens and twelve (52,1%) had NSAID intolerance. Airway hyperresponsiveness was not associated with eosinophil count, skin prick test with inhalation and nutritive allergens, NSAID intolerance. Conclusion: Significant number of patients with chronic urticaria show bronchial hyperresponsiveness, and performing of this test is reasonable in patients with chronic urticaria. An audit of referrals from public sector primary care physicians to an allergy clinic in Singapore Bernard Thong, Khai-Pang Leong, Chwee-Ying Tang, Yew-Kuang Cheng, Faith Chia, Justina Tan, and Hiok-Hee Chng. Tan Tock Seng Hospital, Singapore, Rheumatology, Allergy and Immunology, Singapore, Singapore. Background: An electronic registry for all new referrals (New Case Registry) to our adult Allergy Clinic was set up in July 1998 to determine the sources of referrals, referring diagnoses and final diagnoses. Sources of referrals include primary care physicians in the public and private sectors, specialists, national dermatology centre, military services and self referrals. Aim: To describe the pattern of referrals from primary care physicians in the public sector to an Allergy Clinic in Singapore. Methods: All new referrals were prospectively captured in the New Case Registry during the study period 1 July 1998 to 31 December 2006. Results: There were 227 referrals from public sector primary care physicians comprising 8.4% of all referrals. More than half (57.7%) were females and mean age was 36 T 18 years. The majority (79.7%) were Chinese. The most common referring diagnoses were drug allergy [DA] (35.7%), urticaria and angioedema (23.3%) and food allergy [FA] (10.1%). In contrast, the final diagnosis of the attending allergist was DA in 9.7%, NSAID intolerance in 22.5%, and FA in 7.5% respectively. Among cases with primary care physician diagnosed DA, the allergist`s diagnosis was DA in 23.5%, NSAID intolerance in 40.7% and not DA in 35.8%. Among cases with primary care physician diagnosed FA, the allergist`s diagnosis was FA in only 31.6%. Chronic idiopathic urticaria comprised 19.1% of all cases of urticaria/angioedema. The following conditions were under-diagnosed by family physicians compared to allergists: anaphylaxis (1.8% versus 4%), asthma (2.6% versus 6.6%), atopic eczema (1.8% versus 6.6%), allergic rhinitis (7.5% versus 19.8%). Among all referrals for suspected allergy, 5.7% were deemed not to have any allergic disorder upon completion of evaluation. Conclusion: DA, urticaria/angioedema and FA were the most common reasons for referral. NSAID intolerance accounted for the majority of cases of Fdrug allergy`referred by primary care physicians. Allergic rhinitis, asthma, atopic eczema, and anaphylaxis were under-diagnosed, potentially leading to patients being deprived of appropriate therapy for allergic airway disease and life-saving epinephrine autoinjectors. Increasing the awareness and education may be useful in improving the diagnosis and treatment of allergic diseases in the community. Chronic urticaria: alternative treatment with beta-agonists Summit Shah 1 , Lekha Nair 2 , and Jonathan Bayuk 3 . 1 Tufts-New England Medical Center, Allergy Department, Boston, United States; 2 Tufts-New England Medical Center, Pharmacy, Boston, United States; 3 Hampden County Physicians, Allergy, Springfield, United States. Background: To describe a 46 year old patient with chronic urticaria, unresponsive to standard therapy, who responds to oral beta agonists and to propose possible mechanisms of action. Methods: We performed basic bloodwork and checked eosinophil levels, ANA, ESR, CBC, CMP, anti-thyroid antibodies, TSH and complement levels. Skin biopsy was performed as well. Results: Patient with history of diabetes mellitus presents for treatment of chronic urticaria x 15 years. Initially, patient responded to corticosteroids but since then no medications have helped, including Allegra, Doxepin, and Periactin. Test results revealed normal eosinophil levels, negative ANA, normal ESR, CBC, CMP, Anti-thyroid antibodies, TSH and complement levels. As patient was unresponsive to other medications and requiring frequent oral corticosteroids, albuterol sulfate 4 mg tablets were given. Patient took approximately 32 mg of albuterol orally each day for approximately 14 days in addition to her anti-histamine. On return visit, patient`s symptoms had resolved completely. Conclusion: Beta agonists have a good side effect profile compared to oral corticosteroids and may be of great benefit in reducing urticarial symptoms. Possible mechanisms of action include inhibition of anti-IgE induced histamine release thru desensitization of beta adrenoceptors. They may be a promising pharmacotherapy in the treatment of chronic urticaria. Background: Chronic idiopathic urticaria (CIU) is a relatively common disease, and yet not much is known about the causative factors or the pathomechanism, which makes it difficult to cure. Due to its chronic nature, many patients suffer from significant detrimental effects on their quality of life (QOL). The purpose of this study is to assess the impact of CIU on the QOL of Korean patients, and to determine whether a relationship exists between QOL and the severity of disease. Methods: One hundred twenty four patients with CIU, who first visited our out-patient clinic from August 2005 to July 2007, were asked to complete a questionnaire designed to assess the effects of CIU on the daily lives of patients. QOL was divided into six different categories: mental status (MS), daily living activities (DLA), leisure activities (LA), self-perception (SP), treatmentinduced restrictions (TIR), and social restrictions (SR). All of the QOL scores were recalibrated to a 0Y100 scale, with 100 indicating the worst QOL, and 0 the most favorable. Pearson and Spearman correlation coefficients were used to analyze the relationship between the six different QOL categories, as well as the association between disease duration and QOL. One way ANOVA was used to analyze the relationship between disease severity and QOL. A p value less than 0.05 was considered significant. Results: The average QOL scores obtained from the questionnaire were as follows: MS (50.64), DLA (47.58), LA (39.85), SP (37.06), TIR (35.54), and SR (33.23). The results showed that patients were most affected mentally, and that social restrictions were the least significant. The six different categories showed a positive correlation amongst themselves, which demonstrates that CIU exerts an influence on not one, but many aspects of QOL. LA and TIR were significantly affected in longer term CIU cases (p G 0.05), while SP, MS, DLA, and SR did not show a significant relationship with disease duration. Disease severity was found to have a significant effect on the SR, LA, and TIR categories (p G 0.05). Conclusion: Based on these results, it is suggested that CIU has a negative impact on the QOL of Korean patients. Therefore, it is important to recognize the effects CIU can have on QOL and consider them as candidates for treatment as well. Neutrophil activation finding was noted in patients with ASA induced urticaria Sung-Jin Choi, Gyu-Young Hur, Seung-Youp Shin, and Seung-Hyun Kim. Ajou University School of Medicine, Allergy & Rheumatology, Suwon, Republic of Korea. Background and Objective: ASA ingestion could induce acute and chronic urticaria, however their pathogenic mechanisms are not understood. We compared the level of neutorphil activation and related cytokine between ASA intolerant acute urticaria (AIAU) and ASA intolerant chronic urticaria (AICU). Methods: The 88 patients with AICU, the 51 patients with AIAU and the 102 patients, normal control (NC) were enrolled. The clinical and laboratory findings including serum myeloperoxidase (MPO) level, IL-8 and IL-18 levels were compared among AIAU, AICU, and NC groups. The correlations between MPO and, IL-8, IL-18 were observed. Results: The level of serum MPO were significantly higher in AIAU than in AICU and NC (p=0.032, p=0.014 respectively). The level of IL-18 was significantly higher in AIAU than in NC (p=0.006), while no significant difference was noted in IL-8 level. Within the AIAU patients, significant correlation were noted between MPO and IL-8 (r=0.36, p=0.01), not with IL-18. It is known that about 20% of people develop urticaria at least once in their lives. Chronic idiopathic urticaria affects up to 3% of the population. The incidence of acute urticaria is higher in people with atopy, and the incidence of chronic urticaria is not increased in people with atopy. This study shows that most cases of urticaria are related to such gastointestinal disorders as active gastritis, esophagitis and peptic ulcer. One of the most interesting things is that urticaria is likely to be only a single symptom of a severe and maybe lifethreatening gastrointestinal disorder. Methods: 10 patients with acute (3) or chronic (7) urticaria passed gastroscopy after 5Y14 days of the onset of urticaria. In addition to urticaria, 3 had angioedema, 1 suffered from severe pruritus with desquamation and 1 developed palmar eczema. Results: All patients showed to have different gastrointestinal problems, which correlated very well with the anamnesis data. 7 had gastric erosions, among which 3 with esophagitis; 1 had esophagitis with atrophic gastritis, 1 esophagitis alone, 1 patient with severe generalized urticaria had 2 big (1 cm each) duodenal ulcers. The patient with concomitant pruritus had lots of severe gastric, esophagal and duodenal erosions. Only 3 patients suffered from ocasional epigastric pain or heartburn, but none said that symptoms were severe. Gastrointestinal cause was suspected because episodes of urticaria developed or increased after irritating or acid food (spice, citruses, strawberries), drinks (alcohol, soda) or drugs (NSAIDs, acetyl-cysteinum, ascorbinic acid) but trigger factors were always different and dose-dependent. 9 patients that underwent anti-reflux, antacid, antisecretory and some (2) antibiotic therapy along with symptomatic antihistamine drugs recovered (8) or showed much relief (1). 1 patient with severe pruritus didn't pass prescribed treatment and chose his usual intramuscular injection of diprospan instead. As always, he reported a short 2-week relief of symptoms. Conclusion: These data show that urticaria alone or with angioedema very often occurs as a symptom of a gastrointestinal disorder. It is very important to prescribe gastroscopy and adequate treatment not only to stop urticaria, but moreover to prevent gastrointestinal bleeding or perforation, very genlty operating with steroids, especially oral, in such patients. Etiology and prognosis of acute urticaria in a university dermatology clinic in China: a follow-up study Lin-feng Li and Yuan Cao. Peking University Third Hospital, Dermatology, Beijing, China. Background: Etiology and prognosis of acute urticaria has not been reported in China. Objective: To investigate the etiology and outcome of acute urticaria in a university dermatology setting. Methods: A prospective follow up study was performed. All patients with acute urticaria seen by authors between March 2005 and July 2006 in department of dermatology, Peking University Third Hospital were asked to participate. Etiologic agents and triggers were evaluated by comprehensive and detailed history and physical examination, supplemented with selected laboratory testing and follow up. Prognosis was assessed at 9 months after the first visiting. Cure was defined as complete free of symptoms for more than 6 months without relapse after cessation of the treatment. Results: 75 patients participated. There were 22 male, 53 female, with age ranged 1.5 to 83 years. Of those, 60% were between 18 to 40 years; 32% had a history of acute urticaria during childhood; and 6.5% were atopic. Etiologic agents or triggers could be identified in 54.7% of the patients including medications (21.4%), infection (20%), food (8%), physical stimuli (2.7%), insect bite (1.3%) and contactant (1.3%). In 71 patients finished the follow up, 58 cases (81.7%) cured. 13 patients, all etiology and triggers unidentified, progressed to chronic urticaria, but 6 of them free of symptoms at the end of evaluation. The overall cure rate in patients with disease duration less than one week was 94%, which was much higher than that of patients with a longer duration (41.2%, p G 0.05, chi-square test). Sex, age and severity of the disease had no effect on the prognosis. Conclusion: Etiology and triggers can be identified in more than half of acute urticaria patients. Medicaments, infection and food are common triggers. The outcome of acute urticaria is favorable. The longer disease duration (over a week) is an important risk for poor prognosis. Results: Among 152 patients remitted for evaluation of allergy to penicillins, 47(31%) had angioedema, but in only 39 (26%) penicillin was not excluded as the cause (exclusion by controlled administration of penicillins and s-tryptase). Aspirin (11), nonsteroidal antiinflammatory drugs (6), ACE inhibitors (10) and other drugs (10) were seen as the cause mediated by leukotrienes, bradykinin and direct mast cell degranulation. Food was suspected in 28. Insect sting the cause in 16/40 (honeybee 1/9 and wasp 15/31) and occupational agents in 5/43: latex (nurse), fish protein, flour, dyes, nikel. In only one the c1-esterase inhibitor was slight reduced. Angioedema was associated to allergic rhinoconjunctivitis and asthma. Infections and physical stimuli were well known inducers in rather few. Most patients with non-drug induced angioedema had prophylactic anaphylactic medication with corticosteroid and antihistamines at home. 48 had epinephrine (epipen, autoinjector). Conclusion: Urticaria and angioedema are the most frequent manifestations of severe allergic reactions (anaphylaxis). Many different causes may induce angioedema. However, IgE-mediated mechanism is the most frequent suspected cause of angioedema in a department of medicine. Successful treatment of chronic urticaria with low molecular weight heparin Background: Autologous serum skin test (ASST) has been used to detect circulating vasoactive and histamine-releasing factors in patients with chronic urticaria (CU). Recently, it has been observed that autologous plasma skin test (APST) scores positive more frequently than ASST in CU patients, and its positivity is associated with the activation of coagulation cascade. Aim: We report a case of chronic urticaria successfully treated with anticoagulant therapy using low molecular weight heparin (LMWH) to see whether symptom control is associated with reduction of coagulation activation. Methods: A 63-year-old woman, with a 3-year history of unremitting CU, was evaluated in our outpatient allergy clinic. No other diseases were present in her clinical history. CU improved only with prednisolone 0.5 mg/kg/day, plus anti-H1 antihistamine, while therapy tapering led to symptom recurrence. Known causes of CU were ruled out with laboratory and instrumental examinations. ASST, APST, negative control with saline solution and positive control with histamine were performed after a 5-day discontinuation of steroid and anti-H1 therapy, and repeated after 15 days of LMWH therapy. Prothrombin fragment F1 + 2, marker of coagulation activation, and D-dimer, marker of fibrinolysis, were measured by ELISA before starting LMWH therapy and 15 days later. Results: At baseline, ASPT gave an unequivocal positive response (20 mm mean wheal diameter), whereas ASST was negative; prothrombin fragment F1 + 2 value was 833 pmol/l (normal range 69/229 pmol/l) and D-dimer value was 27.15 nmol/l (normal range 0.50Y4.00 nmol/l) confirming an activation of the coagulation cascade and fibrinolysis. LMWH therapy (enoxaparin 4000 UI twice a day sc) was started. After few days CU improved and the patient stopped anti-H1 and steroid therapy, without recurrence of CU. Fifteen days later the patient was still asymptomatic, while receiving only LMWH therapy. APST became negative and we observed a marked reduction of both prothrombin fragment F1+2 (287 pmol/l) and D-dimer (2.84 nmol/l Background: Angiotensin converting enzyme inhibitors (ACE-Is) are known cause of angioedema. Its incidence is between 0.1% and and 2.2%, but its clinical relevance is due to risk of life-threatening involvement of upper respiratory tract. In the present study we reported the clinical features of patients complaining of ACE-Is-angioedema referred to our Unit in a 5 year period. In a subgroup of them we studied the potential mast-cell involvement by measuring serum tryptase. Methods: Descriptive, observational on ACE-Is-induced angioedema during a 5-year period (January 2001Y December 2006 . Serum tryptase level was evaluated by ELISA assay during the angioedema attacks, and in baseline conditions in three different groups: Group 1 = 12 patients complaining of previous ACE-Is angioedema; Group 2 = 15 patients with previous non-ACE-Is angioedema; Group 3 = 10 patients treated with ACE-Is without angioedema. Results: We observed 32 patients (19 male; mean age 62.3 yrs, range 42 Y 88 yrs) with ascertained ACE-I-induced angioedema. Only one patient was African, whereas the other were white. Patients reported an average of 3.8 attacks of angioedema (range 1Y 9). Upper respiratory tract involvement was observed in 2 subjects. Eleven patients reported a concomitant use of NSAIDs or aspirin (five at low dosage). One patient reported previous cough due to ACE-Is. Drugs responsable were: lisinopril (7), fosinopril (7), enalapril (6), quinalapril (6), rampril (4), captopril (2) . The period of time between the beginning of ACE-Is therapy and the first angioedema attack ranged from 10 days to 4 years (mean 1.4 years). Most of patients (72%) had an isolated angioedema (lips 53%, face 21%, tongue 9%). Only two patients were admitted to the medical intensive care unit and one of them was intubated. Follow-up information were available only for 16 subjects and 12 of them had no relapse after stopping ACE-Is treatment. During the attacks only one patient showed a slight increase of serum tryptase (12 ng/mL), whereas no significant change was observed in baseline condition in the three groups (Group A : 5.7 ng/mL; Group B : 4.9 ng/mL; Group C : 3.9 ng/mL of serum tryptase). Conclusion: ACE-Is are a relevant cause of angioedema mainly in adult and elderly population. Physicians should be aware of the potential risk of upper airway involvement. Mast-cells seem not to be involved in the pathogenesis. Keywords: Angiotensin converting enzyme inhibitors, angioedema, tryptase. Christian Bull, Susanne Haug, and Peter Schmid-Grendelmeier. University Hospital Zurich, Allergy Unit, Dep. of Dermatology, Zurich, Switzerland. Rationale: Hereditary angioedema (HAE) is characterised by skin swellings in various parts of the body, laryngeal edema or abdominal pain. Besides the deficiency or malfunction of plasma C1-inhibitor as cause of HAE, bradykinin is considered to be a key mediator of symptoms in HAE attacks. A new treatment for acute HAE attacks might be Icatibant, a synthetic and highly selective bradykin-B2-receptor antagonist which can be administered subcutaneously. We report on the effect of Icatibant in treating an acute attack in one patient in our hospital. In the open-label extension phase of a phase III, double blind, randomized multicenter trial (FAST-2) which compared Icatibant versus tranexamic acid, patients with abdominal or cutaneous attacks were treated with Icatibant 30 mg sc. Symptom score by patient, documentation of time for the onset of symptom relief measured by a visual analog scale and time to complete resolution of symptoms were the main assessments. Results: The patient treated in our hospital was a 22 year old, otherwise healthy male with known HAE since the age of 6 month. He has been having frequent attacks about twice per month which usually were treated with tranexamic acid. The patient experienced in total 11 attacks (7 abdominal, 2 cutaneous and 2 laryngeal) with an average duration of 4 days during the last 6 months. The patient came to the clinic with a severe cutaneous edema of his genitals. He received one single injection of 30 mg Icatibant sc. The onset of symptom relief was reached rapidly after approximately 10 minutes. Complete resolution of symptoms occurred within 5 hours, so the patient could leave the hospital. Icatibant was well tolerated, only mild injection site reactions such as pruritus were observed. No systemic adverse events were observed. Conclusion: In the present case Icatibant provided fast and complete resolution of severe cutaneous HAE symptoms and was well tolerated. The treatment course of the HAE attack in our case complies with the results of the double blind phase of the FAST-2 trial. The time to first improvement was 0.8 hours for Icatibant vs. 7.9 hours for tranexamic acid (p G 0.001). The primary endpoint (median time to onset of symptom relief) was met with 2.0 hours for Icatibant compared with 12.0 hours for tranexamic acid (p G 0,001). The end of attack (almost complete symptom relief) was reached after 10 hours during treatment with Icatibant compared to 51 hours for tranexamic acid (p G 0.001). Evaluation of the receptor occupancy by desloratadine and levocetirizine in allergic subjects Conclusion: In spite of the higher affinity (Ki) and longer t2 of D compared with L, the higher free plasma concentrations of L at 12 and 24 h post-dose produced higher percentages of RO probably accounting for the statistically significant higher inhibition of allergen-induced wheal & flare induced by L. The impact of hereditary C1-inhibitor deficiency on the development of atherosclerosis Methods: In a cross-sectional prospective study we included 57 adult patients and 20 age and sex matched healthy individuals. The prevalence of cardio-and cerebrovascular atherosclerotic diseases as well as distinct atherosclerotic risk profiles were determined. In addition, the thickness of the intima-media (IMT) of the common carotid artery was also determined. As severe HAE patients receive long-term danazol prophylaxis Yan attenuated androgen-, patients were divided into two subgroups according to prophylactic treatment. Results: Low prevalence of atherosclerosis was found in HAE patients: only 2 subjects were affected (1-1 in the danazol treated and not treated group, respectively) according to the medical history. Patients with long-term danazol prophylaxis had an increased risk profile for atherosclerosis: significantly higher body-mass index, LDL/HDL ratio, creatine kinase activity, creatinine, GPT and hemoglobin levels were found compared to the other HAE patients and healthy controls as well. However carotid IMT did not differ relevantly in study subgroups. Conclusion: Long-term danazol use results in a highly proatherogenic risk profile in patients with HAE. However, it seems that it does not lead to early atherosclerosis. The prevalence of cardio-or cerebrovascular diseases is low in HAE patients irrespectively to their prophylactic treatment and carotid IMT is comparable to healthy controls. We hypothesize, that the functional deficiency of C1-INH might be a protective factor in these patients against atherosclerosis Y which outlines the importance of the plasma enzyme systems in the process which are regulated by C1-INH. Patient perception of levocetirizine in urticaria: a multicenter study in Taiwan and 30 (31%) chronic urticaria. The use of concomitant antihistamines was reduced from 58% (baseline) to 27% at end of treatment. Conclusion: Results confirm western studies that levocetirizine potent, effective and well tolerated also in Taiwanese patients with urticaria. Most of the patients considered it as better than their previous treatments. Levocetirizine has a better overall efficacy than desloratadine in chronic idiopathic Urticaria: A double-blind, randomized, clinical trial Annick Barbaud and on behalf of CUTE Study Group. Hô pital Fournier, Service de Dermatologie, Nancy, France. Background: Levocetirizine (Levo) and desloratadine (Deslo) are H1antihistamines of the latest generation. Levo has been shown to be significantly more potent than Deslo in either histamine or allergen-induced wheal and flare studies. So far, no head-to-head clinical studies in Chronic Idiopathic Urticaria (CIU) have corroborated these findings. Methods: A multicenter, double-blind, 2-parallel groups (438 adults on Levo, 448 on Deslo), randomised trial over 4 weeks (w) of treatment to compare the clinical efficacy and safety of Levo 5mg vs. Deslo 5mg, both once daily. Primary endpoint: pruritus severity score (0 = absent to 3 = severe) assessed daily over the first week (w1) of treatment. Secondary endpoints: pruritus severity over all 4 study weeks (w4); pruritus duration, number and size of wheals over w1 and w4 (all on a 0Y3 scale); global satisfaction with treatment (0 = very dissatisfied to 10 = very satisfied), and safety. CIU composite score, representing the drug's overall efficacy, was also assessed. Conclusion: In this first head-to-head comparison of Levo and Deslo in CIU, patients satisfaction with Levo was significantly higher, onset of action of Levo was faster and the overall efficacy of Levo was significantly better than Deslo in relieving CIU symptoms. Urticaria and angio-oedema due to the daily 80 mg dose of acetylsalicylic acid Hong D. Oei, Wilsa L. Kartanegara, and Richard L. Oei. Albert Schweitzer Hospital, Allergology, Dordrecht, Netherlands. Background: Low dose acetylsalicylic acid have been widely prescribed to prevent or reduce the risk of strokes and heart attacks. There are many reports concerning its known side effects, such as: acute urticaria. However, reports of intermittent urticaria and angio-oedema due to the daily 80 mg intake of acetylsalicylic acid are not common. Methods: We have seen 4 patients (56y old man, 64y old man, 77y old woman and a 79y old woman) who used a daily dose of 80 mg acetylsalicylic acid for more than 6 weeks. They complained about multiple episodes of swelling of the tongue, cheeks and urticaria. The frequency varied from once a week or once in 2Y4 weeks, within a period of 6 to 12 months. Some patients consulted a dermatologist and another physician without any results. Routine screening investigations were negative. Results: The symptoms disappeared in 3Y6 weeks after they stop taking acetylsalicylic acid and taking clopidogrel instead. Urticaria and angio-oedema will reappear within 3Y5 weeks, after the patients continue taking the 80mg daily dose of acetylsalicylic acid, instead of the replacement clopidogrel. Overall efficacy and tolerability were assessed as Bgood/excellent[ by 75% and 68% of patients, respectively. At least 60% of all patients and 75% of those with moderate/severe symptoms reported complete recovery or marked improvement of any individual allergic symptom. 65% of subjects reported the onset of action as very rapid or rapid. Levo was reported as better than their previous therapy by 72% of patients. Good to excellent improvement in quality of sleep and daily activities was reported by 61% and 74% of patients without concomitant medications, respectively. Global satisfaction with Levo was high for both physicians and patients (VAS=7.4). Most frequently reported AEs were somnolence (7%) and fatigue (3%). Conclusion: We conclude that the intake of low dose acetylsalicylic acid may have caused urticaria and angio-oedema in these patients. The reason behind the attack of urticaria and angio-oedema that only happened in once a week or once in the 2Y4 weeks is still unclear. The intermittent nature of urticaria and angio-oedema attacks may also be the reason why many physicians are not aware that the daily 80 mg dose of acetylsalicycic acid may cause of this problem. We suggest replacing the daily 80 mg use of acetylsalicylic acid with clopidogrel, for those patients with urticaria and angio-oedema possibly caused by acetylsalicylic acid. Further studies are needed to investigate the mechanism of this disorder. Distribution of HLA DRB1 specifities in patients with chronic urticaria Gruzdeva Maria 1 , Inna Danilycheva 1 , and Margarita Boldyreva 2 . 1 Institute of Immunology, Basic Allergology and Immunetherapy, Moscow, Russian Federation; 2 NRC Institute of Imunology FMBA RF, Immunegenetics, Moscow, Russian Federation. Background: In 50Y80% of patients with chronic urticaria (CU) the origin of disease is unknown. In most of CU cases autoimmune mechanisms are involved. In CU patients auto antibodies such as aDNA, p-ANCA, ANF, RF often detected along with the clinical manifestations of autoimmune pathology (arthralgia, myalgia, photodermatitis, sub febrile body temperature). Predisposition to autoimmune diseases is connected to certain variants of HLA genes. Relative risk index Q2 demonstrates that individuals bearing this HLA gene variant have higher risk of autoimmune pathology development than other individuals. The goal of the study was to investigate association between certain HLA specifities and CU development. Methods: In our study patients were divided into 3 groups. First grouppatients with acute atopic urticaria (N = 30), second group -patients with CU with clinical manifestations of autoimmune pathology (N = 31), third groupnormal individuals (N = 135). HLA-genotyping was performed with SSPvariant of PCR. Results: In the first group half of chronic CU cases revealed from childhood. Weals persisted less than 6 hours in all cases, atopic background in 37% of cases, 100% efficacy of AH drugs application. In the second group: urticaria debut at the age of 24 to 51,urticaria persistence for 2Y4 years in average, existence of autoimmune pathology manifestation in reumo-tests. In the second group statistically significant increase of autoantibodies was found in comparison to the first group. According to HLA-genotyping data patients with CU demonstrated statistically significant increase in the level of HLA DRB1*04 in comparison to normal individuals. We demonstrates that relative risk in patients with CU for DRB1*04 specifity is 2,33. Data obtained points out possible autoimmune mechanism of urticaria development. Desloratadine reduces symptoms and improves quality of life in patients with chronic idiopathic urticaria: a multicenter, practice-based study in China Zheng Wang. Schering Plough (China) Ltd., Shanghai Schering Plough Pharmaceutical Co., Ltd., Shanghai, China. Background: Chronic idiopathic urticaria (CIU) is associated with a heavy symptom burden, which translates into significant impairment in quality of life (QOL). Non-sedating antihistamines are first-line therapy for CIU. Desloratadine (DL), a non-sedating, selective H1 receptor antagonist has been shown in multiple placebo-controlled studies in Europe and the United States to be effective in reducing the severity of pruritus and wheals in CIU, while improving QOL measures. The impact of DL on CIU has not been studied in a Chinese population. Methods: This open-label, multicenter study evaluated the efficacy and safety of DL 5mg QD administered for 28 days in patients with CIU treated during everyday practice in China. Patients aged Q18 yr and of either gender, with a Q6 week history of CIU were eligible. Patients had to have wheals present for Q3 days/week, and pruritus, wheals and overall CIU severity scores had to be Q2 (0=none to 3= severe). At baseline and days 8, 15, and 29 patients rated the severity of pruritus, wheal size/number, total CIU symptoms, interference with sleep and daily activities and overall CIU condition using a 4-point scale. QOL was assessed at the same time-points using the Dermatology Life Quality Index (DLQI). At day 29 the global response to therapy was rated (1=complete relief to 5= treatment failure); adverse events (AE) that occurred during the trial were collected and rated for severity/relation to study drug. Results: The study included 460 subjects (54.1% female), with a mean age of 37.2 years and a mean duration of diagnosed CIU of 25.7 months. Pruritus, wheal size, wheal number, total CIU score, interference with sleep and daily activities and overall condition of CIU were all significantly improved from baseline at days 8, 15 and 29 (P G 0.0001 for all). DLQI also improved significantly from baseline over the course of the study (P G 0.0001). The percentage of patients with complete or marked relief of CIU rose from 64.8% at day 8 to 80.3% at day 15 and 82.3% at day 29. DL was well tolerated and no serious AEs were observed. The most frequent AEs rated as possibly/probably related to treatment were drowsiness (3.7%), fatigue (1.7%), headache (1.5%), thirst (1.5%) and dizziness (1.3%). Conclusion: This practice-based study in Chinese subjects confirms that DL is effective and well tolerated in the treatment of CIU and is associated with improved QOL. Fresh frozen plasma for the treatment of acute hereditary angioedema Djahna Thomas 1 , Huamin Li 2 , Natalie Ball 2 , and Mark Scarupa 2 . 1 Washington Hospital Center, Internal Medicine, Washington, DC, United States; 2 Institute For Asthma & Allergy, Allergy and Immunology, Wheaton, United States. Background: Chronic prophylactic therapies for hereditary angioedema (HAE) have existed for some time but in the United States acute therapies are lacking. Fresh frozen plasma (FFP) has been used to treat acute attacks but its use is controversial because reports of FFP worsening HAE symptoms exist. Methods: Twenty-three patients with HAE answered survey questions regarding past experiences with FFP. The purpose of the survey was to determine whether FFP administration during acute HAE attacks ever lead to a worsening of symptoms. Additionally, patients were asked whether FFP was perceived to be beneficial. Demographic data and data on the types of attacks treated was also collected. Results: Six of 23 patients received FFP for acute attacks and an additional three patients received FFP for surgical or dental prophylaxis. Greater than 71 acute attacks were treated with FFP. One individual accounted for approximately 50 treatments. Attack locations included abdominal, peripheral, facial, genital, and laryngeal. There were no episodes of acute worsening after administration of FFP or spread to the larynx. There was perceived benefit from FFP in all but five instances. Conclusion: Until more specific therapies for the acute treatment of HAE are widely available, FFP should be considered as a treatment option particularly in non-laryngeal attacks. Hereditary angioedema prognosis in non treated Algerian patients Kamel Djenouhat 1 , Hassen Messaoudi 2 , and Mohamed Cherif Abbadi 1 . 1 Institut Pasteur d'Algérie, Department of Immunology, Algiers, Algeria; 2 CHU Mustapha Hospital, Internal Medecine Department, Algiers, Algeria. Background: Hereditary angioedema (HAE) is an autosomal dominant disease that is characterized by quantitative or qualitative deficiency of a plasma protein called C1 esterase inhibitor(C1INH). The disease is manifested by episodic attacks of nonpitting, nonpruritic, localized oedema that progresses rapidly without urticaria or erythema. Swelling of the intestine can cause intense abdominal cramping associated with vomiting and diarrhoea. Laryngeal oedema may prove fatal. The main of the present retrospective study is to report the follow-up care of HAE non treated symptomatic patients. Patients and Methods: For each patient clinical score was established and complement system investigation including antigenic quantification of C1 INH, C3, C4 and C1q by nephelometry (Dade Behring, Germany). And determination of C1 inhibitor function by chromogenic assay (C1 Y INH, IMMUNO AG, Vienna, Austria) were performed. Results: The two most striking matters of HAE spontaneous course in our patients comparing to literature data before androgenic therapy are, firstly a very low mortality rate (about 5% versus 30%) and secondly, only few of them (less than 10%) had developed moderate attack following dental care even without preventive drug. Conclusion: Our population seems to be more resistant to C1 inhibitor deficiency than European and American patients. We suppose that treated patients would become more sensitive to minor disorders than non treated ones, and also this difference could be due to environmental factor (s) such as microbes. Effect of cetirinax on the skin reactivity in patients with chronic urticaria Sonya Genova, Vania Tzvetkova, Hristina Dardanova, Desislava Yolcheva, Ludmil Terziev, and Temenuga Yosifova. Medical University -Pleven, Clinic of Allergy, Pleven, Bulgaria. Background: Cetirinax\ (Actavis), a derivate of piperazine, belongs to the family of the second-generation antihistamines. It is a fundamental pharmacological active metabolic product of hydrazine Y a first-generation antihistamine. Beside its antihistamine activity, Cetirinax\ posseses also other varied qualities that augment its antiallergic properties. Suppression of erythema and papula is traditional pharmacodinamical mechanism used to determine activity of H1 receptor antagonists. The purpose of that survey is to get in touch with suppressive effect of Cetirinax\ on erythema and papula caused by histamine in patient with chronic urticaria. The aims that we put were to survey suprressive effect according to time, clinical condition of patients, sex and age. Methods: The study involved twenty patients with chronic urticaria, 18Y63 years of age. Histamine sensitivity of all patients was given by prick-test with 10 mg/ml histamine 20th minutes before treatment with 10mg Cetirinax\. After that the same sensibility was determined on 30th minutes, 1st, 3rd, 6th, 9th, 12th and 24th hour. Results: The suppressive effect of Cetirinax\ set in 30th minutes and is highest marked in 3rd hour on the erythema and in 9th hour on the papula. The difference were statistic important compared to basic size of papula and erythema. Depression of erythema was equal like papula in various hours. Depressive effect was marked clearly in 24th hour.We determined that histamine papula and erythema were similar at patients independently ages, sex, and clinic condition. The clinic effect correlates to the excellent suppressive functions. Conclusion: Cetirinax\ is a medicine with excellent quality on the skin reactivity. Background: The impact of childhood asthma on the health care system is considerable. As one of the most chronic diseases of children, asthma is frequent caused for emergency room visits and hospitals admissions. Clinical observations indicate that atopic diseases, particularly bronchial asthma and allergic rhinitis are also common in Saudi Arabia. However, there were no formal studies until recently on the prevalence and etiology of childhood asthma in the country. Methods: Prevalence of Bronchial Asthma in school children were studied in different part of the country using a standardized protocol of 32 questionnaires. The questionnaire was designed by a committee of international expert and first used in 1985Y1986, before ISAAC launched its first phase of limited age prevalence studies. Our questionnaires, however is very similar to ISAAC questionnaires but include children from 06 to 16 years of age. For comparative purposes, we have continued using the same protocol for other parts of the country. The questionnaires were translated in local language and distributed through the Ministry of Primary Education to different urban schools of the Kingdom. Results: Their studies were completed at different intervals within the past several years. The combined data revealed varying prevalence of asthma with highest 24% being in a coastal city bordering Yemen called Gizan (n = 362) followed by Taif 23% (n = 594) and Hail 22% (n = 507) an agricultural city. The prevalence rate of asthma for other places where Al-Gazim 16% (n = 384), Abha 13% (n = 485), Dammam 12% (n = 889), Hofuf 14% (n = 923), Jeddah 12% (n = 531) and Riyadh 10% (n = 988). The prevalence for allergic rhinitis was much higher than asthma. Conclusion: Bronchial Asthma emerged to be one of the prevalent diseases in Saudi Arabia and showed regional diversity. The socio-economic differences between asthmatic and non asthmatic children were none striking. Environmental and geographical influences were considered to be the reasons of the variations in the rate of prevalence. Aim: A negative association between the number of siblings and older siblings especially with allergic diseases symptoms through the impact of infections on Th1/Th2 immune response is suggested. The study was aimed to examine the influence of the presence of siblings and the presence of older siblings on asthma in young adolescents. Materials and Methods: The self-reported data by 5507 adolescents aged 12/ 16 years from 8 cities in The Republic of Macedonia obtained through the ISAAC phase 3 questionnaires on asthma and environmental risk factors in 2006 were analysed. The presence of siblings and older siblings separately (adjusted for sex, passive smoke exposure at home, gas/wood cooking at home, wood/coal/oil heating at home, duration of TV watching daily, cat ownership) were correlated to wheeze ever, current wheeze, current speechlimiting wheeze, exercise-induced wheeze and ever-diagnosed asthma. Odds ratios (OR, 95% CI) in binary logistic regression were performed for statistical analysis of data. Results: A significant association of the presence of siblings in general and the presence of older siblings especially with asthma and its symptoms was not found (p 9 0.05 for all the parameters). Conclusion: It seems that the presence of siblings and older siblings do not influence asthma in young adolescents. Environmental tobacco smoke: a risk for bronchial hyperreactivity and pediatric asthma Mona Elganzoury 1 , Tharwat Deraz 1 , Karima Abd El 2 , and Mona Elganzoury 2 . 1 Ain Shams University, Pediatrics, Cairo, Egypt; 2 Ain Shams University, Pediatric Pulmonology, Cairo, Egypt. Background: Fifty passively smoking chiidren of whom at least one parent was a chronic smoker were included in the present study their ages ranged between 5Y15 years old.Exposure to cigrette smoke was recognized from personal history and was confirmed by measurement of urinary cotinine(the major metabolite of nicotine in urine). Aim: To evaluate the effect of household environmental tabacco smoke on bronchial reactivity and subsequent lung functions in passively smoking children. We also aimed to correlate pulmonary functions in passive smokers children with the number of smoked cigarettes of thier parents as well as the duration of thier exposure to cigrarettes smoke. Urinary cotinin was studied as a mirror image to the number of smoked cigarettes. Its level was also correlated to bronchial hyperreactivity and lung functions in the studied children. Methods: To asses bronical hyperreactivity,metacholine challenge test(MCH) was done by using dosimeter, Pulmonary functions test were performed before and after metacholine challenge test. Challange test was stopped when there is a reduction in forced expiratory volume in 1st second by 20% from base line,or when the maximum concentration of metacholine was reached (25mg/ml). pulmonary function tests was performed by Med Graphics TM SPIROME-TERY 1070 series 2,hard disc computer system. Urinary cotinin was measured by high -performance liquied tomography. Results: 68%(34 children) were found to have bronchial hyperreactivity,14 children out of the 34 did not perform challange test as they had basel bronchial hyperreactivity. 20 chidren showed postive metacholine challange test at various concentrations. There was a significant relation between bronchial hyperreactivity (BHR) AND THE NUMBER of smoked cigrattes.Also there was a significant relaton between mean forced expiratory volumm one second(FEV1) reduction as well as cumulative dose(CD) OF MCH and number of smoked cigarettes by the parents.duration of tabacco smoke was found to be of great influence on the chiidren pulmonary function tests. Conclusion: Passively smoking children has subclinical bronchial hyperreactivity with various degrees, and some of them may turn to be asthmatics. Pulmonary functions of those children depends on multifactors in particular, number of smoked cigarettes and duration of exposure to tobacco smoke. Health education programs to explain the hazardous effect of passive smoking are recommended. Association of skin allergy test with bronchial asthma patients MD. A. Shakur Khan. Nidch, Mohakhali, Dhaka, National Asthma Center, Dhaka, Bangladesh. Purpose: Asthma is a major public health problem in Bangladesh. Allergy plays an important role in asthma. Asthmatics tend to be more atopic and thus more allergic skin test reactivity than non-asthmatics. Hence skin prick test may serve as a useful adjunct in the diagnosis and management of bronchial asthma. Results of Skin Prick Test are an important component in the formulation of preventive plan for the management of bronchial asthma. Methods: This was a prospective case control study carried out in the Asthma Centre, National Institute of Disease of the Chest and Hospital, Dhaka, Bangladesh. 90 bronchial asthma patients were taken as case and 80 subjects having no history of asthma or allergy were taken as control. The mean age of cases was 25.06 T 1.28 with male female ratio 1.57: 1 and of control 23.09 T 1.31 with male female ratio 1 , positive skin test was detected in mite (7.50%), House dust (5%), Cockroach (2.5%), Mosquito (3.75%), Ladies finger (2.5%), Tomato (1.25%), Chicken (2.5%), Egg white (1.25%) and Prawn (5%) In control non-allergic subjects' 1.25%Y7.5% shows positive skin test result. Cause of this positive reaction in non-allergic control may be due to differences in selection of subjects, standardization of extracts and recording of results. Conclusion: Skin allergy test is the most sensitive method for detecting specific IgE antibody. Positive skin test along with clinical relevance determined the consideration of asthmatic patient for immunotherapy. High prevalence of positive immediate hypersensitivity skin test reactions in the asthmatics population is a strong argument that it should be included in the initial evaluation of patients with asthma. Identification of allergen by positive skin test may help in the preventive plan for the management of bronchial asthma patients. The additional reasons of cardiovascular risk increase in asthma exacerbation Tatyana Brodskaya, Vera Nevzorova, and Boris Geltser. Vladivostok State Medical University, Department of Therapy, Vladivostok, Russian Federation. : A vascular disturbances has a great importance in asthma pathogenesis. Research of mechanical properties of vessels, in particular of arterial stiffness has special interest for the estimation of cardiovascular system function. It is not clear whether increase arterial stiffness observes in patients with asthma. Methods: We examined 54 patients with severe and moderate asthma by noninvasive arteriography (arteriograph TensioClinic TL1 (TensioMed, Hungary)). Control group included 25 age-and sex-matched healthy volunteers. Results: The aortal stiffness in asthma exacerbation was significant more, than in healthy persons. It was expressed in increase of aortic pulse wave velocity (aPWV) and augmentation index (IA). APWV in patients with severe asthma a two fold surpassed aPWV in healthy persons and was 10,5 T 1,3m/s. IA in such patients a 6 fold surpassed control level and was 14,4 T 5,8%. Significant raised (8 G 0,001) arterial stiffness can take part in increasing cardiovascular risk during asthma exacerbation. Independently of increasing arterial stiffness in asthma exacerbation, aPWV and IA essentially improved and came nearer to control level in asthma remission. That suggests functional character of these changes. Augmented arterial stiffness in asthma correlates to lung ventilation dysfunction and hypoxia, disease duration and severity. Conclusion: Described changes can explain cardiovascular events rising in patients with exacerbation of asthma. Our data suggests transitory character of arterial stiffness increasing in asthma exacerbation. Tatyana Brodskaya, Vera Nevzorova, Boris Geltser, and Elena Motkina. Vladivostok State Medical University, Department of Therapy, Vladivostok, Russian Federation. : Arterial stiffness is an independent factor of cardiovascular risk. Exacerbation of asthma associates with increase arterial stiffness. The mechanisms responsible of this association are not clear. Methods: We examined 54 patients with exacerbation of severe and moderate asthma and 25 age-and sex-matched healthy volunteers. Arterial stiffness was estimated by noninvasive arteriography (arteriograph TensioClinic TL1 (TensioMed, Hungary)). Plasma IL-10 and TNF-a levels were determined by the immune-enzyme analysis. Inflammation index calculated as ratio TNF-a/ IL-10. Systemic oxidative disbalance was estimated by ratio of oxidative and antioxidative plasma activity, assessed by spectrophotometry. Hypoxemia was assessed by digital pulsoximetry. NOn-plasma level was determined by Greiss's method. Results: Central arterial stiffness was higher in patients with exacerbation of asthma, than in healthy persons. It was expressed in increase aortic pulse wave velocity (aPWV) and augmentation index (IA). As assessed by correlation analysis both (aPWV and IA) was strongly correlated with hypoxemia (r = Y0,921 and r = Y 0,876 (8 G 0,01)) and inflammation index (r = 0,902 and r = 0,868 (8 G 0,01)). Less strongly but clearly aPWV and IA correlated with NOn-level (r = j0,543 and r = Y0,367 (8 G 0,01)) and systemic oxidative disbalance (r = 0,611 and r = 0,459 (8 G 0,01)). Conclusion: Our results suggests that hypoxemia, systemic inflammation, hyponitrooxidemia and oxidative disbalance can be involved in raising arterial stiffness in asthma exacerbation. This question requires future researches. Mojgan Safari 1 , and Reza Amin 2 . 1 Hamedan University of Medical Sciences, Pediatric Ward, Hamedan, Islamic Republic of Iran; 2 Shiraz University of Medical Sciences, Pediatric Ward, Shiraz, Islamic Republic of Iran. Background: Exposure to cockroach was reported to a cause of asthma in many parts of the world. Objective: To determine the prevalence of cockroach sensitivity in asthmatic children less than 5 years of age in Iranian children with asthma. Methods: Ninety two (33 female /59 male) patients were asked to complete a questionnaire covering demographic characteristics, and were subjected to skin prick testing for cockroach allergen. Blood samples were also withdrawn for the assessment of total serum IgE and eosinophilia. Results: Twenty seven percent (n = 25) of subjects had positive skin test results for cockroach allergen. There were no significant correlations between the prevalence of cockroach skin test positivity in male and female children, rural and urban areas or infants under 2 years and olders. The age of youngest subject with positive skin test for cockroach was 2 months. More over, the parent`s levels of education and the types and ages of children`s residence had no effects on the prevalence of cockroach skin test positivity. There was also no significant correlation between total serum IgE and eosinophilia and the prevalence of cockroach skin test positivity. Conclusion: Cockroach is an important source of domestic infestations in the city of Shiraz. Sensitization to cockroach allergens may develop early in life. Skin test for cockroach allergen in children with infantile asthma may provide useful information for institution of environmental controls. Relationship between sensitization and duration of asthma associated with allergic rhinitis Ion Gabriel Stoica. Spitalul de Pneumologie, Amabulatoriul TBC Sector 3, Bucharest, Romania. Objective: The aim of the study was to assess sensitization to common inhalant allergens among adult patients with asthma associated with allergic rhinitis (AR) and to find a relationship with the medication and the duration of asthma/AR Methods: A screening survey questionnaire including questions assessing prevalence of symptoms of asthma over 12 months and questions relating 24 months prevalence of AR symptoms, were distributed to 763 of 16Y87 year old (median age 46 yr) patients presented in the outpatients department during of 1 years. The response rate was 83%. Asthmatic patients were diagnosed according to the GINA definition of asthma. AR was diagnosed due to the clinical symptoms (sneezing, itching, rhinorhoea, nasal congestion) and sensitization to allergens. Results: Hypersensitivity to inhalant allergens (house dust mite, animal dander, pollen) was assessed by skin prick test (positive: 9 3 mm) in 32 responders having a clinical history. 73% of patients with asthma and 64% with AR were sensitized to house dust mites (D.pteronyssinus and D.farinae). Hypersensitivity documented by positive skin prick test in patients with asthma and AR was noted to pollen in 47% and 49%, to cat and dog dander in 27% and 11%, respectively. Conclusion: There was a significant correlation (r = 0.29, p G 0.05) between frequency of sensitization to house dust mites and duration of asthma. The house dust mite sensitivity is the most prevalent allergen sensitization in adult patients with asthma and AR, establishing control of this allergen as critical for optimizing their treatment. Svetlana Mazurina, Valery Kaznacheev, and Yuri Gervaziev. Mechnickov Research Institute for Vaccines and Sera, RAMS, Moscow, Russian Federation. Background: Bronchial asthma (BA) is a chronic disease associated with elevated IgE levels. It was demonstrated that IL-13, as well as IL-4, plays a crucial role in the regulation of IgE production. The aim of our study was to reveal the association between C-1055T, Arg130Gln (G + 2044A) IL-13 gene polymorphism and level of total IgE in BA patients. Methods: The polymorphisms in IL-13 gene were analyzed by the PCR-RFLP in 136 asthma patients and 64 healthy control subjects. Results: Total IgE and IL-13 levels were measured by ELISA. The total serum IgE level was found to be elevated in BA patients group: 290(47,5;495) *kU/L, and within normal range in the control group: 25,5(19;100)kU/L. The levels of total IgE in BA patients and healthy donors differ significantly, p G 0,001. The frequency of alleles combination -j1055T and + 2044A is 6.9% and reliably is different from that in the control group (p = 0.008). Studying the influence of the polymorphisms on the total IgE level we revealed that this level was considerably elevated (up to 315(290;1077)kU/L) among the patients with -1055T and +2044A alleles compared with a group who has allele +2044A and j1055C, and the group who has allele +2044G and j1055C. Total IgE level in this groups was 230(200;440)kU/L and 205(190;377)kU/L, respectively. Patient with allele j1055T and +2044G were not identified. The link between disease and studied traits (j1055T and +2044A) determined by the Wolf's coefficient of association , which was 0.9. Conclusion: Thus in our study we revealed the association between C-1055T, Arg130Gln IL13 gene polymorphism and total IgE levels and atopy. *Data are presented as interquartile range: median (25% quartile; 75% quartile). Background: Despite its high prevalence, elderly asthma remains unrecognized and misunderstood because its symptoms may not be typical as in young patients. And because other cardiac or respiratory diseases may be frequently associated. Only a few studies on the clinical characteristics of elderly asthma have been reported till now. Methods: To investigate whether the characteristics of elderly asthma were different according to the onset of asthma, one hundred eighty asthmatic patients over the age of 65 were enrolled who attended the Asthma and Allergy Clinic, Seoul National University Bundang Hospital. Results: Of 180 patients, ninty nine patients have developed asthma over the age of 65. This group with late-onset asthma (LOA) had a mean duration of disease of 3.4 T 3.5 yr. The remaining group with early onset asthma (EOA) had a mean duration of illness of 18.8 T 17.1 yr. These groups were indistinguishable by the symptoms, the medication requirements and the serum level of total IgE. These with EOA had a greater degree of airway obstruction in FEV1 of predicted value (p G 0.028) and a greater likelihood of neutrophils in induced sputum. Moreover, the novel indicator, namely, neutrophil/eosinophil ratio of induced sputum was inversely related to several PFT indices, including FEV1, FEV/FVC, FEF25-75%, and FEF25-75%/FVC. These findings suggest not only eosionphil but also neutrophil could play an important role in the airway inflammation and airflow limitation. Conclusion: Our study sggested that cellular compositions of airway inflammation, and lung function depends on the onset and possibly duration of disease in elderly asthmatics. These findings suggest that early onset and longstanding asthma may lead to chronic persistent airway obstruction and thereby mimic chronic obstructive lung disease. APAPARI: questionnaire based assessment of monitoring and treatment of asthma by physicians in infants, preschool and older children Krishan Chugh. Sir Ganga Ram Hospital, Pediatrics, Delhi, India. Background: The APAPARI questionnaire was designed to assess the treatment patterns of childhood asthma in the Asia-Pacific region. Methods: A total of 262 doctors treating asthma were invited to respond to the APAPARI questionnaire evaluating treatment patterns of childhood asthma. Results: Of the 262 doctors, 80 completed the questionnaire while 2 did not complete it as they did not treat children. 59 were pediatricians, 14 were pediatric pulmonologists/ allergists, 2 were pulmonologists and 5 belonged to other specialties. 37(45%) were in private practice or hospitals, 22(27%) were in teaching hospitals while 23(28%) worked in other settings. Responses to the use of symptom cards to monitor asthma were as follows: Responses for the criteria to monitor childhood asthma are tabulated below: For treating an acute attack, 62.8% used nebulised salbutamol/ terbutaline while 15.8% used inhaled salbutamol/terbutaline with spacer and 30.2% used oral corticosteroids. Criterion used for hospital admission in an acute attack of asthma is given below: Pulse oximetry as a basis for oxygen therapy was used by 31 (37.8%) in all patients, 40 (48.8%) used oximetry only in severe asthma while 10 (12.1%) did not have pulse oximetry facilities. A favourable outcome was reported by 31 (37.8%) doctors while 8 (9.75%) reported unfavorable results in severe acute asthma with ventilatory support as compared to those treated at a tertiary/university hospital. 36 (43.9%) did not have access to ventilatory/ ICU facilities to treat severe acute attacks. Maintenance therapy in asthmatic infants was always used by 13 (15.8%), frequently used by 34 (41. 4% choice of maintenance therapy for asthma among infants, preschool and older children were as follows: 58 of the 82 respondents (70.7%) knew but had never used any form of immunotherapy, 11 (13.4%) occasionally used it while 9 out of 82 (10.9%) had no knowledge regarding the availability of immunotherapy. Conclusion: A significant segment of the doctors do not follow the current management guidelines for childhood asthma. Expectancy of Bronchial Hyperreactivity (BHR) in children, simply by the use of a short questionnaire Anxhela Qirko Gurakuqi, and Sokol Agolli. UHC, Allergy Department, Tirana, Albania. Background: The provocation test for assesing BHR is time consuming and sometimes impossible to be performed. Does our questionnaire offer an high expectancy for BHR? Does this questionnaire has any practical value, to avoid the provocation test in certain cases? Methods: 200 children 9Y11 years old from Tirana performed bronchial provocation test with 4.5% saline, according ISAAC protocol. These children answered in the same day to our short and original questionnaire of 4 questions: Have you ever had any shortness of breath or cough in these situations: 1-During or after physical effort (gym hour, going up the stairs, etc) 2-In exposure of tobacco smoke? 3-In exposure of car emanations, heating and cooking gases? 4-By laughing loudly? Every positive answer was coded with scores 1,2,3,or 4, with 0 score if no positive answer, and with 5 score if all positive answers. Results: Subjects with positive result of saline provocation (change of FEV1 9 15% pre-post provocation) are divided according the scores 0,1, 2, 3, 4, or 5. The same is done for subjects with negative BHR. Not surprisingly, 65% of subjects with negative result after saline provocation had score 0, comparing to 10% of subjects with positive saline, also only 10% of subjects with negative saline resulted with score 5, when more than 50% of subjects positively reacting to saline provocation had score 5. When the subjects with negative result to saline, regardless to result of score questionnaire (nr = 183) are compared with the sub-category of subjects which, beside the negative saline, had also score 5 (nr = 19), we found in the last group a higher rate of personal and familiar history of asthma and allergy. Sensibility, Specificity, Positive Predictive Value and Negative Predictive Value of this questionnaire to forecast the BHR are calculated: 60%, 89%, 24% and 97% for the score 5 (presence of all symptoms) and for score 0 (absence of all symptoms). Conclusion: 1-This short questionnaire is able to forecast a negative BHR in 97% of cases, if all symptoms are missing (score 0). 2-Subjects with score 5, regardless the provocation result, have an intriguing clinical profile, with positive personal and familiar history for allergy, requiring more attention by doctors and deeper investigations in the future. 3-By only asking 4 simple questions, the time consuming bronchial provocation test could be avoided in everyday medical practice most of the cases, if all the answers are negative. Gabriella Galffy, Lilla Tamasi, Marta Orosz, and Gyorgy Losonczy. Semmelweis University, Dept. of Pulmonology, Budapest, Hungary. The 2006 Global Initiative for Asthma (GINA) reports provide a new asthma classification which is based not only on the severity but also on the level of asthma control. Bronchial asthma is a word-wide problem, with an estimated of 300 million affected individuals. There are more than 200,000 asthmatic patients in our country. Classification of asthma by severity is useful when the doctor has made a decision about management and treatment of a patient. The goal of asthma treatment is to achieve and maintain clinical control. Asthma Control Test is one of a validated measurement to analyze the controlled, partly controlled and uncontrolled levels of asthma. Aim: The aim of the study was to investigate the level of asthma control in our country. Methods: In our outpatient clinic, we studied a group of 133 patients (female: 91, male: 42; mean age: 44y) with bronchial asthma. Asthma Control Test was done by all patients and they also have an other eleven questions regarding smoking habits, asthma medications, side effects, exacerbations, exercise induced dyspnoe. Results: Only 50% of our patients (n = 67 pts) had controlled and partly controlled asthma level, an other 50% (n = 66 pts) had uncontrolled asthma based on Asthma Control Test. The total control level has been achieved only by 18% (n = 24 ) of our patients. There were unable to work and to have a job the 16 % of the patients and 12% were not able to work even in their household because of suffering asthma. Seventy eight % of patients have reduced quality of life. However, 64% of our pts believed and felt they have total asthma control and only 10% of pts subjective opinion were the uncontrolled asthma. Conclusion: In our study, we found that patient's opinion about their own condition of asthma did not correlate to an objective test. Asthma Control Test show us the reality of the level of bronchial asthma. The test is very easy to use even alone by the patient. The Asthma Control Test help to achieve and maintain the clinically controlled asthma level. Obesity -a risk factor for asthma severity? Recently obesity has reached epidemic proportions in many countries. Evidence is now mounting that obesity is also a risk factor for asthma. The aim of our study was to assess relation between obesity and asthma severity in children. We conducted cross-sectional study in the out-patient department at M. Guramishvili Pediatric Clinic in 2005Y2006yy. In the study were enrolled 63 children aged 12Y15 with a mean age of 13.75, 41 boys and 22 girls. We used body mass index (BMI) to detect overweight and obesity. Weight categories were defined by BMI number position on BMI-age growth chartrisk of overweight -85th to less than the 95th percentile, obesity -equal to or greater than the 95th percentile. Asthma severity definition was based on patients clinical and/or spirometeric parameters according to GINA guideline. Children with a history of other lung disease were excluded. The prevalence of obesity in our study population was 31.7%. Among girls 55% and among boys only 19.5% were obese. Mean BMI of females with asthma were significantly higher than males (p G 0.0001). The obese children had a difference in baseline asthma classification (more likely to have persistent asthma). Despite similar severity of illness at admission, they had a longer length of stay at hospital (9.8 +/j 7.0 vs. 6.5 +/j 3.4 days, p G 0.01). We can conclude that there is relationship between body weight and asthma severity and obesity may be a potentially modifiable risk factor for asthma or asthma exacerbation. Background: Recent data from animal studies indicate that fat tissue is a source of eotaxin, which may play an important role in asthma. However, the link between obesity and airway inflammation in asthma is not well understood. Objective: We aimed to investigate the impact of obesity on eotaxin levels in patients with stable mild/moderate asthma. Methods: 20 steroid-naive asthmatics (mean age: 57.2 T 2.2 yrs) and 9 control subjects without lung diseases (mean age: 59.4 T 9.7 yrs) were investigated. According to the body mass index (BMI), subjects were divided into the 2 groups: subjects with normal weight (BMI G 25kg/m 2 ) and subjects with overweight (BMI Q 25 kg/m 2 ). Spirometry, bronchial provocation test with methacholine (PD 20 ), sputum induction and bronchoscopy were performed. Eotaxin-1, -2, -3 concentrations in serum, sputum supernatant and BAL fluid were measured using commercial ELISA kits. Results: In asthma patients with overweight eotaxin-2 concentration in induced sputum was significantly higher comparing to asthma patients with normal weight (118.8 T 46.6 vs 20.1 T 9.3 pg/ml) and control subjects (118.8 T 46.6 vs 16.7 T 7.2 pg/ml; p G 0.05). Whereas, eotaxin-1, eotaxin-3 did not significantly differ between studied groups, and there was no correlations between eotaxin levels and number of eosinophils in induced sputum and BAL fluid. Conclusion: Elevated sputum eotaxin-2 concentration in asthmatics with overweight let suggest, that increased body mass may upregulate inflammatory markers in asthma patients. Fariborz Zandieh. Isfahan university of medical sciences, Asthma, Allergy and Immuology, Isfahan, Islamic Republic of Iran. Background: Interest in the impact of illness on day to day function is leading investigators to include both disease specific and generic health related quality of life (HR QOL) questionnaires in a broad range of clinical studies and to gain a full picture of the impact of asthma on the lives of children with this condition, it is necessary to make direct measurement of health related quality of life. Methods: In response to this need, we used the Juniper`s pediatric asthma quality of life questionnaire (PAQLQ) and Juniper`s Pediatric Asthma Caregiver`s Quality of Life Questionnaire (PACQLQ) that has been developed based on guidelines for construction of over a dozen validated disease specific quality of life instruments. The PAQLQ consists of 23 items that in children with asthma have been identified as troublesome in their daily lives and PACQLQ that contains 13 items in two domains of emotional and activities disturbances. The study design consisted of an 18 month single cohort study. Patients participating in the study were 113 children, 7Y17 years of age, with a wide range of asthma severity and their caregivers. For each patient a PAQLQ and for each caregiver a PACQLQ was completed. One week before visit patients recorded morning peak flow rates, medication use and symptoms in a diary. After complete physical examination, for determining of asthma severity, spirometry was performed. Results: The questionnaires after statically analysis showed good levels of both longitudinal and cross sectional correlations with the conventional asthma indices and with general quality of life. We found that consistently QOL in boys were more disturbed than females, a good relevancy between severity of asthma and QOL scores and more disturbances of QOL in caregivers of male asthmatic patients than caregivers of female asthmatic patients. We could not find any significant relevancy between FEV1 percentage of predicted and overall scores of QOL. Conclusion: Smaller airways, and higher airway resistance and more activity of males than females may explain why boys have more disturbed life style than females. Shadan Pedram. Thehran University of Medical Science, Medical-Surgical-Nursing, Thehran, Islamic Republic of Iran. : Asthma and quality of life in iranian asthmatic patient asthma is a chronic disease,that make many health problems in every where in the world. The aspect of quality of life in iranian asthmatic patient are variable. Objective: To determine factors influence to quality of life in patients with asthma, who reffer to lung clinic of general hospital in tehran. Methodology: This research is a descriptive study.the subjects consisted of 386 patients.samples selected by sample randomized.the method of collecting data was by questionnaire.the data were collected in one stage and analyzed with descriptive and inferential methods and spss soft ware. Findings: The results showed that ,the quality of life inpatients from physical aspect was inappropriateand, in social and economic aspects were rarely appropriate. the most of patients from mental and sleep aspects were inappropriate. Neither sex nor overweight are risk factors for having asthma symptoms in adolescent population of Argentina Background: A positive association between body mass index (BMI) and asthma has been recently suggested; however, the sex-dependence of this association remains controversial. The aim of the present study was to explore the relationship between BMI and asthma, as well as its sex-dependence in young adolescents. Methods: Self-reported data obtained from the standardized International Study of Asthma and Allergies in Childhood (ISAAC) Phase Three. Written questionnaires of 5579 young adolescents aged 13Y14 years old from randomly selected private and public schools in Salta and Cordoba (Argentina) were used. The BMI for each individual was calculated; international cut-off points for BMI for overweight and obesity by sex and age were used (Normal: 18 until 24.99 kg/m2, Overweight: 25 until 29.99 kg/m2 and Obesity 9 30 kg/ m2). Because of the very low prevalence of obesity (0.86 %), they were included in the overweight group (4.6%). Each variable of the ecological questionnaire considered as probable confounder was confronted in 2x2 tables with having wheezed in the last 12 months(13,1%) and BMI, and those variables found significantly associated were included in the final analysis. The data were statistically analyzed by the chi-square test, P and odd ratios (OR, 95 % CI) in binary logistic regression. Results: 51,4% of responders were male. Being a boy was associated with overweight (OR 1,35. 95%CI 1,06-1,71; P 0,013), and exercising one or less/ week was associated with obesity (OR 2. 95%CI 1,07-3,70; P 0,025). After adjusting for each one, both were significantly associated to an increased BMI (P 0,02 and 0,01 respectively). Watching TV one or less hour/day was related to have a lower BMI (OR 0,72. 95%CI 0,55-0,93; P 0.01). Exercising once or less per week was the only factor associated to wheezing after adjusting for BMI and sex (P 0.0001). Conclusion: In our population, neither sex nor overweight were associated to wheezing symptoms; however sex was significantly associated with overweight. Maybe the scarce obese population in our cities contributed to these findings, however sedentary profile contributed to an increased BMI and to wheezing symptoms also, as was expected. 543 Abstract withdrawn 544 Abstract withdrawn 545 A three-year follow-up study of asthma, airway symptoms and self-reported allergy among pilots and cabin crew at commercial aircraft Gunilla Wieslander 1 , Torsten Lindgren 2 , and Dan Norback 2 . 1 Dep of Medical Sciences, Occup and Env Med, Uppsala, Sweden; 2 Department of Medical Sciences, Occupational and Environmental Medicine, Uppsala, Sweden. Background: Cabin air quality is important especially to employees with sensitive airways. There are few studies on incidence of asthma and allergy in pilots and cabin crew. The aim of the study was to study the relations between occupational factors and personal factors (age, gender, atopy, smoking habits) and incidence of asthma and allergy. Methods: A standardized questionnaire (Ö rebro, MN040 NA) was answered in 1997 by pilots (n = 577) and cabin attendants (n = 936). Three years later, in 2000 the same questionnaire was sent to pilots (n = 436) and cabin attendants (n = 698) on duty. The new onset of asthma and allergies (three year incidence) were calculated by logistic regression related to personal factors with logistic regression. Results: The prevalences of airway symptoms and asthma were two to three times lower in pilots than in cabin attendants. The three year incidence of doctors diagnosed asthma was 1.2% among pilots, and 0.6% among cabin attendants. The incidence of any airway symptom (wheeze, attacks of breathlessness) was 4.6% among pilots, and 12.5% among cabin attendants. The incidence of chronic bronchitis (daily productive cough) was 1.1% among pilots, and 3.5% among cabin attendants. Further, the incidence of pollen allergy and furry pet allergy was 5.4%, and 1.6% respectively among pilots and 5.7% and 2.4% respectively among cabin attendants. The incidence of food allergy/ intolerance was 1.8% among pilots and 3.2% among cabin attendants. The incidence of airway symptoms was lower in pilots (p G 0.05). The incidence of doctors diagnosed asthma and food allergy was higher in atopics (p G 0.01). Younger cabin crew and pursers had a higher incidence of pollen-/ furry pet allergy (p G 0.01). No relations was seen as to gender and own tobacco smoking. Conclusion: Age, atopy, and type of occupation was related to the incidence of asthma and self-reported allergies. The incidence of doctors diagnosed asthma among pilots (0.4%/year) was somewhat higher than expected. Airway symptoms in genral were two to three times more common among cabin attendants. The study started before the stop of smoking on intercontinal flights. Our results indicate that Parietaria pollinosis is more and more frequent, from 1987 to 2006, in Greece among patients with SAR/A. Parietaria pollinosis appears to be an increasing problem during the last 20 years, in Greece. This phenomenon may represent the influence of environmental factors. Prevalence of asthma, rhinitis and eczema among Brazilian and Japanese adolescents (AD) in the city of São Paulo, Brazil Inês Camelo-Nunes 1 , Ricardo Yamada 1 , Luis Guilherme Pimentel 1 , Thales Shibata 1 , Flávio Sano 2 , Dirceu Solé 1 , and Charles Naspitz 1 . 1 Federal University of São Paulo, Division of Allergy & Clinical Immunology, São Paulo, Brazil; 2 Hospital Nipo-Brasileiro, Division of Pediatric Allergy, São Paulo, Brazil. Objective: To evaluate the importance of the genetic background on the prevalence of asthma (A), rhinoconjunctivitis (R) and atopic eczema (AE) among AD born in Brazil and from different genetic backgrounds. Methods: ISAAĆ s written questionnaire was applied to two groups of 12Y15year-old AD of similar socioeconomic status living in the city of São Paulo: Brazilian (N = 381) and Japanese (third generation, born in Brazil, from families without mixed marriages, N = 221). Affirmative response to BHave had wheezing in the last year^identified AD with A, to BNasal and ocular symptoms in the last year^those with R, and to BEczema in the last year evidenced in flexural areas^those with AE. Questions^Speech problem in last 12 months during acute attack^, BNasal symptoms interfering with daily activities^, and BKept awake at night by this itchy rash in the last 12 monthsĉ haracterized severity of A, R, and AE respectively. Results: There were no differences according to gender in both groups except for the prevalence of R that was higher among Brazilian females. The prevalence of A (12.7% x 19.4%, p G 0.05) and the prevalence of R (10.4% x 24.4%, p G 0.05) were higher between Brazilian AD when compared to Japanese AD, respectively. There were no difference in A severity (0% x 1.1%) but R`s severity was significantly higher between Brazilian AD (10.4% x 24.4%, p G 0.05). The prevalence of AE was higher between Japanese AD (7.7% x 3.1%, p G 0.05) but without differences in its severity (1.8% x 1.8%, p 9 0.05). The prevalence of these diseases is not different from those observed in Japan and in São Paulo (ISAAC, phase 3). Conclusion: The prevalence observed among the third generation of Japanese AD without miscegenation born in Brazil, showed similar results to those obtained in ISAAC phase 3 recently published in Japan. Considering that both AD groups evaluated have been exposed to the same environment we might speculate that the genetic background in this Japanese population is more important than environmental influences on the development of asthma and allergic diseases. Asthma in Korea (COREA), a large multi-center long-term cohort study of whole spectrum of asthma: Clinical characteristics according to severity of asthma disease, many researchers used to recognize them as a spectrum of 'chronic obstructive airway disorder' (COAD). Accordingly, it is most important that we can reveal prognostic factors influencing on natural course of asthma and prohibit from progressing to severer and irreversibly uncontrolled asthma. It is thought that this is possible only in case asthmatic patients should be observed through long term follow up such as cohort study. Objective: To define the important prognostic factors contributing to asthma severity and suggest new clinical understanding and paradigm of COAD. Methods: Researchers from 11 university hospital have established asthma cohort named 'Cohort for Reality and Evolution of Adult Asthma in Korea' (COREA) since year of 2005 in Korea, which will be continued up to 2013. We have collected patients having chronic persistent respiratory symptoms such as dyspnea, cough, sputum or wheezing more than 3 months and airflow limitation by FEV1/FVC less than 0.7 or AHR confirmed by methacholine bronchial provocation test (MBPT). As the first study of our cohort, we analyzed various clinical factors influencing on the severity of asthma for registered 644 patients at the time of first-visit. Severity was assessed by special allergists by the definition of 2002 GINA guideline. Results: Severity of asthma was associated with skin prick test positivity (p G 0.006), history of hospital visit due to asthma exacerbation (p = 0.001), symptom onset (p = 0.037) and duration of diagnosis (p = 0.041). Severity also was significantly correlated with FVC, FEV1, and bronchodilator response (all p G 0.001). However, smoking didn't affect the severity of asthma, even in case of passive smoking. : In Venezuela, asthma causes more than one million crises per year, a rather significant impact to the health system compared to USA reports. Our demographics situation are similar to the rest of Latin America, with a young and mostly urban population living under variable conditions of poverty; application of massive vaccination programs and other public health measures will ensue that non-communicable diseases occupy major roles in the near future. For Venezuela ISAAC data shows an eighteen percent of wheezing in the past year with an overall prevalence of thirty two percent is reported. Methods: In order to explore such previous high prevalence, we carried out a multidisciplinary and ongoing study (approved by the ethical committee of the Institute of Biomedicine, ratified by the academic council of the medical Faculty of Central University of Venezuela) in 1000 unselected school children, 6Y12 years old, from low to middle socioeconomic background covering different areas from Caracas, Venezuela. After an informed consent was signed from parents or guardians, all children from randomly selected schools were evaluated with a validated modified Graffar's socioeconomic questionnaire and an allergic rhinitis, asthma and atopic dermatitis validated questionnaire after ARIA, GINA and Hannifin criteria; skin prick testing (ALK-ABELLO) for common food and inhalant allergens plus total IgE (ELISA), complete blood count (COULTER) and serial feces examination for ova and parasites were performed in all children. Pre and post bronchodilator FEV1 and PEF spirometric measurements values were obtained (MICROLOOP), and included in data analysis. Results: We found that 60% of the child evaluated had allergic rhinitis, 40% had asthma, and 21.9% Atopic Dermatitis with or without other allergic pathologies. We also found that 64.8% of the child evaluated had parasites and 41.1 had respiratory infectious disease associated. Conclusion: Our data confirms the high prevalence of allergic diseases in Venezuela employing questionnaires, a clinical exam and immune diagnostic evaluation methodology. Hence, our health system and medical faculties should be highly aware of these and similar findings for establish a proper education programs, both at primary health care personnel and also in communities. Developing nations are waiting for simpler but effective approaches in the management of a public health problem who`s time has come. Herberto Jose Chong Neto 1 , Nelson Rosario 1 , Ana Carolina Dela Bianca 2 , Dirceu Solé 2 , and Javier Mallol 3 . 1 University of Parana, Pediatrics, Curitiba, Brazil; 2 University of Sao Paulo, Pediatrics, Sao Paulo, Brazil; 3 University of Santiago, Pediatrics, Santiago, Chile. Background: Large international studies on asthma and allergies in childhood have found notorious variation in prevalence and temporal trend among countries. However, there is no international studies on the epidemiologic characteristics of wheezing in the first year of life. The aim of this study is to validate a questionnaire to assess the prevalence of recurrent wheezing infants. Methods: This study was undertaken in pediatric emergency rooms (PER). A randomized sample of parents visiting PER for lower respiratory illness, answered a questionnaire on wheezing developed by the International Study of Wheezing in Infants (EISL, from spanish Estudio Internacional de Sibilancias en Lactantes). During visit, all infants were examined by one of the authors (blind) who reported, or not, the presence of wheezing on chest auscultation. Sensitivity, specificity, predictive positive value, predictive negative value and concordance (0) level were calculated from parent and physician reports. Results: Two hundred and nine infants aged 12 to 15 months participated in the study. Fifty six parents reported current wheezing and 43 were confirmed by physician; 153 parents did not report current wheezing and 146 had not wheezes at physical examination (0 = 0,74, CI 95% 0,64 Y 0,85). This questionnaire showed high sensitivity (86%), specificity (91,8%), positive predictive value (76,8%) and predictive negative value (95,4%). Conclusion: Regardless of previous experience with wheezing episodes, parents can reliably inform when their infants are currently wheezing. A simple and convenient questionnaire confirmed by physical examination produce an accurate tool to asses the prevalence of asthma symptoms in infants. Different seasonal (monthly) variation in asthma exacerbations in patients with allergic and non allergic asthma Christos Grigoreas, Dimitrios Vourdas, and Konstantinos Petalas. 251 Air Force Hospital, Allergology Department, Athens, Greece. Background: The recognition of clear seasonal (monthly) patterns of Asthma Exacerbations (AE) in a particular area may allow preventive strategies to be developed. Objective: The aim of this study was to determine the seasonal (monthly) variation in AE in patients with allergic and non-allergic asthma. Methods: Data were obtained from our department in Athens, between October 1986 and September 2006 (240 months). We evaluated all AE occurring this period for patients with Allergic Asthma (AA) and Non-Allergic Asthma (N-AA). The diagnosis and classification in AA and N-AA were made according to the results of skin prick testing (SPT) to common aeroallergens. Patients with AA defined as those with 9 1 positive SPTand patients with N-AA defined as those with negative SPT. A positive SPTwas defined as a wheal size at least 3 mm greater than the negative control. AE were defined as a deterioration in asthma resulting in an unscheduled visit (i.e patient-initiated) leading to change in asthma treatment or the need for oral steroids for 9 3 days and/or emergency room visit/hospitalization. Data are presented as monthly averages of these years of combined data, as a percent above (+) or below (j) the average monthly value (%) for the 240 months under study. Results: There were 3.570 AE (2.313 in males, 1257 in females) in 2.345 patients with AA (1.569 males, 776 females). Also, there were 2.585 AE (1.335 in males, 1250 in females) in 1.518 patients with N-AA (736 males, 782 females). Percent above or below an average monthly value (%). Conclusion: These findings suggest that AE have a different clear seasonal (monthly) variations between patients with AA and N-AA. Patients with AA showed an increase in AE in April, May and June (spring and early summer). Especially in May occurred a great increase (peak) in AE (+307,1). Patients with N-AA had an increased AE from January until April (winter and spring) and from September until November (autumn). These results may offer significant opportunities for improved disease management. Seasonal ( defined as a wheal size at least 3 mm greater than the negative control. AE were defined as a deterioration in asthma resulting in an unscheduled visit (i.e patient-initiated) leading to change in asthma treatment or the need for oral steroids for 9 3 days and/or emergency room visit/hospitalization. Data are presented as monthly averages of these years of combined data, as a percent above (+) or below (j) the average monthly value (%) for the 240 months under study. Results: There were 590 AE (370 in males, 220 in females) in 342 asthmatic patients (214 males, 128 females). From 590 AE, 193 were identified in patients 0Y14 years, 231 in patients 15Y29 years, 116 in patients 30Y44 years and 50 in patients 9 45 years. Conclusion: These findings suggest that AE in the Athens region have a seasonal (monthly) variation in patients allergic to mites (D. Pteronyssinus, D. Farinae) in Greece. An increase in AE occurred in the spring months (March, April, May) and the June (peak in the May + 51%) and also in the autumn months (September, October, November) with a peak in September (+ 49%). A decrease in AE occurred in the rest of the months with a nadir in August (j79,6%). Although AE may be triggered by a number of other factor (i.e respiratory viral infections), this study suggests that asthmatic patients allergic to mites represent a seasonal (monthly) variation in AE, in Greece. These results may offer significant opportunities for improve disease management. Conclusion: In Sverdlovsk region BA is spread in the urban area wider than in the rural region. The BA atopic form prevailes both in urban and rural areas. In the city the important asthma RF are bad factors in production facilities and presence of cockroaches and birds. Asthma RF common for urban and rural areas are: allergic rhinitis, a strain of asthma and allergy, presence of fish aquariums in living houses/ apartments. Outcome of childhood asthma in 205 asthmatic children Mohammed Herrag, and Mustapha Iraqui. Ibn Sina University Hospital, Pulmonology-Allergology, Rabat, Morocco. The aim of this study was to investigate the outcome of childhood in asthma. 205 asthmatic children aged 2 to 14 years seen between the years 1990Y91 were followed prospectively till 2005. The severity of asthma was classified: mild, moderate or severe. The severity was evaluated every three years by changes in respiratory symptoms and spirometry. Initially 32p.100 of children have mild asthma, 25p.100 moderate asthma and 43p.100 ware considered as severe asthma. 67p.100 of children who had initially mild asthma have the same severity ten years later, 21p.100 have moderate asthma and 2p.100 have severe asthma. 70p.100 of children who had mild asthma did have mild asthma ten years later. 60p.100 of children who had severe asthma are considered severe ten years later. The final prognosis was influenced by the initial severity of the asthma and presence of associated atopic disease. Effects of nitric oxide synthase inhibitors on airway hyperresponsiveness and inflammation in a murine asthma model Introduction: Excessive airway production of nitric oxide (NO) has been suggested to play a role in bronchial asthma. It is still not clear which iNOS isoform is involved in eosinophilic airway inflammation. Objective: A selective Inducible nitric oxide synthase (iNOS) inhibitor 1400W (N-3-(aminomethyl) benzyl) acetamidine) or a nonselective NOS inhibitor L-NAME (N-nitro-L-arginine methyl ester) have been tested in a murine acute asthma model. Methods: Six week-old female BALB/c mice were sensitized and exposed to aerosolized ovalbumin (OVA). BALB/c mice (each group, n = 4) were treated with indicated dose of 1400W (2 mg/kg, 4 mg/kg, 8 mg/kg, intraperitoneal) or L-NAME (10mg/kg, intraperitoneal) for five times before allergen challenge. Methacholine bronchoprovocation test and bronchoalveolar lavage (BAL) fluid analyses were performed. Results: Airway inflammation was decreased only in 1400W-treated groups. BAL fluid total cell counts (positive control 428 x 10 3 /2l, 2 mg/kg 273x10 3 /2l, 4 mg/kg 210x10 3 /2l, 8 mg/kg 154 x 10 3 /2l, each p = 0.012, p G 0.001 p G 0.001) and eosinophil counts (positive control 318x10 3 /2l, 2 mg/kg 150 x 10 3 /2l, 4 mg/kg 142x10 3 /2l, 8 mg/kg 105x10 3 /2l, each p = 0.016, p = 0.009, p = 0.003) were decreased in 1400W-treated groups compared with positive control group. But there were no significant differences in airway hyperresponsiveness (Log PC200 p = 0.719) regardless of treatment. The results demonstrate that a selective iNOS inhibitor 1400W is effective in improving the airways inflammations through reducing total inflammatory cell counts in the OVA-sensitized BALB/c mice. Levan Bolokadze, Inga Fedotova, and Volodimir Fedotov. Kharkov Allergological Centre, Allergology, Kharkov, Ukraine. Background: To assess the relationship among the sexual, psychological, hormonal, and physical status of women with bronchial asthma (BA). Methods: Twenty-nine women with BA and 18 healthy women as the control group were enrolled in the study. The patients were asked to complete the Female Sexual Function Index, General Health Questionnaire, and Medical Outcomes Study Short Form 36-item Health Survey (SF-36). The mental and physical component summary scores were calculated using the answers on the SF-36. The same questionnaires were given to this group as well. Statistical analysis was performed using the Mann-Whitney U test and Pearson correlation tests. Results: Statistically significant differences were observed for all questionnaire scores (P G0.01). The most common female sexual dysfunction was diminished arousal (n = 21, 72.41%) in women with BA. In the correlation analysis, the total Female Sexual Function Index score had a statistically significant and positive correlation with the mental component summary score (r = 0.517, P = 0.001) and a negative correlation with the General Health Questionnaire score (r = j0.368, P = 0.01). Conclusion: The results of our study have shown that bronchial asthma can be a cause of female sexual dysfunction with mental and psychiatric mechanisms. Ekaterine Karseladze 1 , Maia Davituri 2 , Rusudan Karseladze 3 , and Tinatin Bigvava 4 . 1 Tbilisi State University, Stomatology Department, Tbilisi, Georgia; 2 Tbilisi Children's Hospital #1, Allergology Department, Tbilisi, Georgia; 3 Tbilisi State University, Pediatric Department, Tbilisi, Georgia; 4 Institute of Pediatrics, Neonatology Department, Tbilisi, Georgia. In the difficult pathogenesis of Bronchial Asthma is important nerve inflammatory mechanism and unbalance of the various group of autonomic nervous system. The process made by bronchial asthma cause lipid phase cell membrane changes. The importance is the oxidation process reinforcement and to development an antioxidant syndrome. The study aimed to asses the last product of lipid peroxide oxidation (LPO) -enzyms: Malon Dealdehyde (MDA), ceruloplasmin (CP) and superoxidismutas (SOD) and the function of autonomic nervous system. The situation and personal nervous status were fulfilling the diagnosis tests of C. Spilberg, the emotional status were studied with Lusher`s color test. The investigation of some index of oxidation stress and psychosomatic characteristics in children with bronchial asthma. 90 patient (median age of patients was 7Y15 years) with bronchial asthma and practically, 100 healthy children were investigated. The mathematical treatment of dates had made with program package SPSS,v.12. Results: In the blood serum of the patients the contemt of mda was 63% high than in control group. (p G 0,05). Mda level in the patient with eytonic type of vegetative regulation was lower than in vagotonyc patients (p G 0.05). Especially important was the coefficient dates. LPO product quantity in the patients with 0,05). At G vagotonyc tape is straight contact with emotional stress (r = 0,63; p the end of the study was carried out, that bronchial asthma is conducted the high level of LPO products in blood serum. The LPO activate has shown at almost everytime Y from 1 to 5 year and more. Results of the investigation show us the LPO prosess activateing at the time of bronchial asthma. It`s established the coordibation of the LPO level in the blood serum and the severity of disease of the patients with bronchial asthma, which confirmed the antioxidant decrease in the patients. Low antioxide guard in the patients with eytonic type of vegetatic regulation give us a chance to seperate them as biochemical ?nontrustworthy X lipid peroxide oxidative system. Relation of BCG-Scar diameter and asthma in children There are conflicting ideas about the inverse relationship between delayed type hypersensitivity reaction of BCG and atopic state. Stronger response to PPD test as an indicator of more potent TH1 response is supposed to influence TH2-modulated allergic reactions, but it is still doubtful whether scar of BCG vaccine can also be supposed as an indicator of TH1-immune response or not. In our previous study PPD test response was significantly smaller in asthmatic patients, now in this study relationship between BCG scar and asthma is investigated. Methods: 100 patients younger than 5 years old, vaccinated at birth, with variable severity of asthma were compared to the same number of ageadjusted healthy control group in a case-control study. Mean of vertical and longitudinal diameter was determined. At the same time, the severity of asthma in the case group was evaluated (according to classification of national institutes of health, national institutes of heart lung and blood), by asking mothers about the frequencies of night and day symptoms or intervals of attacks. The patients were divided to three groups: Mild, Moderate and Severe groups (Mild intermittent and mild persistent groups were classified as one group). Results: In 97 case group the mean size of BCG scar was 4.93mm +_1.89mm, again in 97 control group the mean size was 4.94mm + _1.46mm (P = 0.949). Conclusion: Against the result of our previous research which showed a significant reverse relation between PPD-skin test response and asthma, but no significant relation was found between BCG scar and asthma, this may indicates that BCG-scar is not a good indicator of TH1 response and is probably related to other mechanism such as wound healing. Bilun Gemicioglu, Imran Ozdemir, and Bilge Ozgur Yuksel. Istanbul University, Faculty of Medicine Cerrahpasa, Department of Pulmonary Diseases, Istanbul, Turkey. Backround: The ACT is a validated, 5-item questionnaire with a total score ranging from 5 (poor control) to 25 (total control) (Nathan RA et al, JACI 2004). Aim: The correlations of ACT, FeNO and FEV1 were evaluated in not well controlled asthma patients for a three month follow-up period. Methods: ACT, FeNO and FEV1 are measured in 77 female and 20 male persistent asthma patients followed in outpatient clinics of asthma. In 32 of them with ACTG20 and/or FeNO 925ppb and/or FEV1G80% add-on therapy were given and after 3 months, all parameters were remeasured. The correlation of those parameters before and after add-on therapy were evaluated. Results: Mean ACT scores, FeNO and FEV1 before (period 1) and after (period 2) add-on therapy of 23 female and 9 male asthma patients with a mean age of 40,1T13,3 are in the table. In 16 (50%) of the patients all three parameters, in 6 (19%) FeNO and ACT, in 3 (9%) only FeNO, in other 3 (9%) ACT are under normal values. In the remaining 4 patients comorbid diseases are found as the reason of uncontrolled asthma. As a result; increases in all three parameters are significantly well correlated. Conclusion: We conclude that because these three parameters are well correlated with each other, ACT by itself may be sufficient to observe the patients alone although it would be best to measure all 3 parameters if they are available. Global approach to the psychological aspects of the child with asthma Anselmo Sanchez Palacios. Hospital Universitary Insular of Gran Canaria, Allergology Unit, Las Palmas GC -Canary Islands, Spain. Background: Although a significant incidence of various psychological variables has been recognized for a long time in asthmatic children, its true role is the subject of strong controversies. Stress, suggestion and external conditioning can affect the respiratory tract. The chronic aspect of the disease, its disruptive effects on the quality of life and the sensation of threat to their life, can condition the appearance of different psychosocial alterations in the patient, or in their immediate family environment. Experimental tests have shown that the severity of the disease, multiple hospitalizations and the absence of specific IgE mediated symptoms, upon exposure to allergens, could contribute to increased emotional disorders in children. Objective: To search for an efficient intervention procedure which includes the improvement of these children in school and personal, familial and social adaptation, and to help them modify their inadequate attributions and attitudes towards their disease. Materials and Methods: This was a random stratified prospective study including a population of 168 pediatric patients with ages comprised between 9 and 11 years of age (100 boys and 68 girls); 74 had asthma, 56 rhinitis and 38 asthma and rhinitis. Patients were divided into active and control groups. The following tests were used: multifactorial autoevaluation test for infantile adaptation (TAMA), questionnaire for attributions and attitudes toward the disease (ATAC), and a questionnaire for the evaluation of the physical aspects of the disease (EVEO). The program applied to the case group consisted of an emotional/instructive program (PIE_ASA). Results: The psychotherapeutic approach to the treatment of these asthmatic children using the program PIE_ASA achieved a global improvement of the children, fulfilling the objectives of the program. The modification of the negative attributions and attitudes towards their disease was accomplished. No changes in other attitudes such as increase food intake and compliance with medication were obtained. Better results were obtained in boys than in girls, regarding social and school adaptation aspects. However, girls showed a reduction in the number of emergency room visits. Conclusion: We believe that the use of this type of evaluation en asthmatic children is recommended, since it contributes to an overall improvement in the asthmatic symptoms of children. Autoimmunte uveitis, 16 years experience (1988) (1989) (1990) (1991) (1992) (1993) (1994) (1995) (1996) (1997) (1998) (1999) (2000) (2001) (2002) (2003) (2004) 4%) and posterior uveitis in 7 (11.9%) patients. The process was bilateral in 36 (61%), Unilateral right eye affection was established in 13 (22%) patients, and unilateral left eye affection in 10 (17%) patients. Familiar history was present in 4 (6.8%). Uveitis was associated with systemic disease in 18 (30.5%), non-associated with systemic disease in 4 patients (6.8%). Systemic diseases were juvenile chronic arthritis in 9 (15.3%), spondylitis in 3 (5.1%), Vogt Kayanagi Harada disease, nodosa poliarteritis, Wegener granulomatosis, sarcoidosis in 1(1.7%). Infectious etiology agent was identified in 4(6.8%) patients. Clinical signs described were diminution of the visual sharpness in 42 (71.2%), ocular pain in 11 (18.6%), fotofobia in 13 (22%), red eye in 28 (47.5%). We found antibodies antinuclear positive in 17 patients, but with weak pattern. Immunosupresor treatment used was cyclophosphamide in 50 patients, azatyoprin in 8 patients; methotrexate was used in 11 children. Complications included synechias in 8, cataracts and keratopathy in 6 patients. 11 patients left the treatment (18.6%). Conculsion: The clinical and demographic characteristics of the presentation of this disease in latin children in a hospital of third referral hospital are described. Respiratory allergic diseases: Italian monitoring study of GINA and ARIA guidelines (ARGA) S Maio, S Baldacci, M Borbotti, A Angino, F Martini, B Piegaia, P Silvi, M Simoni, F Di Pede, and G Viegi. CNR Institute of Clinical Physiology, Pulmonary Environmental Epidemiology Unit, Pisa, Italy. Background: Asthma is a chronic inflammatory airway disease closely associated with atopic diseases like allergic rhinitis. Both diseases are increasing to epidemic proportion, with increasing medical costs, with a reduced patients' quality of life (QoL) and lower productivity. The correct management of asthmatic and rhinitic patients would be ensured by the use of international World Health Organization (WHO) guidelines as GINA (Global Initiative for Asthma) and ARIA (Allergic Rhinitis and its Impact on Asthma). Many studies prove that the guidelines recommendations are not or only partially applied within the clinical practice. Objective: To evaluate the applicability of the GINA and ARIA guidelines and their impact on patients_ QoL in General Practice; to evaluate the short and long term efficacy of a Continuing Medical Education (CME) intervention for General Practitioners (GPs) in Italy. Methods: Prospective observational study in different Italian areas (North, Centre, South-Islands). 168 GPs (71 attended an educational intervention and 97 didn_t attend it) will enrol their patients with diagnosis of asthma/rhinitis and with prescription of anti-asthmatic or anti-rhinitic drugs or asthmatic/ rhinitic-like symptoms in the previous 12 months. The GPs and their patients will fill in a questionnaire about respiratory allergic diseases and adverse drug events. The patient will fill in also the Rhinasthma questionnaire about the QoL and the Asthma Control Test. A follow-up will be done after 12 months. Expected Results: 1. epidemiological data on asthma and rhinitis burden; 2. evaluation of applicability, utility and adherence to GINA and ARIA guidelines by GPs and patients; 3. short term and long term efficacy of a CME intervention; 4. evaluation of prescriptions appropriateness and treatment cost/efficacy ratio. Respiratory and psychiatric comorbidity in an Italian primary care population: *EPIDEA study S Baldacci 1 , M Borbotti 1 , S Maio 1 , A Angino 1 , F Martino 1 , F Di Pede 1 , M Balestrieri 2 , A Bellomo 2 , M Nardini 2 , and G Viegi 1 . 1 CNR Institute of Clinical Physiology, Pulmonary Environmental Epidemiology Unit, Pisa, Italy; 2 of Epidemiologic Research in Primary Care -EPIMED, -, Pisa, Italy. Aim: To evaluate the relationship between psychiatric disorders and respiratory diseases (chronic obstructive pulmonary diseases (COPD) and asthma) in an Italian primary care population. Methods: Cross-sectional survey carried out in 6 Italian geographical areas. 2083 subjects (68.1% of expected sample, mean age 44 + 13 yrs), randomly selected by 143 (70.1%) general practitioners, participated in the study. Each subject answered to a physician-administered interview, based on the standardized National Research Council questionnaire, for the detection of psychiatric disturbances, respiratory diseases and symptoms, relative risk factors and pharmacological treatments, and filled out the General Health Questionnaire (GHQ). Results: Prevalence of current and lifetime psychiatric disturbances (affective disorders and anxiety) were 27.5 and 46.0%, respectively. COPD and asthma were significantly associated with the presence of anxiety alone and, above all, with the co-presence of anxiety and depression. By logistic regression models adjusted for independent effects of age, area, smoking, education, working position, GHQ score, and sex, COPD resulted associated with anxiety and depression in combination (OR: 2.8, CI: 1.8Y4.6) and with anxiety alone (OR: 2.6, CI: 1.7Y4.1). Asthma was also associated with co-presence of anxiety and depression (OR: 2.2, CI: 1.3Y3.8) and with anxiety alone (OR: 2.1, CI 1.2Y3.5). Conclusion: EPIDEA study confirms the large diffusion of psychiatric disorders in an Italian primary care population. The association between respiratory diseases and affective and anxiety disorders suggests to take into account these aspects in the clinical practice of pneumologists, allergologists and psychiatrists. Exercise-induced anaphylaxis related to Cannabis sativa smoke exposure José Antonio Navarro 1 , Olga Villarreal 2 , Natividad Longo 3 , Alejandro Joral 1 , Jose Francisco Garmendia 1 , Susana Lizarza 1 , and Ascension Aranzabal 4 . 1 Hospital Donostia, Allergy Section, San Sebastian, Spain; 2 Hospital de Mendaro, Allergy Unit, Mendaro, Spain; 3 Hospital de Santiago, Servicio de Alergia e Inmunologia, Vitoria, Spain; 4 Hospital de Zumarraga, Allergy Unit, Zumarraga, Spain. Background: Exercise-induced anaphylaxis is a life-threatening condition that can be food-dependent (specific or non-specific) and drug-dependent; sometimes the only trigger is exercise. We report here a case in which the trigger of the episode was the previous smoking of marijuana (Cannabis sativa) cigarettes. The patient, a 23 years-old-man, had previosly been diagnosed of cereal-dependent, exercise-induced anaphylaxis. In spite of strict avoidance of these foods, he had recurrent episodes of anaphylaxis during exercise (jogging). When he was reevaluated he recognized he used to smoke marijuana cigarettes very often, sometimes before exercise. 2 years before this evaluation he begun presenting sudden onset rhinoconjuctivitis each time he smoked a marijuana cigarette. He gave up marijuana smoking; since then he hasnát had any other anaphylactic reactions. He has been jogging, eating cereals before exercising, without any problem for more than one year. Methods: Skin tests (prick-prick test) with fresh Cannabis sativa leaves, skin tests to a leave extract, IgE immunoblotting to a Cannabis sativa leaves extract. Results: Skin tests were positive, while they were negative in 10 controls On the immunoblotting, the patient's IgE recognized a 35 KD band. Conclusion: We report here the first case of exercise-induced anaphylaxis in which the trigger is the inhalation of Cannabis sativa smoke. The skin tests and in vitro results, as well as the clinical picture and evolution after smoke cessation are consistent with a BCannabis sativa smoke dependent, exerciseinduced anaphylaxis[, in a patient that also has a IgE mediated immediate rhinoconjuctivitis due to marijuana smoking. The mechanisms of the anaphylactic reaction are obscure; there are a few previous reports that confirm that Cannabis sativa smoking can cause immediate hypersensitivity symptoms, like bronchospasm, rhinoconjuctivitis or generalized itching in sensitized individuals, but to our knowledge there are no previous reports of an exercise-induced reaction that appears to be dependent on marijuana smoking. The usefulness of serum tryptase in the diagnosis of shrimp anaphylaxis in children Patcharaporn Wongkaewpothong, Punchama Pajarn, Chaweewan Sripramong, Siriporn Boonchoo, Nualanong Visitsunthorn, Pakit Vichyanond, and Orathai Jirapongsananuruk. Siriraj Hospital, Allergy and Immunology of Pediatrics Department, Bangkok, Thailand. Background: Anaphylaxis is a life-threatening allergic reaction, and foods are one of the most common culprits. Serum tryptase is marker of mast cell activation and could use to confirm anaphylaxis to shrimp. Objective: To determine the utility of serum tryptase in the diagnosis of shrimp-induced anaphylaxis. Methods: Twenty-one patients with previous allergic reaction from shrimp were recruited into a prospective study for shrimp challenge. Twelve patients developed mild allergic reaction and nine patients developed anaphylaxis. Serum tryptase were obtained before shrimp challenge and 1 hour after the onset of symptoms. Results: In both groups of patients, median tryptase levels were significantly elevated after the onset of shrimp challenge as compared to baselines (baseline tryptase -1.08, peak tryptase -2.33 2g/L in anaphylaxis group; baseline tryptase -1.49, peak tryptase 1.56 2g/L in non-anaphylaxis group, p G 0.05). The delta tryptase (peak minus baseline) values in the anaphylaxis group was significantly higher than non-anaphylaxis group (1.33 VS 0.125 2g/L, p = 0.0004). The tryptase ratio (peak divided by baseline) values in the anaphylaxis group was also significantly higher than non-anaphylaxis group (2.11 VS 1.055, p = 0.0001). Using the recommended cut-off range (peak tryptase G 12.0 2g/L) the sensitivity of such cut-off was 0.11 with specificity of 1.0. Conclusion: We recommend using serial tryptase values, including tryptase ratio and/or delta tryptase values, for the diagnosis of food-induced anaphylaxis. The latter two values may be helpful than peak serum tryptase. Suleiman Eisa Al-Hammadi. UAE University, Pediatric, Al-Ain, United Arab Emirates. Food is considered the most common cause of anaphylaxis in children. The common cause of food allergy and anaphylaxis are cow's milk, hen's egg white, wheat, peanut, tree nuts, fish, shellfish and soya. Fresh fruits is also common especially peach (prunus persica). Case: 8 years old boy presented to emergency room with generalized itchy erythematous skin rash all over the body after 30 min. of eating peach. There was no history of having medication, new food or an insect bite. There were no history of shortness of breath, wheezing, stridor, abdominal pain, vomitting, swelling of tongue or mucus membrane or loss of consciousness. the was significant hypotension noticed during obtaining the vital signs as low as 92/27 mmHg, lasted for about 24 hrs instead of 2 appropriate doses og adrenaline injections, chlorpheneramine maleate IV and intravenous fluids. Other history: No atopic dermatitis, allergic rhinitis, asthma. Breast-feded for 2 years and recieved goat milk as a child, mother had allergic rhinitis, father and other 5 siblings are healthy. This child continue to have oral allergy symptoms to accidental peach ingestion in small amounts after the anaphylaxis incident, but never anaphylaxis again to peach or other fruits or highly allergenic foods. Conclusion: Rosaceae fruit allergy (typically peach) is a true fruit allergy. The spectrum of symptoms range from oral allergy syndrome to rarely anaphylaxis case, as in our patient. Anaphylaxis can be the initial presentation of fresh fruit allergy even if it was consumed before. Avoidance and epinephrine autoinjectors can be an appropriate management. Background: Wheat is not an uncommon cause of food-dependent, exerciseinduced anaphylaxis. The aim of this study was to describe common clinical characteristics, laboratory manifestations and natural history of the disease. Methods: Children with history of wheat-dependent, exercise-induced anaphylaxis were identified. Skin prick test and specific IgE for wheat were done. A three-day challenge program including open challenge for wheat, exercise and exercise challenge test after a meal containing wheat was performed. Results: Five children, aged 5-14 years (mean: 8 + 3.74 years) were evaluated. Atopic history was found in 40% of patients. All patients had symptoms involved skin and respiratory systems and two had hypotension. Sera specific IgE for wheat were determined in 3 patients among which two were positive. Three patients completed the three-day protocol. Anaphylaxis occurred in 2 out of 3 patients with the amount of wheat consumed prior to exercise being more than 100 grams. Conclusion: Wheat-dependent, exercise-induced anaphylaxis is more common in male. Skin and respiratory symptoms are major manifestations. A three-day wheat challenge protocol is a definitive diagnostic tool. However the amount of wheat required for challenging should be high. Kazuki Sato 1 , Tomoko Numata 1 , Yoko Nezu 1 , Takayasu Arima 2 , Yuzaboro Inoue 2 , Minako Tomiita 2 , Naoki Shimojo 2 , and Yoichi Kohno 2 . 1 National Shimoshizu Hospital, Department of Pediatrics, Chiba, Japan; 2 Chiba University Graduate School of Medicine, Department of Pediatrics, Chiba, Japan. Two cases of anaphylaxis in exclusively breastfeeding are reported. Case1: 5-month-old boy with exclusive breastfeeding suffered from eczema since 2-month-old and his mother restricted intake of egg and cow's milk since 4 month. He was hospitalized with tarry stools at 5 month of age. Within an hour after breastfeeding after his mother had taken raw fish, he developed whole-body rash three times. Skin prick test was positive to horse mackerel, mackerel, salmon and codfish. After his mother restricted fish ingestion no symptom was observed while breast feeding was continued. Case2: 9-month-old boy with exclusive breastfeeding presented severe eczema at 1 month of age. Thereafter his mother and he avoided eggs, cow's milk, soy, and fishes. At 9 month, he was fed almost breast-fed and ate little solid food. One day he developed rash and cough soon after mother fed him breast milk. He was breast-fed again and 3 hours later presented vomiting and rash over whole body and was hospitalized. Blood examination revealed high titer of specific IgE to egg, cow's milk, sardine and codfish in his serum. Skin prick test was also positive to sardine and eggs. Only small amount of allergens was detectable in breast milk. Although food their mother took was not determined, anaphylaxis due to food allergens in breast milk was most suspected. Systemic immediate type food allergy due to small amounts of food allergens included in breast milk is rare and so far a few case has been reported. Carolina Díaz, Jessica Salinas, Antonieta Guzmán, Paola Toche, and Angelica Marinovic. University of Chile Clinical Hospital, Allergy and Immunology Unit, Santiago, Chile. Background: Anaphylaxis is a severe acute allergic reaction, quickly progressive and potentially fatal, triggered mainly by food antigens and drugs. No prevalence studies of this condition have been conducted in our country, neither of the clinical presentation and/or etiology. Objective: Determine the clinical characteristics of patients with anaphylactic reactions seen at the Clinical Hospital of the University of Chile. Patients and Methods: A retrospective review of medical records spanning 10 years (May 1997-May 2007) allowed us to obtain data of 127 patients who where derived to the Allergy and Immunology Department of the Clinical Hospital of the University of Chile with a diagnosis of anaphylaxis of unknown origin. Three patients were excluded from this study, two with diagnosis of systemic mastocytosis and the other with hereditary angioedema. We evaluated: sex, age, number of episodes before consulting, interval between first and last episode, history of atopy, time elapsed between exposure to the suspected triggering factor and the appearance of symptoms, form of presentation, severity of the reaction, treatment received and clinical/laboratory work up. Results: 127 patients, 48 men and 79 women, were included. Age ranged from 11 months to 81 years, with a mean of 27.5 years; 61% were atopic. The average delay before consulting was 4 years, and median number of previous episodes was 2. Etiology was identified in 83% of cases; the main causes were drugs (31%), followed by foods (27%) and Hymenoptera stings (16%). The average time of onset of symptoms was 42 minutes post-exposure. Ninety six percent of patients presented urticaria and/or angioedema and severe manifestations (Grade III and IV) were seen in 70% of the individuals. From the individuals who had indication of management with epinephrine, only 42.1% received it in the Emergency room. Conclusion: The clinical characteristics of the Chilean patients with anaphylaxis are similar to those reported in other countries. It is relevant to emphasize the great percentage of atopics patients in our series as well as the delay in the consultation. Anaphylaxis in Singapore children SMY Wong, WK Liew, WC Chiang, M Kidon, and A Goh. KK Women's and Children's Hospital, Paediatric Allergy, Immunology and Rheumatology, Singapore, Singapore. Background: To study the epidemiology and review the treatment of anaphylaxis in Asian Singapore children. Methods: This is a 2-year retrospective study, which includes all patients with anaphylaxis seen in the KK Children's Hospital, Singapore. Cases were identified by the relevant discharge codes from the Children's Emergency (CE) and hospital's inpatient records, and cross-checked with referrals to the Allergy clinic. Results: 30 children with anaphylaxis were identified. 19 (63%) were male. 3 (10%) were less than 1 year old at the time of presentation, 8 (27%) were aged 1Y5, 11 (36%) were 6Y11, and 8 (27%) were 12Y16 years. Race distribution corresponded to population demographics. 25 (83%) had features of atopy. Food triggers were reported in 17 (57%), but food specific IgE sensitization was demonstrated only in 11 children (37%). [Egg (4), peanut (4), cow's milk (1), clam (1), chinese pear (1)]. There were 11 (37%) cases secondary to drugs: [NSAIDs (4), paracetamol (3), traditional Chinese medication (1), asparaginase (1), desferrioxamine (1) and blood product (1)]. Two patients had no identifiable triggers and were diagnosed as idiopathic anaphylaxis. Most patients presented with dermatologic (87%) and respiratory features (83%). 8 (27%) children had vomiting or abdominal pain, and 8 (27%) had hypotension. There were no fatalities in this study. Of the 23 (77%) patients that presented to the CE, 74% were admitted for observation. 3 patients required admission to the intensive care unit because of hypotension requiring fluid resuscitation at presentation. 12 (40%) children were treated with epinephrine, but this was administered mostly by the subcutaneous route. 18 (60%) were given antihistamines; 22 (73%) steroids; 17 (57%) bronchodilator therapy; and 9 intravenous fluid boluses. 6 children were equipped with self-injectable epinephrine. Those who did not receive self-injectable epinephrine included cases with drug triggers, patients who defaulted, and parental refusal due to anxiety over self-injection or cost concerns. Anaphylaxis is predominantly food triggered in Singapore children, with a noticeable increase in peanut food allergy. There is also a significant number of cases attributed to ibuprofen and paracetamol. Whilst epinephrine is the cornerstone therapy for anaphylaxis only 40% of our cases received this treatment. There is an urgent need to improve the awareness and treatment of anaphylaxis in our population. The angiotensine converting enzyme activity in patients with an anaphylactic reaction after Hymenoptera sting event The renin angiotensine system seems to play a significant role in the response to anaphylactic reaction. The aim of the study was to evaluate the serum activity of angiotensine converting enzyme (ACE) in patients with anaphylaxis after Hymenoptera sting event depending on the polymorphism of ACE gene and specific IgE. Materials: The study group consisted of 31 patients, mean age 50T12 years. There were 21 patients with specific IgE to wasp venom, 6 to bee venom and 4 to both venoms. Methods: The ACE activity was measured according to the method described by Liberman I/D ACE polymorphism was determined based in PCR technique employing insert specific-primers. Specific IgE was measured with Allergopharma specific IgE ELISA Kit, Germany. Results: The ACE activity in studied group was 29,11T11,5 IU. Table 1 contains the ACE activity depending of the ACE gene polymorphism. *p G 0,01 Table 2 contains the ACE activity depending of the specific IgE to Hymenoptera. **p 9 0.01 Conclusion: 1. The serum activity of ACE in the patients with anaphylactic reaction to Hymenoptera is the same as in general population 2. There are significant statistical differences between ACE activity in groups according to the ACE gene polymorphism 3.The specific IgE does not influence on the ACE activity Andrzej Dbrowski 1 , Renata Rubinsztajn 1 , Jaros 3 aw Góra 2 , Renata Walkiewicz 1 , Zbigniew Gaciong 2 , and Ryszarda Chazan 2 . 1 Medical University, Dept of Pneumonology and Allergology, Warsaw, Poland; 2 Medical University, Dep of Internal Medicine Hypertension & Angiology, Warsaw, Poland. The activity of the ACE may play an important role in the anaphylactiv reaction. The ACE activity is genetically determinated. The DD genotype is correlated with arterial hypertension and other vascular disorders. The ID polymorphism in the Caucasian population is 0,53. The aim of the study was to analyze the frequency of angiotensine converting enzyme gene polymorphism in patients with documented history of anaphylactic reaction to a Hymenoptera sting. Materials: The study group consisted of 48 patients (38F, 16 M), mean age 48T14 years. There were 29 patients with specific IgE to wasp venom, 7 to bee venom and 12 to both venoms. The control group was 23 healthy person (9F, 14M), mean age 41T7 years. Methods: I/D ACE polymorphism was determined based in PCR technique employing inser-specific primers. Specific IgE was measured by Allergopharma specific IgE ELISA, Germany. Results: The ACE gene polymorphism in the control group were: DD n=5 (21.7%), ID n=15 ( 65.2%), II n=3 (13.1%) The ACE gene polymorphism in the study group were: DD n=16 (33.3%), ID n=22 (45.85%), II n=10 (20.85%) Conclusion: In the group of patients with documented anaphylactic reaction to Hymenoptera sting there were no differences between the ACE gene polymorphism and general population. We did not observed differences of this polymorphism between the wasp and bee group, but in the wasp/bee one the allel I was more frequent, and therefore in larger groups of seems justified. Milk and wheat: the most common cause of anaphylaxis in children Zahra Pourpak, Rosita Akramian, Raheleh Shokouhi Shoormasti, Leyla Ghojezadeh, Hasan Bemanian, and Mostafa Moin. Immunology, Asthma & Allergy Research Institute, Immunology Department, Tehran, Islamic Republic of Iran. Background: Anaphylaxis is a medical emergency requiring immediate recognition and treatment. We report common findings of the first case series of anaphylaxis in Iran, which carried out in children who referred to the center of Immunology, Asthma and Allergy. Materials and Methods: Children referred to the clinic of allergy with a previous diagnosis of anaphylaxis were considered in this study (2003 -2006) . A specific questionnaire was completed for all of the patients and a detailed clinical history and demographic data were recorded. For all of the patients, Skin prick tests and specific IgE measurement were performed for suspected allergens. Determination of causative allergen was based on patients' history and IgE mediated tests. Challenge test was not performed for any patient. Results: 39 children ages ranged from 1m to 13 year old with a previous diagnosis of anaphylaxis were referred to the clinic of allergy during 2 years. (48/1% male and 24/1% female respectively). Skin prick tests was positive in 78/9% and specific IgE was positive in 88/2%. The most probable causative agents in this population were foods (92/40%). Milk (55/ 6%) and wheat (18/5%) were the most frequent causes of anaphylaxis in this study.58% of children had more than 2 episodes of anaphylaxis in their life; also 6 cases of multifactorial food anaphylaxis were identified for the first time. The most common causative allergen for recurrence anaphylaxis was milk in 20 patients Dermatologic signs and symptoms were the most common (92/2%) vs. 76/%, 62/5%, 21/6%, 20/8% for respiratory, gastrointestinal, neurological and cardiovascular signs and symptoms, respectively. Flushing was seen in 66/7%, prurities in 56/9%, urticaria in 54/9%, preorbital edema in 52/1% of patients, rash in 50%, erythema in 43/1% and angioedema in 41/2%. Conclusion: These data confirm that foods specially milk and wheat are the most common causes of anaphylaxis in children and dermatologic signs and symptoms are the most common finding, thus parents and physicians should be educated to recognize the most common causative allergens and common presenting clinical manifestation of this disease to avoid the progression of this life threatening syndrome and be able to manage anaphylactic patients. Food anaphylaxis: a clinical study Isabel Carrapatoso, Alexandra Santos, Emília Faria, Celso Pereira, Carlos Loureiro, and Celso Chieira. Coimbra University Hospital, Immunoallergology Department, Coimbra, Portugal. Background: Food allergy is one of the most common causes of anaphylaxis. Objective: The aim of this study was to characterize a group of patients with food anaphylaxis followed-up in a food allergy outpatient department during a one-year period. Methods: Twenty-two consecutive patients observed in our department from June 2006 to May 2007 with a suggestive clinical history of food anaphylaxis were selected. Foods involved in the severe systemic reactions were identified and atopic co-morbidities, occurrence of anaphylaxis in the study period and the medication used were investigated. A standardized questionnaire and skin prick tests to airborne allergens (mites, moulds, cockroach, cat and dog dander, pollens, feathers, latex) and food allergens (egg, milk, seafood, mammals and birds meat, fruits, nuts, cereals, legumes, spices and other vegetables) were performed. Serum specific IgE to airborne and food allergens and/or prick-to-prick with natural food extracts were done in selected cases (if there was evidence of clinical reactivity). Results: All the patients selected had a past history of an immediate severe systemic reaction after ingestion of food. The mean age of the population studied was 37T10 years. Fourteen patients were female. The mean age at the first anaphylaxis episode was 24T11 years. Foods involved were fruits in 4 patients, crustaceans in 4 patients, cow's milk in 4 patients, nuts in 3 patients, legumes in 2 patients, cereals in 2 patients, fish in 2 patients and egg in 1 patient. All patients were sensitised to the culprit food. Sensitisation to airborne allergens occurred in 19 patients (10 to mites, 7 to grasses, 5 each to weeds and trees, 3 to latex and 1 each to cockroach, dog dander and feathers). Sensitisation to more than 1 food occurred in 16 patients. Eighteen patients had atopic co-morbidities related to airborne allergens (12 patients had asthma). Recurrence of anaphylaxis occurred in 2 patients with cow's milk allergy in the study period. Self-administered adrenaline was used by 1 patient twice. Conclusion: In the population studied a high proportion of atopic asthmatic patients was found. Different plant and animal foods were implicated. Hidden milk allergens can be extremely difficult to avoid and this fact could explain the recurrence of anaphylaxis in 2 cow's milk allergic patients, in spite of a strictly restrictive diet. Gil-Soon Choi, Han-Jung Park, Sung-Jin Choi, Gyu-Young Hur, Seung-Youp Shin, Seung-Hyun Kim, Dong-Ho Nahm, and Hae-Sim Park. Ajou University School of Medicine, Allergy and Rheumatology, Suwon, Republic of Korea. Octopus variabilis, a kind of Mollusca, is a favorite food in oriental countries. There have been no published report of anaphylaxis caused by octopus and the mechanism of this food allergy has not been understood. We experienced two cases of anaphylaxis developed afteringestion of raw octopus. The first patient, a 52-year-old man without any previous allergic diseases, experienced generalized urticaria, dizziness, and loss of consciousness at one hour after ingestion of raw octopus. The skin prick test showed weakly responses to octopus extract, D. pteronyssius, D. farinae, oyster, shrimp (A/H ratio was all 2+). Serum specific IgE antibody to octopus extract was undetectable by ELISA, whereas increased level serum specific IgG4 antibody to octopus extract was noted by ELISA. The second patient, a 51-year-old woman with allergic rhinitis and allergic conjunctivitis had suffered from recurrent abdominal pain, chest discomfort and dizziness for 5 min after ingestion of steamed or raw octopus. The skin prick test showed negative response to octopus extract, but positive responses to D. pteronyssius and, D. farinae. Serum specific IgE, IgG1 and IgG4 antibodies to octopus extract were not found by ELISA. These finding suggest that octopus could induce anaphylaxis via non-IgE mediated mast cell activation. Background: Anaphylaxis is an emergency condition. Attempts should be made to find the causative agents for further prevention. Methods: We prospectively studied 28 patients with anaphylaxis at Siriraj Hospital, Thailand from January 1st, 2003 to July 31st, 2004. Direct patient interviews and physical examinations were done. Skin prick tests and challenges to the agents according to history, were carefully performed to identify the causes of anaphylaxis. Results: The causes of anaphylaxis were identified by positive skin test in 24/28 patients. In these 24 patients, oral challenge test and specific IgE wereperformed and found positive in 3/3 and 4/4 patients, respectively. The agents causing positive skin tests were ant (7 patients), seawater shrimp (4patients), immunotherapy (3 patients), wheat (2 patients), wasp (2 patients), seawater mollusk (2 patients), seawater crab (1 patients), wheat-dependent exercise-induced anaphylaxis (WDEIA, 1 patient), cow's milk (1 patient), andciprofloxacin (1 patient). Four cases had negative skin test. In these patients, the causative agents could not be identified in 3 cases and the patientswere diagnosed with idiopathic anaphylaxis. One patient had skin test negative to erythromycin but positive oral challenge to erythromycin. The causes of positive oral challenges in 3 patients were seawater shrimp, seawater mollusk and WDEIA (1 patient each). The causes of positive specific IgE wereseawater shrimp, wheat, cow milk and ant (1 patient We present the case of a woman, 36 years old, who smokes around 20 cigarettes a day, health worker, with a family history of atopy and a personal history of tonsillectomy, left renal colic and habitual consumption of oral contraceptives. From the last five years she has been presenting episodes of urticaria and occasionally angioedema, immediately after the solar exposure (at any season of the year), located only in the exposed areas. The injuries remain for 2 to 3 hours and disappear without leaving any residual injury. During three times she suffered head instability and afterwards loss of consciousness, twice because of a heavy solar exposure (despite of having applied sunscreen cream previously) and another time after reciveing an UVA's bronzed session. Laboratory findings, including complete blood cell count, erithrocite sedimentation rate, lactate dehydrogenase, renal and liver function, glucose, immunoglobulins A, G, M, tryptase, complement C3, C4, rheumatoid factor, total proteins and C-reactive protein did not reveal any pathological values. Test for antinuclear, anti YENA, anti-mithocondrials, anti-muscular smooth, anti-parietal cell antibodies were negative. The total IgE was 1000 UI/ mililiter. The 24 hours urine and faeces test analysis in order to detect porphyrins were negative. Skin prick test with a set of common air-allergens were positive for house dust mites. Photobiological examination: urticarial lesions were caused after unique exposure to UVB and to UVA irradiation, but not with the visible light spectrum. Although the minimal urticaria dose was not determined, the evolution of this case show an extreme seriousness. She continues with clinic symptoms in the area of exposure few minutes after minimal solar exposures in winter (although she has not presented new episodes of anaphylaxis) in spite of the continued treatment: topical broad spectrum sunscreen daily and all through the year, different systemic H1 antihistamines (cetirizine 30 mg/day, hidroxicina, ebastine, desloratadine), beta carotene supplementation, natural desensitisation approach by regular sun exposure. We suspect this is a case of solar urticaria that, clinically changed into an anaphylaxis when receiving very high doses of radiation. Kanika Piromrat. Bhumibol Adulyadej Hospital, Pediatrics, Bangkok, Thailand. Stinging insects comprised approximately 10 percent of the causes of anaphylaxis in Thailand. We live in a tropical climate, so some of the temperate inhabited insects do not exist in Thailand, such as yellow jackets (Vespula spp.), white faced hornet (Dolichovespula maculata), and yellow hornet (Dolichovespula arenaria). The poster presented pictures and brief description of common insects that have a potential of causing anaphylaxis in Thailand, such as tropical wasps, paper wasps, Giant bee, Indian bee, Small bee, bumble bee, venomous ants, and kissing bugs. In conclusion, the study described the species and characteristics of common stinging insects that can cause anaphylaxis in Thailand. Surena Vahabi. University, Perio, Tehran, Islamic Republic of Iran. Background: Mast cells play an important role in allergic reaction, host defense, local homeostasis, inflammation and angiogenesis. The objective of this study was to evaluate the relationship between mast cell numbers and different types of periodontal diseases. Methods: Gingival specimens were taken from 20 moderate to advanced chronic periodontal and 19 moderate to advanced aggressive periodontal sites as case groups and 18 healthy/gingivitis sites as control group in routine periodontal surgeries (flap and crown lengthening) and were examined after toluidine-blue staining for mast cells counting and hematoxylin-eosin staining for assessing inflammation. Inflammatory and mast cells in 5 micron sections were assessed by two observers 3 times, utilizing light microscope at 100X and 400X magnification. ANOVA and T tests with an alpha error level less than 5 percent were used to analyze data. Results: Mast cells numbers were higher in chronic versus aggressive periodontitis and healthy/gingivitis (p=0.000). The aggressive periodontitis didn't have more numbers of mast cells as compared to healthy/ gingivitis (p90.05). There were no relationship between mast cell numbers and degree of inflammation in 3 groups. Conclusion: The present study indicates more mast cell numbers presence in the chronic periodontitis sites than other sites. The results of this study suggest more studies to evaluate dynamic aspects of host defense in conjunct with other aspects of immune system, simultaneously. Exogenous nitric oxide regulates cyclooxygenase-2 expression and prostaglandin D 2 generation in mast cells Tae Chul Moon and A. Dean Befus. University of Alberta, Department of Medicine, Edmonton, Canada. Background: Mast cells (MC) are important effector cells in allergic and inflammatory responses through secretion of various mediators following activation. Nitric oxide (NO) is important signaling molecule that regulates MC function. It can depress MC allergic responses such as leukotriene (LT), cytokine and chemokine production, as well as MC degranulation. However, the involvement of NO in prostaglandin (PG) D 2 production, an important lipid mediator produced in MC, is unclear. In PG synthesis, cyclooxygenase (COX) is an important enzyme and two isozymes of COX, constitutively expressed COX-1 and inducible COX-2, have been reported. It is well established that mouse bone marrow-derived mast cells (BMMC) exhibit biphasic PGD 2 biosynthesis; COX-1-dependent immediate and COX-2dependent delayed PGD 2 production, when BMMC are stimulated with SCF, IL-10 and IL-1$. Methods: The effects of NO on COX-2 expression and PGD 2 generation in BMMC were investigated using NO-donors, S-Nitrosoglutathione (SNOG) and S-Nitroso-N-acetylpenicillamine (SNAP). PGD 2 production by stimulated BMMC was assayed using PGD 2 -MOX enzyme immunoassay kits. Western blot and real-time RT-PCR were used to measure COX-2 protein and mRNA expression. Results: Exogenous NO augmented COX-2 protein expression and increased COX-2-dependent PGD 2 generation in response to SCF, IL-10 and IL-1$. The increased expression of COX-2 by NO-donors was reduced by the p38 MAPK inhibitor, SB202190. Downstream of p38 MAPK, NO-donors augmented not only COX-2 mRNA transcription but also its stability. By contrast, NO-donors did not affect COX-1 protein expression. However, in contrast to the augmentation of COX-2 expression and activity, SNOG (100 to 500 6M), but not SNAP (up to 500 6M) inhibited COX-1dependent PGD 2 generation. Conclusion: Thus, exogenous NO regulates PGD 2 production by MC through regulation both of COX-1 and COX-2. Furthermore, these findings help us to understand the role of NO in MC function and the regulatory mechanisms of lipid mediator generation in MC in inflammatory diseases. Cigarette smoke suppresses the production of cytokines but not chemokines by IgE/Ag activation in mast cells Esmaeil Mortaz, Gerts Folkerts, Ferdi Engels, Danielle Raats, Mehdi Vaezi Rad, Frans Nijkamp, and Frank Redegeld. UIPS, Immunopharmacology, Utrecht, Netherlands. Background: Chronic obstructive pulmonary disease (COPD) is a major incurable global health burden and will become the third largest cause of death in the world by 2020. It is currently believed that an exaggerated inflammatory response to inhaled irritants, in particular cigarette smoke (CS), causes progressive airflow limitation. This inflammation, where macrophages and neutrophils are prominent, leads to oxidative stress, emphysema, small airways fibrosis and mucus hypersecretion. Mast cells are important effector cells in anaphylactic reactions and are involved in a variety of immunological and non-immunlogic processes. However, the role of mast cells in pathogenesis of emphysema not yet documented. Recently, we have demonstrated that CSM induces protease expression and chemokine release by primary mast cells. In this study, we further investigated the effects of CSM on the cytokine production of mast cells respect to activation with IgE and antigen. Materials and Methods: BMMC were cultured from BALB/cBy mice for 3 weeks. Cells were exposed against CSM and activated with IgE and antigen. Degranulation of cells was assessed by monitoring the release of the granular enzyme beta-hexosaminidase. The amount of cytokines were determined in supernatants and IkB-"\ degradation, P65 and CREP phosphorylation were measured by Western blot. Results: CSM attenuated degranulation and of cytokines releases of mast cells by a concentration-dependent and non-cytotoxic manner. CSM did not effect on cytokine releases induced by LPS. Moreover, CSM induces phosphorylation of CREB and ATF-1 when activated with IgE and antigen. Conclusion: Thus, some of the inhibitory effects of CSM on degranulation and cytokine releases may different than chemokine and leukoterine releases. The suppressive effect of Houttuynia cordata extract on HMC-1 cell migration in response to stem cell factor Hottuynia cordata Thunb. (Saururaceae; HC) is known as a therapeutic drug that has been used in traditional oriental medicine for the treatment of allergic diseases. Mast cells function as regulatory cells in a variety of inflammatory diseases, in particular, asthma and atopic dermatitis. In this study, we examined the effect of HC extracts on the chemotactic activity of the human mast cell line, HMC-1, induced by stem cell factor (SCF). The survival rate of HMC-1 cells was not altered by treatments with the HC extracts at a concentration of 10 2g/ml for 24 h. SCF showed the typical bell-shape curve for the HMC-1 cell chemoattraction with the peak of the curve at the SCF concentration of 100 ng/ml. HC-1, which was the whole plant (Houttuynia cordata) extracted with 80% EtOH, and HC-3, which was the residue successively partitioned with EtOAc, both had inhibitory effects on HMC-1 cell movement. After the treatment with 10 6g/ ml HC-1 extract for 6 h and 24 h, the chemotactic index (CI) of HMC-1 cells decreased up to 73% and 63%, respectively. HC-3 extract significantly inhibited the cell movement (72% T 6% and 44% T 2%). The HC-1 and HC-3 extracts had no inhibitory effect on the mRNA and protein expressions of c-kit, SCF receptor. SCF tranduces the chemotaxis signaling via NF-kB translocation, and both extracts blocked the activation. Taken together, our results indicate that HC-1 and HC-3 extracts inhibit the chemotactic activity of HMC-1 cells in response to SCF by blocking the NF-kB activation. These substances may be helpful for treating the inflammatory diseases associated with mast cells. The effect of IL-3 and SCF mixture on cell proliferation and rat mast cell protease-lT synthesis of rat bone marrow derived mast cells Background: Mast cells are the important immune cells to produce cytokines chemokines and histamine, and closely related to allergic reactions processing. The investigation of mast cell activation enhances the understanding of allergies and requires easy supply of large number of mast cells of culture. Bone marrow derived mast cell (BMMC) are the mast cells to provide sufficient number of cells and are affected by the kinds of cytokine combination and dose both in proliferation and in differentiation. This investigation was about the effect of culture condition on rat BMMC cultivation in serum supplement and cytokines. Methods: Rat bone marrow cells were cultured with the media conditioned differently by the addition of individual, or mixed cytokines of IL-3 IL-4 IL-6 and SCF. Rat BMMC proliferation was measured using CCk-8 assay. Rat BMMC differentiation was analyzed by the level of RMCPl T. Serum supplement effect on rat BMMC culture was investigated by comparison of the media including horse sera with the media carrying rat sera. Results: In the culture with horse serum supplement, rat BMMC proliferation in the presence of IL-3 (10 ng/ml) mixed with SCF (10 ng/ml) was the highest among the applied cytokine mixture. Rat BMMC with the mixture of cytokine concentration showed relatively high level of RMCPl T. In the culture with rat serum supplement, the cytokines condition of IL-3 10ng/ml with SCF 10ng/ml did not induce RMCPl T production. However the adjustment of cytokine concentration with the mixture of IL-3 10ngml and SCF 1.25ng/ml enhanced RMCPl T synthesis, comparable to the level produced by IL-3 10ng/ml and SCF 10ng/ml in the media supplemented with horse serum. Conclusion: These results indicated that rat BMMC preparation requires IL-3 and SCF, and that rat BMMC differentiation be affected by the ratio between IL-3 and SCF and by the kind of serum. Mar Guilarte, Victoria Cardona, Maria Vicario, Carmen Alonso, Cristina Martinez, and Javier Santos. Hospital Universitari Vall d'Hebron, Allergy. Digestive Diseases Research Unit, Barcelona, Spain. Background: Stress may facilitate intestinal inflammation by promoting epithelial barrier dysfunction allowing luminal antigens to enter the mucosa that may be crucial for the development of food allergy. Methods: Male WKY rats were submitted to crowding stress (CS) (8 rats/ cage) or sham-crowding (2 rats/cage) for 15 days. In addition, ketotifen, a known mast cell stabilizer or saline was injected i.p. at day 14th (1 mg/kg, 2 doses, 12 h interval). At the end of the stress or sham protocols, jejunal segments were mounted in Ussing chambers to measure macromolecular permeability to horseradish peroxidase (HRP). Rat mast cell protease II (RMCPII), an indicator of mast cell activation, was measured in jejunal perfusates and in jejunal homogenates at the end of stress or sham exposure. Plasma corticosterone and CRH (corticotropin releasing hormone) receptor 1 (R1) and 2 (R2) expression in jejunal tissue was measured by an ELISA assay by RT-PCR respectively. Results: Plasma corticosterone was tree fold increased after the stress protocol in the CS group (CS: 138T27 ng/mL vs. control: 52T16; p=0.0001; n=8/ group). CRH-R1 but not CRH-R2 expression was increased in the CS when compared to control. Crowding stress (CS) induced an increase in HRP permeability (CS: 18,2T5 pmol/cm2/h vs. control: 6,2T2.8; p=0.0004;), that was diminished when ketotifen was administered (12,1T5,2 pmol/cm2/h; p=0,015 vs CS; n=8/group ). Furthermore, jejunal RMCPII was higher in perfusates from stressed rats (CS: 23.6 T 5.1 6g/L vs. control: 14.8 T 9.9; p=0.021; n=8/group) and in tissue homogenates (CS: 456T123 ng/mg of tissue vs. control 274T84; p=0.002: n=8/group) suggesting ongoing mast cell activation. Ketotifen reduced the increased RMCPII in both jejunal perfusates (16,5T7,2 6g/L; p=0,019 vs. CS; n=8/group) and in tissue homogenates (198T72 ng/mg of tissue; p=0,002 vs. CS; n=8/group). Ketotifen did not induce any change in the parameters evaluated in the control groups. Conclusion: Stress evoked an epithelial barrier defect that may facilitate the uptake of luminal macromolecules and is mediated by mast cells. This synergistic effect may play an important role in food allergy. Importance of matrix metalloproteinase -9 (MMP-9) and its inhibitor (TIMP-1) in development of postoperative complications of patients with degenerative-dystrophic diseases (DDD) of hip joints Roman Vinchel R and Elena Markelova E. Vladivostok State Medical University, pathophisiology, Vladivostok, Russian Federation. Purpose of our work was to estimate of prediction possibility of development of complications in postoperative period by means of monitoring of MMP-9/TIMP-1 level in patients' blood serum before and after hips replacement. Materials and Methods: We tested blood of 27 patients before and after hips replacement operation, undergoing, 11 patients (40.74%) from them had postoperative infectious complications being developed, and 16 patients (59.26%) had smooth postoperative course without infectious complications. Blood of 30 healthy donors was as control. It was drawn fourfold blood sampling, as the following: the first blood sampling was made before operation, the second -during first 24 hours after operation; the third -during fifth 24 hours after operation and the fourth -during fourteenth day after operation. Estimation of MMP-9/TIMP-1 was made by immunoenzyme method using of the test-system of BR & D System Inc.[, USA. Results: It was detected the following: before operation patients from group with smooth postoperative course had level of MMP-9/TIMP-1 equal 10.90T0.83 ng/ml that was essentially higher than patients of control group had, being 0.22T0.07 ng/ml (pG0.05) typical for healthy people. There was increase of MMP-9/ TIMP-1 (12.59T1.02 ng/ml, pG0.001) on the first days after operation with subsequent continued increase up to 12.75T1.25 ng/ml, pG0.001 on the 5th day. Essentially decrease of MMP-9/ TIMP-1 up to level of 12.40T0.98 ng/ml was observed on the first day after operation that for certain did no differ from values of first day after operation. In the patient group with complicated postoperative course the index MMP-9/ TIMP-1 before operation was 9.54T1.37 ng/ml, this index was higher in the control group as well (0.22T0.07 ng/ml (pG0.01). On the first day after operation it was observed rapid growth of MMP-9/ TIMP-1 (13.37T1.41 ng/ml, pG0.05) in above-mentioned patient group, and on the 5th day the level still persisted high (14.82T1.74 ng/ml). On the 14th day the level of MMP-9/ TIMP-1 decreased slightly and was 14.27T1.07 ng/ml (p90.05). Conclusion: Divergence of MMP-9/ TIMP-1 ratio towards its significant increase already before operation for patients with DDD testifies to essential pathogenetic role of MMP-9 in pathogenesis of dystrophic diseases of hips. The role of hyalgan at inhibition of IL-1beta-stimulated production of matrix metalloproteinase Natalya Yatsyshyn, Roman Yatsyshyn, and Yevgen Neyko. Medical University, Internal Diseases, Ivano-Frankivsk, Ukraine. Background: Hyalgan is now widely used in the treatment of osteoarthritis (OA) by intra-articular administration into the affected joints. Although a clinical benefit of Hyalgan has been demonstrated with respect to pain relief in patients with OA, the level at which the drug acts remains unclear. Objective: This study was aimed to investigate the mechanism of inhibitory action of Hyalgan on interleukin-1beta (IL-1beta)-stimulated production of matrix metalloproteinases (MMPs) in human articular cartilage. Methods: IL-1beta was added to human articular cartilage with or without changes of osteoarthritis (OA) in explant culture to stimulate MMP production. Articular cartilage was incubated or preincubated with Hyalgan to assess the effect of Hyalgan on IL-1beta-induced MMPs. Secreted levels of MMPs-1, -3, and -13 in conditioned media were detected by immunoblotting, while intracellular MMP synthesis in chondrocytes was evaluated by immunofluorescence microscopic analysis. Penetration of Hyalgan into cartilage tissue and its binding to CD44 were analyzed by fluorescence microscopy using fluoresceninated Hyalgan. Blocking experiments with anti-CD44 antibody were performed to investigate the mechanism of HA action. Results: Treatment and pretreatment with Hyalgan resulted in significant suppression of IL-1beta-stimulated production of MMPs in normal and OA cartilage explant culture. Fluorescence histocytochemistry revealed that Hyalgan penetrated cartilage tissue and localized in the pericellular matrix around chondrocytes. Hyalgan -binding blocking experiments using anti-CD44 antibody demonstrated that association of HA with chondrocytes was mediated by CD44. Preincubation with anti-CD44 antibody, which suppressed IL-1beta-stimulated MMPs, reversed the inhibitory effect of Hyalgan on MMP production induced by IL-1beta in normal and OA cartilage. Conclusion: This study demonstrated that Hyalgan effectively inhibited IL-1beta-stimulated production of MMP-1, MMP-3, and MMP-13, which supports the clinical use of HA in the treatment of OA. Such Hyalgan action on IL-1beta may involve direct interaction between Hyalgan and CD44 on chondrocytes. Background and Purpose: It is well known that after respiratory syncytial virus (RSV) bronchiolitis during infancy and early childhood, a number of children develop a persistent wheezing. The study of eosinophil cationic protein (ECP) suggests that, as with asthma, eosinophilic inflammation mechanisms may also play a role in RSV infection. The aim of our study was to prove the existence of eosinophilic inflammation in infant with RSV infection, by measuring local/systematic ECP. Methods: ECP was measured in the nasal lavage fluid and serum of 41 infants with RSV infection and 29 other respiratory infections, all who were admitted to our hospital between October, 2005 and March, 2007 (they ranged from 0 months to 24 months of age, except for four pair of twins). Results: Nasal lavage fluid ECP levels were significantly higher in infants with RSV infection than in the infants with other respiratory infections. (3.148T0.527 vs 2.723T0.461 microgram/L, pG0.05). But serum ECP levels did not indicate such a tendency. Moreover, a subanalysis showed that wheezing infants also had significantly higher levels of nasal lavage fluid ECP levels than in the case of non-wheezing infants, (3.240T0.512 vs 2.833T0.491 microgram/L, pG0.05), but serum ECP levels did not indicate such a tendency. With regard to both nasal fluid lavage and serum, no significant difference in ECP levels were found whether there was a family history of atopy or not. This suggests that RSV infection may induce local eosinophilic inflammation even during infancy. Conclusion: Eosinophils are more strongly activated in RSV infection than in other respiratory infectious disease. Theoretically, this result supports the hypothesis that RSV infection may cause reactive airway disease. Mechanisms of eosinophil infiltration in the middle ear of patients with eosinophilic otitis media Yukiko Yokoyama 1 , Ruby Pawankar 2 , Chika Ozu 2 , Tetsuo Ikezono 2 , Manabu Nonaka 2 , and Toshiaki Yagi 2 . 1 Nippon Medical School Chiba Hokusoh Hospital, Otorhinolaryngology, Chiba, Japan; 2 Nippon Medical School, Otorhinolaryngology, Tokyo, Japan. Background: Patients with intractable otitis media (OM) with co-existent bronchial asthma have an extensive accumulation of eosinophils in the effusion and mucosa of the middle ear; called eosinophilic otitis media (EOM). Previously, we reported that mast cells can regulate eosinophilic inflammation in nasal polyps. In order to elucidate the mechanisms of eosinophil accumulation in the middle ear, we analyzed eosinopil chemoattractants like RANTES, eotaxin and their ligand CCR3 as well as mast cell mediator tryptase in middle ear biopsies and effusion (MEE) of patients with OM with and without asthma. Materials and Methods: By ELISA, we analyzed the levels of RANTES and eotaxin in MEE and mucosa of patients with OM with and without asthma (control). The number of tryptase+ cells (mast cells), MBP+ cells (eosinophils), RANTES+, eotaxin+ and CCR3+ cells were analyzed by immunohistochemistry in middle ear biopsies. Results: Levels of RANTES and eotaxin were higher in patients with OM with asthma as compared to controls. Whereas in the mucosa of controls, MBP and eotaxin+ cells were not detected, they were markedly increased in patients with asthma in concert with high numbers of tryptase+, RANTES+ and CCR3+ cells. Background: Eosinophils preferentially accumulate at sites of airway inflammation of asthma. For circulating eosinophils to participate in the asthmatic airways, it is necessary to interact with adhesion molecules expressed on endothelial cells and then expose to inflammatory mediators such as cysteinyl leukotrienes(cysLTs). There is evidence that cysLTs including leukotriene (LT)D 4 regulate the functional status of eosinophils. Objective: To investigate whether interaction with adhesion molecules modifie eosinophil functions induced by cysLTs. Methods: rh-VCAM-1, rh-ICAM-1, or rh-P-selectin was dissolved in 0.05 M NaHCO 3 coating buffer, added to 96-well EIA plates and incubated at 4 -C overnight. Residual fluid was decanted and HBSS/0.1% gelatin was added to reduce non-specific activation of eosinophils. Eosinophils were isolated from blood of healthy donors, incubated in the EIA plates, and then exposed to LTD 4 . The generation of superoxide anion (O 2 j ) and release of eosinophil-derived neutrotoxin (EDN) were evaluated by cytochrome C reduction assay and ELISA, respectively. Finally, a combination of LTD 4 and VCAM-1, but not ICAM-1 or P-selectin, induced the release of EDN. Conclusion: The combination of VCAM-Ior ICAM-I and cysLT effectively induce effector functions of eosinophils. Eosinophil adhesion to and migrate across endothelial cells via these speficic adhesion proteins and subsequent exposure to cysLT may be involved in the manifestations of eosinophil activation in the airways of asthma. Anker Hansen 1 , Lars K. Poulsen 1 , and Kresten Skak 2 . 1 Rigshospitalet, AllergyClinic, Copenhagen K, Denmark; 2 Novo Nordisk, Department of Pharmacology, MålLv, Denmark. Introduction: Eosinophils are recruited to late phase inflammation and elicit host defense against parasitic infections. However, in allergy eosinophils contribute negatively by eliciting tissue damage. Our aim was to evaluate the expression of interleukin-21 (IL-21) and IL-21 receptor (IL-21R) in eosinophils. Methods: Eosinophils where purified from blood samples taken from allergic and healthy individuals using CD16 and CD3 microbeads to deplete neutrophils and remaining T-cells from the granulocyte population. RNA was purified using ABI prism 6100 robot and real-time PCR was performed using applied biosystems Taqman Gene Expression Assays. ELISA assays were performed for IL-21 receptor. Eosinophilic apoptosis was measured in a flow cytometric assay using AnnexinV and propidium iodide for 200,000 cells at 24, 48 and 72 hour time points. Results: The yield of eosinophils from healthy and allergic persons was 64x10 3 and 137x10 3 eosinophils per ml full blood respectively. Using realtime PCR no IL-21 mRNA could be detected; however IL-21 receptor mRNA was detected with a mean Ct-value of 31.16 from 6 donors. No difference was seen between allergic and healthy individuals. The naBve Bcell line Ramos was used as a positive control and yielded a Ct-value of È20, which is roughly a 1000-fold higher expression of IL-21R mRNA in Ramos compared to eosinophils. The IL-21R mRNA expression in eosinophils was comparable to the eosinophilic cell line HL-60 clone 15. A sandwich ELISA for detection of IL-21R in eosinophil cell lysate was established using a polyclonal rabbit anti-IL-21R antibody and a mouse monoclonal anti-IL-21R. The ELISA had a detection limit of 1 ng/ ml or roughly 1000 receptors per cell and no IL-21R expression was detected in either eosinophils or Ramos cell line. The effect on eosinophil apoptosis in presence of IL-21 or the other Type I cytokines (IL-19, IL-20, IL-22, IL-24, IL-26, IL-28 and IL-29) was examined, however, no apparent effect of either increased survival or increased apoptosis could be seen. Conclusion: Eosinophils express IL-21R mRNA. Real-time PCR showed a mean Ct-value 31.16 from 6 donors. IL-21R protein expression was examined using both flowcytometry and ELISA, but no IL-21R could be detected. Functional apoptosis assay showed that IL-21 or the other Type I cytokines had no effect on eosinophils, so it is unlikely that IL-21 stimulation of eosinphils is of major significance. Background: IL-5 is critically involved in proliferation, activation, migration and survival of eosinophils. As such, IL-5 may play a pathogenic role in hypereosinophilic syndromes (IHES), which are a heterogeneous group of disorders, characterized by sustained peripheral blood and/or tissue eosinphilia. We assessed the safety and efficacy of a humanized monoclonal anti-IL5 antibody in a patient with IHES with respiratory involvement. Methods and Results: The patient is a 25 year-old woman, with a 3 year history of IHES. Biopsies confirmed on various occasions the involvement of lungs and paranasal sinuses. The onset of the disease was an eosinophilic pneumonia (56% eosinpohils in bronchial lavage) with asthma and respiratory failure. The persistent eosinophilia and respiratory symptoms were successfully treated with doses of prednisone (25Y75 mg/daily), variable according to eosinophilia. Attempts to shift to inhaled steroids plus montelukast, invariantly lead to relapse. However, withdrawal of steroid treatment was forced by severe systemic side effects and bleeding gastric ulcera. An attempt with initimab was not effective. A treatment with mepolizumab (5 mg/kg intravenously) monthly was then started. Mepolizumab was well tolerated and rapidly reduced both eosinophilia and symptoms. Eosinophils decreased from 998/mmc to 210 after the first infusion and remained below 250/mmc at the subsequent controls. After 8 weeks the oral steroid could be discontinued. Conclusion: Mepolizumab is safe, and effective in lowering eosinophil count , has a glucorticoid sparing effect in patients with IHES. Keywords: hypereosinophilic syndrome, pneumonia, mepolizumab. Lin-Shien Fu. Taichung Veterans General Hospital, Section of Immunology & Nephrology, Dep. of Pediat, Taichung, Taiwan. Background: Recent studies suggest that several antihistamines can modulate various inflammatory reactions besides their H1-receptor antagonism. We investigated the effect of cetirizine (Ceti) and levocetirizine (Levo) on granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-8 (IL-8) secretion of A549 human airway epithelial cells. Methods: A549 cells were pre-incubated with Ceti (0.1, 1, 2.5, 5, 10 6M) or Levo (0.1, 1, 2.5, 5, 10 6M) individually for 16 hours and were then stimulated with IL-1$ for 8 hours. The levels of GM-CSF and IL-8 in cultured supernatants were measured by enzyme-linked immunosorbant assay (ELISA). Results: Our data showed that Ceti 5 and 10 6M and Levo 2.5, 5, and 10 6M significantly suppress GM-CSF secretion by 29.29%, 38.45%, 39.94%, 38.09% and 42.85%, respectively. While Levo 5 and 10 6M significantly suppressed IL-8 secretion (22.43% and 34.58%, respectively), this was possible only with Ceti 106M (24.96%). Conclusion: These results suggest that both cetirizine and levocetirizine at higher concentrations reduce the release of GM-CSF and IL-8 from A549 human airway epithelial cells stimulated with IL-1$. These observations indicate that these antihistamines may exert anti-inflammatory effects beyond their antagonistic histamine H1-receptor activity. Our results suggest that Levo could be more potent than Ceti in terms of anti-inflammatory activity. A new interesting property of hydroxyzine, a drug used in the treatment of chronic idiopathic urticaria and anxiety disorders Background: Human Vgamma9/Vdelta2 (V,9/V&2) T lymphocytes are known to participate in the defense against microbial pathogens and have antitumor activity. The cellular immune system of newborns is generally considered to be immature and hypo-responsive in comparison to that of adults. In this study we made a quantitative and qualitative comparison of the V,9/V&2 T lymphocytes, their activation status and production of cytokines in cord blood mononuclear cells (CBMC) versus those in adult peripheral blood mononuclear cells (PBMC). Methods: We employed flow cytometric analysis to compare numbers and phenotypic characteristics of V,9/V&2 T lymphocytes between CBMC and adult PBMC. Results: Numbers of V,9/V&2 T cells are lower in CBMC than those in adult PBMC while TCR expression levels were similar in both groups. Stimulation of CBMC V,9/V&2 T cells with isopentenyl pyrophosphate (IPP) did not induce IF-,, in contrast with adult PBMC positive production of this Th1 cytokine. The addition of IL-2 to IPP did not enhance IFN-, in CBMC V,9/ V&2 T cells. PMA/ionomycin stimulation did not lead to INF-, production in CBMC V,9/V&2 T cells, inversely again, adult PBMC activation produced this cytokine. Conclusion: In addition to the immaturity of the adaptive immunity in newborns, such as "$ T cells; CBMC V,9/V&2 T cells are hypo-responsive in comparison with adult PBMC V,9/V&2 T cells and this may be associated with the predisposition to infection with virus, bacteria and fungus, as well as the development of allergic inflammation in newborns. The matrix metalloproteinase 9 and its first type's inhibitor type (hMMP-9/TIMP-1) in bronchalveolar lavage fluid of pulmonary tuberculosis patients Andrey Safronov, Svetlana Sotnichenko, and Elena Markelova. Vladivostok State Medical University, Pathophysiology, Vladivostok, Russian Federation. Significant components of the territorial matrix are the matrix metalloproteinase that is a group of proteolytic enzymes, which are especially active in conditions of an inflammatory response. Matrix metalloproteinase 9 (MMP 9) is produced by normal alveolar macrophages, granulocytes, and neutrophiles. Expression of MMP 9 is increased in tissues affected by the process of remodeling and angiogenesis. Negative influence of this process is restricted by tissular inhibitor of metalloproteinase (TIMP-1). Main function of TIMP-1 consists in regulation of distraction of collagen and basic membrane components. We have researched a quotient of MMP 9 and its first type's inhibitor contained in the bronchalveolar lavage fluid of 23 teenagers suffered from pulmonary tuberculosis. Received results have been processed through statistical analysis; an arithmetic mean (M) and a mean error (m) have been also determined. Fidelity of results has been reckoned through Mann -Whitney nonparametric test. Evaluation was made in accordance with the clinical and laboratory findings and bacteriological test of sputum. The researches have shown that quantity of hMMP-9/TIMP-1 in the bronchalveolar lavage fluid of pulmonary tuberculosis patients comes to 2803, 85 T 270, 72 pg/ml. It was also elicited a tendency for increase of hMMP-9/TIMP-1 in Koch`s bacillus cases (+) (3127, 23 T 555, 20 pg/ml vs. 2735, 77 T 315, 58 pg/ml; p90,05). Differences in hMMP-9/TIMP-1 of patients with local pulmonary tuberculosis and infiltrative phthisis cases haven`t been revealed. The received findings are not sufficient for making unambiguous conclusions concerning pathogenetic value of the researched mediators and denote necessity for further searches of diagnostic characters for inflammation in cases of teenagersp ulmonary tuberculosis. The cytokines production at the experimental pneumonia Anna Kostyushko and Elena Markelova. Siberian Branch of Russian Academy of Medical Science, ¡The Fareastern Scientific Centred, Vladivostok, Russian Federation. Activation of cytokines system at different variants of the immune inflammation occurs without dependence from the etiologic factor. At the same time there is the data, testifying that microorganisms can develop the products being similar cytokines, and thus influence on the cytokines cascade at the immune answer. The purpose of the present investigation was to reveal influence of the etiologic factor on parameters of the local and systematic structure of the opposite cytokines (IFN, and IL-10) at the experimental pneumonia caused by S. aureus and E. coli. Infection of mice of Ñ Â À line was spent intranasally in the doze of 1x103 mt/ml for S. aureus (I experimental group) and 1x105 mt/ml for E. coli (II experimental group). Production of cytokines by blood cells and mice lungs was investigated for the 10th day after infection in reaction of IFA. Results and Conclusion: At infection of mice S. aureus level of IFN, considerably (8 G0,05) decreased (up to 0,84+0,1 pg/ml in blood and up to 36,38+2,38 pg/ml in supernatant of lungs). IL-10 level at mice of tentative group also tended to decrease and made 14,15+1,3 pg/ml in blood and 70,8+2,88 pg/ml in supernatant of pulmonary tissue. At infection of mice E. coli content of IFN, in blood changed little (3,97+0,28 pg/ml), and decreased in supernatant of lungs, but not so prominently, as at mice of tentative group (70,84+2,17 pg/ml). IL-10 level in blood of mice of the second experimental group was authentic (8 G0,05) below than in group of the control and made 10,8+0,87 pg/ml. And IL-10 level in supernatant of pulmonary tissue at infection of mice by E. ñoli increased practically in 2 times in comparison with the group of the control (186,08+5,13 pg/ml, 8 G0,05). Thus, carried out investigation revealed features of local and systemic production of cytokines depending on etiology of the experimental pneumonia. IFN, level under influence S. àureus prominently decreased. Development of inflammatory process in the lungs, caused by E. coli, rendered significant influence on local production of IL-10 increasing its level more than in 2 times, moreover IL-10 authentically decreased in peripheral blood. Ophtalmoherpes is a grand medical and social problem. Herpetic infection of eyes is relapsed in 50% of cases and results to reducing acuity of vision, disablement and deteriorates life activity. Enlargement of conception regarding inflammatory mechanism let us opportunity to optimize respective therapy. Objective: To work out cytokines content (TNF-á, GM-CSF, IL-12p70, IL-12p40 and IL-2) in blood serum and lacrimal liquor of the patients. Methods and Materials: Serum and lacrimal liquid of 60 ophtalmoherpes patients were tested conformably (women 38, man 22). Control group embraced 30 practically healthy volunteers without disease of organum visus. Cytokines were quantified with IFA method using specific reagents BR and D Diagnostics Inc.[ (USA). Consequences and Discussion: We found polysemantic modification of local and systemic cytokine profile of patients as against reference group. In blood serum were determined acceleration of IFNá 30-50 times more (ñG0,001), IL-2 level 2-10 times more (pG 0,5), IL-12p40 content decreased 1,5-3 times (pG0,05), whereas GM-CSF and IL12p70 were near to standard. The abovementioned indices in lacrimal liquor appeared in the following way. Authentically increasing of IL12p70 level (10,08+1,29 pg/ml against 4,98+1,1 pg/ml, pG0,01) and GM-CSF (7,57+1,73 pg/ml against 2,15+0,64 pg/ml, pG0,01). TNFá and IL-12 modifications had a tendency to heightening (pG0,2), whereas IL-12p40quantity reduced (pG0,05) in the same way as in blood serum. Obtained results allow us to draw a conclusion, that at ophtalmoherpes disease neutrophiles and macrophages mainly are more active locally and produce GSM-CSF and IL-12p70. System response is characterized by hyper production of TNF! and IL-2. Compensative anti-inflammatory reaction distressed as locally as systematically. Streptokinaza indused pulmonary infilltrates with hipereosinophilia-case report Biserka Kaeva, Oliver Jovkovski, and Sasa Kaeva. Medical Faculty, Pulmology and Allergology Clinic, Skopje, Macedonia, Fyrom. The various drugs and enviromental agents capable of producing acute pulmonary infiltrates with eosinophilia. We present a man on 48 with acute infarct of myocardi who was treated with streptokinaza besides the usual cardiological treatment. From the anamneza we faund aut that the patient did not have problems with his heart, neither any kind of alergic or some other kinds of ilneses. By the first impresion the patient was conscious, skared,dispnoic, afebril,with strong pain on his chest. By auskultation he had anormalvezicular breathig and heart rate of 110/min. The other phisical finds were in order. Imidiately after the posed diagnoza, a usual cardiological therapy was given and in the same time a streptokinaza (1500 000IE) was included. The patient beard this treatment very well. On the third day of the therapy his health condition deteriorated. He got frequent chaemoptisis, increased temperature(38C), decreased blod presure, and rich find of rales on clinical examinaton. Laboratory examination showed:ESR 42mm/1st hour, strong blod reduction, eosinophilia 49%, leukocytosis. On the x-chest ray both upper lobes were caracteristicaly involved and were described as the Bphotographic negative[ the radiographic shadows seen in pulmonary oedema. Chest CT showed bilateral pulmonary infiltrates mainly in upper and middle lobes. Bacterial examination and cultures of sputum were all negative. The search for parasistic infestation was negative as well as for BK infection. The lung volumens and diffusion capacities werw decreased and hipooxemia was most prominent. We started the therapy with 40mg prednisone daily with successive decrising in the folowing 6 weeks and also antibiotic and oxigen therapy. Some improvement was seen in less than a week but the radiograpfic shadows had completely resolved for 6 weeks. The number of eosinophils olso came back to normal. Biserka Kaeva, Gorica Breskovska, Zoran Arsovski, and Kamelija Busljetic. Medical Faculty, Pulmology and Allergology Clinic, Skopje, Macedonia, Fyrom. Background: Pulmonary thromboembolismus can be presented through pulmonary infiltration, which manifestation depends on extensiveness of the pulmonary embolisation and the presence of pleural effusion (PE). In a period of 3 years 29 pts with pulmonary thromboembolism were studied with regard to serum concentrations of IgE, D-dimer and Eosinophils in the pleural fluid during acute phase (on hospital admission) and after treatment (recovery phase). The aim of the study was to estimate the correlation between these parameters during acute and recovery phases. Methods: The number of eosinophils in the pleural fluid was determined from smear of pleaural fluid colored by May-Grunwald Giemsa. Serum IgE was measured with Radioimmunoassay diffusion (RID). Results: The serum IgE concentration increased during the acute phase to 422+/-307 IU/ml and decreased afterwards in all pts. The increase in serum IgE concentration lagged a few days behind that of the serum D-dimer concentration, indicating later IgE production then thrombus formation and lysis. Most of the pts developed a small chaemoragic PE, while 7 of them had more then 30% of eosinophils and highest levels of IgE measured in acute phase. Conclusion: These results indicated a relationship between serum IgE concentration and patophisiology of pulmonary thromboembolismus. Serum IgE and Eo count may be a good mark of severity for pulmonary thromboembolismus indicating on pathogenesis and prognosis. Modulation of peripheral and cord-blood derived gamma/delta T cells immune response after stimulation with newly designed compounds: possible new therapeutic approach Mancino Giorgio 1 , Auricchio Giovanni 1 , Battistini Luca 2 , Galli Elena 1 , Colizzi Vittorio 3 , Brunetti Ercole 1 , and Placido Roberta 2 . 1 San Pietro Hospital -Fatebenefratelli, AFaR Research Centre, Roma, Italy; 2 Santa Lucia Foundation -IRCCS, Neuroimmunology Unit, Roma, Italy; 3 University of Rome -Tor Vergata, Dept. Biology, Roma, Italy. Immune system, in response to infective or allergic agents, makes use of different cellular populations, such as alfa/beta and gamma/delta (g/d) T cells. The latter are present in both peripheral (PB) and cord (CB) blood, being the second in a immature stage, and respond to a variety of non-peptidic antigens, through a HLA-independent system, by secreting pro-inflammatory and immunomodulant cytokines, expressing membrane receptors and costimulatory molecules, and producing cytotoxic effectors such as perforin and granzyme. Recent studies have suggested that also the less common g/d T cells may play a role as effectors and immunoregulatory cells in the development and perpetuation of allergic inflammation and also in bronchial asthma, rhinitis and eczema We aimed at analysing the capability of g/d T cells derived from PB or CB to respond to non-proteic antigens, in view of a putative therapeutic use in newborn or infant pathologies. We firstly found that CB-derived g/d cells strongly respond to stimulation with aminobiphosphonate (ABs) compounds, such as pamidronate and zoledronate, in terms of expansion, whereas stimulation with pyrophosphates (PPs) only lead to no or little expansion. The population expanded with ABs is also perfectly functional, since cells are able to produce large amounts of cytokines (TNFalfa e IFNgamma) if secondarily stimulated with he same antigens. Interestingly, also PPs can induce ABs-expanded cells to cytokine production after a secondary stimulation, suggesting the existence of different responsive pathways concerning expansion and cytokine production in CB-derived g/d cells. Once established that CB g/d T cells are functional and can be efficiently stimulated, we focused our study to the stimulation or down-regulation of g/d T cells with newly designed and synthesised compounds (NCs). We therefore characterised a small-compounds library, and selected few molecules for their capability to stimulate or down-modulate g/d T cells from PB. We than used these molecules also for CB g/d stimulation. We found that, following stimulation with NCs, CB-derived g/d T cells strongly respond in terms of both cellular expansion and cytokine production, and this result was confirmed though the use of CB-derived gamma/delta clones. Studies are in progress to evaluate the toxicity of these compounds, and consequently their employment in vivo as g/d T cells immunomodulators in new therapeutic approach on infectious or allergic diseases. A novel low molecular weight thiol compound, N-acetylcysteine amide, attenuates allergic airway inflammation and hyperresponsiveness Recent studies have demonstrated that antioxidants are able to reduce airway inflammation and hyperreactivity in animal models of allergic airway disease. A newly developed antioxidant, small molecular weight thiol compound, Nacetylcysteine amide (AD4) has been shown to increase cellular levels of glutathione (GSH) and to attenuate oxidative stress related disorders. However, the effects of AD4 on allergic airway disease such as asthma are unknown. Objective: This study aimed to determine the effects of AD4 on bronchial inflammation and airway hyperresponsiveness. Methods: We have used a mouse model for allergic airway disease. The effects of AD4 on bronchial inflammation and airway hyperresponsiveness studies by measurement of intracellular ROS, GSH, glutathione disulfide, vascular permeability, proteins, tissue inflammation, and airway resistance. Results: The increased levels of Th2 cytokines, nuclear factor-.B (NF-.B) and hypoxia-inducible factor-1! (HIF-1!), and vascular endothelial growth factor, the increased ROS generation, the increased vascular permeability, and the increased mucus production after ovalbumin inhalation were significantly reduced by the administration of AD4. Conclusion: These results suggest that AD4 attenuates airway inflammation and hyperresponsiveness by regulating activation of NF-.B and HIF-1! as well as reducing ROS generation in allergic airway disease of mice. Clinical implication: These findings provide an important molecular mechanism for the use of a novel antioxidant, AD4 to prevent and/or treat asthma and other airway inflammatory diseases. Th1-dominant shift of T cell cytokine production, and subsequent reduction of serum immunoglobulin E response by administration in vivo of beta-carotene in a mouse model Mykola Korzh. Kharkov National University, Fundamental Medicine, Kharkov, Ukraine. Background: Th1 and Th2 cells, resulting from antigenic stimulation in the presence of IL-12 and IL-4, respectively, are implicated in the pathology of various diseases including allergic and autoimmune diseases. In this study we have asked whether administration of beta-carotene brings about a Th1/Th2 shift in vivo of the mice, and subsequent reduction of circulating IgE. Methods: Feed containing beta-carotene was administered orally to BALB/c mice immunized intraperitoneally with ovalbumin (OVA) for approximately 1 month. The titers of OVA-specific IgE, OVA-specific IgG1 and OVA-specific IgG2a in the mouse sera were determined. Cytokine productions by the spleen cells and serum Ig concentrations were studied by ELISA. We also examined the effect of fed beta-carotene on active systemic anaphylaxis. Results: Feeding beta-carotene to mice immunized with OVA inhibited the immediate reduction of the body temperature induced by antigen stimulation. Furthermore, the increase in serum histamine in the mice fed beta-carotene under active systemic anaphylaxis was lower than in controls. We then examined the pattern of cytokine production by spleen cells from mice followed by restimulation with OVA in vitro. The spleen cells showed enhanced Th1-specific cytokine production; spleen cells from the betacarotene administered mice produced more IFN-gamma as compared with those from control mice in an antigen-specific manner. IL-2 and IL-4 secretions of the spleen cells were comparable between the two mouse groups. Beta-carotene administration did not reduce serum IgG concentration. It markedly reduced total IgE level and an IgG1/IgG2a ratio, reflection of Th1/Th2 balance, in sera. Furthermore, beta-carotene administration reduced ovalbumin (OVA)-specific IgE levels in sera of the OVA sensitized mice. Conclusion: Thus, beta-carotene enhances IFN-gamma secretion and thus modulates Th1/Th2 cytokine balance, leading to reduction of serum IgE. Chiara Folli 1 , Desideria Descalzi 1 , Francesca Scordamaglia 1 , Anna Maria Riccio 1 , Cinzia Gamalero 1 , and Marco Barbieri 2 . 1 University of Genoa, Department of Internal Medicine, Genoa, Italy; 2 University of Genoa, ENT Department, Genoa, Italy. Background: Statins are serum cholesterol-lowering agents used for the prevention of atherosclerotic vascular disease but there is, a growing evidence they might have immunomodulatory activities. Methods: To evaluate the effect of these drugs in allergic diseases we treated PBMC from healthy and allergic patients in vitro with ParJ1,2 (5 2g/ml), Fluvastatin (0,1 2M), Atorvastatin (1 2M) and Simvastatin (1 2M) alone and in combination. NK derived from healthy subjects were activated with IL-2 (300 U/ml) alone or with the addition of statins. We also evaluated their effect on fibroblast cultures derived from human nasal polyps and turbinates stimulated as follows: not treated and treated with FGF (5ng/ml) alone or in combination with statins. Results: After different days in culture, cells were analyzed by flow-citometry for the evaluation of the following cell surface receptors: CCR3, CCR4, CXCR3 and CCR5 for PBMC; NKp30, NKp44 and NKp46 for NK cells and CD106 and CD54 for fibroblasts. Our results show that statins have no modulatory effect on cell surface protein expression from both PBMC and NK. CD54 was not modulated by the treatments in nasal polyp fibroblasts, whereas in turbinate fibroblasts its expression was upregulated by FGF and downregulated after the addition of Fluvastatin or Atorvastatin. Interestingly, also CD106 expression was not modified in nasal polyp fibroblasts stimulated or not with FGF and statins. Fibroblasts from turbinates showed an increased expression of CD106 after stimulation with FGF and only the addition of Atorvastatin was able to downregulate it. Conclusion: In conclusion, the different behaviour of statins on the diverse kind of cells in vitro is prompting news in the studies about their use for the treatment of inflammatory diseases. Latex allergy in operating room; not frequent but severe Gulbin Karakoc, Mehmet K]l]c, Ayfer Inal, Seval Kendirli, Derya Altintas, and Mustafa Yilmaz. Cukurova University, Faculty of Medicine, Pediatric Allergy-Immunology, Adana, Turkey. Objective: Latex allergy is becoming a major health concern among healthcare workers, of whom approximately 2.8 to 18% are reportedly sensitized. The aim of this study to determine the prevalence of latex allergy and potential crossreacting foods in operating room stuffs. Methods: One hundred and four operating room stuffs (aged 24 years to 58 years) including doctors, nurses and technicians completed a latex allergy questionnaire. They were questioned about symptoms of latex reactivity and about other allergies particularly to foods that may crossreact with latex. Informed consent was obtained and skin prick tests were performed with natural rubber latex.and with five potentially crossreacting foods (banana, kiwi, melon, tomato and potato). Specific IgE antibodies against latex and these foods were evaluated (Pharmacia CAP RAST system). Results: Five personnel (4.8%) described allergic symptoms they attributed to latex exposure. All latex allergic stuffs had skin symptoms, 2 had severe anaphylactic reaction and 2 had severe asthma. All these 5 personnel had positive reactions to both latex and crossreactive foods. Specific IgE against to latex was found to be positive in this group. There was no significant difference between the latex SPT-positive and -negative health care workers according to age, sex and total exposure time to latex. Conclusion: In this study we found the prevalence of allergy 4.8% among the operating room personnel. Although latex allergy was not more frequent in our population its presence may lead to severe allergic reactions such as anaphylaxis and severe asthma. Background: The effect of dog ownership during childhood on the development of allergy has been investigated in few studies with conflicting results. Objective: We investigate the association between childhood dog ownership, regular contact to dogs, and indoor endotoxin exposure during infancy and the development of allergic sensitization and atopic disease up to age 6 in 2 German cohort studies. Methods: Data from two ongoing birth cohorts GINI (n=1962) and LISA (n=1193) were analysed. In both studies, information on children_s contact with dogs and their allergic symptoms and doctor diagnosed allergic disease were collected at each follow-up using questionnaire. Specific IgE antibodies to common aeroallergens were measured at age 6. House dust samples were collected when the children were 3 months old and the amount of endotoxin in house dust was determined. Results: Dog ownership in early childhood was associated with a significant lower rate of mixed pollen and inhalant sensitization but had no effect on dog sensitization and the prevalence of allergic symptoms and diseases up to age 6. Regular contact with dog during childhood without ownership was not associated with any of the health outcomes. No associations were found between house dust endotoxin exposure during infancy and sensitization to dog, mixed pollen, and inhalant allergens. Conclusion: Dog ownership in early childhood protects against the development of inhalant sensitization and this effect cannot be attributed to the simultaneous exposure to endotoxin. School-a source for exposure to furry-pet allergens Lena Elfman, Yahong Mi, Dan Norbäck, and Greta Smedje. Occupational and Environmental Medicine, Department of Medical Sciences, Uppsala, Sweden. Background: The school environment is an important source of exposure to furry-pet allergens for most children in Sweden. This will have an impact on their health and possibly on development of allergy. The aim of this study was to compare two methods of allergen sampling, and to investigate the association between percentage of animal owners and allergen levels. Methods: Totally 120 classes in 35 primary and secondary schools in Uppsala, Sweden, were randomly selected. A questionnaire, including questions on pet ownership, was answered by 2 355 pupils aged 7Y13 years. Samples of settled dust were collected from floors and furniture throughout the classroom with a vacuum cleaner with a special sampling filter (ALK Abello). Airborne particulates were collected passively in Petri-dishes placed in the class-rooms over a week. Allergen levels for cat, dog and horse were determined using ELISA. Correlations between allergen levels in settled dust and air, and number of cat-and dog owners as well as those who ride in each class were analysed by Spearman_s rank correlation, with a two-tailed significance level of 5%. Results: The geometric mean (GM) allergen level in settled dust was 1530 ng/g for Fel d 1, 1420 ng/g for Can f 1, and 1340 U/g for Equ cx. In Petri dish samples, the GM allergen level was 1.8 ng/sample for cat, 1.3 ng/sample for dog and 4.7 U/sample for horse. The percentage (GM) of cat-owners was 27%, dog-owners 14% and horse contact 8%. Cat allergen level in dust correlated with cat in air (0.33, p=0.000) and percentage of cat-owners (0.30, p=0.001). Dog allergen level in dust correlated with dog allergen in air (0.27, p=0.003), and percentage of dog-owners (0.40, p=0.000). The same was valid for horse allergen level in dust, which correlated with horse allergen in air (0.37, p=0.000) and percentage of those who ride (0.44, p=0.000). Conclusion: In all cases, we found a correlation between percentage of animal-owners or riders and allergen levels in dust and air samples. A correlation between dust and air allergen levels has seldom been shown before. We believe that sampling over the whole floor, benches and chairs gives a more representative value. However, dust levels are proxy variables, and air levels may be a better measure of allergen exposure. Nipasiri Voraphani, Suparat Pohnu, Pantipa Chatchatee, and Jarungchit Ngamphaiboon. Faculty of Medicine, Chulalongkorn University, Pediatrics, Bangkok, Thailand. Background: Cockroach is the second most common aeroallergen in all over the world. However, there were few studies about the correlation of socioeconomic status or environmental factors and cockroach sensitization in Southeast Asia. The purpose of our study was to determine the socioeconomic and environmental factors affecting cockroach sensitization in atopic children. Methods: One hundred and twenty children aged 3Y15 years attending the allergy clinic, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, with symptoms of asthma, allergic rhinitis or atopic dermatitis, and with skin test reactivity to at least 1 allergen were enrolled. Questionnaires and skin prick testing results were analysed. Results: There were 80 boys and 40 girls (average age 9). Eighty one percent of patients had allergic rhinitis, 23% had asthma, and 9% had atopic dermatis. Cockroach sensitization was found in 76% of patients, of which 22%, 12%, 42% were sensitized to German cockroach, American cockroach, and both, respectively. There was no statistical correlation between socioeconomic status or environmental factors, including household income, home's character, home environment, amount of cockroaches seen in home, and cockroach sensitization. Conclusion: There was high incidence of cockroach sensitization in atopic children. However, socioeconomic status, amount of cockroaches seen in home, and other environmental factors were not correlated with degree of sensitization. Asthma severity is influenced by indoor dust mites but not endotoxin or nitrogen dioxide exposure in Hong Kong children Background: Endotoxin exposure has dual effects on protecting against wheezing disorders in early life but worsening control in Caucasian asthmatics. Exposure to house dust mites (HDM) and nitrogen dioxide (NO 2 ) were also known risk factors for severe asthma in this population. However, such data is unclear in Asians. This study investigates indoor exposure to endotoxin, HDM and NO 2 in Hong Kong asthmatic children, and its relation to their disease severity. Methods: Asthmatics aged 6 to 18 years old were eligible, and spirometry and exhaled nitric oxide (eNO) were measured in the clinic. Home visits were done within 10 days. Dust was collected from patients' mattress, and carers completed ISAAC written questionnaire. House dust Der p 1 and endotoxin levels were measured by enzyme-linked immunosorbent and chromogenic limulus amoebocyte lysate assays, respectively. Average NO 2 levels in kitchens were measured over 48-hour periods by Ogawa sampler badge. Results: 115 asthmatic patients, aged 12.4 T 3.2 years, were recruited. Their median FEV 1 and eNO were 92% predicted and 50.5 ppb. Fifty-eight percent of these patients wheezed in past 12 months, and 44% of asthmatics received inhaled steroid. During home visits, 39.2 (24.6/65.9) mg mattress dust was collected. The median (IQR) mattress Der p 1 and endotoxin levels were 0.61 (0.24/2.46) 2g/g and 12.4 (6.4/19.5) EU/mg, respectively. A trend was observed for the correlation between mattress Der p 1 and endotoxin load (> = <0.176, P = 0.060), but neither factor was associated with indoor NO 2 (P 9 0.15). Der p 1 levels = 2 2g/g was found in 29% of families. Kitchen NO 2 levels were 27.1 T 10.4 ppb, and which exceeded 21 ppb in 78% of families. Indoor Der p 1 levels were significantly associated with night awakening (P = 0.049) and eNO (P = 0.010), whereas mattress endotoxin load was higher in patients with low peak expiratory flow rate (P = 0.031). No association was found between indoor NO 2 and any clinical or spirometric parameter. Conclusion: Our results suggest that asthma severity in Chinese children is affected by current exposure to HDM allergen in mattress but not indoor endotoxin or NO 2 exposure. Effect of indoor mold concentrations on daily symptom severity of children with asthma and/or rhinitis monosensitized to molds Little is known about the contribution of indoor molds to the symptoms of asthma and/or rhinitis in children monosensitized to molds. We aimed to investigate the effect of indoor mold spore concentrations on daily symptoms of asthma and/or rhinitis in children monosensitized to molds. Nine-teen children with asthma and/or rhinitis sensitized only to molds recorded their daily symptoms and PEF values to the diaries, from February 2005 to January 2006. In this study period, indoor mold concentrations were measured monthly from the living/bedrooms. The median indoor mold concentration was 37.5 CFU/m3. Most commonly recovered indoor molds were Cladosporium (26.4 %), Penicillium (24.7%) and Aspergillus (7%). There was not found significant correlation between indoor mold concentrations and daily rhinitis score (r = <0.021, P = .932), daily asthma score (r = 0.155, P = .554), daily morning PEF (r = <0.056, P= .475) and evening PEF (r = <0.057, P= .471). The effect of indoor molds is not evident on the symptoms of our patients with asthma and/or rhinitis monosensitized to molds. Influence of air quality in schools upon the children with chronic bronchopulmonary diseases Rodica Selevestru, Svetlana Sciuca, Grigore Friptuleac, and Cazacu Angela. State of Medical Univercity, Pediatry, Chisinau, Moldova, Republic of. Aim: To emphasize the influence of air quality in study rooms of schools in children with chronic bronchopulmonary diseases. Methods: The study planned to interview 1384 of pupils of I-IV class from different schools, for selection of children with chronic respiratory diseases. The lot of study included 54 of children with chronic bronchopulmonary diseases. The clinic examination of these children emphasized 18 cases with bronchial asthma, 9 cases with obstructive chronic bronchitis and in 27 cases simple chronic bronchitis. The hygienic condition of occupation conditions in study rooms of pupils was carried out by means of Air Quality Monitor (AQ-5000, USA), that determines the values of microclimate factors (temperature, air humidity, CO2 concentration). Results: Hygienic evaluation of occupation conditions in study rooms detected a subnormal thermal regime 15,54 T 0,1-C, at the beginning of first lesson 16,8 T 0,2-C and at the end of last lesson (optimal t +20 +25-C). The values of air humidity in study rooms were increasing from 51,11 T 1,8% at the beginning of first lesson to 56,71 T 2,3% at the end of last lesson (optimal humidity 30/60%). More significant are the modifications of CO2 in the air, that varies during the day 0,07 T 0, 009% at the beginning of first lesson and 0, 21 T 0,02% (p G 0,001) at the end of last lesson (optimal concentration of CO2 G 0,1%. The respiratory functional condition of children with chronic and recurrent respiratory diseases certifies the presence of obstructive disturbances (PEF j 72,3 T 1,7%; FVC/FEV1 j82,6 T 2,3%; FEF25j75 j91,7 T 2,1% and minimal restrictive changes FVC Y 76,5 T 2,1% ; FEV1 j82,6 T 1,9%. Conclusion: The reduced temperatures, increased humidity and excessive concentrations of CO2 in child microambience lead to installation and perpetuation of respiratory symptoms in children with bronchial asthma and chronic bronchitis. Background: The prevalence of bronchial asthma has increased worldwide over the last twenty years. This tendency is particularly valid for the adolescent population. The aim of the presented study is to evaluate the prevalence of adolescent smoking patterns, parental smoke, exposure to environmental tobacco smoke and the asthma-like symptoms in their relation with active and passive smoking among teenage school-children. Methods: It is done a cross-sectional questionnaire study (a modified version of Compendium of European Respiratory Standard Questionnaires-CERSQ) with 304 students, aged 15Y16 from three secondary schools in Sofia-Bulgaria inquired. Results: Active smokers are 85 students (28,7%), 45% of them smoke regularly. The average age for starting smoking is 13,2 years and average number of smoked cigarettes per day is 7,4. Regular exposure to tobacco smoke is reported by 201 students (66,3%) and the exposition is between less than 5 years and 15 years. Respiratory symptoms (during the last 12 months): wheezing and whistling in the chest are reported by 22,3% of the students; with in addition shortness of breath in 38.2% of them. Night cough is reported by 39,1% and asthma attack by 2,6% of the examined students. Antiasthma drugs have been taken by 1.6%. The statistical analysis of the questionnaires showed a significant association between Bwheezing and whistling in the chest [ and Bactive smoking[ (p G 0, 01) ; between Bnight cough[ and Bactive smoking[ (p G 0,01); Bnight cough[ and Bactive smoking[ (p G 0,05); Bnight cough [ and Bpassive smoking[ (p G 0, 01) . Conclusion: The founded associations support the idea that smoking (both active and passive) plays a role in the initiation, development or maintenance of asthma-like symptoms. The intervention strategies for smoking control should include both public policies elements and preventive measures implemented by health professionals, in particular the general practitioners and school health staff. Rajan Bhandari 1 and Shambhu D. Joshi 2 . 1 Community Health and Environmental Society Nepal, Community Health, Kathmandu, Nepal; 2 Nepal Medical College Teaching Hospital, Medical Sciences, Kathmandu, Nepal. Background: Developing country like Nepal has poor health status in school students. Objective: To know and evaluate the indoor environmental condition of government and private-schools and the health impacts on students. Methods: A cross sectional studied of representative samples of 35 schools of selected region of Nepal including government and private schools. Onsite observation and health check up & interview with students and teachers were done. Specific scores was given in each criteria. The data were analysed and edited in EPI info program. Results: The results shows that 89% of government school and 45% of privateschools have poor environmental condition and 69% of government schools students are suffering from environmental-health problem while only 22% of private school student are suffering from some kind of diseases especially allergic. Government-school doesn_t have the standard classroom, adequate sports facility, safe drinking water, light and ventilation in comparison with private schools. Conclusion: We conclude that the main causes are poor socio-economic status, illiteracy of parents, negligence, hard housework for children, diseases, malnutrition, incomplete immunisation and lack of health education. The poor environment condition includes crowded students in a classroom, poor ventilation, shortage of clean drinking water, untidy clothes of students, poor nutrition and lack of greenery in the school area, school near by road, air pollution and lack of environmental awareness among teachers and parents. The government schools have limited budget, resources with compared to private schools and most of the lower and lower middle class family children are studying in government schools which covers nation 82% of total students. Recommendation: The government should allocate the special budget to the government schools and it should be utilised from the available resources such as good ventilation, limited student in a class, awareness among teachers and parents. Last but not the least, this type of programs are helpful to prevent from environmental health hazards also. Air-conditioning bedroom and allergic sensitization in atopic patients Tachakwan Intapuntee, Panithi Boondumnern, Pantipa Chatchatee, and Jarungchit Ngamphaiboon. King Chulalongkorn Memorial Hospital, Pediatrics, Bangkok, Thailand. Background: One of the recommendation of environmental control for allergic prevention was air-conditioning bedroom. The purpose of our study was to determine the relationship between air-conditioning bedroom and allergic sensitization. Methods: Patients with asthma and/or allergic rhinoconjunctivitis attended pediatric allergy clinic, King Chulalongkorn Memorial Hospital age 3/25 years old and had positive prick skin test for aeroallergen at least 1 antigen were studied by questionnaire. Results: A total of 180 patients were included, 116 (64.4%) males, 64 (35.6 %) females, 96 (53.3%) lived in households with air-conditioning bedroom. American cockroach sensitization was significantly more prevalent in patients lived households without air-conditioning bedroom (58 %) compared with the other who lived in air-conditioning bedroom (44 %) (p = 0.048, Odds ratio 1.726). In contrast, Alternaria sensitization was significantly less prevalent in households without air-conditioning bedroom (3.4 %) compared with air-conditioning bedroom (15.6 %) (p = 0.006, Odds ratio 0.2). Polysensitization (skin prick test positive at least 3 antigen) was no significantly related among households without and with air-conditioning bedroom (p 9 0.05). Cockroach is the second most common aeroallergen sensitization from mite but there was no correlation between amount of cockroach seen in the house and skin test results. There was no correlation between cat and dog exposure or cigarette smoking and allergic sensitization. Conclusion: Patients living in households without air-conditioning were at increased risk of cockroach sensitization but decreased risk of in mold sensitization. Efficiency of neem tree extract (Milbiol) in decreasing the level of house dust mite allergens Tiia Voor, Ylle Kuld, Tiina Rebane, and Kaja Julge. Tartu University Hospital, Childreńs Clinic, Tartu, Estonia. Background: Sensitization against house dust mite (HDM) allergens is common in Estonia and the level of D. pteronyssinus (Der p1) and D. farinae (Der f1) is high in the houses. Diminishing contact with HDM reduces allergy symptoms and therefore it is important to decrease the level of HDM allergens. Milbiol (Hexal AG, Holzkirchen, Germany ) is an oil extract produced from neem tree (margosa) seeds. Milbiol is harmless for humans and animals, although it hinders growth and reproduction of mites. Aim: To estimate the efficiency of Milbiol in decreasing the level of HDM allergens. Methods: Study period was Dec 2003 -Dec 2004. Study group comprised 44 beds from 19 homes. Twenty-five beds were vacuum cleaned and treated with Milbiol (intervention group) and 19 beds were only vacuum cleaned (control group). Frequency of cleaning was identical and it was performed at the same time for beds of both groups. The dust samples were collected on the 1st, 3rd and 12th month and treatment with Milbiol was performed on the 1st and 2nd month according to the manufactory instructions. The levels of Der p1 and Der f1 in the dust samples were measured by ELISA method. Data about the living conditions were also collected. Results: In all beds the level of Der f1 (8614 T 2654 ng/g) was higher than the level of Der p1 (520 T 260 ng/g, p G 0.001). HDM allergens were more common in old brick houses, on lower floors, in rooms with carpets and in beds older than 5 years. Before and after treatment with Milbiol the mean level of Der f1 was 4180 T 1502 ng/g and 2784 T 868 ng/g, respectively; p 9 0.05, however, after one year the level increased up to 6032 T 2092 ng/g. The level of Der p1 increased after treatment with Milbiol from 556 T 353 ng/g up to 784 T 408 ng/g; p = 0.005; although after 12 months the level was lower than before treatment (282 T 163 ng/g). The mean level of HDM allergen decreased also in the control group beds due to vacuum cleaning -Der p1 from 471 T 394 to 177 T 121 ng/g and Der f1 from 14447 T 5628 to 8733 T 3700 ng/g, p G 0.05; respectively. Conclusion: Milbiol alone was not effective for decreasing HDM allergens in beds during one-year period after treatment; however, more frequent treatment with Milbiol could give better results. Lower level of HDM allergens was gained also by regular vacuum cleaning. The analysis of Alpha1-Antitrypsin serum level among people exposed to industrial dust Background: In healthy persons, alpha1 antiprotease serves as a protective screen that prevents alveolar wall destruction. Long-term industrial pollutants exposure may lead to imbalance in the oxidant-antioxidant or proteasesantiproteases systems in the pulmonary tissue witch is followed by unimpeded neutrophil elastase digestion of elastin and collagen in the alveolar walls and progressive emphysema. Methods: We were interested in comparing the level Alpha1-antitrypsin (AAT) between patients with occupational pulmonary pathology, healthy men, working under industrial pollutants exposure more than 15 years and healthy donors. The subjects in the study were 84 men between the ages of 45 and 60. The main group of study included patients with chronic occupational bronchitis (n = 25), electric welder pneumoconiosis (n = 10), silicoses (n = 29); the first (1) control group included 10 patients without occupational lung disorders and the second (2) control group -10 healthy donors. We measured the level of AAT by immunoturbidimetric assay for quantitative in vitro determination of AAT in human serum on automated clinical analyzes. We used reagents produced by BRoche-Diagnostics[. Normal level 0,9/2,0 g/l. Results: We found that the level of AAT was under normal among patients with electric welder pneumoconiosis (0,78 T 0,23). It was significantly lower (p G 0,05) than the level in both control groups (group 1/1,23 T 0,16; group 2 Y 1,26 T 0,16). We did not find significant differences of serum concentration of AAT between silicoses (1,09T0,37), chronic occupational bronchitis (1,15 T 0,27) and control groups. It was even normal. It was a tendency of AAT concentration increasing due to the term of pollutants exposure. Conclusion: Long-term industrial pollutants exposure may lead to chronic airway inflammation and caused the imbalance in proteases-antiproteases systems. There was an activation of alpha-1-antitrypsin secretion as the most important proteinase inhibitor among men working under industrial pollutants exposure. The level of alpha-1-antitrypsin was normal among patients with chronic bronchitis, silicoses and healthy workers. We found that AAT level could be decreased among patients with electric welder pneumoconiosis. We supposed that it could be the result of oxidative inhibition of the welding aerosol components on AAT secretion. Environmental dust granulometry and microbial load in the presence of tobacco smoke Gianni Macrina 1 , Michele Baroffio 1 , Barbara Bruno 2 , Carla Tazzer 2 , Paolo Crimi 2 , and Giorgio Walter Canonica 1 . 1 University of Genoa, Department of Internal Medicine, Genoa, Italy; 2 University of Genoa, Department of Health Sciences, Genoa, Italy. Background: Several studies describe damage from passive smoking in humans and animal models. However, how different chemical and physical components relate to mechanisms of damage is not completely understood. Cigarette smoke is composed by a gas phase and a particulate phase, in which a lot of toxicologically relevant compounds are present. We focused our investigations on the particulate phase of environmental cigarette smoke, as the amount and dimensions of suspended particles have been only partially studied so far. It is noteworthy that the size of particles is very important in the inhalation process and in their subsequent deposition into the respiratory tract. Methods: Differences in granulometry of suspended dust and bacterial load were studied in a 120-cubic meter no-smoking room during and after one cigarette smoke in ten different experiments. To evaluate granulometry in suspended dust we used a 6 channel particle counter, which analyzes particles according to their granulometry. We evaluated suspended bacterial load by using a multipierced sampler on a fixed replaceable plate set to aspirate air. Evaluation of sedimented bacterial load was performed by using the Air Microbial Index technique which expresses the total number of microrganisms settled by gravity onto a Petri plate. Results: An increase of medium and small, but not large suspended particles number was observed. Bacterial and mycotic load decreased during and up to 60 min after cigarette smoking. Gram-positive cocci never decreased while chromogen-bacteria and mycetes decreases at 30 min. Furthermore, we observed an increase of sedimented bacteria and mycetes at 60 min, a decrease at 120 min and a stabilization at 180 min. Conclusion: We hypothesize that tobacco burning accelerates sedimentation of larger agglomerates of mean-size smoke particles and bacteria which may be resuspended by movements of people in the room and consequently be inhaled. They could have a stronger affinity with airways surfaces, modifying the normal mechanisms of non-specific and immunologic defenses against microbes, above all in children. This could partially explain the increasing of respiratory infections in children exposed to second-hand smoke. Proteomic analysis of birch and ragweed pollen artificially exposed to NO 2 Roberta Aina 1 , Elisa Berra 1 , Giada Marino 1 , Elide A. Pastorello 2 , and Sandra Citterio 1 . 1 University of Milano Bicocca, Dept. of Environmental Science, Milan, Italy; 2 Niguarda Cà Granda Hospital, Unit of Allergology and Clinical Immunology, Milan, Italy. Background: Air pollutants seems to play a fundamental role in promoting allergies and in worsening the health conditions of allergic people. New interesting findings suggest that traffic-related pollutants do not only irritate the airways, but can also make allergens more aggressive, especially pollen allergens. In particular, air pollutants can induce and/or increase the expression of some allergenic proteins. They can also induce post-translational modifications (PTMs). These changes can make proteins more reactive towards the IgE. Our objective has been to investigate the effect of nitrogen dioxide (NO 2 ) on pollen from birch (Betula pendula Roth) and ragweed (Ambrosia artemisifolia L.), two very common and important sources of allergens in Europe. Methods: Pollen samples were artificially exposed to different concentrations of NO 2 . Sera of birch and/or ragweed allergic patients were also collected. Pollen proteins were extracted from control (C) and treated (T) samples, separated by 2D electrophoresis, blotted onto nitrocellulose membranes and incubated: (a) with patient's sera to identify pollen allergens; (b) with an anti-nitrotyrosine antibody to estimate the entity of protein nitration; (c) with a staining solution specific for glycosylated proteins. Finally, a treatment with lambda protein phosphatase (ëPPase) were performed to identify changes in phosphorylation pattern. Results: NO 2 treatments did not induce new allergen expression, neither in birch, nor in ragweed pollen. Regarding PTMs, the treatments induced modifications in both birch and ragweed phosphorylation patterns. On the contrary, no changes in glycosylation profiles were detected in the analysed plants. Finally, few nitrated proteins were induced in birch treated samples. Conclusion: Exposure of pollen to NO 2 induced PTMs in allergens. However, future experiments are needed to confirm these first results. Vipul Shah. Allergy Clinic, Allergy, Surat, India. Introduction: A new pollen allergen was suspected in patients who while receiving allergen immunotherapy showed recurrence of symptoms during the period Jan-March. Methods: Fifteen patients having various allergic disorders were subjected to skin allergy testing, were found to be sensitive to various allergens and they were on immunotherapy. While on IT they developed recurrent symptoms during Jan-March. To confirm presence of the new allergen, safranin stained glycerin coated slides were exposed in the patients_ surroundings with Bpersonal volumetric air sampler[. With this Bsampler[ pollen collection on the slide is much better then simple exposure. Mangifera pollen was recorded in abundance. Antigen of mangifera pollen was tested on all patients and strongly positive reaction was recorded. These patients were subjected to allergen immunotherapy course with mangifera pollen allergen. Results: Patients who showed positive results with mango pollen allergen and then underwent immunotherapy course with this pollen antigens showed significant improvement in their symptoms during the next pollen season of mangifera indica. Conclusion: 1) Mangifera Indica pollen could play an important role in nasobronchial allergy. 2) BAir sampler[ makes the detection of causative allergen possible & easier when sometimes simple slide exposure may fail. Atsushi Fukushima 1 , Tin-Tin Win-Shwe 1 , Shoji Yamamoto 1 , Naoki Kunugita 2 , Yasuhiro Yoshida 3 , Keiichi Arashidani 2 , and Hidekazu Fujimaki 1 . 1 National Institute for Environmental Studies, Research Center for Environmental Risk, Tsukuba, Japan; 2 University of Occupational and Environmental Health, Japan, Health Sciences, Fukuoka, Japan; 3 University of Occupational and Environmental Health, Japan, Medicine, Fukuoka, Japan. Background: Since outdoor and indoor air pollutants are environmental risk factors that have been shown to contribute to the development of respiratory infections and allergy, it is speculated that there is a positive relationship between allergic inflammation and inhalation of volatile organic compounds. Effect of low-level toluene inhalation on Th1/Th2 balance in allergic response remains unknown. Methods: To study the effect of exposure to low-level toluene on transcription factors for T-cell differentiation, male C3H/HeN and BALB/c mice were exposed to filtered air (control), 5, 50 and 500ppm toluene by whole body inhalation for 6 hours per day, 5 days per week, for 3 and 6 weeks. The allergic model group of mice was injected intraperitoneally with 10 2g ovalbumin (OVA) plus 2 mg alum, and each of these mice was then challenged with nebulized OVA as a booster once every 2 weeks (weeks 2, and 4) during the exposure period. Then, by using a quantitative real-time PCR method, we investigated the expression of GATA3, T-bet and FoxP3 genes in spleen. Results: Exposure to 500ppm toluene for 3 weeks significantly increased the expression of GATA3 and FoxP3 mRNAs in spleen of C3H mice. In C3H mice exposed toluene for 6 weeks, the expression of GATA3, T-bet and FoxP3 mRNAs was increased at 50ppm toluene as compared to that of filtered air control. However, we did not observe any such pattern for the expression of T-bet and FoxP3 mRNAs in spleen of BALB/c mice. The expression of GATA3 mRNA was significantly suppressed by exposure to 500ppm toluene in BALB/c mice. Conclusion: This is the first study to show in vivo modulation of the expression of transcription factor genes in the different strains of mice by exposure to low-level toluene, in concert with antigenic stimulation. Background: Patients with asthma is prone to experience recurrent exacerbation of symptoms especially when they are exposed to environmental triggers. But it is not well defined what kind of clinical characteristics is associated with this predisposition. Methods: The Cohort for Reality and Evolution of Adult Asthma in Korea (COREA) study is a prospective, observational multi-center study and will be conducted for nine years (2005/2013) in Korea. Cross sectional data of baseline clinical characteristics were compared between asthmatics susceptible to asthma exacerbation by environmental triggers and asthmatics tolerable to those triggers based on self reported questionnaire at the time of enrollment. Results: Common cold (22.9%) was most frequent trigger followed by polluted or dusty air (16.6%), second hand smoke (11.5%), emotional stress (9.5%), animal (3.8%), and drug (1.0%) in descending order. The environmental triggers-sensitive asthmatics (TSA) showed female dominance (60.2% vs. 50.5%, P = 0.027) and higher prevalence of rhinosinusitis (63.2% vs. 53.6%, P = 0.041) than environmental triggers-tolerable asthmatics (TTA), while there was no significant difference in obesity, duration of symptom, atopic status, and lung function. More patients among TSA reported aggravation of asthmatic symptoms after moving to current house (16.1% vs. 3.9%, P = 0.001) and TSA had been living in the current house for significantly less time than TTA (P = 0.002). Conclusion: Female sex, having rhinosinusitis, moving into a new house was significantly associated with the susceptibility to asthma exacerbation by environmental triggers. A survey of the children often exposed to Passive Smoking (PS) had relatively higher risk of developing obstructive pulmonary & allergic diathesis M. Ishaq, I. Khan Sameera, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Introduction: Is to pursue the deleterious effects of (PS) on the health of the exposed children. Methods: In a retrospective study undertaken from1995/2004, including 600 nonsmoking asthmatic children ages 9/15years (mean age 12 years) with history of house hold smoking as passive smokers (PS). They were likely found to have 60/76% increased tendency of allergic & obstructive pulmonary manifestations (eczema, allergic rhinitis, ocular allergies, middle ear suffusion & shortness of breath) than those residing in a nonsmoking household. Boys were more sensitive to (PS) than girls, and likewise manifestations increased with age and duration of exposure. Children of smoking parents had an increased prevalence of respiratory symptoms (cough, sputum, wheezing) & lower pulmonary function tests (PFTs) values than children of non smoking parents, the association of changes in (PFTs) may persist into adulthood. The significance of the maternal smoking was marked as adding to the population of childhood asthma. The later on follow up has evidenced that passive smoking in early childhood increased the risk of lower respiratory tract infection by 1.5 to 2.2 folds. In the same study/follow up children with congenital pulmonary malformations & family history of allergies were likely to be more sensitive to & exacerbated by (PS) than non smokers. On further follow up into adult life (PS) were. showing relatively increased prevalence of recurrent infection & less productivity of life. Results: Underestimation of the significance of (PS) in a house was followed by the grave health related hazards, some of which were irreversible. Conclusion: The impact of Smoking in a community has variable degree of adverse reactions with paramount morbidity index on terminal ages. Marginlization of smoking to restricted places is followed by a positive socio economic impact. The significance of the graded improvement in pulmonary function & physical wellbeing amongst college students with smoking cessation M. Ishaq, I. Khan Sameera, and Munir Imran Khan. Al-Junaid Hopsital, Allergy/Pulmonology, Nowshera, Pakistan. Purpose: Significant change in respiratory function/physical well being were documented with stepwise cessation of smoking that had socio-economical & prospective impact upon the college students. Methods: In the study College students ages 17/21 years both sex having had 1/4 years history of smoking, had manifestation like hacking cough, breathlessness & wheezing more so at night. Some had raised temperature & weight loss altered taste etc. Smoking cessation followed under supervision from weeks to months with advise & encouragement. The outcomes had been as under. Results: Assessment of the pulmonary function Physical wellbeing, Conclusion: Immediately after quitting, ex-smokers had been advised breathing exercises with steep line increase in tempo. Improvement in pulmonary function/physical well being had been inversely proportional to the period & history of smoking. Clinical Implication: Cessation advises followed under supervision with incentives for smoker students in the form of credits in study, awards & certificates for quitters of the week, months etc. khan et al aljh nsr nwfp pk. 627 Ocular, nasal and skin signs of aircrews in relation to before and after the ban on smoking on flights -a follow up study Torsten Lindgren, Dan Norbäck, and Gunilla Wieslander. Uppsala University Hospital, Dep of Occupational and Environmental Medicine, Uppsala, Sweden. Background: The main aim of this 3-years follow-up was to gain further insight into ocular nasal and skin symptoms, and to determine whether the incidence was related to personal, psychosocial, and occupational factors. Methods: To evaluate the differences of ocular nasal and skin symptoms, a self-administered questionnaire was mailed, in 1997 (n = 1857). Three years later, in 2000 (n = 1446), the questionnaire was sent again to airline crew on duty. The participation rate was 81% 1997 (n = 1513) and 79% 2000 (n = 1145). A cohort of 1134 subjects is used in this study. At the time of the questionnaire study 1997, smoking was allowed on intercontinental flights. Results: Atopy showed a significant reduction, and hay fever showed a significant increase. During both 1997 and 2000 ocular nasal and skin symptoms were more common in the cabin crew than in the pilots. The incidence of ocular nasal and skin symptoms was more common in cabin crew compared to pilots. The new-onset nasal and skin symptoms were more common in cabin crew compared to pilots. Younger age, work satisfaction, and stress due to excess of work were significantly more related to new-onset ocular nasal and skin symptoms. Cabin crew had significantly more new-onset of nasal symptoms compared to the pilots. Airline crew with hay fewer reported significantly more complaints on new-onset ocular and nasal symptoms. Airline crew with doctor's diagnosed asthma reported significantly more complaints on newonset nasal symptoms. Conclusion: Age, hay fever, asthma, psychosocial working environment and type of occupation are related to new-onset of symptoms. Atopic diseases, allergic sensitisation and exposure to traffic-related air pollution in children Background: In vitro studies, animal experiments and human exposure studies have shown increased risks for atopic outcomes. However, results derived from observational studies are inconsistent. To assess the relationship between individual based exposure to traffic-related air pollutants and allergic outcomes in a prospective birth cohort study during the first six years of life. Methods: We followed-up 2860 children at the age of 4 years and 3061 at the age of 6 years to investigate atopic diseases and allergic sensitisation. Longterm exposure to particulate matter (PM2.5), PM2.5 absorbance and nitrogen dioxide (NO2) was assessed at residential addresses using geographic information systems based regression models and air pollution measurements. The distance to the nearest main road was used as a surrogate for traffic-related air pollutants. Results: Strong positive associations were found between distances to nearest main roads and asthmatic bronchitis, hay fever, eczema, and sensitisation. A distance-dependent relationship could be identified with the highest Odds Ratios for children living less than 50 m from busy streets. For PM2.5 absorbance statistically significant effects were found for asthmatic bronchitis, hay fever and for allergic sensitisation to inhalant allergens due to pollen sensitisation. NO2 exposure was associated with eczema, while no association was found for allergic sensitisation. Conclusion: There is a strong evidence for an increased risk of atopic diseases and allergic sensitisation when children are exposed to particulate matter. Air cleaners or polyurethane bedcovers for asthmatic children with cat or dog allergy Background: A comparative multidisciplinary and inter-regional study on allergy/asthma and indoor exposures could give important information leading to an increased understanding of the aetiology of this morbidity. However, the existing national studies followed different methodologies, which hampered the use of the obtained data for potential comparison. Such drawback can be overcome by following identical protocol with regard to questionnaires, inspections and measurements. Objective: The present paper proposes a methodology for performing crosssectional and case-control studies on the impact of indoor home environment on allergy and asthma among children based on the recently performed ALLHOME (Bulgaria) and BDampness in Buildings and Health[ (DBH, Sweden) studies. The proposed approach includes a strategy for performing an epidemiological study, selection of target population, identification of the sample size. All key steps in performing the cross-sectional and case-control studies are listed. Suggestions for improving the quality of the obtained data are given. The DBH and ALLHOME studies identified cultural behaviour as an important factor when performing epidemiological studies. Information gathered from the descriptive cross-sectional study could provide clues leading to the formulation of epidemiologic hypotheses, which could be tested in the analytic nested case-control study. A target group of pre-school children was proposed, since children at that age spend most of their time at home. A crosssectional questionnaire, developed for the DBH study and containing 87 questions on demographic data, health symptoms, housing characteristics and lifestyle factors was proposed. The use of parental questionnaires for information on housing characteristics could be an effective tool in conducting studies on housing and health. The proposed nested case-control study includes dwelling inspections of a non-destructive type (building checklist, sensory and visual evaluation of dampness, dust sampling for HDM, moulds, pollens, phthalates), engineering measurements (air temperature, humidity, CO2 level) and medical examinations (general examination and skin prick test). Special attention should be paid on storage of collected data, statistical analyzes and dissemination of the research results. Conclusion: The paper can be used as a guideline for performing studies on indoor environment and allergy/asthma. Multiplex technologies -expanding horizons for assessment of allergic sensitization and exposure Background: Current methods used for the assessment of allergic sensitization and allergen exposure require separate testing for each allergen-specific antibody or allergen. We have applied fluorescent suspension array technology to allow the simultaneous detection of total and allergen-specific IgE in serum or multiple allergens in environmental samples in a single quantitative test. Methods: Fluorescent multiplex arrays were developed to allow the simultaneous detection of indoor allergens in environmental samples or total and allergen-specific IgE in serum. A 9-plex suspension array measuring Der p 1, Der f 1, Der p 2, Der f 2, Fel d 1, Can f 1, Mus m 1, Rat n 1 and Bla g 2 was developed using monoclonal antibodies covalently coupled to fluorescent microspheres. A 10-plex array for the detection of total IgE and IgE specific to Der p 1, Der p 2, Fel d 1, Can f 1, Bet v 1, Phl p 5, Bla g 1, Bla g 2 and Asp f 1 was created using monoclonal antibodies and purified allergens covalently coupled to fluorescent microspheres. The multiplex arrays were validated by comparing results between the multiplex methods and established enzyme immunoassays. Results: There were highly significant correlations between results obtained by the multiplex methods and established enzyme immunoassays with p G 0.001 (n = 970). In addition, the sensitivity, limit of detection, reproducibility, intra assay coefficient of variance (CV), and inter assay CV of the fluorescent multiplex array was shown to be equal to or better than the ELISA methods. Conclusion: Our results suggest that fluorescent multiplex technology will facilitate epidemiological studies of exposure and allergic sensitization such as birth cohort studies, population surveys and studies of gene-environment interaction. The arrays can, in principle, be expanded to include other allergens and allergen-specific antibodies. Reduction in indoor aeroallergen by using an xtreme 3000 air purifier The quality of the environment within buildings is a topic of major importance for public health. Presently Indoor Air Quality (IAQ) is a major concern at various work places. Here is an important quote on air quality from a journal: BWith all the publicity, more and more people are realizing that pollutants in the indoor air could make them sick. The worst thing that has happened to the indoor air quality marketplace in the last year or so is also mold. This is because much of the media coverage is designed to sensationalize the topic and frighten the public -so much so, that the word Fmold' always seems to be preceded by the adjective Ftoxic._ Thus, homeowners and building managers are scared to death of any minor infestation that might possibly be toxic mold, and they often ignore other health issues, such as combustion byproducts, Volatile Organic Compounds, second-hand tobacco smoke and poor ventilation.[ This study was aimed at examining the efficiency the air purifier 3000 Xtreme in reducing the indoor spore and pollen counts of allergenic taxa present in different office facilities and residential buildings. Sets of the Petri plates and coated slides were placed with distances of 1foot, 2 feet, 4 feet, 6 feet and 12 feet away from the air purifier. Assay was done with the Petri plates prepared from Brain Heart Infusion agar (VWR) and Gelvatol coated slides with air purifier off for the control and on for the treated. The control Petri plates and slides were placed in the room without using the air purification system. The treated sets were assayed after 24 hour, 48 hour and 72 hour treatment. The slides were examined, analyzed, and photographed using a BX-40 Olympus microscope attached to a DP-70 Digital Camera. Petri plates were examined with an SZ-40 stereo-scope to observe and count the colonies. The control set of plates showed vigorous growth of the microbial colonies after incubation in an incubator at 37-C for 24 hours. Petri-plates closer to air purifier (2 ft. and 4 ft.) produced least number of colonies after 24 h., 48h. and 72 hours of treatment of the indoor air with the air purifier. There was very minor trace of inoculums from the Petri-plates from 2 ft. 4ft. and 8 ft. after 24 hours. After a 2-month continuous exposure of the room air with the Xtreme 3000 at BHigh[ setting we found complete eradication of aeroallergens. Background: Wide variations in the prevalence of asthma, rhinitis and eczema have been reported between differents countries. The ISAAC (International Study of Asthma and Allergies in Childhood) was thought in order to maximize the value of epidemiological research into asthma, allergic rhinitis and atopic eczema in childhood, stablishing a standarized methodology and facilitating collaboration and comparison between severe countries. Methods: A standard questionnaires was distributed through schools and selfcompleted by 13/14 years old children and by the parents of 6/7 years old children. Results: 1675 children aged 6/7 years ansewered the questionnaire and 3082 in the group aged 13/14 years. In the group of children age 6/7 years, the prevalence of wheezing in the last year was 8.4% and 5.6% in the 13/14 years group. The prevalence of rhinitis in the last year was 55.5% in the 6/7 years group and 31.4% in the 13/14 years group. The prevalence of eczema in the last year was 9.5% in the 6/7 years group and 6.2% in the 13/14 years group. Conclusion: Prevalence of allergic disease was similar to reported in other sdudies, except for rhinitis with hight prevalence rates with respect to other studies performed worlwide. Alarming results of the comparison of the ISAAC Study in 6/7 and 13/14 years old children in Islamabad, Pakistan Background: Allergic diseases and asthma are registering an alarming increase all over Pakistan, especially in children. Pakistan has a population of over 160 million, in which almost 40% are children below the age of 15 years. The ISAAC Study conducted in schools in 1997 and in 2001/2 is the only reliable documented study looking at the prevalence of these illnesses in this country. A comparison of the two study groups has been evaluated to assess the rising danger of respiratory allergy and asthma, in Islamabad, the capital of Pakistan. Methods: The standard Questionnaire based ISAAC protocol was used for this study. Schools were randomly selected for the study, and all children falling within the study age group were included in the study. Children of age 13/14 years answered the questionnaires themselves, while they were answered by parents of the 6/7 year age group. Results: Total Number of Children Studied. Conclusion: The ISAAC study conducted in Islamabad has revealed that the prevalence of nasal problems and wheezing is more than double in the 13/14 year age group, as compared to the younger 6/7 year children. The very large number of 13/14 year olds reporting nasal problems is also very alarming, as there is a 3 fold more danger of these children developing asthma later in life. The study also highlights the need for better diagnosis of asthma in both age groups, as only 1 in every 4 wheezing children have been diagnosed to be asthmatic. The results indicate the need for increased awareness amongst the general public and the physicians regarding allergic diseases and asthma. Jeong-Whun Kim, Chul Hee Lee, and Jeong-Hun Jang. Seoul National University College of Medicine, Otorhinolaryngology, Seongnam, Republic of Korea. Background: ARIA guidelines are in the way of general criterion in classifying allergic rhinitis. Objective: The objective of study is research on Korean patients who are underwent allergic rhinitis according to their symptoms, severity, frequency and distribution patterns. Materials and Methods: Statistical analysis was done in 610 patients who had been diagnosed as allergic rhinitis at Seoul National University Bundang Hospital and 540 patients at local clinics. All patients were categorized into 4 groups, mild intermittent, mild persistent, moderate to severe intermittent and moderate to severe persistent groups according to their questionnaire research and allergy laboratory results. Results: The incidence of moderate to severe persistent group was 34.7 % and moderate to severe intermittent group was 17.1%. In the allergy laboratory results, there was a significant difference between groups in eosinophi count (P = 0.004). In questionnaire research, there were significant differences at each group concerning olfaction (P G 0.001), self-awareness of rhinitis (P = .013), previous history of rhinitis (P G 0.001), self-awareness of asthma (P = 0.001) and conjunctivitis (P G 0.001). There were significant differences in frequency of symptoms for allergic rhinitis and severity between groups (P G 0.001) and moderate to severe persistent group showed highest score. Conclusion: According to ARIA guidelines in categorizing patients, moderate to severe persistent group stands out as the biggest part. Consequently differentiated therapeutic approach between groups will be useful. Associations between ocular and nasal symptoms, atopy, personality traits and stress Roma Runeson 1 and Dan Norbäck 2 . 1 Akademiska sjukhuset, Occupational and Environmental Medicine, Uppsala, Sweden; 2 Akademiska Sjukhuset, Occupational and Environmental Medicine, Uppsala, Sweden. : The aim was to study associations between ocular and nasal symptoms, atopy, personality characteristics measured by Antonovsky_s Sense of Coherence (SOC), Karolinska Scales of Personality (KSP), and stress measured through projective drawing. Totally 194 subjects initially working in 19 buildings with suspected indoor air problems were studied. Information on a history of atopy, hay fever, ocular and nasal symptoms was gathered by a postal questionnaire, including KSP, SOC and projective drawings of Ba person in the rain[. Results on SOC: There were no associations between SOC and history of atopy or hay fever, however those with low SOC had more ocular (p = 0.005) and nasal symptoms (p = 0.04). On KSP: Those with atopy had higher social desirability (p = 0.022), lower irritability (p = 0.016), and lower guilt (p = 0.047). Those with hay fever had lower guilt (p = 0.045). Those with eye symptoms had lower social desirability (p = 0.018), and higher indirect aggression (p = 0.025). Those with nasal symptoms had higher: somatic anxiety (p = 0.002), muscular tension (p = 0.007), and psychic anxiety (p = 0.019). On projective drawings: Those with atopy draw clouds (p = 0.023) and long rain strokes (0.039) more often and short rain strokes (p = 0.005) less often. Those with hay fever drew water puddles (p = 0.009) more often. Drawing of long rain strokes was associated with ocular symptoms (p = 0.005). Drawings of strong wind and rain was associated with nasal symptoms (p = 0.045). Conclusion: History of atopy is associated with personality traits measured by KSP and projective drawings. Ocular and nasal symptoms during non-pollen season are associated with SOC, as well as KSP and projective drawings. Different scales were associated with atopy and symptoms. Long rain strokes were consistently associated with both atopy and ocular symptoms. Sensitization profile analysis in children with either asthma or eczema alone using a panel of mite allergens from Dermatophagoides pteronyssinus and Blomia tropicalis Run Mei Angeline Chong 1 , Nge Cheong 2 , Su Min Audrey Teo 3 , Fong Cheng Yi 2 , Pei-Chi Lynette Shek 2 , Yoke Chin Giam 4 , Kaw Yan Chua 2 , and van Bever Hugo 2 . 1 Nanyang Technological University, School of Biological Sciences, Singapore, Singapore; 2 National University of Singapore, Paediatrics, Singapore, Singapore; 3 A*STAR,, Genome Institute of Singapore, Singapore, Singapore; 4 National Skin Centre, Dermatology, Singapore, Singapore. Background and Aim: The prevalence of multi-allergen sensitization to 2 mite species in patients with asthma from Singapore and Malaysia has been reported earlier. It is important to discern whether a specific disease symptom of allergy is correlated with different mite species. The present study is a comparison of the sensitization profiles in children suffering from either asthma or eczema alone using a panel of mite allergens from Dermatophagoides pteronyssinus (Der p) and Blomia tropicalis(Blo t). Methods: A total of 60 children (age between 4 to 16 years old), 30 with asthma and 30 with eczema respectively were recruited in Singapore for the present study. The IgE levels specific to a panel of house dust mite-allergens from two mite species were measured using ELISA. The panel of allergens used is Der p extract, Der p 1, Der p 2 and Der p 5 from Der p mites and Blo t extract, Blo t 4, Blo t 5, Blo t 11 and Blo t 12 from Blo t mites. Results: Children with asthma and rhimitis exhibited the highest sensitization to Blo t 5 (77%) with relatively lower sensitization frequency to Der p 1 ( 50%) and Der p 2 ( 67%) while children with eczema showed high sensitization to Blo t 5 (73%), Der p 1 (50%) and Der p 2 (70%). Generally, more eczema patients were sensitized to this panel of Der p allergens with much higher measured IgE titres as compared to the asthma group. Conclusion: These two patient groups tend to show differential sensitization profiles to Der p and Blo t allergens. Patients with asthma and rhinitis show high sensitization to Blo t mite allergens, especially Blo t 5 whereas eczema patients showed high sensitization to Blo t 5, Der p 1 and Der p 2. This suggests the possible correlation between the manifestation of different disease symptoms of allergy and the mite species. Sensitization and clinical relevance of ragweed pollen in patients with intermittent rhinoconjunctivitis Results: The total number of patients with ocular allergy pathology: 41% belongs 9 40 year old, 39% between the 16/39 years old and 20% under 15 years old; 61% females and 39% males. As for the allergy sensitivity (+ ) for d.pte and d.fa (86%), alternaria and or cladosporium (60%), gramineae (37%), trees (33%), cat (33%) and dog (12%). 45% of patients presented elevated IgE in tear with normal serum IgE and 34% presented both high. The purpose of consultation and/or associated pathology was perennial allergic rhinitis (85%), bronchial asthma (23%), eczemas (14%) and pure allergic conjunctivitis (5%). Conclusion: We observed through our work that the majority of patients consulted our centre for other allergic entities which aroused the suspicion of this ocular affection by means of a complete interrogation and ophthalmology examination and then confirming with the laboratory and/or SPT. We therefore consider that it is a sub diagnosed entities as it is not the most common consulting, but must be suspected by the Allergist in other allergy pathologies and by the trained ophthalmologist by managing it with discipline to improve the patients quality of life and to avoid after effects. The Results: Average total incidence during the investigated period was 160,9. The highest peak was found in the age group of 3-6 years (361,8), the second one was in the age group of 6-14 years (296, 9) . Adolescents (14-18 y.o) and young adults had nearly the same incidences (average: 289 Comparative studies are necessary to determine/analyse regional distribution patterns worldwide. The prevalence of allergic diseases in rural and urban regions of Poland -preliminary report Methods: The project consisted of two stages: survey using a questionnaire and diagnostic tests in outpatient clinics. The questionnaire study (adapted ISAAC and ECRHS II) was carried out in a randomly selected groups of children, adolescents and adults (age groups: 6-7, 13-14 and 20-44 ). One third of the subjects underwent medical examination directed towards allergy recognition. The study was carried out in two regions: in Warsaw -the capital city of Poland and the largest urban agglomeration, with high density of population and dense traffic and in the rural region of southeastern Poland -two districts in the province of Lublin (the districts of Zamosc and Krasnystaw), both characterized by a very low level of environmental pollution, low density of population and natural farming methods. Results: In 2006 (June -October) 3833 questionnaires were completed. In the rural area the number of answered questionnaires reached the level of 2237 (1105 children and 1132 adults) while in the urban area -1596 (398 children in age group 13-14, 378 children in the age group 6-7 and 811 adults). Bronchial asthma was declared by 2,6% of all respondents from rural area and by twice as many (6%) from urban area. 14,5% of respondents in Zamojszczyzna and 24,8% in Warsaw declared themselves to suffer from allergic rhinitis. The question concerning skin allergy was answered positively by 23% of respondents from rural region and by twice as many of Warsaw inhabitants. Table presents questionnaire-based results in relation to age and place of residence. Conclusion: Preliminary questionnaire-based results may indicate higher prevalence of allergic diseases in Warsaw residents in comparison with residents of rural area of the southeastern Poland. Initial analysis of data from medical examination does not confirm this finding, therefore further analysis is essential. 629 subjects living in rural area and 474 residents of Warsaw underwent verifying medical examination. Bronchial asthma was diagnosed in 5,1% of examined subjects from rural region and in 3,2% from Warsaw. 14,8% of examined rural area inhabitants and 16,5% of Warsaw inhabitants appeared to suffer from intermittent allergic rhinitis, while persistent allergic rhinitis was diagnosed in 12,7% and 6,7%, respectively. Allergy for cat and sources of the allergens in the urban versus rural areas of Poland Liliana Trzpil, and Boleslaw Samolinski. Medical University of Warsaw, Prevention of the Environmental Hazards, Warsaw, Poland. Background: The aim is the comparison of a frequency of positive skin prick test for cat allergen, and the frequency of animal presence at home in various areas of Poland (rural versus urban). The presented data are a part of the earliest outcome of large epidemiological study in the field of allergies which is now run in Poland. Methods: Randomized probe of various polish populations differing in the age range (20Y44 13Y14, 6-7 years old) were included in to the study. Analyzed urban population was gathered in Warsaw (capital of Poland). The second selected region was Zamojszczyzna, being the typical rural area with low industrial density. 3833 respondents completed questionnaires ECRHS II and ISAAC (1596 at urban and 2237 at rural region). 1103 of them were subjected to medical examination directed towards allergy recognition, and had made a skin prick test (474 at the urban and 629 at rural region). Results: 65,5% of persons from urban area revealed the positive skin results (for at least one allergen), while of the rural one the prevalence was significantly lower and equal to 34,5% (p = 0.0001). An frequency of positive cat prick test is significantly higher for urban region when compared to the rural region (18,4% vs. 10.8) , and the intensity of the reaction measured as the average diameter of the observed skin prick test bubble is also significantly higher (4,01 vs. 3,63 mm) . It was found that a significant difference between urban and rural areas was observed for the cat frequency of presence at home 15% vs. 30%. This data can be also compared with the general frequency of the cat presence in the home and surrounding. For rural areas cat is present in about 59.8% of homes and surroundings, while in urban areas cat is present in 15.4% of homes and surroundings. About 50% of people with positive skin tests for cat have cat at home. In urban area these values are significantly lower and equal to 19.5%. Conclusion: For cat allergies one can observe that frequency of positive skin prick test is higher on the urban region than on the rural region, contrastively to the lower presence of the animal allergy sources. Additionally the frequency of cat presence in homes of sensitive persons is much higher in rural areas than in urban ones. This observation can lead to the conclusion of protective, immunotherapeutic role of the high environmental allergen concentration in rural region. Background: Allergy is influenced in clinical characteristics by genetic and environmental factors. We assessed the clinical characteristics of new-onset respiratory allergy in immigrants in Brescia, Italy. Methods: We collected the data of all the immigrants referred to our allergy service within the last two years, focusing those patients without family or clinical history of allergic diseases. Results: Two hundred and 39 patients were evaluated (108 male, mean age 35 years). Their countries of origin were uniformy distributed among the 4 macroareas (Pacific southwest, Africa, South America, East Europe). None of them had previous symptoms of allergy and their family history was negative. The mean time of onset of symptoms after immigration to Italy was 5.21 years, and the first clinical presentation was rhinitis+asthma in 68%, whereas the remaining had rhinitis only. The percentage of monosensitized subjects was 25% and 20% had also sensitization to cockroaches. Only 16% and 6% had a concomitant positivity for foods or drugs respectively. Conclusion: In immigrants to Italy the new-onset respiratory allergy displays a) long latence, b) preferential onset with rhinitis and asthma associated, c) poor occurrence of sensitization to non inhalant allergens, d) high prevalence of cockroach sensitization. Keywords: immigrants, respiratory allergy. Roos Bernsen 1 , Nico Nagelkerke 1 , Carel Thijs 2 , and Johannes van der Wouden 3 . 1 United Arab Emirates University, Community Medicine, Al Ain, United Arab Emirates; 2 Maastricht University, Epidemiology, Maastricht, Netherlands; 3 Erasmus MC-University Medical Center Rotterdam, General Practice, Rotterdam, Netherlands. Background: Pertussis infection has been suspected to be a potential causal factor in the development of atopic disease because of the effect of pertussis immunization on specific IgE antibodies. Although several studies found a positive association between pertussis infection and atopic disorders, this relationship has not yet been studied in a population stratified by vaccination status. Objective of the present study was to assess the association between pertussis infection and atopic disorders in pertussis-unvaccinated children and in pertussis-vaccinated children. Methods: Using data from a previously conducted study on the relationship between the diphtheria-tetanus-pertussis-(inactivated) poliomyelitis vaccination in the first year of life and atopic disorders, the study population of 1872 8Y12 year old was divided into children pertussis-unvaccinated and children pertussis-vaccinated in the first year of life. Within each group the association between pertussis infection and atopic disorders (both as reported by the parents) was assessed. Results: In the unvaccinated group there were no significant associations between pertussis infection and atopic disorders. In the vaccinated group all associations between pertussis infection and atopic disorders were positive, the associations with asthma (odds ratio (OR) = 2.24, 95% confidence interval (CI95%): 1.36Y3.70), hay fever (OR = 2.35, CI95%: 1.46Y3.77), and food allergy (OR = 2.68, CI95%: 1.48Y4.85) being significant. Conclusion: There was a positive association between pertussis infection and atopic disorders in the pertussis vaccinated group only. From the present study it can not be concluded whether this association is causal or due to reverse causation. Background: Allergic diseases are fairly common in all parts of the world and involve all ethnic groups with bronchial asthma, allergic rhinitis, conjunctivitis and eczema being the commonest manifestations. Despite our efforts, their incidence is on an escalating path. There appear to be no authentic data on the prevalence of allergic diseases in children in Pakistan, therefore the need was felt to carry out the prevalence studies in the country. Methods: To estimate the prevalence of allergic diseases in our country, this first ever study was carried out in the largest coastal city of Karachi which houses people from all ethnic groups and all socio-economic classes. As there were no statistics available for any age group in this regard, a Saudi Arabian Protocol using questionnaires for school children were used instead of ISAAC Protocol. Questionnaires were distributed to 1200 school children, age 6-15 years, in three different areas for proper proportional representation of high, middle, and low socioeconomic levels. A total of 700 (58.3%) questionnaires returned by the parents were found valid and thus included in the survey. The data was analyzed by using SPSS software. Results: The result shows that the prevalence of allergic diseases is quite high in Pakistan. The frequency of wheezing was found to be 15.2%, while the diagnosed cases of asthma were 9.5%. The frequency of allergic rhinitis was found to be 34.3%. The frequency of those having allergic rhinitis as well as wheezing episodes was 8%. There was no statistically significant difference between asthmatics and non-asthmatics by sex (P-value:0.402). Socioeconomic status was found to affect significantly (p value 0.001) as the prevalence of diagnosed asthma cases was 6.17% in high socioeconomic class, 13.11% in the middle-class and 2.4% in the low socioeconomic class. Family history of atopy was also found to be significantly higher in asthmatics. Conclusion: Allergic diseases are quite common in Pakistan but they remain under-diagnosed. The prevalence data of allergic rhinitis shows an alarming situation. These diseases were found to affect middle-class the most, while the lower and lower-middle class remains the least affected. In addition to diagnosis and management, prevention and education may help the country to fight back against the disease. Our study is in progress to increase the sample size and compare with the data from other cities being collected under the same protocol. Allergic sensitization among Surabaya suburb school children and undergraduate students Objective: To identify major allergic sensitization among school children and undergraduate students in Surabaya, a cross-sectional epidemiologic study was done. Participants were recruited by multistage simple random sampling to represent students of Surabaya_s suburb areas. Methods: Only 499 out of 550 invited students were agreeing to be involved in this study in voluntary basis. They were 128 students of primary schools, 221 of secondary schools, and 150 of a university. Complete personal and family history of allergy, allergic symptoms or diseases, environmental or home exposure of various common allergens were obtained, physical examination and skin prick tests were done using a set of 36 different inhalant and ingested allergen extracts. Total serum IgE and specific IgE RAST for selected allergens were also measured. Results: There was a trend of increasing skin prick test (SPT) positivity among study subjects, being 21.90% among primary school children, to 29.05% among secondary school children, and to 45.30% among undergraduate students. Cockroach (42.85%) and mould (42.85%) were the most common allergens in primary school children, followed by house dust mites (28.57%), grass pollen (17.86%), and crab (14.29%). The order of most common allergens for secondary school children were: house dust mites (63.16%), cockroach (37.85%), mould (28.04%), grass pollen (23.38%), and crab (22.77%). The order for undergraduate students were: house dust mites (58.82%), cockroach (47.05%), crab (41.17%), mould (36.76%), and grass pollen (20.59%). There were some multiple SPT positivity among them, toward 2 or 3 allergens in general. Urticaria and rhinitis were the commonest clinical manifestation of allergy. History of atopy in the family was positive for only 36.7% of respondents. Conclusion: The prevalence of allergic sensitization among school children and undergraduate students in Surabaya suburb areas were increased, being above the previous estimates for Indonesia in 2003. House dust mites and cockroach were the most common allergen; while urticaria and rhinitis were the commonest allergic manifestation. Keywords: allergy, sensitization, schoolchildren, skin prick test, epidemiologic study Marianne Empson, and Julie Smith. Auckland City Hospital, Clinical Immunology, Auckland, New Zealand. Background: A high prevalence of atopic disease and asthma has been documented in NZ based primarily on studies in children. No prevalence data is available for anaphylaxis although anecdotally the incidence appears to be high. This study obtains baseline data for anaphylaxis rates in a well-defined adult population in Auckland, NZ. Methods: A retrospective audit of adults admitted to Auckland City Hospital (ACH) was performed. Patient charts for the audit were identified using a broad range of diagnostic codes that may have been used in cases of anaphylaxis. Four major diagnostic categories were included in the chart retrieval: allergy/allergic reaction, anaphylactic shock or reaction, angioedema/urticaria, and insect sting. Two time periods were selected for the audit, Files were reviewed and data extracted almost exclusively by one author. A cohort of patients that satisfied the Olmstead County definition of anaphylaxis was identified for each time period. Where the diagnosis of anaphylaxis was uncertain the case was excluded. Interval A included community-onset anaphylaxis only while B included all cases of anaphylaxis. Detailed data on the population served by ACH is available for both time intervals. This was used to determine anaphylaxis rates in adults (older than 14 years). National population data will be used to determine agestandardised rates for New Zealand. Results: 367 and 664 admissions were reviewed for intervals A and B respectively. 129 episodes of community-onset anaphylaxis were identified in 121 individuals in A and 113 episodes in 111 individuals in B. The number of individuals per 100,000 adults suffering at least one episode of anaphylaxis in the community per year was 41 (95% CI 34 Y 48) in A and 34 (95% CI 28 Y 40) in B. An additional 37 episodes of anaphylaxis occurring in hospital were identified in B. The total rate of anaphylaxis in all settings in B is therefore 46 (95% CI 39-53) per 100,000 adults per year. Conclusion: The rate of anaphylaxis in NZ adults is one of the highest in reported literature. However this is still an underestimate as it does not include cases managed solely in the community or those misclassified as asthma. In addition in this retrospective study some cases of anaphylaxis may have been excluded due to uncertainty. Despite this it provides us with an important assessment of the potential size of the problem for further studies. Knowledge and concerns of treatment and care among Thai allergy patients in Rayong Province, Thailand Patient education and health promotion are important to prevent and decrease morbidity and mortality in allergic diseases. Despite improvement of health care system, improper knowledge, attitude and incorrect beliefs on allergy have been worried. To evaluate the knowledge and concerns of treatment and care of allergy would give the information for health care personnel and physicians to develop a good campaign on prevention and care among allergy patients in the future. Self-assessment by questionnaire on knowledge and opinions about allergy and allergist was performed among 125 patients in Rayong Province, which is one of leading centers for tourist and machinery industry. 44.8% know that allergy is an important global health problem and 52.8% also think that it is not severe and life threatening. 39.2% agree to be treated only to relieve symptoms not for continuing treatment and 59.2% need continuing therapy. So 56.8% used the over-thecounter medicine from pharmacy and 4.8% had never used any drug to relieve their allergic symptoms. 80% prefer oral drugs to inhaled drugs but 85% of them will follow the physician advice of using inhaled drug. 53.6% still believe that allergy is permanently cured and about one-forth to third of the patients think that allergic symptoms can be alleviated by nutrient (24%) supplements and air cleaners (34.4%). Though 69.6% know the existence of allergy specialist but few (9.6%) have visited the special clinic and only 7 patients (5.6%) were tested and known their own specific allergens by allergen skin test. However, 89.6% indicated that allergen avoidance is a key role of treatment and 84% know that house dust mite is the most common allergen in Thai population. Only one-fifth (21.6%) satisfy an updated information of allergy and all of them give their concerns on insufficiency of the information provided to their community. Allergy and migraine -a significant comorbidity M.V. Francis. Eye and Migraine Centre, Ophthalmology, Cherthala, India. Background: Molecular mechanisms of peripheral sensitisation of nociceptors could be identical for painful and pruritic (allergic) diseases. Therefore therapeutic approaches for pain treatment may also be beneficial for allergy. Montelukast, cyproheptadine and ketorolac tromethamine are found to be effective in migraine and allergy. This study is to document the association between migraine and allergic disorders. Methods: Study spanning 5 years. 10240 migraine patients were questioned about various allergic manifestations. Results: 7995 (78%) were suffering from various allergic manifestations either at the time of consultation or sometime in the past. Nasal allergy was the commonest 6395 (80%), ocular allergy in 4797 (60%), bronchial allergyin 3198 (40%), and skin allergy in 640 (8%). 1439 were suffering from all 3 allergies(naso oculo bronchial). Conclusion: Allergic disorders increased the risk for migraine and migraine increased the risk for allergy. This bidirectional association immensely helps the clinician to explain the causative molecular mechanisms and the genetic origin of these two conditions in the most simple way to their patients. House dust are considered to be a source of potent allergen responsible for different allergic manifestations. A total of 54 species of mites belonging to 33 genera, 18 families under 3 orders have been recorded from house dust samples collected from asthmatic patients residing in and arround rural areas. All the allergenic mites viz. Dermatophagoides pteronyssinus, D. farinae, Austroglycyphagus sp., Blomia tropicalis., Acarus siro, Glycyphagus domesticus, G.destructor, Tyrophagus putriscentiae except Euroglyphus sp. have been recorded. Among them Dermatophagoides alone contituted more than 60% of the total mites isolated. Patients bed contained significantly higher mite population than the control subjects bed dust (pG0.02) and among two different habitats examined i.e. bed and corresponding bed room floor dust, the former contained higher ( p G 0.01) mite density than the later. Analysis of skin prick test against dust related allergens revealed that 82.4% of the patients showed marked preference towards Dermatophagoides mite allergens. Estimation of total serum IgE level indicates that patients sera contained significantly higher level than the control sera (p G 0.001). RAST result further confirms the higher degree of sensitivity towards house dust mites (84.76%). The clinical history of the patients, the physical appearance of allergenic mites in higher concentration in the patients bed, positive skin test results with mite extract and identification of mite specific IgE antibodies in patients' sera by RAST technique established the role of dust mites on the aetiopathogenesis of bronchial asthma in developing country. Background: There was a strongly controversy between the protective or inductor effects of parasites infections and the risk of development allergic diseases in several countries. Moreover, It had been proposed that shared antigens between helminthes parasites and environmental allergens could modulated allergic immune responses. However, evidences for this process has not been consistently establish. Methods: The aim of this study was to evaluate the allergic reactivity and helminthes parasites coexistence in a group of unselected 61 scholar Children, reflecting a population with a profound socioeconomic depression, a high rate of common childhood infectious diseases such as diarrheas (8344 x 100.00 inhabitants), acute respiratory infections (1528 x 100.00 inhabitants) and high prevalence of helminthic infections (53%). Additionally by ELISA, we detected the levels of th1 (INF+), and th2 (IL-4) cytokines as well as the levels of total and specific IgE against Dermatophagoides pteronissynus, Blomia tropicalis and Ascaris Lumbricoides. We performed an inhibition specific IgE test between mites and Ascaris lunbricoides in order to establish the cross reactivity between these allergens. We also performed clinical evaluation, hematological studies and skin prick test with common food, mites and parasites allergens. Results: Sixty seven percent of children (group 1) showed respiratory symptoms or skin manifestations. Fifty % had high leukocyte count and had high INF+ levels (4190-5320 UI/ml) than control group. Skin prick tests were positive in 43% of children to at least one of the common allergens evaluated: 77% were positive to Ascaris lumbricoides, 27% to Blo t and 27 % to D.pt. Fifty % had high total IgE levels, (3812 -910000UI). Eighty one percent showed levels of specific IgE higher than 17.5 UI/ml to Ascaris and 67% to D.pt. Before the inhibition test the median levels of specific IgE to Ascaris diminished from 14.17UI/ml to 3,43UI/ml % and to D.pt from 12,94 to 3,37 UI/ml. These results indicates that high allergic reactivity, (evidenced by clinical, skin prick test and specific IgE), and high levels of Th1 cytokines were both been present in sera of children evaluated. We also found that there were an inhibition in the level of specific IgE in the sera of these children, suggesting a cross reactivity between mite and Ascaris allergens. The approach to improvement diagnosis of lambliasis in patients with atopic bronchial asthma Dmitry Dolbin, Yuriy Tyurin, and Sergey Kulikov. Kazan Scientific Research Institute of Epidemiology and Microbiology, Rospotrebnadzor, Kazan, Russian Federation. Backround: Since 1859 when has described first time G. lamblia as the activator of disease of a gastroenteric path, the huge material of epidemiology and pathogenesis of lambliasis and as effective enough ways of therapy of the given disease have been developing and collecting. But also at the moment, concerning this disease, there are the numerous questions expecting the decision. Most important of this group of problems, intensively discussed problem of diagnostics and prevalence G. lamblia among people is. Methods: We use commercial ELISE-test and the immunochromatography methods revealing specific antibodies in whey of blood and antigen_s in excrements. They possess more sensitivity (66,3 Y 98,9 %) and high specificity (92,6 %) unlike routine microscopic methods in diagnosis of lambliasis. In connection with before be the aforesaid, till now the most widely applied test in laboratory diagnostics of lambliasis, such as a microscopy of sedimentary components of excrements or researching contents at duodenal sounding for the purpose of revealing cysts of G. lamblia. In the literature as the method enrichment formalin-radio is recommended. Unfortunately, the specified tests of laboratory diagnosis of lambliasis possess low sensitivity, subject to influence of some subjective and objective factors among which, such as changeable allocation cysts of G. lamblia, difficulty for microscopic identification. Results: We used the combined method of research of excrements This method allowed to improve essentially delectability cysts G. lamblia in comparison with a routine method microscopy of fecal. We will especially notice, that at comparative research of tests in which activators lambliasis have been revealed and using three components a method and microscopy research of fecal in the first case the big degree invasion came to light. It has paramount value at definition of necessity of carrying out of therapy lambliasis. Conclusion: As using combined method using three components for flotation of research of excrements, we managed to increase essentially delectability G. lamblia and as to reduce quantity of repeated researches necessary for the exact diagnosis of lambliasis. Accuracy of definition of degree invasion an organism this elementary has besides, increased, that has allowed to correct therapy. Titanium used for fabrication of obturators after maxillectomy -an alternative therapeutic option in cases of incompatibility of polymer based denture materials Rita Depprich 1 , Eva Kusch 1 , Alexandra Meissner 2 , and Guido Meissner 2 . 1 Heinrich-Heine-University Duesseldorf, Department of Cranio-and Maxillofacial Surgery, Duesseldorf, Germany; 2 Private Clinic, Cranio-and Maxillofacial Surgery, Wuppertal, Germany. Background: Microbiological colonization and subsequent biofilm formation on denture materials play an important role in the development of denture stomatitis often triggering allergic incompatibilty of denture materials as the penetrability of the mucosa is changed. Despite the high standards in reconstructive surgery and even with the use of free vascularized flaps postmaxillectomy rehabilitation with obturator prosthesis is still the mainstay reconstructive procedure in several cases. With current use polymer based silcone-or methacrylate-obturators bear the risk of an increasing settlement and penetration of bacteria and thus the risk of local or even systemic infections. Titanium-obturators [TO] for oral rehabilitation offer favorable characteristics concerning growth of microbial biofilms. The aim of our study was to compare the prevalence of microbial contamination on titanium-and polymer based -obturators [PO] used for post-maxillectomy rehabilitation. Materials and Methods: We investigated the obturators of 36 patients (17 TO, 19 PO), all had undergone maxillectomy due to malignancy in the period 1999-2204. Microbiological samples of the obturator surfaces, the adjacent mucous surfaces and of the interior of the PO (when new fabrication was planned) were collected with a sterile swab. Microbiological analysis was performed by conventional methods, including microscopy and culture on different agars. For statistical data analysis Fisher_s exact test [FT] (P-value P e .05) and odds ratio [OR] (95% confidence interval) were used. Results: PO showed a significantly higher quantitative contamination and a shift to non physiological bacteria compared to TO (p = 4.944e-06 [FT], 6.84 [OR] ). There is a high statistical risk (factor 1.4) for penetration of bacteria to the interior of PO when the pathological bacteria are seeded on the surface of PO (p = 0.01966). Conclusion: The use of titanium-obturators for the oral rehabilitaton of post-maxillectomy patients significantly reduces the risk of contamination with pathological oral bacteria and therefore prevents patients from local oral or systemic infections. Especially in cases of incompatibility of polymer based denture materials titanium-obturators offer an alternative therapeutic option. Wirach Maek-a-nantawat 1 , Sarawan Maythaarttaphong 2 , Rawiwan Suphan 2 , Hathairat Chueathai 2 , Urairat Phantong 2 , Doungngoen Khaminthakul 2 , Kanokon Boontanom 2 , Suranya Sarapat 2 , Sukanya Tomkum 2 , and Udomsak Silachamroon 1 . 1 Mahidol University, Allergy Clinic, Clinical Tropical Medicine, Bangkok, Thailand; 2 Faculty of Tropical medicine, Mahidol University, Vaccine Trial Center, Bangkok, Thailand. Allergy has been estimated about 10Y20% of Thai population. Most studies conducted in Bangkok and other regional centers showed a increase trend of allergy prevalence in a recent decade. This study is aimed to determine the prevalence of allergic diseases in an urban area, Rayong province, which has high growth of industry and oil/gas refineries. 524 Rayong residents, with mean age (T SD) of 26.26 (T 3.77) years old and male 59. 4% , were asked about their history of allergy regarding rhinitis, asthma, conjunctivitis, dermatitis including urticaria, food allergy and drug allergy. Prevalence of overall allergy is 15.6 %. Persistent rhinitis, asthma and dermatitis are 12.3 %, 3% and 2.3%, respectively. About urticaria, 9.5% reported history of acute (78%) and chronic type (22%). 4.4% revealed history of food allergy of which the most common culprit was sea-food allergy (73.9%). 2.7% showed history of insect sting allergy in which 46.2% of them had large local reaction and 3 of 13 had anaphylactic reaction. Drug allergy is 2.3% with history of allergy to betalactams (25%), NSAIDS (25%) and paracetamol (16.7%). Family history of atopy is significantly associated with history of rhinitis (P G 0.0001), asthma (P G 0.0001), urticaria (P = 0.001), conjunctivitis (P = 0.029) and food allergy (p G 0.0001). There is no association between history of allergy and sex, age or occupation. There is also a close association among chronic rhinitis, asthma and food allergy (P G 0.0001). In conclusion, allergy is not uncommon even in an urban area like an eastern part of Thailand. History of atopy in family is useful to be a predisposing factor of allergic diseases. Effect of prenatal and postnatal exposure to low-level toluene and peptidoglycan on systemic Th1/Th2 immunity in infant mice Shoji Yamamoto 1 , Atsushi Fukushima 1 , Tin Tin Win Shwe 1 , Rieko Hojo 1 , Yasuhiro Hojo 2 , Naoki Kunugita 3 , Keiichi Arashidani 3 , and Hidekazu Fujimaki 1 . 1 National Institute for Environmental Studies, Research Center for Environmental Risk, Tsukuba, Japan; 2 University of Occupational and Environmental Health, Japan, Medicine, Fukuoka, Japan; 3 University of Occupational and Environmental Health, Japan, Health Science, Fukuoka, Japan. Background: Toluene, a widely used aromatic organic solvent, has been well characterized as a environmental toxic chemical. Volatile organic compounds including toluene are the possible substances causing chemical sensitivity in human. Our laboratory has previously showed that the exposure of low-level toluene causes immunological and neurological modulation in adult mice. Recently, it has been suggest that TLR2/4 ligand such as peptidoglycan (PGN) or LPS modulates Th2-driven allergic immune responses in animal studies. The aim of the present study was to investigate the effect of low-level toluene exposure and PGN stimulation on systemic Th1/Th2 immunity in infant mice. Methods: We exposed pregnant BALB/c mice to filtered air (0 ppm), 5 ppm, and 50 ppm toluene for 6 h/day on 5 days/week for 1 week (from day 14 of gestation until delivery) in the whole body exposure chamber. The offspring were further exposed to toluene of same schedule for 3 weeks (from the day of birth up to day 21). Some of the pregnant mice were repeatedly exposed to aerosolized PGN 2002g/10 ml (3 times per week) during the toluene exposure period. Offspring from the toluene and PGN exposed pregnant mice were further repeatedly administered PGN 1002g (5 times per 2 weeks) intraperitoneally from day 7 up to day 21 during the toluene exposure period. One day following the last toluene exposure, we collected blood, spleen, and lung from each mouse. Then, total IgE, IgG1, and IgG2a antibodies in plasma and the production of cytokines (IL-12, IFN-+, IL-4, IL-5) in spleen and lung were assayed by ELISA method. Also, the expression of cytokine and transcription factor T-bet mRNAs in spleen was analyzed by real-time RT-PCR method. Results: Exposure of mice to low-level (5 or 50 ppm) toluene resulted in (1) increased IgG1 and decreased IgG2a antibodies in the plasma; (2) decreased T-bet mRNA significantly in spleen; and (3) decreased IFN-+ and IL-12 mRNA in spleen but not statistically significant. Next, exposure of mice to low-level toluene by combination with PGN stimulation compared to toluene alone resulted in decreased IgG1 and IgG2a antibodies in the plasma and IL-12 mRNA in spleen. Conclusion: These findings suggest that low-level toluene exposure from fetal stage to neonatal stage may enhance the systemic Th2 function in infant mice. However, the exposure to combination of low-level toluene with PGN stimulation shifted Th1/Th2 immunity to suppressive direction. Herberto Jose Chong Neto and Nelson Rosario. University of Parana, Pediatrics, Curitiba, Brazil. Background: Although recurrent wheezing is frequent, there are few data regarding risk factors associated with this condition. The aim of study is to verify the risk factors for recurrent wheezing in infants, in the south of Brazil. Methods: Cross-sectional study using a standardized and validated questionnaire with 45 questions about recurrent wheezing and familiar, enviromental, socio-economic and respiratory infections risk factors in the first twelve months of life, from International Study of Wheezing in Infants (EISL, from spanish: Estudio Internacional de Sibilancias en Lactantes). Parents of infants that attended the Health Centers between August/2005 and December/ 2006 for regular immunization were interviewed. Among 107 Immnunization Centers 35 were randomly selected in order to maintain a homogeneous selection of population sample. Factors associated to recurrent wheezing in a univariate analysis were studied using multivariate regression of Poisson. Prevalence ratio and confidence intervals of 95% were calculated due to a high prevalence of wheezing infants and statistic was considered significant when p G 0.05. Results: Three thousand and three infants, aged 12-15 months-old were studied. The prevalence of recurrent wheezing was 45.4%. There are associations between recurrent wheezing and male gender, mother, father and siblings asthma history, motheŕs instruction (secondary and advanced), others pets in domicile during pregnancy (birds), 6 colds episodes or more, personal history of dermatitis and molds in home. Updated immunization was a protection factor for recurrent wheezing. Conclusion: Prognostic risk factors for recurrent wheezing in the first year of life were male, familiar history of asthma, high motheŕs instructions, other pets during pregnancy and molds in home, a high number of colds episodes, and personal history of dermatitis. Interventions acting on these factors may decrease the occurence of recurrent wheezing in infants in Curitiba, Brazil. Monitoring of cytokines in breast milk from allergic and non allergic mothers: prospective birth cohort study Galli Elena 1 , Palmesi Francesca 2 , Nisini Rita 2 , Agostino Rocco 2 , Carpino Valentina 1 , Gabriele Ida 1 , Mora Nadia G.M. 1 , Brunetti Ercole 1 , and Mancino Giorgio 1 . 1 San Pietro Hospital -Fatebenefratelli, AFaR Research Centre, Roma, Italy; 2 San Giovanni Calibita Hospital -Fatebenefratelli, Neonatology Unit, Roma, Italy. Background: It is well documented that human milk contains several immunomodulatory components which are important during the newborn_s immune system development. In this context, breastfeeding could provide the best possible nutrition. Its role in the development of allergies remains although controversial, due to possible individual differences and variations in the composition of the breast milk. Furthermore, other stimuli could interact with immune system during perinatal-neonatal period, interfering with breast feeding and hence with the immune system development. For these reasons, we are interested to investigate the different components (environmental, diet, familiarity etc.) that could be involved in the allergic diseases apperarence. Aim: The aim of this prospective birth cohort study is to investigate the concentration of cytokines involved in allergic reactions, IgA antibody production, soluble (s) CD14 and fatty acids in cord blood, colostrum and mature milk, from allergic and non allergic mothers. Moreover, the aim is to investigate whether breastfeeding practice, correlated with immunological profile, is determinant for the increased risk of atopic disease development in children. 32 mothers (17 allergics, 15controls) and their full term newborns were included in the study, until now. We measured cytokines in colostrum samples (2 days after birth), and in transitional breast milk samples ( Results: Different cytokines levels in colostrum and milk samples, were evidenced between allergic and non allergic mothers. In particular, our preliminary data showed that colostrum from allergic mothers has a significant increased level of IL-4, IL-13, IL-5, TGF beta 1 e 2 compared to samples from non allergic mother, while no differences in cytokines level were revealed in the milk samples from the same two groups. Follow-up is in progress, first round of clinical evaluation 6 months after birth, does not reveal yet any allergic symptoms in all the children. Kawasaki disease and allergic disease -a sibling control study 60 years old) . The adult-sample was considered representative since it consisted of all staff of the Greek Air Forces who were examined (after written consent was obtain from all) during their obligatory annual medical check up. A mean wheal diameter of 3mm or more, in the absence of any reaction in the negative control SPT, was considered as a positive reaction. Multiple logistic regression was performed to reveal any statistical significant association between type of respiratory allergy (AR and/or asthma) and all known parameters (gender, age, type of sensitization, tsIgE levels). Thereafter the diagnostic properties of tsIgE and type of sensitization in discriminating between rhinitis and asthma were analyzed using the Receiver Operator Characteristic (ROC Results: 22 patients reported severe allergic rhinitis and 6 with severe ocular itching precipitating headpain attacks resembling migraine 24 of them were also getting migraine attacks when exposed to common migraine triggers in this region. These common triggers are exposure to sunlight, travelling by bus, sleep disturbances and missing meals at the right time. Travelling by bus triggered allergic symptoms in 21 of these patients most probably due to exposure to dust and smoke. Conclusion: Molecules involved in allergic inflammatory processes definitely trigger migraine episodes by exciting peripheral nociceptors. Familial allergy in Surabaya's suburb schoolchildren Results: The maximum sensitivity was recorded for D. farinae (94%), grass pollen (76.55%) and Fusarium spore (53.92%) among the three groups of allergens tested. The prevalence of at least one positive prick tests was 37.12%. A large difference by age was found: 58.9% were sensitized in the age group of 20Y29 years, 42.8% in the age group of 40Y49 years and 28% in the age group of 50Y60 years (p G 0.001). Sensitization to multiple allergens was common in the age group of 20Y29 years with 42.1% of the sensitized responding to at least four allergens, while this portion was only 12.8% of the sensitized among those aged 50Y59 years. The prevalence of physician-diagnosed asthma, allergic rhinitis (AR), conjunctivitis and wheeze increased significantly with increasing number of positive skin responses. Having a family history of AR or conjunctivitis was a significant risk factor for allergic sensitization and for sensitization to any of the pollen grains. Further, urban living in childhood added an increased risk for pollen sensitization. Conclusion: The prevalence of allergic sensitization was high in the urban adult population of Calcutta. More than half of those aged 20Y29 years were sensitized and 42.1% was sensitized to at least four allergens. Sensitization to multiple aeroallergens was associated with a high prevalence of asthma, AR or conjunctivitis and wheeze. HLAG, a nonclassical MHC class Ib molecule, plays a significant role in immuno-tolerogenic processes. HLA-G expression through alternative splicing generates membrane-bound and soluble isoforms secreted by activated (by IL-10) peripheral blood CD14+ monocytes. Genetic diversity of HLA-G consists of sequence variations in promoter, exonic and 3_untranslated (3_UTR) regions, some of which have been shown to affect HLA-G mRNA stability. Polymorphisms in the HLAG promoter region had previously been associated with asthma. Of interest is also the fact that the gene encoding the effector IL-10 also exhibits genetic polymorphism in populations. Given the plausible effect one over the other, in this study, we investigated the potential role of a 14-bp insertion/deletion polymorphism in the HLAG-3_UTR region in conjunction with IL10 gene promoter polymorphisms (-1082 A/G, -819 C/T and -592 C/A) on respiratory allergy development. The association study involved 125 patients suffering from asthma and/or rhinitis, all monosensitized against Derp 1 and 74 matchedcontrols, all originating from Algeria. PCR-based techniques were used to assess the HLAG and IL-10 genotypes. While the IL-10 promoter allele or genotype frequency did not differ between patient and controls, the 3_UTR +14-bp insertion allele was significantly more frequent in the patient group than in the control group (Pc = 0.01) with the homozygous state of the 3_UTR -14-bp deletion allele being more prevalent in the healthy controls (Pc = 0.04). Our findings suggest that HLA G gene may participate in the pathogenesis of respiratory allergic diseases in our population. A possible mechanism that could be envisaged is that the diminished immunosuppressive activity, mediated by HLA-G, may contribute to initiation/persistence of chronic airway inflammation in asthma. Sensitization to ascaris spp. influences total and specific IgE levels to dermatophagoides pteronyssinus and blomia tropicalis Sylvie Lafosse-Marin. Cabinet d'Allergologie, Allergologie, Fort de France, Martinique. Background: Although sensitization to Ascaris may not play a role in the severity of asthma, it may be implicated in total and specific IgE levels to common aeroallergens. Objective: To determine sensitization to the mite species D. pteronyssinus and B. tropicalis in patients who have a positive, or negative specific IgE determination to Ascaris spp. Methods: We analysed 852 consecutive patients (417 males and 435 females; mean age 18.5 years; ranging from 9 months to 82 years. 518 children (302 boys and 216 girls) with allergic asthma and/or rhinoconjunctivitis with or without eczema were included. Patients were separated into 2 groups. Group A consisted of Ascaris spp. positive patients and Group B of patients with a negative specific IgE determination (Phadia) to Ascaris spp. Conclusion: Sensitisation to Ascaris spp. seems to play an important role in rising total, as well as specific IgE levels to D. pteronyssinus and B. tropicalis. Statistically, more patients were sensitized to mites in the Ascaris spp. positive than in the Ascaris spp. negative group. The clinical significance of these findings remains to be established. In vitro sensitization to Ascaris spp. in a population of allergic individuals in the caribbean island of Martinique Enrique Fernández-Caldas. Dr. Beckmann Pharma GmbH, Research & Development, Seefeld, Germany. Background: Sensitization to allergens of the nematode Ascaris spp. is common in the tropics, where a large proportion of the population has been in contact with this, and other intestinal parasites. It has been suggested that sensitization to Ascaris spp. allergens may influence the allergic response to other allergens. Objective: To study specific IgE levels to Ascaris spp. in a large population of individuals consulting for allergic respiratory complaints in an Allergy Clinic in Fort de France, Martinique. Materials and Methods: This study has a prospective, cross-sectional design and was conducted from January 2003 to March 2007. A total of 1,060 consecutive patients (507 males and 553 females; mean age 20.41 years; 6 months to 88 years), evaluated for allergic respiratory complaints were entered in the study. Specific IgE to Ascaris spp. was measured by the CAP method (Phadia). Results were stratified by age and sex. Results: A total of 431 individuals; 239 men (47.14%) and 192 women (34.72%) were positive to Ascaris spp. When analysing differences by age, a total of 599 children with G15 years of age were included (349 boys and 250 females). In this population, 271 children (45.24%) were positive, including 169 boys (48.42%) and 102 girls (40.8%). In the group of patients with 915 years of age (461 individuals; 303 females and 158 males), 160 (34.92%) were positive, including 70 men (44.3%) and 90 females (29.7%). Overall, there was a steady decline in the percentage of positive results in males and females until the age of 44. After this age, however, there was a sharp increase in the prevalence of positive results in males. Conclusion: This study demonstrates a high prevalence of sensitization to Ascaris spp. in the Caribbean island of Martinique and confirms that approximately 40% of patients seeking diagnosis, and or treatment, for allergic respiratory diseases are sensitised to Ascaris spp. allergens. In younger children, especially in boys, this prevalence is closer to 50%. Although antiparasitic treatment may be required in these patients, we have not confirmed if specific IgE to Ascaris spp. represents current infestation or the effect of previous antiparasitic treatments, since the majority of children are systematically treated against intestinal parasites in Martinique. Methylation status in the promoter region of IL-12 gene might be correlated with allergy Yasuko Kobayashi 1 , Hirokazu Arakawa 1 , Akihiro Morikawa 1 , and Takumi Takizawa 2 . 1 Gunma University Graduate School of Medicine, Pediatrics and Deveropmental Medicine, Maebashi, Gunma, Japan; 2 National Institutes of Health, National Cancer Institute, Bethesda, Meryland, United States. Background: Helper T cells can be classified into Th1 and Th2 subtypes, which have been shown to contribute to cellular immunity and humoral immunity, respectively. It is currently thought that impairment of the balance between Th1 and Th2 cells may cause immune diseases, including allergic diseases. Th1 and Th2 cells are polarized from naive T cells following their contact with mature dendritic cells (DCs), which are antigen-presenting cells resident at the frontline of immunity. Only mature DCs can activate naive T cells. When the contacted DCs secrete interleukin (IL)-12, the naive T cells become Th1 cells. In contrast, when the contacted DCs secrete IL-4, the naive T cells become Th2 cells. Therefore, the type of cytokine secreted by the DCs is very important. DCs generated from cord blood were previously shown to secrete lower amounts of IL-12 than DCs from adult peripheral blood. The authors suggested that this finding was correlated with the immune status of infants, who have weak cellular immunity and develop allergic diseases easily. Objective: We hypothesized that reduced secretion of IL-12 by DCs from both allergic patients and cord blood would be due to methylation of the IL-12 gene promoter region, and that the allergic status may be attributed to delayed demethylation of this region. To clarify this hypothesis, we examined the methylation status of the IL-12 gene promoter region in monocytes (DC precursors) prepared from peripheral blood from healthy adults (HA) and allergic adults (AA) and from cord blood from normal infants (NI). Methods: Monocytes were purified from each of the three groups using a MACS system and an anti-CD14 antibody. Genomic DNA was extracted from the monocytes. The methylation status of the IL-12p35 gene promoter region was investigated using a bisulphate-sequencing analysis method. In addition, real-time RT-PCR amplifications were carried out to determine the IL-12p35 mRNA levels in cultured and lipopolysaccharide-stimulated monocytes from peripheral blood from HA and AA. Results: Among the three groups, there were significant differences in the methylation ratios at 7 CpGs in IL-12p35 gene promoter region. Contrary to our expectations, the methylation ratios were significantly lower in AA and NI than in HA. The IL-12p35 mRNA levels in HA and AA did not differ significantly. Conclusion: These results suggest that hypomethylation of the promoter region may be involved in the development of allergic diseases. The membrane-spanning 4-domains, subfamily A, member 2 (MS4A2) +6960A/G genetic polymorphism and the risk of allergic disorders Background: The atopic triad of asthma, allergic rhinitis, and atopic dermatitis are multifactorial diseases with a considerable genetic component. Several genes, through association and linkage analyses, have been found to serve as susceptibility factor of atopy. A polymorphism in the membranespanning 4-domains, subfamily A, member 2 (MS4A2 or FCER1B) has been shown to be associated with increased IgE levels. The role of MS4A2 +6960A/ G (E237G) polymorphism in modulating atopic disorders in the Filipino population was evaluated in this study. Methods: One-hundred two pairs of allergic individuals and controls were phenotyped for total serum IgE level using enzyme-linked immunosorbent assay (ELISA). Atopic status was defined by serum IgE concentration 9100 IU/mL. DNA was extracted and genotyped for the MS4A2 +6960A/G polymorphism by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Allele A was identified after PCR by a 280 bp band on 4% agarose gel and G allele by a 238 bp band. Results: The study population was in Hardy-Weinberg Equilibrium (Â 2 = 0.007521, df = 1, p G 0.05). There was no significant difference in the total serum IgE levels between subjects with AA, AG, and GG genotypes (p90.05). Likewise, allele frequencies of +6960A/G polymorphism did not differ statistically between cases and controls (Â 2 = 2.624, df = 1, p 90.05). Frequency of the G allele was 0.103 in cases and 0.157 in controls (OR = 0.617, 95% CI = 0.342 Y 1.111). Conclusion: The lack of association between MS4A2 +6960A/G polymorphism and total serum IgE levels among allergic individuals in the Filipino study population indicates that the single-nucleotide polymorphism may not directly be involved in the development of allergic disorders. Alpha-1-antitrypsin genotype and allergy A.G. Palma-Carlos and M.L. Palma-Carlos. Clinical Allergy Immunology Center, CAIC, Allergology, Lisbon, Portugal. Background: Alpha-1-antitrypsin deficiency (alpha-1-AT) is usually correlated to COPD or to hepatic disease. However alpha-1-AT deficiency can be a risk factor for allergy, co-existing or interacting with asthma or other allergic diseases. Methods: 2 patients with confirmed alpha-1-AT deficiency and allergic diseases and complete identification of genotype have been studied. Genetic studies have been done in the Genetics section of J. Chaves Clinical Pathology Clinic. Results: Case 1: C.J.J. 62 male heavy smoker during 30 years, complaints of cough, sputum from 7 years, more than 3 months each year. No diagnosis. Previously treated with fluticasone salmeterol + tiotropium objective examination respiratory sounds diminished. PFR 300 (normal 550T100). CT. Centro lobular emphysema mainly in superior lobes. Hyperinsuflation. Electrophoresis decrease of alpha-1 fraction (0,6% of 6,9) alpha-1 antitrypsin 64,5. Genetic study genotype Pi SZ mutations GLU 342 lys and GLU 264 val. Case 2: C.M.C. 40, female, non-smoker, story of rhinitis and urticaria without lower respiratory symptoms. Objective examination normal. On electrophoresis alpha-1 fraction was 0,6% of 7.45. Alpha-1 antitrypsin assay 25-Genotype Pi ZZ mutation GLU 342 lys. Thyroid auto antibodies increased TG 72,3 TPO, 1.673. Conclusion: Alpha-1-AT Z alleles are more frequent in northern Europe and in Iberian Peninsula. The S variant gives alpha-1 antitrypsin levels of 60% and Z variant 10% of normal (M allele). Therefore a MS heterozygote must have levels of alpha-1-AT around 40% (30% from S allele and 10% from Z allele) and a homozygotic ZZ had levels of 10%. In our cases the patient with the slightest form of alpha-1-AT deficiency has respiratory symptoms and an emphysematous pattern in CT. The theoretical more severe form ZZ homozygotic presented only rhinitis and urticaria without any lower respiratory complaints. That discrepancy could be explained by the nonsmoking habits of the homozygotic patient in contrast with a long history of smoking habits of the patient SZ. In patients even without lower respiratory complaints but presenting less than 1% of alpha-1 fraction in electrophoresis assay of alpha-1-antitrypsin must be done and in cases of low values a genetic study completed. Changes of "-casomorphins content in human milk during lactation Background: Milk is the best, complete food important for development and nourishment of neonate. Except nutrients milk contains biologically active opioid peptides derived from "-casein named "-casomorphins which can exert effects in neonatal gastrointestinal tract as wall as in all organism. It has been suggested that opioid peptides may play the role in regulation of infants' gastrointestinal functions and development and be involved in the process of immune maturation. The content of "-casomorphins in human milk during maturation phases has not been studied so far. Therefore, the aim of this study was to determine the content of "-casomorphin-5 ("C5) and -7 ("C7) in human milk in different phases of lactation. Methods: There have been chosen thirty healthy newborns and healthy mothers who plan to breast feed. The milk samples were collected every two months starting from delivery. The peptides were extracted according to Halwarker and Elliot method, purified by SPE with the use of C18 columns and identified using HPLC. Results: A significantly highest "-casomorphin concentration was found in colostrums "C5 = 5,03 T 1.02 2g/ml; "C7 = 3.10 T 0.892g/ml ) than in mature milk. The concentration of "-casomorphin in milk collected in the second month of lactation "C5 = 0,98 T 0,352g/ml; "C7 = 0.39 T 0.072g/ml) was similar to the level obtained from the fourth month of lactation ("C5 = 0,58 T 0,21 2g/ml; "C7 = 0.33 T 0,132g/ml). The level of "-casomorphin-5 in individual phase of lactation was not significantly different than the level of "-casomorphin-7. Conclusion: "-Casomorphins content in human milk is changing with the period of lactation. The level of opioid peptides may depend on the function of these peptides in neonate?s organism and change with the maturation process. The concenttation of "-casomorphines 5 and 7 in women with history of allergy and food hypersensitivity in their babies Maciej Kaczmarski 1 , Katarzyna Sidor 1 , Elzbieta Kostyra 2 , Beata Jarmolowska 2 , Krzysztof Bielikowicz 2 , Malgorzata Iwan 2 , and Henryk Kostyra 3 . 1 Medical University of Bialystok, IIIrd Department of Pediatrics, Bialystok, Poland; 2 University of Warmia and Mazury, Chair of Biochemistry, Olsztyn, Poland; 3 Polish Academy of Sciences, Institute of Animal Reproduction and Food Research, Olsztyn, Poland. Background: Breast milk provides the optimal nutritional supply which is critical for the postnatal adaptation of newborns. Although human milk is considered a hypoallergenic food, the proportions of breast-fed children demonstrating symptoms of atopic dermatitis is increasing. The allergy preventing effect of breast -feeding remains controversial and there are no consistent data regarding the degree to which breastfeeding prevents, reduces, delays or increases the development of allergic disease. A number of studies reports suggested the differences between allergic and non-allergic mothers in relation to milk concentration of immunological agents involved in allergic reactions. "-casomorphine seems to be a milk ingredient associated with pseudo-allergic reactions. Methods: Samples of human milk were collected from 12 volunteer breastfeeding women with a history of allergy and clinical manifestation of allergy to bovine milk proteins and from 30 healthy women. Milk samples were collected in two phases of lactation -first sample -colostrums, the second -one month after delivery. In the allergic group 9 of 12 women had asthma, 4 were diagnosed with allergy to bovine milk proteins; one women had both diseases. All infants mothers of allergic group have presented skin allergic symptoms i.e. atopic dermatitis. The peptides were extracted according to Halwarker and Elliot method, purified by SPE with the use of C18 columns and identified using HPLC. Results: "-casomorphins concentration in colostrums in the allergic group was roughly similar ("C5 = 1,58 T 0,402g/ml; "C7 = 1,67 T 0.422g/ml) and there was no statistically significant difference in "-casomorphins content between phases of lactation for both investigated peptides (mature milk: "C5 = 1,63 T 0,642g/ml; "C7 = 1,26 T 0.642g/ml). Results in the control group were more diversified -the content of "-casomorphins ? 5 and -7 in colostrums ("C5 = 5,03 T 1,022g/ml; "C7 = 3,10 T 0.892g/ml) was significantly higher than in mature milk ("C5 = 0,98 T 0,352g/ml; "C7 = 0,39 T 0,072g/ml): p G0,001 and p G0,05 respectively). Conclusion: The content of both investigated "-casomorhins: 5 and 7 in healthy breast-feeding women is decreasing during lactation but remains roughly constant in the group of mothers with allergy symptoms. The use of a (!1-3)-Fuc-core-modified N-glycans-containing glycoepitope from a seed glycoprotein for a novel approach to the diagnosis and therapy of vespids and honey bee-allergic patients Background: Mucuna pruriens seeds are an oral prophylactic for snake bite in humans. We demonstrated that GpMuc, a 29 kDa glycoprotein found in these seeds, is the immunogen of the antibody that cross-reacts with snake venom proteins. The immunogenic properties are postulated to reside in its core ("1-2)-linked xylose-and core !(1Y3)-linked fucosemodified N-glycan chains. To verify this hypothesis we investigated fragmented gpMuc for immunoreactivity. Since the same core modified Nglycans in honeybee and vespid venoms have often been considered immunogens responsible for venom allergies, we also investigated cross reactivity between gpMuc and Hymenoptera venom proteins. Methods: 1 mg of gp-Muc was trypsin digested. The digested sample was fractionated by reverse-phase HPLC and an aliquot of each fraction was analyzed for immunoreactivity to anti-gpMuc IgG by ELISA and dot blot tests, and the positive glycopeptide analyzed by Maldi TOF MS. Serum samples from patients with IgE positive to Hymenopera venoms were investigated by immunoblot and/or ELISA for IgE and IgG against the venom proteins, gpMuc and the corresponding glycopeptide. Anti-gpMuc IgG were analyzed for reactivity against the proteins of venoms. Results: The anti-gpMuc epitope was found to be a 1760 Da glycopeptide characterized by N-glycans containing fucose. Anti gp-Muc IgG only cross reacted with a protein having an apparent MW of 29 kDa when the venom proteins of Vespula polistes, Vespa cabro, Vespula germanica were analyzed and with a protein of 20 kDa in the protein fraction of Apis mellifera venom. Twelve of eighteen patients with IgE positive to one or more Hymenoptera venoms showed IgE and IgG positive to the 29 kDa venom protein of vespids and/or to the 20 kDa protein of Apis venom and to the gpMuc and corresponding glycopeptide. Conclusion: Anti-gpMuc glycoepitope is included in a 1760 Da peptide and the glycoepitope is shared by a similar venom protein in vespids and by a different one in Apis. Second finding: IgE and also IgG cross reaction between vespid and Apis venoms is restricted to one protein. The third finding was the existence of IgE and IgG cross reaction between the cross reacting protein of Hymenoptera venoms and gpMuc, specifically with the peptide containing the glycoepitope. This raises the possibility of using the gpMuc glycopeptide in place of whole venoms for the diagnosis and immunotherapy of Hymenoptera allergies, at considerable saving and with reduced risk. Insect sting allergies of emergency department visits in Al-ain Hospital, United Arab Emirates Shirina Alsowaidi 1 , Abdulrahman Mohamed 2 , Awad Alessa 1 , and Abdishakur Abdulle 1 . 1 Faculty of Medicine, UAE University, Department of Internal Medicine, Al Ain, United Arab Emirates; 2 Al Ain Hospital, Internal Medicine, Al Ain, United Arab Emirates. Background: Insect sting is a common public health hazard in the United Arab Emirates. However, nothing is known about the demographic, clinical manifestation and management of insect sting allergic patients. Objective: To assess the clinical manifestation of insect sting allergies and its management in Al-Ain hospital. Methods: In this retrospective observational study, we analyzed emergency department (ED) records of insect stung patients in Al-Ain hospital. A total of 783 patients (Mean age T SD; 29.2 T 12.6 yrs) were seen at the emergency unit of Al-Ain Hospital. Results: Of the total sample, 425 (54.3%) were female. Only 39 (5%) patients of the total were UAE nationals and 744 (95%) were heterogeneous expatriate workers. The site of the sting of 130 (16.6 %) patients was on the leg, 70 (8.9 %) on the arm, 21 (2.7%) on the head, 21 patients (2.7 %) on the trunk, and 12 patients (1.5 %) on the neck. However, a significant majority of the patients 528 (67.4 %) there was no available data with regard to the site of sting. In this study 41% of the subjects had local reactions, 40.4% had urticaria, 4.3% had angioedema, 47.5% had dyspnoea or wheezing, while only 3 patients were presented with unconsciousness. According to Muller classification, 17.8% were of class I severity, 4.2% class II, 36.8% class III and 2.6 % class IV. At ED 68.2% of the patients received antihistamine, 90.5% systemic antihistamine, 53.3 % systemic corticosteroids and only 1% received epinephrine treatment. Conclusion: Insect sting is common particularly among the expatriate community, often under documented, and of sever presentation. Since most of the insect sting was on the legs and arms, culturally covered clothing among the UAE nationals may have had significant protection impact. Increasing awareness, appropriate diagnosis, and adequate management of insect sting allergies is required. Robert Heddle 1 , Pamela Hudson 2 , and Simon Brown 3 . 1 Flinders Medical Centre and Flinders University of South Australia, Respiratory Medicine, Adelaide, Australia; 2 Flinders Medical Centre, Respiratory Medicine, Adelaide, Australia; 3 University of Western Australia, Emergency Medicine Research Unit, Fremantle, Australia. Background: In south eastern Australia the Jack Jumper Ant (JJA) is a dominant cause of sting anaphylaxis. We have described the epidemiology of JJA sting anaphylaxis on the island of Tasmania in a prospective study; approximately 12% of allergic subjects were stung each year, 70% of these experiencing further systemic reactions. Age over 35 years was a risk factor for severe reactions. Aim: To examine the natural history of JJA sting allergy in South Australia (SA), 1000 km NW of Tasmania. Methods: Subjects with a history of systemic reactions to ant stings were recruited through the Australian Ant Venom Allergy Study (AAVAS). Demographic details, blood samples for allergy testing, ant specimens and details of sting reactions were obtained. Subjects were followed prospectively for further sting reactions. Results: In 116 subjects with a history of systemic reaction to JJA sting there have been 15 further JJA sting events in 12 subjects during a mean follow-up period of 10.8 months; theses stings led to a total of 12 systemic reactions in 9 subjects. Only 4 of the 12 who were stung were less than 35 years of age, all reporting a lessened (2) or local only (2) reaction. Four of 8 older subjects experienced reactions similar to their worst previous sting event. Conclusion: Despite wide geographic separation the frequency of re-sting (10% of subjects per annum) the proportion of systemic reactions (~75% of subjects, 80% of sting events) and the tendency of those aged 935 to have further serious reactions bears a striking similarity to the Tasmanian data. Older subjects allergic to venom of this aggressive ant have a high risk of further systemic reaction to future stings. Reactions to hymenoptera sting in adult patients: experience in a clinical allergy/immunology service in Monterrey Mexico Maricela Hernandez 1 , Sandra Gonzalez 2 , Gabriela Galindo 2 , Alejandro Diaz 1 , and P Rodriguez 2 . 1 University Hospital, Allergy and Clinical Immunology, Monterrey, Mexico; 2 Univerisity Hospital, Allergy, Monterrey, Mexico. Introduction: The most commonly implicated insects in stings to humans are the honeybee, yellow jacket wasp and paper wasp. Stinging ants include the red and black fire ants. In Mexico, there are no data on the degree to which the general population is affected. Methods: A retrospective study, in which a questionnaire was given to all the patients who came to our clinic for the first time for different allergic symptoms, from January 2001 to December 2001 the questionnaire, asks about personal history of hymenoptera sting. We found 72 patients with positive history. Results: The distribution by sex was male (26) 36.1%, female (46) 63.9%. The distribution by insect_s sting was; Ant (58) 80.6%, honeybee (22) 30.6%, yellow jacket wasp (13) 18.15%, paper Wasp (2) 2.8%, none identified (1) 1.4%. The average age was 15 a 59 years SD (32.97 + 1.25) (63) 87.5% did not have work risk, number of sting trough life (51) 70.8% 1 a 3 stings episodes, (21) 15.1% 2 a 5, (19) 26.4% had an interval of 1Y6 months between sting episodes, (16) 22.2 % had a 3Y5 years interval between stings. (36) 50 % had itch or local pain, (6) 8.3%, had intense itching, (2) 2.8 % urticaria or angioedema (2) 2.8 % cardiovascular symptoms, (2) 2.8 % referred to have late reactions, (6) 8.3% had large local reactions. 69 (95.8%) did not seek for medical attention, 4 (5.6 %) New the severity of the problem, 3 (4.2 %) New what to do, (27) 37.7 % had allergic rhinitis and or allergic conjunctivitis, (16) 22.2% had urticaria and or angioedema, (15) (18.9%) had contact dermatitis, (7) 9.7% had asthma, (4) 5.6% have had at least one adverse reaction to drugs, (3) 4.2% had atopic dermatitis, (1) 1.4% had food allergy, (1) 1. Background: An animal model resembling the human clinical manifestations of allergy is needed for various aspects of allergy research, e.g. for investigating mechanisms and pathway of immunopathology, measuring allergenicity of a protein, testing drug and vaccine therapeutic efficacy. Objective: To develop a murine model of cockroach allergy. Materials and Methods: Individual BALB/c mice were intraperitoneally (i.p.) injected with three doses, i.e. on days 0, 7, and 14, of alum adjuvanted-crude extract of American cockroach (CR), Periplaneta americana, which is the predominant CR species in Thailand. The animals were subsequently aerosolically challenged on days 21 and 22 with 10 ml crude CR extract (1.0% w/v in PBS), using a home-made air-pressure nembulizer for mice. The mice were bled and sacrificed on day 23. Their serum samples, broncho-alveolar lavage fluids (BALF) and lungs were collected. Control mice received only alum i.p. and aerosolic PBS via the nembulizer. Results: All CR extract treated mice showed features of allergy. Their BALF contained inflammatory cells that out-numbered those of the non-allergic counterparts as determined by differential cell counts after cyto-spinning and hematoxylin-eosin staining. Most of the cells were eosinophils. Eosinophil infiltration was also pronounced in the lung tissues of the CR extract treated mice and was negligible in the tissue of the controls. Levels of serum IgE specific to the American CR extract rose significantly above those of the control levels. Conclusion: A murine cockroach allergy model with features resembling the human allergic manifestations was successfully developed. The model can be used for study of various attributes of CR allergy, e.g. allergenicity of a protein of interest, immunopathology, and drug and vaccine therapeutic efficacy. Crude Forcipomyia taiwana (biting midge) extracts stimulate PBMC proliferation, IFN-!, IL-6 and IL-10 secretion from midge-allergic subjects Yi-Hsing Chen 1 , Mei-Fang Lee 2 , Hsin-Ru Wu 1 , and Guang-Yuh Hwang 3 . Tunghai University, Department of Life Science, Taichung, Taiwan. Background: Forcipomyia taiwana (F. taiwana) is a tiny (1Y1.5 mm) bloodsucking midge that is widely distributed in urban and suburban Taiwan as well as in southern China. F. taiwana attacks in groups to exposed parts of the human body during the day, causing intense pruritis and swelling in sensitive individuals. There are two types of reactions after midge bite: 1) immediate reaction: large local swelling develops within one hour of bites 2) delayed reactions: intense itching papules and vesicles occur 6Y24 hours after bites. The delayed lesions may turn centrally necrotic and last for weeks or even months. We have previously reported that about 59.1% of the exposed subjects develop hypersensitivity reactions to F. taiwana bites. Among which, about 14% developed immediate reaction only, 43% developed an immediate reaction followed by delayed reactions and 43% developed solely delayed reaction. Our previous study suggested that the immediate large local reaction is IgE-mediated and the 24-, 35/36-and 64-kDa proteins are major IgE-binding allergens. However, from our previous studies, we also found that delay reactions to F. taiwana bites may not react through the IgE pathway. Delay itching skin papules or vesicles usually last for weeks or even months that causing an even greater impact on patient's quality of life than the rapidly resolved immediate reaction.Except some limited studies focused on the immunologic mechanisms to mosquito bites, there has been no report in the literature regarding the immunologic mechanisms to biting midges. Materials and Methods: Peripheral mononuclear cells (PBMC) from midgeallergic patients with delayed reactions were cultured with media only, Con A or F. taiwana crude extract at different time points. Cell proliferation was measured by MTT assay. Change of cytokines in culture supernatants was measured using fluorescent bead immunoassay. Results: In the present study, we demonstrate that peripheral mononuclear cells (PBMC) from delayed typed midge-allergic subjects proliferate and secret more interferon-! (IFN-!), interleukin-6 (IL-6) and interleukin-10 (IL-10) in response to crude F. taiwana extract. IL-4 and IL-5 were not detected in the culture supernatants. Conclusion: The delayed reactions of F. taiwana bites may be mediated by a more Th1 type cells. The role of IL-10 in midge-bite allergy requires more investigation. Local allergic reaction on bone wax in a patient with bee venom allergy Carlotta Wirz 1 , Heidrun Mueller 1 , Regina Treudler 2 , Gero StrauQ 1 , Sylvia Meuret 1 , and Andreas Dietz 1 . 1 University of Leipzig, ENT Department, Leipzig, Germany; 2 University of Leipzig, Clinc for Dermatology, Venerology and Allergology, Leipzig, Germany. Introduction: A 67 year-old farmer was transferred to our clinic after an outpatient operation on his right middle ear (stapes reconstruction) that showed an intraoperative strong bleeding. During the exploration of the ear in our operation room a damaged raised bulb of the jugula vein could be detected. The lesion was closed by conventional bone wax, fibrin glue and perichondrium. Follow Up: Ten days after the removal of the endaural tamponade the patient complained of chronical otorrhoea of the operated ear that showed resistance to any treatment. The examination showed a distinct inflammation of the external and middle right ear with a clear secretion. During the operative revision of the situs an extensively grown granulation tissue around last bits of the applied bone wax was seen. After its removal the defect was closed by cartilage of the tragus. Diagnostic Investigation: After the second operation the patient mentioned a previously known bee venom allergy he had not mentioned in the first examination. Recurrent anaphylactoid reactions had happened on bee stings. We decided to get into contact with the producer of the bone wax. We experienced that the used common bone wax consists of 80% of bee wax and 20% of isopropylpalmitate. The allergy diagnostic on the patient confirmed the bee venom allergy (prick test, RAST), furthermore the epicutaneous testing on the applied bone wax batch showed a positive result. The epicutaneous testing of the single components of the bone wax (bee wax, isopropylpalmitate) was negative. Conclusion: For the producer could not guarantee a 100% pureness of the product, we conclude that the applied charge of bone wax containing bee wax was probably contaminated with bee venom. Bee wax represents a great part of the base of common ointment and creams. The patient never mentioned reactions on those substances. Therefore, the probabiltiy of an allergy against uncontaminated bee wax (allergic reaction type I on propolis) is very low. In future, a detailed allergy anamnesis should be performed before using bone wax in ear or any other ent surgery. Alternatively, hypoallergenic substances should be used in patients with known bee wax allergy undergoing ent surgery. Carpenter bee anaphylaxis: description of two cases and identification of the responsable allergens with anaphylactic systemic reaction after Xylocopa violacea sting. All the two patients were also stung by yellow jacket before and after anaphylaxis, well tolerating the stings. At the clinical visit we performed serum venom specific IgE antibodies and venom skin prick test and intradermal tests with yellow jacket, paper wasp, european hornet, honeybee and Polistes dominulus venoms. Five patients with negative allergenic history were used as negative controls. Xylocopa violacea extract: Xylocopa violacea extract was supplied by Anallergo (Florence, Italy). SDS-PAGE and Immunoblotting: SDS-PAGE was performed with Xylocopa violacea and honeybee venom extract. The gel was blotted on a nitrocellulose membrane, cutted into strips and matched with the sera of the two patients. After incubation with anti-IgE antibodies, the allergenic bands were detected by a chemiluminescens revealing system. Results: In the two allergic patients skin tests and serum IgE detection were positive only for yellow jacket extract. In vitro analysis did not show any IgE binding in the honeybee immunoblotting, while IgE reactivity was shown with the band at 25,2 kDa, 18,8 kDa and 6,5 kDa in the Xylocopa violacea extract. No response was found in control sera. The allergenic bands at higher molecular weight were specific of Xylocopa violacea venom, while the band at 6,5 kDa corresponds to mellitin. Conclusion: This is the first report of two cases of anaphylaxis caused by Xylocopa violacea, a member of the Apidae family. Two allergenic bands (25,2 kDa, 18,8 kDa) are specific allergens of this venom extract, not shared with honeybee venom extract. Tryptase, mastocytosis and Hymenoptera venom allergy Petr Kucera 1 , Milada Cvackova 1 , Martina Heroldova 1 , Helena Vavrova 2 , and Jan Ort 2 . 1 3rd Medical Faculty, Charles University and FNKV, Dept Allergy Immunology, Prague, Czech Republic; 2 FNKV, Dept Allergy Immunology, Prague, Czech Republic. Background: Elevation of basal serum tryptase as a one of signs of systemic mastocytosis (SM) can be a risk factor for the reaction severity in patients allergic to Hymenoptera venom (HV). The exact pattern of the relationship between elevated basal tryptase and anaphylaxis is not completely understood. We focused on the relationship between serum basal tryptase and patient_s clinical data, severity of systemic allergic reactions (SAR) and safety of venom immunotherapy. Patients and Methods: We analyzed data from 69 consecutive patients with SAR due to HV allergy. 27 men and 45 women were included; mean age 48.26 years, most of them with sensitization to relevant venom allergen (prick tests or specific IgE positive). The IgE and tryptase level was measured by Phadia ImmunoCAP FEIA assay. Results: We found significant differences between mean tryptase values in groups of different severity of SAR: Grade I (mean 4.77 ug/l), Grade II (5.56), Grade III (5.48) and Grade IV (12.67, p=0.035). 10 patients (14.5 %) had elevated basal tryptase (mean value 28.52 versus 5.28 ug/l, p G 0.0001). Patients with elevated tryptase were significantly older than those with normal value (61.3 and 46.2 years, p=0.0049). Two patients were diagnosed by systemic mastocytosis, one by non-Hodgkin lymphoma, in two patients SM was not confirmed. Venom immunotherapy was well tolerated. Conclusion: The elevated basal serum tryptase level (not systemic mastocytosis only) can be a risk factor for more severe SAR caused by Hymenoptera venom allergy, particularly at the higher age. Background: Increased serum tryptase (ST) levels and Mastocytosis are described in patients with severe anaphylactic reactions after hymenoptera sting (from 7,3% to 10.5% depending on the study). Aim of this study was to determine basal value of ST and its possible clinical relevance in hymenoptera venom allergy. Methods: One hundred six patients with a history of venom anaphylaxis (79 yellow-jacket, 10 paper-wasp, 5 european hornet, 12 honey-bee) were included. The grade of allergic reactions was considered according to Müeller classification. ST levels were measured by UniCAP, Pharmacia-Uppsala, (normal range G11.4 2g/L) at baseline and in 25 patients after 1 year immunotherapy. Venom serum specific IgE levels (sIgE) were measured at baseline by CAP System, Pharmacia-Uppsala. Results: Mean basal ST level were 3.86 (IQR 2.61Y5.39). Only 4 patients (3.77) have elevated ST levels. There were no significant difference in ST levels between HB and other Vespid patients (3.47 vs 3.91) . Mean basal venom-specific IgE (sIgE) was higher in HB than in all the other venom allergic patients (19.4 vs 4.14). In Vespid patients, but not in HB ones, higher ST levels were statistically correlated with a higher Müeller grade of reaction (p = 0.049), while in HB patients there is an inverse correlation between basal ST and sIgE (p = 0.053). No difference in ST levels was observed after 1 yr immunotherapy in respect to baseline values. Conclusion: In our population cutaneous or systemic mastocytosis appear not to be so recurrent. The only patients with high ST levels were allergic to Vespid venoms. HB patients, despite a higher specific venom IgE levels, did not present statistically more severe reactions than Vespid patients. To note that HB patients lower ST levels correlate with higher sIgE levels. These data suggest that in HB patients severe anaphylactic reactions are due to the high sIgE level and not to a larger number of mast cells which a higher ST level would indicate. is known about the quantitative weight of the different work activities. Aim of study was to evaluate the distribution of work habits among subjects with HVA. Methods: Subjects referred to our unit for suspected HVA underwent the standard diagnostic workup. According to the modality of exposure to hymenoptera stings, they were classified: i) at high risk (subjects working usually outdoors or in places were hymenoptera live): farmers, gardeners, firemen, truck drivers, masons, beekeepers, dustmen; ii) occasionally at high risk (working sometimes outdoors): plumbers, electricians, railswaymen, at low risk (working indoors) students, housewives, clerks, professionals. The distribution of occupations in the general population was provided by the Central Office for the Work Insurance of Regione Veneto. Results: Three hundred and 46 subjects (238 male, mean age 44 T 15 yrs) seen at our unit between 2000 and 2007 were diagnosed as having HVA. All had a clinical history of systemic reactions to stings (20% bees, 17% polistes, 63% wasps), of various grades of severity (56% grade III-IV). Concerning exposure, 23,4% of patients were at high occupational risk (43 farmers, 15 masons, 14 truck-drivers, 4 gardeners, 3 beekeepers, 2 dustman), 8,38 % had occasionally high risk (3 warehousemen, 3 soldiers, 9 factory workers, 1 plumber, 5 electicians, 2 railwaymen 1 agronomist, 5 engineers) and 68,2 % were at low risk. In the general population, wokers not at risk were 91.3%, at occasional risk 0.9%, and at high risk 7,8%. Among the patients at high risk or occasionally high risk (110 patients), 71% were stung only during their work activity. Beekeepers, considered typically at high risk, in our database were less represented than, for instance, masons and truckdrivers. Conclusions. The risk of having HVA is higher than expected in some categories. Also taking into account the limit of such a survey, where the data of the general population may be imprecise, it is clear that some occupations are much more frequent in patients with HVA. Thus, for some categories (farmers, truckdrivers) allergy to hymenoptera venom could be considered an occupational disease. Keywords: Hymenoptera venom allergy, work occupation, risk. Results of a modified ultrarush immunotherapy in hymenoptera venom allergy Julia Klecker, Heidrun Mueller, and Andreas Dietz. University of Leipzig, ENT Department, Leipzig, Germany. Background: Patients allergic to hymenoptera venom may develop local or generalized sting reaction or they may show systemic reactions of differing severity. If venom immunotherapy (VIT) is indicated, increase doses are usually given during hospitalisation, because severe adverse reactions may occure. There is a wide range of rush-and ultrarush protocols known. We have tested a 3-day ultrarush-VIT protocol according to its tolerance. Methods: 20 patients were treated with VIT, 4 of them received bee-VIT, 16 vasp-VIT. We administered 15 injections over 3 days. Incidence and intensity of adverse reactions were evaluated. Besides the coherence between cardiovascular diseases (especially hypertension), severity of systemical reaction to the insect sting challenge, specific IgE-level and skin prick test and the intensity of adverse reactions during VIT were determined. Results: In total 3 of 20 patients showed adverse systemic reactions, all of them got a vasp-VIT. 2 of these patients had a third grade anaphylactic reaction (according to Ring and Messmer) due to a hymenopteron sting in case history, 1 patient reported about a first grade anaphylactic reaction. Local reactions to the injections ranged from mild (G5cm) to moderately severe (up to 10cm). There was no obvious correlation between intensity of local reaction during VIT and degree of anaphylaxis in case history. 2 of the mentioned 3 patients showed positive reactions in skin prick test at 12g-dose, 1 patient just reacted at 1002g-dose. The specific IgE-level also showed no correlation to local or systemic reactions. Conclusion: The 3-day ultrarush protocol was well tolerated by our patients. We noticed 3 systemic reactions. This is in accordance with the data of studies in literature. Due to the small number of current participants we can find no connection between the incidende of systemic reactions during VIT and the other facts examined. The only noticeable fact was that 2 of 3 patients with systemic reaction during treatment reported about third-degree anaphylactic reaction by first sting challenge. Local reactions to the injections showed moderate intensity, there were no skin effects more than 10cm. Leishmanial antigen detecting by dot immunoblot assay in phlebotomus papatasi, in Iran Hassan Nekouie. Pasteur Institute of Iran, Parasitology, Tehran, Islamic Republic of Iran. Introduction: Leishmaniosis is a polymorphic disease of the skin and viscera caused by an intracellular Protozoan. Zoonotic Cutaneous Leishmaniosis (ZCL) is a major health problem in rural areas of Iran. Methods: A simple and highly reproducible dot-immunoblot assay was developed to detect leishmanial antigen in Phlebotomus papatasi that were naturally infected with Leishmania major. The test was sensitive to as little as 10 ng of antigenic protein and also appeared to be specific, in that it gave a positive result with some P. papatasi (the primary vector of L. major in Iran) and L. major but not with P. sergenti or other pathogens. When used to investigate a large number of sandflies collected from areas of the Iran where cutaneous leishmaniasis is endemic Discussion: The assay appeared sufficiently sensitive and specific to detect the naturally infected insects. The simplicity, reproducibility, high sensitivity and high specificity of the assay should make it useful for field studies. Cockroach sensitization in patients with allergic airways disease in reunion island Bashir Omarjee 1 , Bernard Tanguy 1 , and Ahmad Zaidou 2 . 1 Centre Hospitalier Departemental, Insect Allergy, Saint Denis, Reunion; 2 Centre Hospitalier Departemental, Service de Maladies Respiratoires, Saint Denis, Reunion. Background: The objective of this study was to evaluate the prevalence of skin Prick sensitivity and RAST (UniCap-Pharmacia)to Blattela germanica, Blatta orientalis,Periplaneta americanain patients with rhinitis and / or asthma living in Reunion Island (Indian Ocean). Methods: Patients were 51 male, 53 female, aged 5 to 65 years (mean age of 29.6 years) were evaluated in one private clinic and in one public hospital: 18.2% had asthma, 32.5% rhinitis and 49.3% rhinitis and asthma. They were considered sensitized if the prick test induced a papula with at least one half the area of papula induced by histamine, or , if specific IgE was superior to 0,70 KU A/L. Skin tests were conducted with extracts of B. germanica, B. orientalis, P. americana and a battery of common aeroallergens (Allerbio, ALK Abello, France). Results: A positive skin test was recorded in 98 patients (94.5%), 87% of which were positive to mites, 15% to pollens, 9% to moulds, 4.8% to epithelia and 51.9% to at least one of the cockroach species tested; 24.9% of these patients was sensitized to all 3 cockroaches and 27% were sensitized to just one species.The prevalence of sensitization to the respective species was: 41.8% to B. germanica, 33.9% to B. orientalis, 39.8% to P. americana. Mean values of specific IgE for positive RAST were respectively: 34,9 T 36,4;25,1 T 31,0; 24,6 T 32,6. In addition, a significant correlation between results of skin test to cockroach and mite was observed (p.0004). Conclusion: The authors concluded that sensitization to cockroach is frequent in Reunion Island. Our results suggest the recommendation that cockroach extracts should be routinely used for evaluation of patients with asthma and rhinitis,or both in tropical countries. Effective treatment of mosquito bites with rupatadine; an experimental study Timo Reunala 1 , Ari Karppinen 1 , and Henrikki Brummer-Korvenkontio 2 . 1 Tampere University Hospital, Department of Dermatology, Tampere, Finland; 2 National Public Health Institute, HIV Laboratory, Helsinki, Finland. Background: People frequently experience whealing and delayed papules from mosquito bites. Whealing is mediated by antisaliva IgE antibodies and histamine. Rupatadine is a new antihistamine and PAF antagonist effective in allergic rhinitis and urticaria but the effect on mosquito-bite allergy is not known. Methods: A double-blind, placebo-controlled, cross-over study was performed with rupatadine 10 mg and matched placebo in 30 mosquito-bite-sensitive adults. The mean age was 37 years and the subjects had suffered from harmful mosquito bites for a mean of 15 years. Either rupatadine or placebo was taken at 8 am for 4 days, followed by a 5 day wash out period and then alternative treatment was given for 4 days. On day 3, in the both drug periods the subjects received two Aedes aegypti mosquito-bites on the forearm. The size of 15 min bite lesions and intensity of pruritus (VAS) were measured. Results: 27 subjects could be analysed for the efficacy. Rupatadine decreased the size of 15 min wheals by 43% compared to placebo. This effect was significant (P = 0.0065) whereas the effect on accompanying pruritus did not reach statistical significance although there was a favourable trend (P = 0.0621). Conclusion: The present placebo-controlled study in mosquito-bite-sensitive adults shows that prophylactically given rupatadine 10 mg is an effective treatment for the mosquito-bite whealing. Background: Atopic dermatitis represents a chronic inflammatory skin disease which results from complex interactions between genetic and environmental mechanisms. Referring to this the colonization and infection with staphylococci is a serious issue in skin disorders especially atopic dermatitis (AD). Human skin epithelium provides a mechanical barrier to invading bacteria and also participates in innate immune defence by producing cationic antimicrobial peptides, e.g. "-defensins (hBD) and the cathelicidin LL-37. The aim of our study was to determine the potential virulence of staphylococci from AD patients in bacterial skin infections and to examine the innate immune response to infection through expression of antimicrobial peptides. Methods: Identification of the staphylococci was performed by using T-RFLP analysis of the gap gene. Pathogenicity profiles of Staphylococcus spp. isolated from AD patients were determined by screening genes for staphylococcal enterotoxins (sea-see, seg-sej), toxic shock syndrome toxin 1 (tst) and Panton-Valentine leukocidin (luk-PV) by PCR. A model of bacterial skin infection was used to test the pathophysiology of these isolates in a human keratinocyte cell line (HaCaT), including the induction of hBD-1, hBD-2, hBD-3, LL-37, IL-6 and IL-8 (real time PCR, ELISA) and cellular invasion (fluorescence staining, confocal laser scanning microscopy). Results: Toxin genes with various pathogenicity profiles were studied for S. aureus. No superantigenic exotoxins by coagulase-negative staphylococci were detected. Exposure of keratinocytes to Staphylococcus spp. triggers the production of cytokines IL-6 and IL-8 and of "-defensins (hBD-1 mRNA (2-to 8-fold), hBD-2 mRNA (2-to 270-fold), hBD-3 mRNA (2-to 6-fold)) and the cathelicidin LL-37 (2-to 6-fold), whereas the levels of this activity were different for all clinical isolates. Adherence and invasion to keratinocytes was shown for clinical isolates of S. schleiferi and S. aureus, but not for S. hominis, S. epidermidis and S. capitis. Conclusion: Staphylococci from AD patients showed various pathogenicity profiles, invaded keratinocytes, the production of cytokines and antimicrobial peptides. Thus, they are potential pathogens in skin infections, especially atopic dermatitis. Vinit Mehrotra 1 , G. Awasthi 2 , and S. Singh 2 . 1 Himalayan Institute of Medical Sciences, Biochemistry, Dehradun, India; 2 Dolphin Institute of Biomedical Sciences, Biochemistry, Dehradun, India. Background: Pemphigus Vulgaris (PV) is an autoimmune, intraepithelial, blistering disease affecting the skin and mucous membranes. It is mediated by circulating autoantibodies (IgG) directed against keratinocytes cell surfaces and potentially life-threatening. PV antibody binds to the keratinocyte cell surface molecules Desmoglein-1 and Desmoglein-3 (Dsg-1 and Dsg-3). In India the incidence of PV is 86% with the mean age 30Y40 years while in Western countries is 40% only with the mean age 50Y60 years. Methods: So it is very important to identify the implicated antigens in PV patients and diagnosis the pattern of expression of Dsg1 and Dsg3 in patients suffering from bullous diseases. The levels of cytokines-TH1 and TH2 was also studied for the involvement of T and B cells as inflammatory. Results: It has been seen that the mucocutaneous type of PV is more prevalent in Indian patients as two types of band pattern are observed during immunoblotting with patient sera against 160kD and 130kD antigens extracted from normal human patients. It was also found that cytokine profiling of the patient sera that the level of TH2 like cytokines for example IL-4 and IL-10 were significantly high during the active stages of PV whereas, no relative difference was seen for TH1 like cytokine for example IFN-g. Cytokine profiling for confirmed PV cases from sera showed direct evidences for the involvement of auto reactive T cells responses to Dsg-3, which may be critical for pathogenesis of PV. Among its other biological activities IL-10 also induces B-lymphocytes to proliferate and differentiate to produce antibodies, which may play a important role in production of auto antibodies for PV. Conclusion: This study indicates that PV is an antibody-mediated disease with increase in TH2 cytokines and auto reactive T cells from PV patients produced TH2 cytokine preferentially in active stages of PV disease. This study will help to elucidate the cellular immune abnormalities leading to production of IgG autoantibodies in PV patients. Successful treatment in pulmonary alveolar proteinosis with the combination of intravenous immunoglobulin and pulse methylprednisolone: a case report Nongnapa Jirarattanasopa 1 , Tantikul Chutsumarn 2 , Orathai Jirapongsananuruk 2 , Nualanong Visitsunthorn 2 , and Pakit Vichyanond 2 . 1 Siriraj Hospital, Allergy and Immunology of Pediatrics Department, Bangkok, Thailand; 2 Siriraj Hospital, Pediatrics Department, Bangkok, Thailand. Background: Pulmonary alveolar proteinosis (PAP) is a rare disease in children, characterized by intra-alveolar accumulation of lipoproteinaceous material that positively stained with periodic acid-Schiff (PAS). The susceptibility to pulmonary infection is a salient feature of the disease which could be partly explained by defective alveolar macrophage function. Methods: We report a case of PAP and chronic interstitial pneumonitis in a girl who presented with chronic respiratory distress since the age of 5 months. Her clinical condition progressively worsened with chest radiograph showing diffuse hazy opacities of both lungs. High resolution computerized tomography (HRCT) of the chest demonstrated crazy-paving pattern. Definite diagnosis of PAP was made by open lung biopsy showing marked interstitial lung inflammation with alveolar proteinosis pattern. Results: Bronchoalveolar lavage were attempted twice without any clinical improvement. The respiratory clinical collapse with continued recurrent pneumonia was observed. After pathological diagnosis, a short tried of G-CSF was attempted again to no avail. Low dose methylprednisolone and hydroxychloroquine was then initiated without satisfactory improvement. It was decided to start intravenous immunoglobulin (IVIG) (500 mg/kg, every 4 weeks) with pulse methylprednisolone (30 mg/kg, every 4 weeks) to which she gradually responded. She no longer required oxygen supplement after 21 months of treatment. Currently she is asymptomatic at age 4 years and continues to receive IVIG and pulse methylprednisolone every 4 weeks. Conclusion: Aside from therapeutic lung lavage and GM-CSF therapy, IVIG and pulse methylprednisolone may have potential role in successful treatment of PAP. Cytokine profile of children allergic diseases : Inflammation process is participated by a cascade of cytokines, which predetermines development, direction and power of immune response and regulates cellular interaction. The character of changes in cytokine profile may be a distinguishing sign of a pathologic process. The research objective was to investigate specific features of cytokine spectrum of nasal secretion, saliva and blood serum in the process of children allergic diseases development (allergic rhinitis, atopic dermatitis, bronchial asthma). IL-4, IL-13, IL-8, IFN+, TNF! cytokine content, I IFN+/IL-4 cytokine regulatory index and total IgE level were monitored within a year at different stages of the disease. Cytokine level in biological liquids was determined by immuno-enzymatic method, sandwich variant, with R and D Diagnostics Inc. (USA) reagents. Researchers examined 22 children in the age from 1 to 16 with allergic diseases, control group was made of 30 health children of the same age. Control group examination results: serum cytokines IL-4 = 8,5 T 1,1 pg/ml, TNF! = 12,1 T 2,2 pg/ml, IL-13 = 8,01 T 1,6 pg/ml, IFN+ = 50,6 T 4,2 pg/ml, I IFN+/IL-4 = 5,2 T 1,1; saliva cytokines IL-4 = 0,3 T 0,1 pg/ml, TNF! = 1,47 T 0,4 pg/ml, IL-13 = 2,25 T 0,4 pg/ml, IFN+ = 65,2 T 3,4 pg/ml, I IFN+/IL-4 = 80,5 T 5,4 respectively; nasal secretion cytokines IL-4 = 1,88 T 0,64 pg/ml, TNF! = 6,87 T 1,08 pg/ml, IL-13 = 6,19 T 1,14 pg/ml, IFN+ = 156, 8 T 4, 8 pg/ml, 4 T 4, 8. High IL-4 and IL-13 levels at decreasing INF+ were recorded mostly in nasal secretion of children with allergic diseases in isolated allergic rhinitis (AR) cases (I IFN+/IL-4=0,34 T 0,06). High IL-4 content 87,62 T 2,59 pg/ml in nasal secretion were observed in exacerbation period and in rhinitis and bronchial asthma (BA) combined cases, at the same time children with BA but without AR showed 15,04 T 2,21 pg/ml and I IFN+/IL-4 = 4,6 T 1,2. In the period of AR clinic remission IFN+ content in nasal secretion was 1,1 T 0,96 pg/ml, in exacerbation period Y 9,36 T 2,21 pg/ml. Cytokine levels observed in saliva in children dermatitis cases were as follows: IL-4 = 89,3 T 0,64 pg/ml, IL-13 = 124,41 T 5,9 pg/ml, IFN+ = 11,9 T 1,7 pg/ml, blood serum IL-4 = 55,2 T 4,2 pg/ml, IL-13 = 39,4 T 4,9 pg/ml, IFN+ = 21,2 T 2,0 pg/ml. Investigation of local cytokine secretion and determination of their ration help to specify the localization and rate of allergic inflammation providing data for forecast and treatment effectiveness evaluation. Severe eczema associated with IgG2/G3 subclass and IgM deficiencies successfully treated with intravenous immunoglobulin Somboon Chansakulporn, Nualanong Visitsunthorn, Pakit Vichyanond, and Orathai Jirapongsananuruk. Siriraj Hospital, Pediatrics, Bangkok, Thailand. Background: Patients with severe eczema and recurrent infections should be investigated for immunodeficiency. We report our experience in treating a patient with severe eczema, recurrent bacterial infections, chronic otitis media, IgG2/IgG3 subclass and IgM deficiency with monthly intravenous immunoglobulin (IVIG) infusion. Methods: An 8 year-old boy was a case of long standing severe eczema with multiple food and drug allergies since 2 years of age. He had had chronic otitis media for the past 2 years. His treatment included elimination of suspected foods and drugs, topical corticosteroid, oral antihistamine and topical calcineurin inhibitor without satisfactory improvement. He was admitted several times due to bacterial skin infections with which he developed Acenitobacter septicemia on one occasion. His CBC did not show thrombocytopenia (platelet 300,000 /mm3) nor small platelets. Immunoglobulins were: IgG 1,480 mg/dl, IgG1 1,030 mg/dl, IgG2 115 mg/dl, IgG3 10.7 mg/dl, IgG4 18.3 mg/dl, IgA 142 mg/dl, IgM 18.3 mg/dl, and IgE 8,080 mg/dl. Specific IgE for egg white and cow's milk were positive. Immunophenotype revealed: CD3 692 cells/mm3, CD4 284 cells/mm3, CD8 351 cells/mm3, CD19 378 cells/mm3 and NK cell 193 cells/mm3. Lymphocyte proliferative response to PHA was below normal. The antibody response to pneumococcal vaccine increased 8.1 folds after receiving conjugated pneumococcal vaccine. Results: IVIG infusions at 400 mg/kg were given every four weeks. The patient's skin lesions were gradually improved after the 4th dose of IVIG infusion. Conclusion: The patient who presented with severe eczema and recurrent infections should be evaluated for immunodeficiency. The mechanism of beneficial effect of IVIG therapy in this patient was unknown but could be due to prevention of infection and its anti-inflammatory effects. Long-term effects of treatment on the cytokine production in scleroderma patients Roman Yatsyshyn, Yevgen Neyko, and Natalya Yatsyshyn. Medical University, Internal Diseases, Ivano-Frankivsk, Ukraine. Background: Scleroderma, a debilitating acquired connective tissue disease, is characterized by fibrosis, particularly of the skin and lungs. The cytokine disbalance is one of the main manifestation of scleroderma. The treatment of this pathology is very difficult problem for therapy. Objective: To compare the long-term effects of infusion of alprostadil with those of oral pentoxyphyllin on the production of cytokines in patients with systemic sclerosis (SSc), and evaluate their relationship with effects of the two treatments on clinical parameters. Methods: The production of cytokines by alloactivated circulating mononucleated cells was assessed before and after one year of treatment in a subset of 50 patients enrolled in a one-year study. Twenty five patients were treated with a 10-day (100 mkg) infusion of alprostadil -synthetic analog of prostaglandin E2. Another 25 patients were treated with an oral slow-release formulation of pentoxyphyllin, 600 mg twice daily. Quantitative determinations of interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) in the serum of patients were performed with a ELISA. Results: The production of IL-1beta was significantly lower in the alprostadil group than in the pentoxyphyllin group. Both the cutaneous fibrosis and the capillaroscopic patterns were better in patients treated with alprostadil. There was significant positive covariance between IL-1beta and TNF-alpha changes and the changes in both the skin score and the capillaroscopic score. Conclusion: There are several mechanisms by which alprostadil could exert its clinical efficacy. Vasodilatation and inhibition of platelet aggregation are certainly important, but they are transient. We suggest that the long-lasting modulation of the cytokine network observed in the present study could be another potential mechanism responsible for the persistent efficacy of alprostadil despite its administration. Endothelial dysfunction and inflammatory markers at the chronic heart failure with hypertension Natalya Yatsyshyn, Roman Yatsyshyn, and Yevgen Neyko. Medical University, Internal Diseases, Ivano-Frankivsk, Ukraine. : The chronic heart failure and arterial hypertension has a lot of biochemical manifestation. The ideas of our research work were: 1) to compare high sensitive C-reactive protein (hsCRP) and soluble receptors TNF alpha (sTNF-Rs) with coronary heart disease (CHD) and the chronic heart failure (CHF); in the control group (C group); 2) to compare ultrasound markers of endothelium-dependent vasodilatation (EDV) and endothelium-independent vasodilatation (EIV) in hypertensive (HT) patients in both groups. Patients and Methods: 105 hypertensive pts (mean age 61.3 yrs.), with CHD+ CHF were investigated. In the NYHA classes I/ II/ III/ IV were 20/31/46/8 pts. The C group was represented by 20 healthy volunteers. Ultrasound assessment of flow mediated dilation (FMD) of brachial artery was used for the measurement of EDV and the response to isosorbide dinitrate (ISDN) for the measurement of EIV. sTNF-Rs and hsCRP were examined by the ELISA method. Results: FMD was significant diminished in hypertensive pts. In 60th sec of reactive brachial artery hyperemia the luminal size has not reached the basal parameter (HT pts mean FMD: 92.0 T 9.9%, C group mean FMD: 109.5 T 7.8%, p G 0.001). EIV was significant reduced in HT pts comparing with C group (HT pts mean EIV: 105.2 T 18.3%, C group mean EIV: 118.6 T 10.5%, p = 0.01). In HT group significant elevated levels of hsCRP and sTNF-RI were found (HT pts mean hsCRP: 8.14 T 4.05 mg/l, C group mean hsCRP: 3.81 T 3.02 mg/l, p G 0.01; HT group mean sTNF-RI: 1161.6 T 168.1 pg/ml, C group mean sTNF-RI: 638.5 T 37.5pg/ml, p G 0.001). No significant difference in sTNF-RII levels between the groups was found. Correlation between sTNF-RI and rest luminal size of brachial artery was observed (r = 0.25, p = 0.05). No other correlation between EDVa EIV and inflammatory markers was found. Conclusion: Significant reduced endo thelium-dependent and endo theliumindependent vasodilation of brachial artery in HT pts was found. Significant elevated levels of hsCRP and sTNF-RI in these pts were measured, too. No significant correlation between ultrasound markers of endothelial dysfunction and inflammatory parameters was observed. Ultrasound assessment of endothelial dysfunction and inflammatory markers may give us an information concerning the severity and progression of disease. Pharmacogenetic study for predicting cyclophosphamide side effect in thai systemic lupus erythematosus Pintip Ngamjanyaporn 1 , Ammarin Thakkinstian 2 , Oravan Verasertniyom 3 , Monchand Vanichapuntu 3 , Kanokrat Nantiruj 1 , Kitti Totemchokchyakarn 1 , and Suchela Janwityanujit 1 . 1 Allergy Immunilogy and Rheumatology Division, Ramathibodi Hospital, Mahidol University, Medicine Department, Bangkok, Thailand; 2 Ramathibodi Hospital, Mahidol University, Clinical Epidemiology Unit, Bangkok, Thailand; 3 Ramathibodi Hospital, Mahidol University, Reseach Center, Bangkok, Thailand. Background: Systemic lupus erythematosus (SLE) is the serious autoimmune disease that primarily affect young women. Although cyclophosphamide improves outcome of SLE, frequent ovarian toxicity is a major side effect of this therapy. Cytochrome P450 (CYP), CYP2C19 polymorphism has been shown to significantly influence the pharmacokinetics as well as the ovarian toxicity of cyclophosphamide in Caucasian SLE patients. Objective: To assess association between CYP2C19 polymorphism and ovarian toxicity in Thai SLE patients who have been treated with cyclophosphamide. Patients and Methods: Female patients who were diagnosed as SLE according to ACR criteria and treated with cyclophosphamide were genotyped for CYP2C19 polymorphism. Ovarian toxicity was defined as sustained amenorrhea for more than 12 months occurring before 50 years of age or lack of menstruation for more than 4 months during treatment. Chi-square test was used for assessing association. Logistic regression was applied. Results: Seventy-one patients were enrolled for analysis. The mean age at diagnosis and starting cyclophosphamide was 26.42 T 8.35 and 29.27 T 7.82 years. The mean cumulative cyclophosphamide dosage was 23.36 T 20.81 grams. Thirty-six (59.7%) patients developed ovarian toxicity. CYP2C19*2 allele frequencies were 27.8% and 21.4% in ovarian and non-ovarian toxicity groups. Genotype frequencies in non-ovarian toxicity group were complied with Hardy-Weinberg equilibrium (p = 0.304). ROC analysis was applied to calibration cut-off of cyclophosphamide dosage and found that dosage of 23.75 grams yielded likelihood ratio for developing ovarian toxicity of 3.5. Logistic regression analysis was applied to assess interaction effect between cyclophosphamide dosage and CYP2C19 allele. We found that among CYP2C19*1 allele, patients received cyclophosphamide 23.75 grams or higher were 12.6 (95% CI = 3.3Y46.8) times more likely to have ovarian toxicity than patients received cyclophosphamide G 23.75 grams. This risk was decreased to e 5.1 (95% CI = 1.1Y22.7) times if patients had CYP2C19*2 allele. Conclusion: Cyclophosphamide dosage of 23.75 grams or higher carries a high risk of inducing ovarian toxicity. However, the risk can be decreased if patients have CYP2C19*2 allele. Background: Aspirin-induced asthma (AIA) is a distinct clinical entity and appears to be a specific subtype of asthma. There are lots of efforts to uncover AIA pathogenesis, but adverse reaction by aspirin is still remains unclear. Aspirin is a well known non-steroidal anti-inflammatory drug. The antiinflammatory action of aspirin generally has been attributed to direct inhibition of cyclooxygenases (COX-1 and COX-2). AIA takes part in 10% of patients with severe asthma. Theophylline has been demonstrated to have an antiinflammatory effects as an long-term medication for severe asthma and COPD. It works as a PDE inhibitor and a nonspecific adenosine receptor antagonist. Objective: To assess the therapeutic effects of aspirin, theophylline, and combination of two drugs in murine model of asthma. Methods: To generate a murine model of asthma induced by LPS with OVA, 6-week-old C57BL/6 mice were sensitized intranasally with 75 ug of OVA on days 0, 1, 2, and 7 with or without LPS, and then challenged intranasally with 50 ug of OVA on days 14, 15, 21, and 22. Whenever mice were challenged, aspirin (10mg/kg), theophylline (10mg/kg), and both were injected intraperitoneally. Asthmatic phenotypes such as lung inflammation using bronchoalveolar lavage (BAL) cellularity, cytokine production, and lung histology were evaluated on day 24. Results: Total cell number of BAL fluids in mice treated with each of Aspirin and theophylline was significantly decreased compared with non-treated group of mice. However, inflammatory cells in BAL fluids were increased in mice treated with aspirin and theophylline combination. And similar results were represented in histology. IP-10 protein, downstream of IFN-gamma, was decreased in each aspirin and theophylline treated mice compare with each of aspirin and theophylline non-treated group of mice. However, aspirin and theophylline combination treated mice were represented increase of IP-10 level in BALF. Moreover, only aspirin and theophylline combination treated mice were revealed specially increased level of 15-epi-LXA, which generated from lipid metabolite by aspirin. Conclusion: In murine model of asthma enhanced by LPS and OVA, each of aspirin and theophylline treatment is effective, but combinative treatment resulted in adverse reaction. Reza Amin, Ahmad Amin, and Soheila Alyasin. Nemazee Hospital, Pediatric-Allergy and Clinical Immunology Research Lab, Shiraz, Islamic Republic of Iran. Background: Beta-thalassemia major is one of the major health problems in our country. Many studies have confirmed the fact that, these patients have an increased susceptibility to bacterial infections. Objective: In this study, we have assessed the humoral immune system in 68 thalassemic patients by measuring their serum concentration of Immunoglobulin G (IgG), IgM, IgA, C3 and C4 in order to find out a responsible immune defect. Methods: Sixty eight beta-thalassemia major patients were enrolled randomly from referrals to Dastgheib clinic of thalassemia. The same number of case controls with matched age and sex were selected from healthy-people without any history of recent or recurrent infections. Serum IgG, IgM, IgA, C3 and C4 levels were assessed using Single Radial Immunodiffusion (SRID). Results: Serum levels of IgG, IgM and IgA were significantly higher (P G 0.01) and those of C3 and C4 were significantly lower (P G 0.01) in thalassemic patients than the controls. Considering the result of analytic tests, it was revealed that, thalassemia patients show much more increase in serum immunoglobulin levels as they get older. Splenectomized patients had higher serum IgG and IgA levels than non-splenectomized patients but had no difference in serum IgM, C3, and C4. Serum ferritin level had no correlation with the changes of humoral immunity; however, patients with serum ferritin level 9 2500 ng/ml had higher serum IgM level. Conclusion: These results can be due to continuous exposure to antigens, repeated infections, chronic liver disease and splenectomy but not iron overload. The only probable cause of humoral immune deficiency found in these patients is a defect in serum complement levels. Keywords: Thalassemia Major, IgG, IgA, IgM, C3 and C4 697 A rare cause of acute eosinophilic pneumonia Boris Golzman 1 , Svetlana Turkot 1 , Jakob Kogan 1 , Alex Rozov 2 , Arie Feldman 3 , Arela Rotlev 1 , and Shmuel Oren 1 . 1 Barzilai Hospital, Internal Medicine Department A, Ashkelon, Israel; 2 Barzilai Hospital, Pulmonology Unit, Ashkelon, Israel; 3 Barzilai Hospital, Internal Medicine Department B, Ashkelon, Israel. Introduction: Acute eosinophilic pneumonia (AEP) is a rare disease described as follows: an acute febrile illness with severe hypoxemia, diffuse pulmonary infiltrates, an increase in bronchoalveolar lavage (BAL) eosino-phils, no evidence of infection, asthma or atopic illness and complete resolution after treatment with corticosteroids. It is suggested that AEP is an acute hypersensitivity reaction to an unidentified inhaled antigen. We report a case of AEP that developed after Narghile water pipe smoking (WPS). Case report: An 18-year-old woman was admitted with a cough and high fever for the last 4 days. She had no previous history of asthma or atopic disease. There was no history of smoking, recent traveling or using medications. During the week before her admission she smoked a narghile water pipe. The patient was in moderate general condition, dyspneic, RR 20, Sat O2-90% in room air, T-38-C. The physical examination was unremarkable besides bilateral inspiratory rales. Chest X-ray revealed bilateral ground glass opacities. Peripheral WBC count was 14370 cell/mm 3 with 1465 eosinophils/mm 3 (10 %). IV antibiotics, oxygen support and inhaled bronchodilators were administered. Blood cultures were negative. The eosinophil count rose to 3700 eosinophils. Total IgE level was 943 IU/ml (normal range 0Y100). A chest CT scan revealed bilateral patchy ground glass opacities with no lymph node enlargement. Fiberoptic bronchoscopy was performed and BAL yielded 80% eosinophils. IV corticosteroid therapy was given with remarkable improvement. The peripheral blood eosinophil count returned to normal (1.7%) in two days. The patient was discharged in good condition and 3 months later she was asymptomatic. Discussion: Our case met the 6 diagnostic criteria of AEP: an acute illness of short duration, high fever, severe hypoxemia, bilateral infiltrates on chest radiographs, eosinophilia more than 25% on BAL, and no history of drug hypersensitivity or evidence of infection. There are reports of AEP in new cigarette smokers. Narghile tobacco smoke contains tar, nicotine, carbon monoxide and heavy metals and it is higher than in cigarette smoke. Water-pipe smokers may absorb higher concentrations of these substances due to the mode of smoking (higher frequency of puffing, depth of inhalation). A search in PUBMED revealed no case reports of AEP that evolved after narghile water pipe smoking. In our case it was the only etiologic factor that has emerged through detailed history. Serum concentrations of eosinophil cationic protein and eosinophils of patients with Kimura's disease Nobuo Ohta, Shigeru Fukase, Yusuke Suzuki, Shinnichi Sakurai, Masaru Aoyagi, and Shinichi Okazaki. Yamagata Univ., School of Medicine, Department of Otolaryngology, Yamagata, Japan. To clarify the role of eosinophils in the pathogenesis of Kimura's disease and the values of measuring serum levels of eosionophil cationic protein (ECP) for monitoring disease activity, a total 14 serum and 7 tissue samples from patients with Kimura's disease were studied. The concentrations of ECP and cytokines (interleukin-4 (IL-4), granulocyte-macrophage colonystimulating factor (GM-CSF), and interleukin 5 (IL-5)) in sera from patients with Kimura's disease were measured by enzyme-linked immunosorbent assay (ELISA). The density eosinophils and the degree of activation of eosinophils in the tissue were also studied immunohistochemically. The concentration of ECP in sera from patients with Kimura's disease was significantly higher than that in the control group (p G 0.05). At the time of the remission, a significant decrease of ECP was observed. In interfollicular areas, most infiltrated eosinophils were positive for EG2 antibody (64.0Y94.0%) and the mean percentage of EG2-positive eosinophils was 75.7%. The concentrations of IL-4, GM-CSF, and IL-5 in sera from patients with Kimura's disease were within normal ranges or below the detectable level in all sera examined. Our findings suggest that eosinophils play an important role in the pathogenesis of Kimura's disease and ECP may be used as an additional parameter of disease activity. The evaluation of osteocalcin and osteoprotegerin, osteogenic markers of bone remodeling in rheumatoid arthritis Cioaca Gabriela Ramona, Drugarin Doina, and Paunescu Virgil. Victor Babes University of Medicine and Pharmacy Timisoara, Department of Physiology and Immunology, Timisoara, Romania. : The purpose of this study is to evaluate some osteogenic markers involved in bone remodeling, such as the estimation of osteocalcin (OC) and osteoprotegerin (OPG/RANK) concentrations in the sinovial fluid as well as in the serum of rheumatoid arthritis (RA) patients. These measurements were performed in the initial stage of the disease, without using any antiinflammatory non-steroidal therapy for at least 1 month before our study began. The concentration of OPG in the sinovial fluid represented 14.25 T 0.75 pmol/l while in the serum the concentration was 1.69 T 0.80 pmol/l, which was significantly increased (p G 0.001) compared to the values determined in the serum of healthy controls (1.19 T 0.30 pmol/l). The increase of OC concentration evolves in the same manner in both sites of our measurements, the values being of 6.43 T 4.80ng/ml and 19.9 T 4.8ng/ml respectively, compared to the values determined in healthy controls, in the sinovial liquid (G 0.5ng/ml) and in the serum (3.1 T 0.1ng/ml). Background: Undernutrition in children is associated with increased susceptibility to infections and depressed effect on cell-mediated and humoral immunity. Probiotics can enhance immune responses which can lead to illness prevention or decrease duration and severity. Objective: The study aims (1) to determine baseline immune markers (CD3,CD4, CD8, CD19, CD 20, CD56) of undernourished Filipino children aged 3Y5 years old (2) to determine whether probiotics can enhance immunity by (a) changes in immune markers, (b) changes in weight and (c) decreased incidence and duration of common infections (e.g., colds, pneumonia and diarrhea). Methodology: Undernourished children were randomized into control and experimental group. Probiotics (mixture of Lactobacillus bulgaricus, L casei subsp rhamnosus, L fermentum, L plantarum, Bifidobacteria bifidus, B breve, B infantis, B longum, and B lactis.) were given 2 times a day for eight weeks. Weekly weight, incidence and duration of illnesses were recorded. Blood for immune markers was drawn before and after the study. Results: 29 children were enrolled, 14 in the control and 15 in experimental group. The basal levels CD3, CD4, CD8, CD19, CD 20, CD56 were normal in both groups. The control group showed an increase in CD4, CD8 levels compared to the probiotics group but both groups' levels stayed within normal limits. The probiotics group gained significant weight by 1 kg (p G 0.003). The incidence of illnesses (diarrhea and colds) was the same for both, but there was significant shorter duration for the probiotics group. The median was 2 and 14 days for the probiotics and control group respectively (p G 0.001). Conclusion: Probiotics lead to significant weight gain. They also lead to shorter duration of infections. Habibeh Mozaffari 1 , Zahra Pourpak 2 , Neda Entezari 2 , Chavoshzadeh Zahra 2 , Mohammad Hassn Bemanian 2 , Mohammad Gharagozloo 2 , Masoud Movahedi 2 , Asghar Aghamohammadi 2 , and Asghar Aghamohammadi 2 . 1 Immunology, Asthma and Allergy Research Institute, Medical Sciences/ Tehran University, Tehran, Islamic Republic of Iran; 2 Immunology, Asthma and Allergy Research Institute, Immunology Department, Tehran, Islamic Republic of Iran. Background: Leukocyte Adhesion Deficiency (LAD) is a rare congenital immunodeficiency with fewer than 200 affected individuals in the world which is characterized by sever recurrent infections. The purpose of this study was to describe the survival time of Iranian patients with LAD over a period of 16 years. Methods: Twenty seven patients with definite diagnosis of LAD (based on standard criteria and laboratory tests), attending Allergy, and Immunology clinic in Children Medical Center Hospital-from 1991 to 2006 ranging in age from 3 months to 24 Years old, were studied and followed. Severity of disease was defined as a presentation of CD18 marker on the surface of leukocyte (less than 1% of normal expression). Also a questionnaire was filled for recording all necessary information. Results: All patients (63% male, 37% female) was LAD type I. The prevalence of consanguineous marriage was 81.5%. The mean age at the time of disease onset was 0.78 T 2.03 months and the mean age of diagnosis was 1.33 T 0.55 months. Overall 33.3% of patients (9 patients) had severe form of disease. The overall 5 year survival rate was 67% and mortality rate was 22.2%. The mean of survival time for alive patients was 73 T 27 months (CI:21Y125). The main leading cause of death was bacterial infections. Conclusion: Mortality rate in other studies mentioned about 4% which is very lower than our study (20.2%). Diagnostic delay can result in high mortality and more complications in untreated patients and failure to provide therapy can be a cause of low survival rate. Therefore early diagnosis, good control and appropriate treatment will improve the survival rate. Acute myocarditis after steroid withdrawal in an asthmatic patient Elena Ossi 1 , Claudio Bilato 2 , Annamaria De Pellegrin 1 , Andrea Rossanese 1 , Marilisa Andretta 3 , and Mauro Cancian 1 . 1 Clinica Medica 1, Medical and Surgical Sciences Dept., Padova, Italy; 2 Cardiac Unit, Cardiology Department, Padova, Italy; 3 ENT Clinic, Medical and Surgical Specialities Dept., Padova, Italy. A 64-year-old Caucasian man presented with febrile dyspnoea one month after having withdrawn steroidal therapy previously prescribed to treat asthma of recent appearance associated to a marked increase of serum IgE (2493 U/L). The finding of increased troponin and myoglobin levels on hospital admission, seemed to point to a cardiac ischemia, although ECG showed only sinusal tachycardia and despite of a leucocytosis with 60% eosinophils, Thus, he was directly referred to the cardiac intensive care unit, where he underwent a cardiac catheterization and a stent was inserted in the left anterior descending coronary artery. Three days later, cardiac enzymes started to increase again (troponin up to 64 2g/L) and the clinical picture worsened with severe dyspnoea and fatigue. For this reason, he had a further catheterization done, that showed no problem with the stent. In the meantime, paresthesias and neurological pain, slightly present on admission, worsened to a severe stage. The temperature was still high and cutaneous signs of vasculitis appeared. At this point a cardiac magnetic resonance was performed which revealed a diffuse subendocardic necrosis. Finally, thinking of a Churg-Strauss Syndrome (CSS), a myocardial biopsy was obtained and ANCA were tested, both confirming the working hypothesis. Moreover, EMG showed a severe mixed multineuropathy. High-dose-methylprednisolone was started, with prompt clinical improvement and normalization of cardiac enzymes in 10 days of therapy. CSS is a vasculitis characterized by asthma, hypereosinophilia and involvement of other organs. Being CSS more prevalent in the fifth decade, when a novel asthma presents in an individual aged 60 or more, it is common to think of a previously latent atopy, made overt by the concomitant intervention of precipitating factors, or of a cardio-pulmonary disease. In the present case report, the steroid withdrawal in a novel asthma of an aged patient caused the burst of a severe carditis in an undiagnosed CSS. Does breast feeding protect against asthma and other atopic diseases? Mohammed Herrag, and Mustapha Iraqui. Ibn Sina University Hospital, Pulmonology-Allergology, Rabat, Morocco. Rationale: Breast feeding is associated with the lowest asthma and allergic disorders rates and it significantly reduces the severity of respiratory illnesses during the first year of life. Methods: we had included questions related to breast feeding in the ISAAC questionnaires (the international study of asthma and allergies in childhood) to see the outcome of allergic diseases in both groups: breast feeding and non breast feeding. Results: Mean (S.D) age: 9 years, weight: 30 kg and height: 135.6 cm. Children breastfed more than 8 months had significantly lower prevalence rates of persistent cough in the past year, ever rhinitis, rhinitis in the past year, and itchy-watery eyes in the past year than those who were not breastfed (18% vs 25%; 22% vs 32%; 15% vs 23%; and 6.5% vs 14.2%, respectively). Incidency was lower of asthma and wheeze in breastfed children than those who were not breastfed (asthma: 2.5y vs 4.5 y; 2.2y vs 3.9y, respectively). Significantly higher proportion of children who breastfed more than 8 months were resolved from asthma, wheeze and eczema than those who breastfed less than 8 months (89% vs 76%; 90.7% vs 77.2%, and 97.4% vs 82.2%, respectively). Conclusion: Our Study shows the impact of breast feeding for at least 8 months in protecting and reducing the prevalence of allergic disorders. Methods: Patient JPA male, 5 years, consulting by Asthma and rhinitis perenneal worsening in fall and winter and a previous history of atopic eczema at 2 years. Objective examination was normal and prick skin tests only positive to house dust mites. Blood analysis were requested. Meanwhile an acute respiratory illness treated symptomatically by G.P. On analysis PBC count was normal, IgG 891 mg/dl IgA 105 mg/dl IgM 155 mg/dl. In electrophoresis a peak was detected which leads to an immunofixation showing an IgMK monoclonal gammopathy. Total IgE was a 160 Ul/ml. Specific IgE 1,5 KU/l for D.pteronyssinus and 2,23 KU/l for D.farinae. A battery of antibodies to Virus, Mycoplasma and Leishmania which is slightly endemic in south of Tagus river was requested. One week after new immunofixation showed a marked decrease of monoclonal peak and IgM decreased to 113mg/dl. No light chains in urine. 2 weeks after antibodies to EBV, CMV, Chlamydia pneumoniae Leishmania were negative. Antibodies to Mycoplasma pneumoniae positive: IgG 2,54, IgM 3, 3 (normalG0, 9) . 4 weeks after new immunofixation was done with absence of the monoclonal peak. Conclusion: Secondary gammophaties can be included in the broader group of monoclonal gammopathies of indetermined signification (MGUS). In general and with the exception of Leishmania infection a clear monoclonal peak is not detected after an acute infection. IgM is the immunoglobulin implied in primary response to infections. In children the occurrence of monoclonal gammopathies is very rare and mainly described for CMV and EBV. Transient monoclonal gammopathy is characterized by a fast decrease of the monoclonal peak after the infection. In our case a recent Mycoplasma pneumoniae infection was shown by positive serology for IgG as well for IgM. This young patient has a slight increased concentrations of total IgE and the values of specific IgE for house dust mites were not very high. The fast decrease and disappearance of the monoclonal IgM peak points out to a transient monoclonal gammopathy in an atopic child triggered by a Mycoplasma infection. Mozhgan Moghtaderi 1 , Reza Amin 1 , and Soheila Alyasin 2 . 1 Nemazee Hospital, Pediatrics-Allergy and Immunology, Shiraz, Islamic Republic of Iran; 2 Nemazee Hospital, Immunology and Allergy, Shiraz, Islamic Republic of Iran. Hyper IgE syndrome with recurrent infections is a rare immunodeficiency characterized by recurrent skin infection and pulmonary abcess and extremely elevated levels of IgE in serum, associated facial and skeletal features have been recognized. Their frequency and genetic basis is poorly understood. The patient is a seven-year old girl presented with two years history of productive cough, superinfected facial eczema from infancy and two prior hospitalization for pneumonia and perianal abcess. She had coarse face, crackles on both lungs, clubbing of fingers, laxicity of elbow joints. Immunological work up revealed eosinophilia and markedly raised IgE levels. In view of the clinical features and other finding diagnosis of hyper IgE syndrome was suggested. Chest X-ray revealed multiple large cystic lesions in left total lung which confirmed by spiral CT-scan. Left thoracotomy and pneumanectomy was done and removed severe adhesions. Histological report revealed cystic adenomatoid transformation. CAT is Characterized by the presence of variously sized cysts lined by epithelium. Probably, this is the first report of CAT in a child with hyper IgE syndrome due to repeated infections. Early diagnosis and prophylactic antibiotic therapy is effective in management of these patients. Pulmonary complications and some changes in immune reactivity on the course of acute heroin intoxication Julia Radenkova-Saeva 1 , and Bogdan Petrunov 2 . 1 Emergency Hospital, Department of Clinical Toxicology, Sofia, Bulgaria; 2 NCIPD, Department of Immunology and Allergology, Sofia, Bulgaria. Objective: To examine the complications in pulmonary system and changes in immune reactivity (some parameters of humoral and cell mediated immunity) on the course of acute heroin intoxication. The study includes 16 patients, with acute heroin and mixed with other psychoactive drugs intoxications at average age, 21,5 T 5.04 years (12 men and 4 woman), hospitalized in the Clinic of Toxicology, MHATEM BN.I.Pirogov^, Sofia. We have used clinical, clinicolaboratory, immunological, chimicotoxicological, instrumental methods. Results: Results from the study shows that in severity intoxications with heroin and other psychoactive drugs we observed spectrum of complications in pulmonary system -pneumonia, aspiration of gastric contents, pulmonary edema, and ARDS. Death is registered in 3 persons. It was found some changes of the immune reactivity: statistically significant lower mean levels of IgG, IgA, IgM and tendency to lower mean levels of complement components -C3 and C4 in the studied patients in comparison with the values in healthy people. The changes were more demonstrative in the group with pulmonary complications compared to the group without pulmonary complications. We observed that the CD4 lymphocytes are significantly less in the studied patients and is determined lower level of CD56 bearing lymphocytes -natural killer /NK/ cells, compared to healthy controls. Conclusion: The acute heroin and mixed with other psychoactive drugs intoxications leads to complications in pulmonary system, and changes of some parameters of cell mediated and humoral immunity. Key words: intoxication, heroin, psychoactive drugs, immune reactivity, noncardiogenic pulmonary edema, pneumonia, ARDS 707 Atypical presentation of polyarteritis nodosa: a case report Horacio Del Olmo Tellez, Yamazaki Nakazhimada, Marco Antonio, and Francisco Espinosa Rosales. National Institute of Pediatrics in Mexico, Allergy and Immunology, Mexico City, Mexico. Polyarteritis nodosa is a systemic vasculitis that affects median sized muscular arteries, characterized by multi organ involvement. Presenting GI manifestations include: abdominal pain, nausea, vomiting, diarrhea, acute abdomen and, rarely, GI bleeding in the form of hematemesis, melena and occult bleeding. A 14 year old boy presented five years ago with hemetochezia, 5 to 7 episodes per day. he presented severe anemia and he was hospitalized to investigate mieloproliferative disease or systemic lupus erythematosus He was treated with IVIG and methylprednisolone with clinical and hematic biometry improvement. A colonoscopy showed suggestive data of CUCI with diffuse inflammation, pseudopolyps, adhered fibrin to mucosa, inflammatory lesions involving descendent colon and sigmoides, estenotic parts of the colon and bleeding. Hystologic examination described ulcer and inflammatory infiltrate with polimorphonuclears, limphocytes, plasmatic cells in lamina propia and submucosa, mild fibrosis and eosinophils. He received treatment with methylprednisolone, mesalazine, omeprazole, and azathioprine and he was referred. Physical examination revealed severe emation 45.4 kg, hepatomegaly, splenomegaly, bilateral pulmonar hipoventilation with bilateral crackles. The diagnosis of severe desnutrition, CUCI, and communitary pneumoniae were made, and treatment with cefuroxime and claritromicine was started. He presented fever, rectorragia with hemodinamic compromise that required transfusion of packed red blood cells, and platelets, and involvement of liver, kidney, gut and persistent fever, pancytopenia and elevated ferritin. He required orotraqueal intubation because ventilatory compromise, in the neurology area, he had hypotonic proximal extremities, hiporreflexia, and generalized seizures, which were controlled with phenytoin. The laboratory findings revealed negative, antinuclear antibody, perinuclear antineutrophilic cytoplasmic antibody, hepatitis B surface antigen, and anti hepatitis C virus were negative. Central nervous computed tomography showed isquemic temporal areas. Magnetic resonance and angiography revealed microaneurysm in both renal arteries, and vasculitis in central nervous system, hepatic and intestinal arteries. These symptoms and laboratory findings were compatible with diagnostic criteria for childhood classic polyarteritis nodosa, treatment of IVIG and various methylprednisolone pulses let to clinical improvement. Background: According to Evidence Based Medicine, conclusions from meta-analyses and RCT represent the most solid evidence source to assess the efficacy of an intervention. Specific immunotherapy (SIT) is currently considered as the only allergen-oriented biological response modifier, and it is regarded as an essential part of the therapeutic approach for allergic diseases. Objective: To provide a descriptive review of all available randomized, double-blind and placebo-controlled trials (RCT BD PC) published which evaluated the efficacy of SIT in the treatment of respiratory allergy. Study Design: Systematic review. Methods: A comprehensive search of the EMBASE and MEDLINE databases up to April 30th, 2007 was carried out. The search strategy retrieved citations containing the subject heading specific immunotherapy limited to RCT DB PC combined with asthma, allergic rhinitis or both. Variables recorded were the study design, journal impact factor, patient's and immunotherapy characteristics, clinical and laboratory outcomes and online full-text papers' availability. Results: Two hundred ninety-six articles were identified and reviewed; 179 (61%) were RCT DB placebo-controlled. Of them, 116 evaluated efficacy of subcutaneous IT (SCIT) and 63 sublingual IT (SLIT). Twenty-six percent of studies (n= 46) had a minimum duration of 24 months (21% vs. 30% for SCIT and SLIT, respectively). Characteristics of studies are described in Table. (+ Median-Interquartilic rank; # Number of studies found; * Months; ** Mean; @ At least one immunological mechanism studied) Conclusion: One hundred seventy-nine Ib evidence-level studies on efficacy of SIT in the treatment of respiratory allergy were identified in two electronic databases. Both selection and search bias must be ruled out. The effect of antihistamine pretreatment on side effects during specific immunotherapy Zoran Arsovski, Dejan Dokic, Miroslav Gavrilovski, Biserka Kaeva, Zlatica Goseva, and Olivera Caparoska. Clinic of Pulmonology and Allergy, Scientific Department, Skopje, Macedonia, Fyrom. During one year of our study in a group of 47 patients we have conducted the specific immunotherapy (SIT) with House Dust Mite (HDM) Novo Helisen Depot extracts and pollen Allergovit extracts. 493 shots were applied and 32 local reactions occurred (6,49%). All of them were 9 5 cm and all of them occurred before the maintenance dose was reached. No systemic reaction or life-threatening reactions were observed. In a group of HDM treated patients 5,21% of injections resulted in large local reaction versus 7,45% in a group of patients with pollens. There was no statistical difference between those two groups p=0,31. Reductions in IT-schedule were performed due to severe local reactions. Only 6 patients experienced more than ones a large local reaction. Four of them were on pollen immunotherapy and two were on HDM immunotherapy. All of these patients have had skin reactivity on skin prick test (SPT) more than 3+. We have decided to use loratadine 10mg as antihistaminic premedication in these patients. After the administration of the pretreatment antihistaminic drug, patients did not experience any severe local reaction. We can conclude that in a very limited number of patients who underwent SIT (especially in those with very extensive cutaneous reaction to SPT) it could be useful to use antihistaminic premedication to reduce/secure the number and severity of cutaneous reactions. Shorter dosing intervals of sublingual immunotherapy lead to more efficacious treatment in a mouse model of rhinitis Carola Rask, Jens Brimnes, Sandra Urioste, and Kaare Lund. ALK-Abelló, Experimental Immunology, HLrsholm, Denmark. Background: Sublingual immunotherapy (SLIT) with allergen extract has been shown to be efficacious in treating allergic rhinitis and asthma. Clinically, SLIT is performed with varying doses of allergen extracts and with varying intervals of dosing, and although a dose-response relationship has been established in SLIT, the importance of dosing intervals remains to be documented. The objective of this study was to explore the importance of the dosing intervals in SLIT. For this purpose we used a mouse model of allergic rhinitis, induced by a clinically relevant allergen, timothy grass pollen extract. Methods: Mice were sensitized to Phleum pratense (Phl p) by intraperitoneal injections of alum-adsorbed allergen extract. Sensitized mice were SLIT-treated with different intervals and with different doses of Phl p extract. In order to study the importance of dosing intervals, doses of 25 kSQ, 58 kSQ or 175 kSQ were given sublingually to mice 7, 3 or 1 day per week, respectively, resulting in a weekly cumulative dose of 175 kSQ for all groups. Two additional groups receiving the lower dose (25kSQ) of Phl p one or three days per week were also included. Following nine weeks of SLIT, all mice were challenged intra-nasally with Phl p. Antibody levels, T cell responses and numbers of inflammatory cells in bronco-alveolar lavage (BAL) were used as read-outs for the efficacy of SLIT in the different treatment groups. Results: Increases in the Phl p specific IgA antibody levels showed a strict positive correlation with increased dosing frequency. The levels of Phl p specific IgE were reduced to the same extent in all groups, but both the systemic-(spleen) and local (draining lymph nodes) T cell responses decreased with increased dosing intervals, indicating a positive effect of higher dosing frequency on T-cell tolerance. In addition, the maximal reduction in allergen-induced inflammation in the lungs (BAL) was seen in the group receiving SLIT for 7 days per week. Conclusion: The group receiving SLIT for 7 days per week was the only group with demonstrated positive effects of all immunological parameters measured. Our data demonstrate that although the effect of SLIT is dosedependent when using the same allergen extract, a once daily regimen is more efficacious than regimens where SLIT, with the same weekly cumulative allergen dose, is administered with longer intervals. Faster reach to maintainance pleteau by delayed rush immunotherapy Naren Pandey. Bellevue Clinic, Allergy & Asthma, Kolkata, India. Introduction: Intermittent Rush immunotherapy was tried on some patients to evolve some faster and affordable immunotherapy modality to make the patient achieve the maintenance pleatu within a very short time span which does not also involves hospital stay. Conventional method of immunotherapy is administered with long durations, rush immunotherapy is super fast methodology in attaining the maintenance/boosting module, which requires hospitalization and other precautionary methods, and multiple allergen vaccines to be administered within short span of time. But in this method which was tried it was found that within a duration of around 15-20 days the relief of the immunotherapy was reached. Place of study: 1) Belle Vue Clinic, kolkata, 2) Hope Super Speciality Hospital, Purnea. Materials and Methods: 62 patients out of which 46 with urticaria allergy and 16 with allergic Rhinitis. Were subjected for Intermittent Rush Immunotherapy schedules. The therapy consists of administration of four vials of allergen extracts, 1st vial : 1:25,000 dil, 2nd vial 1:2,500 dil, 3rd vial 1:250 dil, 4th vial 1:50 dil. The 1st & the 2nd concs were administered in daily schedules in a graphically rising manner. The patients had been given pre-medication before the study. Follow-up of the patients included blood examination IgG & IgE level estimation, before & after 8 weeks after the procedure. Results: Some of the patients showed local skin reactions which subsided on duration of time no drugs were required and 7 patients developed systemic reactions which were managed by Deriphylline & Decdan injections. There was substantial decrease in IgE level & increased IgG level was marked, significant & marked satisfactory relief was observed in the patients symptomatology, thus the procedure may be graded as a very fast & affordable immunotherapy. A two year double blind placebo controlled study on immunotherapy with single and mixed allergen preparation Shailendra N. Gaur 1 , Deepsikha Srivastava 2 , Naveen Arora 2 , and Bhanu P. Singh 2 . 1 Vallabhbhai Patel Chest Institute, Department of Respiratory Medicine, Delhi, India; 2 Institute of Genomics and Integrative Biology, Allergy and Immunology, Delhi, India. Background: Studies on immunotherapy with single allergen extract have been performed to determine the efficacy in asthma and rhinitis patients. However, very few studies have been performed comparing single and mixed allergen immunotherapy in a double blind manner. Here, we have carried out a double blind placebo control trial to assess the efficacy of single and mixed allergen immunotherapy based on various clinico-immunlogic parameters. Methods: One-hundred allergic patients were screened by history, skin tests and specific IgE levels. Sixty patients fulfilling the criteria of the study were divided randomly into two groups Y (a) active-single allergen, (b) activemultiple allergen and (c) -placebo containing 20 patients each. Single allergen immunotherapy was given with housefly extract. In mixed allergen immunotherapy, insect, pollen and fungal allergen extract were used in vaccines. In vivo parameters such as skin test and airway reactivity were assessed at baseline and after 2 year`s of immunotherapy. In vitro parameters such as allergen specific IgE, IgG1 and IgG4 were also assessed. Apart from these, symptom/drug score cards were filled by the patient and analyzed every 3 month. Efficacy of the treatment was analyzed in terms of clinical (skin test, symptom score, and airway reactivity) and immunological parameters (IgE, IgG4, IgE/IgG4, and IgG1/IgG4). Results: Patients getting active immunotherapy in both single as well as multiple allergen group showed significant improvement (pG 0.01) in disease state in comparison to placebo patients. Skin tests and IgE values however showed non-significant reduction compared to baseline. There was a significant improvement (pG 0.01) in airway hyperreactivity, symptom score and IgG4 levels after 2 year of immunotherapy in active group. Also good correlation was observed between symptoms reduction and increased IgG4 levels. All the above parameters were non-significant in placebo group. In the group treated with the single extract, fewer patients suffered with asthmatic symptoms after 2 years of IT and required less medication than in the group treated with the multiple extract. However, these differences, as well as differences in other clinical and immunological parameters were not statistically significant. Conclusion: Both single and mixed allergen immunotherapy for 2 years have shown improvement in clinical and immunologic parameters of asthma and rhinitis patients. Vipul Shah. Allergy Clinic, Allergy, Surat, India. Introduction: Allergic conjunctivitis is many times an ophthalmologist's nightmare. Antiallergic eye drops which often contain corticosteroids, may temporarily bring relief of symptoms but may bring an early cataract, among other side effects. Allergy testing and immunotherapy based on the test results can cure a significant number of these patients and provide relief to others. Methods: 24 patients of allergic conjunctivitis were subjected to modified skin allergy testing. Out of these 10 patients also had associated allergic rhinitis and asthma. Common allergens found in these patients were: Dust, Dustmites, Pollen, Fungi & Food. Based on test results, vaccines were prepared for individual patients and immunotherapy given. The patients who did not significantly respond to conventional medicines and vaccine, we reviewed the history to discover any missed out allergen. A specially designed indigenous Bpersonal volumetric air sampler^with safranine stained glycerin coated slides inside, was kept in patients home, office and surrounding atmosphere. Results: Out of 24, 18 patients (75%) started improving within 3 months of starting immunotherapy. After 6 months their eye drops and other medicines were stopped. Restricted food items started one by one after 6 monthes. They are symptoms free for last 12 to 2 years. For 6 patients who did not respond to above therapy, specially designed Bair sampler^brought great surprises. The fungi and pollens, which were not tested, previously were found. Also three new pollens were discovered. Appropriate immunotherapy made 5 out of 6 patients symptoms free. Conclusion: Allergy testing and immunotherapy can cure allergic conjunctivitis. Personal volumetric air sampler is a very useful tool to find out additional allergens in case of resistant cases. Quality of Life and Symptoms assessment in sublingual immunotherapy for patients with house-dust mite related perennial rhinitis: definition of a responder profile Background: The efficacy and safety of sublingual immunotherapy (SLIT) in well-established in seasonal allergic rhinitis. However fewer data were available regarding perennial allergic rhinitis. The aim of this study was to assess the relationship between the effect of SLIT on symptoms and Quality of Life changes in patients with perennial rhinitis. Methods: This exploratory phase IIIb, double-blind, placebo-controlled trial was conducted during years 2003 to 2005 in the Cape-Town area (South Africa). Sixty patients having house-dust mite-induced allergic rhinitis were enrolled. Patients were randomized to receive, during 2 years, Dermatophagoides pteronyssinus SLIT solution (Stallergenes, Antony, France), with a maintenance dose of 300 IR (n = 39), or placebo (n = 21). The primary efficacy endpoint was the mean T5SS [Total Symptom Score for five symptoms: sneezing, runny nose, nasal congestion; ocular redness/itching/ tearing; itchy nose/throat/ears. The limit percentage of improvement was fixed at 60% for the Bresponders^. Rescue medication intake, individual symptom scores and quality of life (QoL-RQLQ) were assessed as secondary endpoints. Results: The intention-to-treat (ITT) population included 55 patients (mean age: 32.93 yrs T 11.31). The mean T5SS change was lower for the 300 IR [-6.57 ] group compared to placebo [-5.02 ], but the difference did not reach significance. The mean percentage of days with rescue medication, as well as each individual symptom score were lower for the 300 IR group compared to placebo. The percentage of good responders is significantly higher in the active group (p=0.0405). For the QoL, the percentage of good responders the ocular score is higher in the active group (p=0.0512), reaching significance. The association between good responders to QoL and good responders to T5SS gives an odds ratio = 15 (pG0.0001) for all patients (OR = 8.75, p=0.0061 in active group) showing a good correlation. The most frequent adverse events were oral pruritus and throat irritation. No related SAE occurred throughout the study. Conclusion: This exploratory study gives promising results with a two-year treatment using Dermatophagoides pteronyssinus SLIT solution at a maintenance dose of 300 IR three times a week. A population of Bresponders^may be defined as patients with an improvement of 60% or more of the clinical symptoms. QoL is strongly correlated to clinical symptoms. Measuring quality of life in the treatment of allergic rhinitis with specific immunotherapy -identifying the best suitable instrument Ulrik Vestenbaek. ALK-Abelló, International Marketing, Hoersholm, Denmark. Background: Quality of Life (QoL) is becoming an increasingly important clinical outcome parameter, also in treatment of allergic rhinitis (AR). In AR QoL is often assessed with Rhinoconjunctivitis QoL Questionnaire (RQLQ)a disease specific instrument. For pharmacoeconomic evaluation a generic QoL instrument must be used. RQLQ only detects symptoms in the upper airways. Due to a strong epidemiologic relationship between AR and asthma suggesting that both are manifestations of the same disease, an instrument also detecting symptoms in the lower airways seems more appropriate. To show the full impairment of QoL caused by AR and to perform pharmacoeconomic evaluation it is important to identify the right generic instrument to be used in clinical trials with specific immunotherapy (SIT). Methods: QoL gain with the grass allergen tablet (Grazax, ALK-Abelló) against AR was assessed in a randomised, parallel-group, double-blind, placebo-controlled trial with 634 subjects using the generic EQ-5D questionnaire. EQ-5D contains 5 dimensions: mobility, personal care, usual activities, pain/discomfort and anxiety/depression, with 3 levels within each dimension. For each of the 5 dimensions the average score was calculated. It was then tested if the difference in scores between active and placebo was significantly different (t-test). Results: The treatment with the grass allergen tablet significantly increased QoL compared to placebo (pG0.001). The difference between groups was mainly due to differences within the dimensions: Usual activities and Pain/ discomfort. Mobility did also produce a borderline significant difference (Table 1) . Although there is a significant increase in QoL for subjects receiving treatment with the grass allergen tablet, the increase was only justified in 2 of the 5 dimensions. EQ-5D is a very crude instrument and might not be optimally suited to detect QoL impairment in a disease like AR. This implies an underestimation of the QoL result with the grass allergen tablet. An instrument with a more detailed focus on usual activities could be more relevant to use within SIT and AR in future studies. Examples of potentially well suited instruments are SF-36 and EQ-15D. Background: There are no studies comparing the effects of immunotherapy and antileukotrienes. The main limit is the long duration required to appreciate the clinical effects of immunotherapy. We compared the effect of montelukast or SLIT, added to standard therapy in moderate persistent asthma up to 4 years. Methods: Open controlled randomized trial. Patients with moderate asthma (and rhinitis) solely due to birch pollen were randomized to receive either monteukast MK (20 mg/day) or birch-SLIT (Anallergo, Florence, Italy) in the pollen seasons, in addition to formoterol 25 mcg+fluticasone 500 mcg twice daily. All patients received also salbutamol and cetirizien as rescue medications. Asthma and rhinitis symptoms were evaluated by diary card February-may at baseline and after 2 and 4 years of study. In-season nasal eosinophils and outseason bronchial hyperresponsiveness were evaluated as well. Results: Thirty-three adult patients were enrolled and 29 completed. The groups were homogeneous at baseline. In the SLIT vs MK e group the total asthma symptom score at baseline, 2 and 4 years were: 178 vs 180 (NS), 76 vs 183 (G0.001) and 42 vs 167 (G 0.001). At the same timepoints the bronchial scores were 80 vs 92 (NS), 48 vs 87 (0.01) and 25 vs 81 (G0.01). In both group there was a significant decrease in the use of bronchodilators but in the SLIT group there was a significant decrease of nasal eosinophils and an increase of outseason bronchial hyperresponsiveness as compared to baseline and to MK group. In patients with grass-pollen induced moderate asthma and rhinitis the addition of SLIT provides a greater clinical benefit compared to MK. SLIT response in children with HDM and cockroach Nagendra Prasad Komaral, and Paranjoty Kanni. Bangalore Allergy Centre, Allergy, Bangalore, India. Background: Recent times the allergy disorders is increased and most of the therapeutic approach is incomplete and yet not promising. In recent time the SLIT emerged with patient friendly Immunotherapy. This became a good boon among the children who were depend only on the symtpmatic drugs. This SLIT became an eye opener to Physicians and to parents to control the allergy symptoms. SLIT is introduce in Indian continent and made it possible at affordable price and showed good response in preventing the allergies. Methods: A total 112 children aged about 2 to 14 yrs from various parts of India who were previously diagnosed with Allergic Rhinitis and Allergic Asthma and reported receiving allergy and asthma medication took part in this study. These children were tested with aeroallergens by SPT and selected for SLIT who were sensitive to House dust mite (body and fecal matter) sensitivity and among insects who were sensitive to Cockroach. Results: Most of the patients' parents reported with good response and the pharmacotherapy drugs relatively reduced over a period of time and the symptoms drastically came down with increase PEFR. The drop outs were more fascinated to the alternative medicine and they continue to search for the relief from one system to another with continued suffering. Some reported frankly and some said they feel better. Conclusion: The results of this study provide insight into how the SLIT greatly influence the therapy and increase the quality of these children and the normal growth pattern which is appreciated with most of the parents. Background: There are few study comparing the effects of immunotherapy and inhaled steroids. The main limit is the long duration required to fully appreciate the clinical effects of immunotherapy. We compared the effect of inhaled budesonide vs SLIT in mild persistent asthma up to 4 years. Methods: Open controlled randomized trial. Patients with mild persistent asthma and rhinitis solely due to grass pollen were randomized, after a run-in season, to receive either budesonide 800 mcg/day in the pollen season or grass-SLIT (Anallergo, Florence, Italy) continuously for 4 years. Rescue medications were prescribed to all patients. Asthma and rhinitis symptoms were evaluated by diary card may through july at baseline and after 2 and 4 years of study. In-season nasal eosinophils and outseason bronchial hyperresponsiveness were evaluated as well. Results: Fifty-one patients were enrolled and 46 completed. The groups were homogeneous at baseline. In the SLIT vs budesonide group the total nasal symptom score at baseline, 2 and 4 years were: 120 vs 92 (NS), 56 vs 99 (G0.01) and 36 vs 108 (G0.001). At the same timepoints the bronchial scores were 180 vs 161 (NS), 85 vs 138 (NS) and 56 vs 122 (G0.01). In both group there was a significant decrease in the use of bronchodilators and no change in FEV1. In the SLIT group there was a significant decrease of nasal eosinophils vs the budesonide group at 2 and T1. Background: Immunotherapy by sublingual route (SLIT) is nowadays largely employed. Currently by drops or tablets. Sublingual spray has been used for other therapies. This route offers a great potential for immunotherapy increasing the area of mucosal contact with allergen and the rate of absorption. A first trial of SLIT-spray has been done in order to validate safety and patients compliance. Methods: 40 patients with respiratory allergy, allergic rhinitis, asthma or both have been included in the trial. Informed consent has been obtained from all patients. SLIT spray has been given in a standardized schedule from 4 vials in progressive concentrations A 1/125, B 1/25, C 1/5, D 1/1. For mites the vaccine was standardized at 5 mcg/ml for Derp 1+Derp2 and Derf1+Derf2. Cummulative dose at the end of induction was for Derp 1 around 490 mcg (DIATER Laboratories, Spain). The composition of SLITspray was choosen in each case by history, skin tests and specific IgE. The procedure has been carefully explained at all the patients. Daily application for VIALS A,B,C 1 to 4 puffs. D manutention 1 puff daily continued 2 months more. A booklet with instructions was given to the patients and a visual scale from 0 to 5 points marked in a second appointment: convenience, practicability, clarity of instructions and general evaluation. Side effects were registered for each dosage of each vial. Symptomatic therapy was allowed during SLIT-spray. Results: 40 patients, 30 adults (16Y45) (18 M, 12 F) and 10 children (5 M 5 F) (9Y15). Allergens used, Dpteronyssinus 28 cases, D.farinae 26, Lepidoglyphus 11, Tyrophagus 2, Blomia 1,cat 1, dog 1, grass pollens mix 6, Parietaria judaica 3, Olea 1, Plantago 3. Adverse effects in 8 patients only one withdrawal. adverse events were reported. Adverse events reported: urticaria 1, bad taste 1, tongue swelling 2, nausea 1 (withdrawal), sneezing 1, fever 1, wheezing 2, rhinorrea 1,(20%). Adverse events disappear with prosecution of immunotherapy. Evaluation by patients: convenience, practicability, comprehension of instructions and general scores of 4,3Y4,2Y4,0 and 3,5 in a 0Y5 scale. Conclusion: SLIT-spray is a new safe and convenient form of SLIT. High dosage is quickly attained, side-effects relatively rare. The evaluation by patients give good level of appreciation. Larger studies must be done in order to confirm these results. Background: Perennial conjunctivitis due to House Dust Mite is the most frequent form of allergic conjunctiviis in urban environments. However, its diagnosos remains difficult for the ophthalmologists. In India, very little work is done in this field. Keeping this in mind, we have tried to present a case report with Sublingual Oral Immunotherapy. Methods: A male patient, Mr. Pravin J. Patel, 34 yrs,residing at Ahmedabad was suffering from Perennial Allergic Conjunctivitis for over 4 years. Routine therapy including local antiinflammatory drops, antihistaminics, etc didn't give him long term relief. The antigenspecific conjunctival provocation test, skin prick tests and ImmunoCAP RAST were carried out with D.pteronyssinus and D.farinae mites. CPT was positive at low dilution allergen of D. farinae and negative for D.pteronyssinus. Skin Prick Tests showed two plus sensitivity to D.farinae. ImmunoCAP RAST revealed Class VI (Very High) sensitivity to D.farinae. Skin prick tests showed one plus positivity and ImmunoCAP RAST showed Class I response to D.pteronyssinus. The extract to be given for SLIT containing D.farinae was tested for Conjunctival Provocation Test and Skin Prick Test to confirm the positivity. The SLIT was given as twice a day schedule,initially in diluted form and later on with increasing concentration. The maintenance dose was then continued. Clinical assessment, Skin Prick Tests and ImmunoCAP RAST were carried out at 6 months and 12 months interval. The patient is still under SLIT therapy. Results: It was observed that the patient was symptomfree at the end of six months and the redness and swelling had totally disappeared. Skin Prick Tests revealed one plus sensitivity to D.farinae at 6 and 12 months interval. ImmunoCAP RAST showed 46.6 kUa/l (grade IV) and 16.5 kUa/l(grade III) at the end of 6 and 12 months respectively. Conclusion: Sublingual ORAL Immunotherapy has shown excellent clinical response with significant improvement in skin prick test and ImmunoCAP RAST test results in case of Perennial Allergic Conjunctivitis due to D.farinae.This is an isolated case and much more cases need to be studied. Safety, tolerability and efficacy of sublingual allergoid immunotherapy starting with a 4-day build-up scheme. Rationale and study design of an ongoing multicentre Portuguese study in allergic patients Rationale: The good efficacy, safety and tolerability of the sublingual monomeric allergoid (allergoid SLIT, Lais\, Lofarma S.p.A., Milan, Italy), either in tablets or drops, has already been shown in many studies. Yet, so far, only one study on allergoid SLIT starting with a 4-day build-up phase to achieve in a shorter time the maximum suggested dose for the maintenance therapy was performed in 39 allergic patients with promising results. The objective of the present study, still ongoing, is thus to confirm, in a broader population, if this approach is really useful and safe. Study design: One hundred and thirty patients, gathered from the normal daily clinical practice of 16 Portuguese doctors, with an history of oculorhinitis and/or rhinitis with or without mild-moderate asthma, were enrolled during the last 24 months. The build-up rush scheme involved the administration of daily increasing doses of the allergoid SLIT: 500 AU the first day, 1000 AU the second, 1500 AU the third and 2000 AU the fourth. The cumulative dose administered in 4 days was 5000 AU. Also during the maintenance period the treatment dose was kept higher than one normally used. Main endpoints of the study were both tolerability and safety, as well as treatment efficacy. For the evaluation of this one, patient's subjective perception has been taken into consideration as well as the variations in the use of additional drugs for symptom control during SLIT. Results: The final results will be presented at the end of the study in a subsequent poster. Sublingual immunotherapy for paediatric allergic rhinitis Woei Kang Liew 1 , Wen Chin Chiang 1 , Mona Kidon 1 , Anne Goh 1 , Jenny Tang 2 , Oh Moh Chay 2 , Ai Hui Teo 3 , Hwee Hoon Lim 3 , Ai Hui Teo 3 , and Bala Abhilash 4 . 1 KK Hospital, Paediatric Allergy, Immunology and Rheumatology, Singapore, Singapore; 2 KK Hospital, Paediatric Respiratory Medicine, Singapore, Singapore; 3 KK Hospital, Paediatric Medicine, Singapore, Singapore; 4 KK Hospital, Paediatric ENT, Singapore, Singapore. Background: No immunotherapy programme existed in KK Hospital prior to 2006. Work commenced in late 2005, to start sublingual immunotherpy (SLIT) as a new clinical service. Methods: Paediatric AR patients, with moderate persistent symptoms that are not responding to conventional pharmacotherapy, were enrolled. Other inclusion criteria include house dust mite sensitisation, children 5 years and above, with committed and motivated parents/patients. Doctor visits are scheduled at baseline, 1st, 2nd months and 3 monthly intervals thereafter. An ENT review is performed to exclude any anatomical lesions. Allergy symptom scores were completed for 2 weeks prior to each visit and the mean score calculated. Mean rhinitis quality of life (QOL) questionaires were collected at 6 monthly intervals using a computerised database at the Allergy laboratory. Medication usage and adverse events were documented in a file diary provided. House dust mites allergen extracts (Dermatophagoides pteronyssinus and/or Blomia Tropicalis) from Stallergenes (Staloral\) were used, and immunotherapy was initiated as per recommended protocol.\ Results: 12 patients, 58% boys, were recruited over one year. The mean age was 12 years old (range 5.5Y19 years). All had moderate persistent allergic rhinitis, 91.7% had concomitant allergic conjunctivitis, 33.3% had asthma, and 33.3% had eczema. Most (91.7%) have sensitisation to 2 different species of house dust mites, and were prescirbed separate HDM allergens for SLIT. There is an improvement in the mean allergy symptom scores in most patients during the first 6 months ranging between 28Y53%. There is also a reduced mean QOL scores from 40.4 to 22.7, though not reaching statistical significance. There were no serious adverse events or anaphylaxis during SLIT treatment. The most common adverse reaction is sublingual itch 66.7%, which is transient and easily treated. In addition, there were complaints of mild abdominal discomfort and nausea in 2 patients; urticaria rash in 1 patient. One patient had deterioration of pulmonary function and cough 6 months after SLIT, requiring addition of inhaled corticosteroids for asthma. Conclusion: SLIT is a safe and effective treatment for paediatric persistent AR. Good patient-medical team rapport is essential to the success of such programmes. Follow up study in patiens with pollinosis after sublingual immunotherapy (SLIT) compared to subcutaneous shorttimeimmunotherapy (SCIT) Bettina Hauswald 1 , Cornelia Wahl 2 , and Thomas Zahnert 2 . 1 University Hospital Dresden, ENT Department, Dresden, Germany; 2 Universal Hospital Dresden, ENT-Department, Dresden, Germany. The purpose of this study was to evaluate clinical efficacy and safety of sublingual immunotherapy (SLIT) in comparison to subcutaneous immunotherapy (SCIT) in patients with allergic rhinitis. Alltogether we evaluated immunotherapy in 150 patients via inquiry scedule. 101 patients were treated with SLIT and 49 patients were treated with SCIT. Patients treated with SLIT experienced a greater reduction of allergic symptoms (80%) than patients treated with SCIT (75%). In both therapies a reduction of antiallergic medication and an increase of quality of life was shown. The main adverse reactions in SLIT were burning and swelling of the mouth base mucosa especially during dose intensification. In SCIT a local swelling and induration at the injection sight was shown which disappeared completely within 4 to 6 weeks after the last injection. Due to the high compliance of patients and the good results in both strategies the decision for the right therapy for each patient should be made in regard of the patients individual needs and ailments. Tolerance of high dose sublingual immunotherapy with pollen extracts administered under rush and ultra-rush schedules Borja Bartolomé 1 , Begoña Madariaga 1 , María Cruz Gómez 1 , Alberto Martínez 1 , À ngel Ferrer 2 , Antonio Rodríguez 3 , and José Vigaray 4 . 1 Bial ArÚ stegui, R&D Department, Bilbao, Spain; 2 Hospital Vega Baja, Allergy Department, Orihuela, Spain; 3 Hospital Virgen del Camino, Allergy Department, Pamplona, Spain; 4 Clínica Alder, Allergy Clinic, Madrid, Spain. Background: High dose sublingual immunotherapy (SLIT) has been demonstrated as an effective alternative to subcutaneous immunotherapy (SCIT). However high doses could lead to adverse reactions, thus the tolerance of every type of extract should be ascertained. Parietaria and grass pollens are two of the most prevalent aeroallergens in southern European countries. In this work, we tried to asses the tolerance of P. judaica and grass high dose SLIT under two induction phases. Methods: Biologically standardized extracts of P. judaica and grass pollen were used to prepare vaccines at a concentration which reached monthly accumulated doses of 20 and 50 fold, respectively, regarding conventional SCIT. Par j 1, Par j 2 and grass group 1, were controlled in the final preparations. Thirty-two subjects received grass SLIT and twenty P. judaica SLIT. Rush schedule consisted in the administration of one drop (50 6l each) the first day, two the second and thus successively until reaching five drops the fifth day. Ultra-rush consisted in the same volumes given all in the same day within 30 min intervals. Adverse events were registered and graded according to EAACI. Results: There was no systemic reaction nor in the patients treated with grass vaccine neither in the patients treated with Parietaria. Only four out of the 22 patients treated with grass vaccine administered by an ultrarush schedule, showed local reactions in the induction phase (3.6% of doses). Over the whole period (6 months), eleven patients experienced some kind of local reaction (itching and/or swelling), that is 0.38% of the doses. There were three withdrawals due to local reactions. In the 30 patients under the rush schedule, 8 local reactions appeared in the build up, that is 5.3% of the doses. In the whole period, local reactions rose to nine (0.25% of the doses). There was only one withdrawal in the Parietaria group of the study due to repeated local reactions. Conclusion: High dose SLIT vaccines with pollen extracts showed an acceptable safety profile administered by accelerated schedules. Short term efficacy of sub-lingual spray immunotherapy A.G. Palma-Carlos. Clinical Allergy Immunology Center, CAIC, Allergology, Lisbon, Portugal. Background: In the last few years sublingual immunotherapy (SLIT) has gained increasing popularity at least in Europe and all allergen producers laboratories have launched sublingual drops. Sublingual sprays have a larger mucosal contact than drops or tablets increasing also the rate of absorption. Therefore this method of dispensing drugs and allergens can also be applied to SLIT. The efficacy of this technique must be assayed. Methods: 40 patients with respiratory allergy, allergic rhinitis, asthma or allergic rhinitis asthma syndrome have been included in the trial. Informed consent has been obtained from all patients. SLIT spray has been given in a standardized schedule from 4 vials in progressive concentrations A-1/ 125, B 1/25, C 1/5, D 1/1. For mites the vaccine was a standardized at 5 mcg/ml for Derp 1+Derp2 and 5 mcg/ml, for Derf1+Derf2 and Derf1. Cummulative dose at the end of induction was for Derp 1 around 490 mcg (DIATER Laboratories, Spain). The composition of SLIT-spray was chosen in each case by clinical history, skin tests and specific IgE. In children less than 10 years half-concentration of SLIT-spray was employed. The procedure has been carefully explained at all the patients. Daily application for VIALS A,B,C from 1 to 4 puffs each day. Vial manutention dosage 1 puff daily. This dosage being attained only after 13 days and continued 2 months more. A booklet with instructions was given to the patients and a visual scale from 0 to 5 points registered in a second appointment to evaluate facility, convenience, practicability, clarity of instructions and general evaluation of treatment. Side effects were registered for all the patients for each dosage. Symptomatic therapy was allowed during trial with anti-histamines nasal steroids, and formoterolbudesonide when necessary. Results: Short-term efficacy of immunotherapy was good in all patients either for house dust mites allergy or pollens. Evaluation by the patients give on a visual scale an average of 3,5 on 5 points (70%). Medical evaluation was of good (12) or very good (24) results in the patients even in the patients treated during pollen season. Only 1 patient withdrawed from the trial due to the tongue swelling and nauseous state. SLIT spray is effective generally well tolerated and very convenient for the patients. Larger series of SLIT-spray must be evaluated in order to confirm these preliminary results. Sublingual immunotherapy with canova, a homeopathic formulation Dr Tripathi DM. Allergy Clinic, Allergy, Mumbai, India. Rationale: Clinical efficacy of sublingual immunotherapy has been established using allergenic extracts, but there are very few studies using plant extracts. Canova a homeopathic formulation consisting of aqueous extract of Aconitum nepellus, Arsenicum album, Bryonia alba and Thuja occidentalis has been used as immunomodulator along with subcutaneous immunotherapy in patients suffering from different allergic disorders. Methods: In a multi centre study, 30 subjects were selected, who had sensitivity to various allergens, confirmed by Skin Prick Testing, and were undergoing allergen specific immunotherapy course. These subjects were advised for sublingual immunotherapy with Canova along with subcutaneous immunotherapy (SIT). An other group was administered only SIT to compare the results. Results: No relevant side effects were observed with the subjects who had undergone with sublingual immunotherapy with Canova. However, there was a significant reduction of clinical scores for nasal obstruction, sneezing, nasal itching and cough in this group in comparison to those subjects who were on SIT. Conclusion: Canova a homeopathic formulation could be used as an adjuvant with allergen immunotherapy for the treatment of allergic rhinitis, allergic asthma etc. Administration of SIT together with an immunomodulator (Canova) may facilitate safe and effective immunotherapy. With the aim evaluate the efficacy of SIT in patients with SAR we selected 25 patients with ragweed sensitization. The patients were divided into two groups. 10 patients of 1 group were treated with modified SIT by Na-DNA salt and 15 patients of the 2 group -SIT with the same allergen and placebo (dilutant). Methods: Microscopy of nasal smears, ELISA, blood tests, skin prick test, intradermal tests, immunologic examinations and tests: (CD3,CD4,CD8, Result relative group 1 value MTm CD16,CD20,CD3;CD95,CD3; HLADR,NST,IgA,IgMIgG,IgE,Th1,Th2). All groups were treated with combinations of sublingual and intranasal mucosal SIT using allergens dilutions beginning from 1:1,000,000 then 1:000,000 then 1:10,000 in doses of 0.1-0.2-0.4-0.8ml daily then 1:1000 given at the same doses 2 times per week; followed by 1:000 and 1:10 in doses of 0.1-0.2-0.3-0.4-0.5-0.6-0.7-0.8-0.9ml one a week until a dose of 0.1 of pure allergen was reached as the maintain dose given one a month. Results: We stopped treatment for the period of seasonal manifestations and examined patients again. After comparing results in turned out that the patients overcame the therapy better in the 1 group. The cases of nasal congestion and ocular irritation during treatment were in the 1 group in 0.1% and in 1.2% of patients in the 2 group. No cases of systemic reactions were fixed. The average number of eosinophilic cells in nasal smears in the period of seasonal manifestations was less in the patients of the 1 group (68% in the 1 group and 95% in the 2 one). Patients of the 1 group noted that severity of symptoms and need in additional medication was less then during previous years and patients in the second group did not noticed major changes. The immune system tests showed changes towards Th1 response. Conclusion: We can speculate that these positive changes in immune system of the patients of the 1 group could be the response of lymphoid tissue to the modified allergen stimulation during SIT and the anti-inflammatory effect that lead to better overcoming of the treatment. This shows the benefit of using of modified allergen SIT with Na-DNA salt. Metanalisis of the efficacy of sublingual immunotherapy in asthma in pediatric patients, 3 to 18 years of age Martin Penagos 1 , Giovanni Passalacqua 1 , Enrico Compalati 1 , Carlos Baena-Cagnani 2 , Socorro Orozco 3 , Alvaro Pedroza 3 , and Giorgio Walter Canonica 1 . Background: Recent studies have documented the efficacy and safety of sublingual immunotherapy (SLIT) in patients with rhinitis, but the value of this treatment in asthma is still debated. We evaluated the efficacy of SLIT in the treatment of allergic asthma in children, by a meta-analysis of randomized, double-blind and placebo-controlled (DBPC) clinical trials. Methods: Electronic databases were searched up to May 31st, 2006, for randomized, DBPC trials assessing SLIT in pediatric asthma. Effects on primary outcomes (symptom scores and concomitant use of rescue medication) were calculated with standardized mean differences (SMD) using a random-effect model. RevMan 4.2.8 program was used to perform the metaanalysis and we followed the recommendations of the Cochrane Collaboration and QUORUM statement. Results: Seventy three articles were identified and reviewed. Nine studies, all published after 1990, fulfilled the selection criteria. 441 patients had a final assessment and were included in the analysis. Two hundred thirty two patients received SLIT and 209 placebo. The results of the present analysis demonstrated a relevant heterogeneity due to widely differing scoring systems. Overall, there was a significant reduction in both symptoms (SMD Y 1.14, 95%CI Y 2.10 to Y 0.18; p = 0.02) and medication use (SMD Y 1.63, 95%CI Y 2.83 to Y 0.44; p = 0.007) following SLIT. Conclusion: SLIT with standardized extracts reduces both symptom scores and rescue medication use in children with allergic asthma compared to placebo. Suzan Hosny. Ainshamsuniversity, Immunotherapy, Cairo, Egypt. Background: To assess the clinical efficacy & safety of local nasal immunotherapy &comparing it with other routes of immunotherapy in allergic rhinitis patients. Methodology: 1000 patients from 6 egyptian governorates were included in this study. Results: Skin tests showed significant improvement after local nasal immunotherapy as well as blood, nasal& sputum eosinophilia were all improved also totel ige & nasal symptoms before & after local nasal immunotherapy. Conclusion: Local nasal immunotherapy showed very good efficacy, safety & a more convenient allergen delivery in pts. With allergic rhinitis also simplified, self adminstrable, method with reduction of local nasal symptoms. Efficacy and safety of sublingual immunotherapy with high dose blomia tropicalis extract in children with allergic respiratory disease: comparison study with drugs Bashir Omarjee, Bernard Tanguy, and Ahmad Zaidou. Centre Hospitalier Departemental, Service de Maladies Respiratoires, Saint Denis, Reunion. Background: Sublingual immunotherapy (SLIT) might become a viable alternative to drug in the treatment of airway allergies. The aim of this study was to evaluate,in a pediatric population monosensitized to Blomia Tropicalis mites, the clinical efficacy and safety of high dose SLIT in comparison with drug treatment (antihistamines plus $ 2 agonists). Methods: 97 children:52 males and 45 females with a mean age of 9.4 years, range 7 to 13 years, were included in the study. Of them 49 were treated with T1. Patients with severe persistent asthma, poliallergies and vernal conjunctivitis were excluded. After the 3 week-induction period, the patients were kept on a daily extract dosage of 20 drops of 300 IR/ml for another 21 weeks. The cumulative dose came to about 41654 IR. Clinical efficacy was assessed using symptom and medication scores. Results: A gradual reduction in daily rhinitis/asthma scores was observed in SLIT group (P G 0.001). In only 3 patients,out of 49, it was necessary to employ antihistamines and in 7 out of 49 salbutamol. Also the symptoms score was statistically lower (P G 0.01). The clinical efficacy was higher in patients with mild/moderate persistent asthma. No patient showed local or systemic side effects of any relevance. Local adverse reactions were oral pruritus, gingivitis and aphthas. Conclusion: In this study on patients with single sensitization to Blomia Tropicalis allergen suffering from allergic respiratory disease, the SLIT improves the allergic symptoms to a greater extent than drug treatment. SLIT even at ultrahigh doses was safe and well tolerated. Immunological characterization of recombinant lactobacillus plantarum expressing Bt5 major allergen protein LM Liew 1 , CH Huang 1 , SV Seow 1 , Ying Ding 1 , HM Wen 1 , IC Kuo 2 , and KY Chua 1 . 1 National University of Singapore, Department of Peadiatrics, Singapore, Singapore; 2 A*STAR, Singapore Institute for Clinical Sciences, Singapore, Singapore. Background: Sensitivity to Blomia tropicalis (Bt) accounts for more than 50% of the incidence of mite allergy in the tropical regions and Bt5 is a major Bt allergen. In recent years, numerous studies have suggested that recombinant probiotics could be effective in primary prevention and management of allergic diseases. The long term objective of this study is to develop recombinant lactobacilli expressing dust mite allergen for clinical application. Methods: Bt5 gene was inserted into an expression vector pSIP412 containing a strong inducible promoter P orfX . Lactobacillus plantarum was electrotransformed with pSIP412 vector carrying Bt5 gene and designated as Lp-Bt5. Murine bone marrow-derived dendritic cells (BM-DCs) were pulsed with heat-killed Lp-Bt5 overnight. The cell surface molecule expression and cytokine production of the pulsed-DCs were examined by flow cytometry and ELISA. The antigen presenting function of the pulsed BM-DCs were analysed by co-culturing with Bt5-specific Th2 cell lines. In vivo study of Lp-Bt5 was performed in C57BL/6 mice fed with the live Lp-Bt5 for three consecutive days per week over three weeks duration. Sera and splenic culture supernatants were collected for antibodies and cytokines analysis by ELISA. Results: High level of recombinant Bt5 expression was achieved in Lp-Bt5 upon induction using 50ng/ml sakacin P inducing peptide. After 4 hours induction, approximately 406g of Bt5 protein was expressed by 10 9 cfu live Lp-Bt5. Lp-Bt5-pulsed BM-DCs showed upregulation of CD86 and MHCII with production of IL-10, IL-12 and TNF-alpha. Lp-Bt5-pulsed DCs were capable of inducing cell proliferation but suppressing Th2 cytokines production by the Bt5-specific Th2 cells. In contrast the wildtype-Lp-pulsed DCs, in the presence of exogenous Bt 5 protein, induced both cell proliferation and Th2 cytokine production by the Bt5-specific Th2 cells. In vivo studies showed that the splenocytes of the mice fed with live LAB-Bt5 produced IL-5, IL-6, IL-10, INF-gamma and TNF-alpha upon stimulation with Bt5 protein. 25% of the fed mice produced detectable levels of Bt5specific IgG1. Conclusion: Recombinant lactobacilli expressing Bt5 allergen is potentially useful for the modulation of allergen-specific T cells. Further studies are required to assess its application in the prevention and treatment of allergic diseases. Role of allergen specific Immunotherapy in allergic bronchial asthma due to Sorghum Vulgare pollens Sanjay Pawar. Shriratna Intensive Care & Hospital, Chest Medicine, Karad (Satara) Mah, India. Introduction: Immunotherapy for specific allergy is extensively used as definitive therapy for bronchial asthma and it is suppose to be only disease modifying treatment. In our area, Sorghum Vulgare crop is extensively cultivated and lot many patients are having allergic manifestation during pollination. Aim: To find out role of sorghum Vulgare pollens in causation of bronchial asthma with reference to pulmonary function test and also Study effect of allergen specific Immunotherapy in comparison with pharmacotherapy. Methodology: Prospective, comparative, Cohort study was planned. Sorghum Vulgare pollens are collected and analyzed. For SV pollens SDS page electrophoresis and ELISA_ inhibition assay done and 14Y100 kDa proteins are separated. The SV extract was highly immune reactive and require approximately 7 ng of proteins for 50% inhibition of specific IgE binding in asthma. Immunoblot identified 8 allergenic brands of which 5 proteins (MWs 94, 70, 40, 35 , and 14 kDa) recognized by more than 90% of serum samples of SV pollen sensitive patients. Skin test were done with Modified skin prick test technique. Histamine diphosphate is used as positive control while buffered saline is used as negative control. Testing is performed on forearm. Immunotherapy of above extracts was given by subcutaneous route. Effect of Immunotherapy was studied for 659 patients. Results: Short term (4mth) improvement in clinical symptoms and pulmonary function test (FEV1,PEFT,FVC,FEF25Y 75) is observed. Eleven percent patient (n=4) showed improvement with immunotherapy and 86% (n=13) with Pharmacotherapy. But long term (3yrs) improvement observed in 47% (n=17) with immunotherapy and 20% with pharmacotherapy. Conclusion: MWs 94, 70, 40, 35, and 14 kDa proteins are recognized in sorghum vulgare sensitive patients. Allergen specific immunotherapy is helpful in long term management allergic bronchial asthma due to sorghum Vulgare pollens. Antigenicity prediction in melittin: approach for fragment based peptide drug development from Apis dorsata Virendra Gomase 1 , and Ganesh Vankhede 2 . 1 Department of Bioinformatics, MGMs, College of Computer Science and Information Technology, Nanded, 431605 MS, India, Bioinformatics, Nanded, India; 2 Department of Zoology, Sant Gadge Baba Amravati University, Zoology, Amravati, India. The bee venom is used for treating a wide variety of conditions from acute tendonitis to chronic back pain to rheumatoid arthritis (RA). In this assay we found peptide nonamers from melittin protein of Apis dorsata, which can be used to select for use in rational vaccine design and to increase the understanding of roles of the immune system in allergic reactions. For development of MHC peptide binder_s prediction, an elegant machine learning technique support vector machine (SVM) method has been used. Antigenic epitopes on melittin protein of A. dorsata are important determinants of protection against Rheumatoid Arthritis. As our knowledge of the immune responses to a protein antigen progressed, it became clear that the whole protein is not necessary for raising the immune response, but small peptide fragments 4-AILKVLSTGLPALIS-18 of protein called the antigenic determinants or the epitopes are sufficient for eliciting the desired immune response. We also found peptide nonamers, which are from a set of aligned peptides known to bind to a given major histocompatibility complex (MHC) molecule as the predictor of MHC-peptide binding. MHCII molecules bind peptides in similar yet different modes and alignments of MHCII-ligands were obtained to be consistent with the binding mode of the peptides to their MHC class, this means the increase in affinity of MHC binding peptides may result in enhancement of immunogenicity of melittin protein. Tertiapin is a neurotoxin from the honeybee venom. It interacts specifically with calmodulin in the presence of Ca2+. Antigenic epitopes on tertiapin protein of Apis mellifera (honey bee) is important determinants sites for protection against disorders. This assay we found peptide nonamers from tertiapin protein, which can be used to select for use in rational vaccine design and to increase the understanding of roles of the immune system in infectious diseases. For development of MHC peptide binder_s prediction, an elegant machine learning technique support vector machine (SVM) method has been used. SVM has been trained on the binary input of single amino acid sequence. In this analysis, we predicted the binding affinity of tertiapin protein having 21 amino acids. We found the MHCII-IAb peptide regions 7-RIIIPHMCW, 13-MCWKKCGKK, 10-IPHMCWKKC, 11-PHMCWKKCG, (optimal score is 0.318); MHCII-IAd peptide regions 3-CNCNRIIIP, 4-NCNRIIIPH, 5-CNRIIIPHM, 6-NRIIIPHMC, (optimal score is 0.202); MHCII-IAg7 peptide regions 7-RIIIPHMCW, 9-IIPHMCWKK, 4-NCNRIIIPH, 8-IIIPHMCWK, (optimal score is 0.805); and MHCII-RT1.B peptide regions 1-ALCNCNRII, 13-MCWKKCGKK, 9-IIPHMCWKK, 11-PHMCWKKCG, (optimal score is j0.730) which represented predicted binders from tertiapin from Apis mellifera protein. These peptide nonamers are from a set of aligned peptides known to bind to a given major histocompatibility complex (MHC) molecule as the predictor of MHC-peptide binding. MHCII molecules bind peptides Background: The indication to specific vaccination in patients with hymenoptera allergy and concomitant mastocytosis is still a matter of debate, since severe adverse reactions have been described. Methods: We treated with immunotherapy 16 patients with systemic mastocytosis. All had positive bone marrow biopsy and elevated serum tryptase and 3 of them had also positive skin biopsy for urticaria pigmentosa. All the patients (12 male, mean age 51.8 years) have had severe adverse reactions following hymenoptera stinging (Grade IV according to Mueller). Of them, one was allergic to bee, 7 to polistes dominulus and 8 to jellow-jacket. They were vaccinated with a slow-induction protocol (7-week induction phase). Results: Only one patient had systemic urticaria in the updosing phase, that was easily controlled slowing the escalation and giving antihistamine premedication. No adverse event was recorded in the maintenance phases with 100 mcg venom. Nine patients were re-stung while on maintenance. Five had only a mild local reaction, two were stung by a different insect without reaction and two had a grade III reaction. In these subjects the maintenance was escalated to 200 mcg and after further re-sting they had no more reaction. Conclusion: Despite the relative rarity of systemic mastocytosis, hymenoptera venom hypersensitivity can provoke life threatening reactions in those patients. Based on our experience, specific immunotherapy with a slow induction is safe and can lead to an relevant, although not complete, clinical improvement Key words: Systemic mastocytosis, hymenoptera allergy, specific immunotherapy 739 A modified latex hevein (Hev b 6.02) retains lymphocyte proliferation response Rationale: We designed a hypoallergenic Hev b 6.02 peptide by changing all cysteine residues to serine residues with the goal to produce a standardized specific immunotherapy reagent. Methods: Sera of 24 latex allergic patients with confirmed IgE reactivity to MBP-rHev b 6.02 were tested for their IgE binding capacity to the modified hevein peptide by ELISA. BALB/c mice were vaccinated either with this modified Hev b 6.02 peptide alone or coupled to tetanus toxoid. The antibody responses of these mice to natural Hev b 6.02 were analyzed by immunoblot and to MBP-rHev b 6.02 by ELISA. Cytokine production was tested in spleen cell cultures of immunized BALB/c mice. Results: The Hev b 6.02 peptide showed markedly decreased IgE binding (9/24) of latex allergic patients' sera. Immunization with each of the modified Hev b 6.02 peptide formulations induced Abs that cross-reacted with the natural and the recombinant Hev b 6.02. Uncoupled Hev b 6.02 peptide induced lymphocyte proliferation and Th1 (IL-2, INF-+) cytokine production in spleen cell cultures indicating that this peptide was able to induce a Th1 biased T-cell response. Tetanus toxoid coupled modified Hev b 6.02 peptide revealed high titers of IgG, mainly of the IgG2a subclass, to MBP-rHev b 6.02. Conclusion: The ablated allergenicity but retained lymphozyte proliferation capacity of the Hev b 6.02 peptide showed that such a reagent can induce a Th1-biased immune response that is also directed against the major latex allergen Hev b 6.02. Background: It is well known that allergen immunotherapy modifies the peripheral immunoglobulin profile down-regulating the Th2 cytokines. Notably CCR5 and CXCR3 are found on Th1 cells, CCR3 and CXCR4 on Th2 cells. Aim of the present study was to identify an immunological marker that correlates with the clinical effects among CXCR3, CCR3, CXCR4, CCR5 expression on PBMC. Materials and Methods: 18 patients were enrolled according to the following criteria: moderate/severe rhinoconjunctivitis, moderate asthma since at least 2 years, sensitization to Parietaria only, age between 18 and 55 years, no previous immunotherapy. After a one-week run-in period where an optimal asthma therapy was prescribed, patients were received either SLIT to parietaria (SLIT-ONE Alk-abellò) or drug treatment for rhinitis. Control visits were carried out at 1, 2, 6, 10 and 16 weeks (follow-up visit). Quality of life was evaluated with a validated questionnaire (Rhinasthma). The immunological parameters assessed were: PBMC proliferation with specific allergen Par-j1, detection of CD4, CD8, CD3 and chemokines receptors CCR3, CXCR3, CXCR4, CCR5 before and after stimulation with Par-j1 Results: The symptom scores decreased in the control group since the first week of study whereas in the SLIT group a significant improvement versus baseline was seen after 6 weeks of treatment. Starting from 6 weeks there was no more difference in total symptom scores between the two groups. Quality of life significantly improved in the SLIT group if compared with controls from week 6. After 6 weeks SLIT group significantly decreased in drug doses for asthma treatment whereas no statistical difference in therapy dosage was seen in the control group until week 6 the SLIT group had a significant higher use of rescue medication if compared with control group after week 6 the uptake of rescue medications had an inverse trend with statistically significant higher intake in the control group. In the same period there was a decrease in CCR3 and an increase of CXCR3 in treated group these variations became significant at the end of the study from the baseline. No difference in CD4, CD8, CD3, CXCR4, CCR5 expression. Conclusions: This study demonstrates efficacy SLIT when compared to conventional therapy for persistent asthma and rhinitis, correlations between clinical scores and both CXCR3 and CCR3 seen after 6 weeks of immunotherapy in treated patients demonstrate an immunological shift towards Th1 lymphocyte pattern. The new paradigm in vaccine design is emerging, following essential discoveries in immunology and development of new major histocompatibility complex (MHC) Class-I binding peptides prediction tools. In this we analyzed MHC binders of neurotoxin M14 protein from Buthus eupeus [Lesser Asian scorpion] are important determinant of protection against cardiovascular disorder. Buthus venom was tested in vivo in anaesthetized rats and in vitro on isolated cardiac and skeletal muscle preparations. In vivo, the venom caused marked rhythmical fluctuations in blood pressure preceding cardiovascular collapse and death. Scorpion toxins constitute a family of homologous proteins that exert potent pharmacological effects on potassium or sodium ion channels. In this analysis, we predicted the binding affinity of B. eupeus protein neurotoxin M14 having 66 amino acids. We found the fragments peptide regions 8-DDRNCVYTCALNPYCDS-24 of protein called the antigenic determinant or the epitope is sufficient for eliciting the desired immune response. The predicted binding affinity is normalized by the 1% fractil. The MHC peptide binding is predicted using neural networks trained on C terminals of known epitopes. In analysis predicted MHC/peptide binding is a logtransformed value related to the IC50 values in nM units. Predicted MHC binding regions acts like red flags for antigen specific and generate immune response against the neurotoxin M14 antigen. So a small fragment of antigen can induce immune response against whole antigen. This theme is implemented in designing subunit and synthetic peptide vaccines. The sequence analysis method is allows potential drug targets to identify active sites against venom action. The method integrates prediction of peptide MHC class I binding; proteosomal C terminal cleavage and TAP transport efficiency. Background: Levels of allergen specific IgE are relatively unaffected by allergen specific immunotherapy (SIT), whereas levels of allergen specific IgG is markedly increased. The effect on antibodies to minor as compared to major allergens is not clear. In this study antibodies towards Bet v 1 and Bet v 2 of Betula verrucosa were investigated in sera of SIT treated patients. Methods: Specific IgE was bound to monoclonal anti-IgE coupled to paramagnetic beads and measured by the binding of 125 I labelled purified natural Bet v 2 and recombinant Bet v 1.2801, respectively. IgE depleted serum was tested for specific IgG and following incubation with the labelled allergen, free and bound allergen were separated by protein G affinity chromatography. Sera were tested in immunoblotting with or without absorption using 1 mg birch pollen extract per serum sample. Results: All sera (n=50) were positive for IgG and IgE against Bet v 1 before and after SIT. The relative binding activities of IgG compared to IgE were up to 30 times higher after SIT. In contrast, 22% of non-treated patients were positive for IgE anti-Bet v 2 and 48% for IgG anti-Bet v 2. At 1.5 years of SIT, the prevalence of IgG-and IgE-anti-Bet v 2 was unchanged, however, after 5 years of treatment, the prevalence of IgG anti-Bet v 2 was 91% but the prevalence of IgE anti-Bet v 2 was unchanged when analysed by quantitative immunoassay. Contrary to this observation, 3 sera became IgE anti Bet v 2 positive after 5 years of treatment when measured by immunoblotting. Conclusion: The dominant binding activity against Bet v 1 and Bet v 2 in sera from birch pollen allergic patients both before and after SIT resides in the IgG fraction. Prolonged SIT treatment induced IgG against minor allergen Bet v 2 in most individuals. New IgE reactivities to Bet v 2 were observed after SIT in 3 patients by immunoblotting, however, this result was not confirmed by quantitative solid phase immunoassay and may possibly indicate differences in assay sensitivity with respect to denaturation insensitive epitopes. The induction of allergen-specific antibodies which inhibit the IgE recognition of allergens is a major mechanism underlying allergen-specific immunotherapy. In order to develop therapeutic antibodies for the treatment of allergy to birch and related allergen sources, we immunized mice with surface-exposed peptides derived from the major birch pollen allergen Bet v 1 and established 14 hybridomas secreting peptide-specific mouse monoclonal IgG1 antibodies. When tested for interference with allergic patients_ IgE by ELISA competition experiments, Bet v 1-specific monoclonal antibodies inhibited polyclonal serum IgE reactivity of allergic patients (n=44) to Bet v 1 and Bet v 1-homologous allergens between 60Y70%. After humanization, the described Bet v 1-specific monoclonal antibodies may be used for passive vaccination or local therapy of birch pollen allergy. Differences in specific cutaneous sensitivity between allergic patients treated with dermatophagoides pteronyssinus depigmented and glutaraldehyde polymerised extracts, nonchemically modifed, and treated only with pharmacotherapy Magdalena Lluch, Alfonso Malet, Pau Amat, Estrella Llamas, Mariela Dall ' Aglio, Marcia Dias, and Casanovas Miquel. Al.Lergo Centre, Clinica, Barcelona, Spain. Background: The evolution of skin sensitivity is one of the objective tolols to evaluate the efficacy of specific immunotherapy. Objective: To compare the evolution of the cutaneous sensitivity of allergic patients with rhinoconjunctivitis with or without asthma due to hypersensitivity against D. pteronyssinus, treated with vaccines containing standardized extracts of this mite (alum adsorbed depigmented and polymeryzed -modifiedor non-modified extracts -retard-) or only with pharmacological treatment. Materials and Methods: It was conducted an open and controlled clinical trial with 3 groups of patients. These patients received at random the modified (n=21), the retard (n=21) or only pharmacological treatment (n=7). After an initial wash-out period, and before receiving inmunotherapy, there were skin prick testeds in duplicate, on the volar surface of the forearm using the same batch of native D. pteronyssinus extract at 0.002, 0.02, 0.2 and 2 mg/ml. The vial of maximum concentration had a potency of 100 HEP/ml and contained 40.70 mg of Der p 1/ml and 17.22 of Der p 2. These tests were repeated after 6 and 12 months of treatment (end of study). The results were expressed as the 10 HEP value (quantity of allergen extract that produces a wheal of the same size as the wheal produced by histamine HCl 10mg/ml). Results: The patients treated with the modified preparation needed to obtain the 10 HEP value 0.233 mg at baseline, 0.508 mg after 6 months of treatment, and 1.1 mg at the end of the study (P=0.008). In the group of patients treated with the retard preparation the values were 0.328, 0.436, 0.880 mg, respectively (p=0.037). The control group experienced an increase on the cutaneous sensitivity (0.220, 0.188 y 0.115 mg, respectively), however the difference was not significant (P=0.513). Conclusion: This open and controlled clinical study demonstrates that the patients sensitized against D. pteronyssinus and treated with specific inmunotherapy experienced a highly significant reduction on cutaneous sensitivity, being the group treated with the vaccine containing the modified extract the first in presenting improvement (after 6 months of treatment), whereas the patients treated with the retard experienced improvement after 12 months. The patients not treated with inmunotherapy did not show reduction on the specific cutaneous sensitivity. Safety evaluation of a fast build-up schedule of immunotherapy using therapeutic vaccines containing depigmented and glutaraldehyde polymerised allergen extracts Alfonso Malet 1 , Magdalena Lluch 1 , Pablo Amat 1 , Estrella Llamas 1 , Isabel Garcia 1 , and Miquel Casanovas 2 . 1 AlILergo Centre, Clinica, Barcelona, Spain; 2 Laboratorios Leti, L, Madrid, Spain. Background: Various administration schedules of immunotherapy have been reported to be beneficial for the treatment of inhalant allergy: conventional, cluster and rush. The use of therapeutic vaccines containing modified allergen extracts (depigmented and glutaraldehyde polymerised), in which the allergenicity is decreased, and the immunogenicity maintained, allows to reach the maintenance dose using conventional schedules with few administrations in a very short period of time. The objective was to evaluate the safety of a very short build-up phase using these hypoallergenic vaccines containing standardised modified allergen extracts. Materials and Methods: One hundred and nine patients (56 female and 53 male, mean age 29.7 years) with rhinoconjunctivitis and/or asthma, sensitised to mites and/or pollens were included in an observational cohort study. All of them received an individualised therapeutic vaccine containing depigmented and glutaraldehyde polymerised allergen extract. The patients reached the maximum dose after 2 injections of the vial of maximum concentration (0.2 as the first dose the first day and 0.5 ml as the maintenance dose after 1 week, followed by 0.5 ml after 1 month). The tolerance was assessed by recording all side reactions related to immunotherapy, which were classified according to the criteria of the EAACI. Results: The total number of injections was 327. All patients reached the maximum recommended dose the first week. No premedication with drugs was used before the administration of the immunotherapy. Eight reactions (2.44 % of the total injections) were recorded, 7 local and 1 systemic. The 7 local reactions consisted of 3 immediate (diameter 9 5 cm) and 4 delayed (diameter 9 10 cm). One systemic reaction of grade 2 was reported. This reaction consisted in a slight urticaria that resolved with treatment with antihistamines (ebastine). Conclusion: The therapeutic vaccines containing chemically modified extracts can be safely administered using a fast schedule in an immunotherapy unit, reaching the maximum dose the first week with 2 injections and without the need of premedication and/or hospitalisation. The influence of different application forms of allergen immunotherapy on the level of allergen specific IgG antibodies in patients with grass pollen allergy Ctirad Andrys, Irena Krcmova, Marcela Drahosova, and Yvona Hanzalkova. University Hospital, Institute of Clinical Immunology and Allergology, Hradec Kralove, Czech Republic. Background: Allergen immunotherapy (AIT) is the only therapeutic approach in the treatment of IgE mediated allergies which is targeting immunopathogenetic process. Allergen could be administered in several application forms preferentially subcutaneously or sublingually. It is difficult task to follow the efficacy of this immunomodulation. The aim of study is comparison of influence of two different routes of application of immunotherapy on the serum level of allergen -specific IgG and IgG4 antibodies. Methods: Together 62 patients (31 male, 31 female) with grass pollen allergy were enrolled. Allergen immunotherapy by subcutaneous injection (Phostal, Stallergenes) was administered to 21 patients. Allergen immunotherapy in the form of sublingual capsules (Staloral, Stallergenes) was administered to 18 patients. 23 patients were treated symptomatically during pollen season. The blood samples were collected before the start of therapy (January, 2003), and after 12, 24 and 36 months of treatment. The serum level of specific IgG and IgG4 antibodies against immunodominant allergen (timothy grass) was evaluated by diagnostic kit (Dr.Fooke, Germany). Results: No significant differences were found between groups of patients before start of allergen immunotherapy in any parameter tested. There is statistically significant increase in allergen specific IgG in both groups of patients after one year therapy (both pG0.0001) and after 3 years of therapy (both pG0.0001). Statistically significant increase was found in allergen specific IgG4 in both groups of patients after 3 years of therapy (subcutaneous group p=0.0007, sublingual group p=0.042). There is statistically significantly higher level of allergen specific IgG and IgG4 (both pG0.0001) after 3 years of therapy in patients treated by subcutaneous form of allergen immunotherapy in comparison with patients treated by sublingual form of immunotherapy. The level of allergen specific IgG and IgG4 in control group remains without any significant change. Conclusion: The subcutaneous application of allergen immunotherapy results in strong increase of allergen specific IgG and IgG4 antibodies in comparison with sublingual administration of immunotherapy. Two different application forms of grass pollen immunotherapythe changes in the levels of cytokines IL-4 and IFN+ after 36 months of immunotherapy Irena Krcmova, Ctirad Andrys, Marcela Drahosova, and Jakub Novosad. University Hospital, Institute of Clinical Immunology and Allergology, Hradec Kralove, Czech Republic. Background: Allergen immunotherapy (AIT) is the only therapeutic approach in the treatment of IgE mediated allergies which is able to target immunopathogenetic process. Allergen can be administered in several application forms preferentially by subcutaneous or sublingual route. Subcutaneous and sublingual forms are differentially interacting with immune system. Data concerning the comparison of these two forms of application favour studies with clinical efficacy over laboratory studies. Aim: Of this study is the comparison of the influence of two different types of AIT on the serum levels of cytokines IL-4 and IFN+. Methods: Together 62 patients with grass pollen allergy were enrolled. Subcutaneous AIT (Phostal) was administered to 21 patients. Sublingual AIT (Staloral, 100 IR/ml)) was administered to 18 patients, 23 patients were treated symptomatically. Blood samples were collected before the start of study (January, 2003) , after 12, 24 and 36 months. The serum level of cytokines IL-4 and IFN+ was evaluated by diagnostic kit (Quantikine, RDS). Results: No significant differences were found between the groups before the study. There was significant increase of IFN+ only in the group of subcutaneous AIT after 12 months of therapy (p= 0.016) and after 36 months (p= 0.0294). The IL-4 demonstrated significant decrease in the group of subcutaneous AIT after 12 months of therapy (p= 0.022) and in both groups of AIT after 36 months (pG 0.0001), but comparing to control group only subcutaneous AIT group reached significance. Conclusion: The subcutaneous and sublingual applications of AIT after 36 months resulted in the significant decrease of IL-4 in both groups, but decrease of IL-4 in the subcutaneous group was manifested earlier -after 12 months. Expecting increase of IFN + was significant only in group of subcutaneous AIT. Immunomodulation by subcutaneous AIT seems stronger than sublingual AIT, especially in the first year of the study. For continual sublingual AIT are needed higher concentrations of allergens then 100 IR/ml as maintenance dosage. Double blind placebo controlled trial of sublingual monomeric allergoid in grass pollen allergy A.G. Palma-Carlos, M. Branco-Ferreira, A.L. Pregal, A.S. Santos, and M.L. Palma-Carlos. Clinical Allergy Immunology Center, CAIC, Allergology, Lisbon, Portugal. Background: Double blind placebo controlled trials of allergy vaccines are considered the better method to confirm his efficacy. An assay of a sublingual monomeric allergoid in tablets was done in patients allergic to grass pollens during 2 years. This monomeric allergoid was obtained by carbamylation of amino groups (lysine) of allergenic proteins. Methods: 33 rhinitic patients allergic to grass pollens as confirmed by clinical history, skin prick tests and specific IgE were included in the trial. Patients were randomized between placebo and active vaccine both given as sublingual tablets administered by the sublingual swallow method (LAIS, Lofarma, Milano, Italy) . A traditional posological scheme was used starting with a very low dose increased progressively in the course of 14 weeks before grass pollen season. Enrolment of the patients was preseasonal followed by a seasonal visit in May and a end of the year visit in October. Patients were followed during 2 consecutive years. Nasal provocation tests with the allergens included in the vaccine was done before treatment and after 1 and 2 years of treatment and symptom scores registered after each challenge. The use of nasal steroids as rescue was authorized for rhinorrea, sneezing and conjunctivitis. Symptom scores for eventual side effects and pharmaceutical scores were registered for each patient. Informed consent was obtained from all the patients. Results: Results were evaluated after 1 and 2 years of treatment. Between vaccines and placebo the score for rhinorrea (pG0,03) sneezing (pG0,03) and conjunctivitis (pG0,02) was significantly decreased for the second wear of treatment. Between the first and second years of treatment the scores for placebo were not changed but for active vaccine, rhinorrea (pG0,03), sneezing (pG0,03) and conjunctivitis (pG0,02) were significantly decreased. Symptomatic score after nasal challenge was decreased (pG0.03) after 2 years treatment. Nasal steroid consumption were significantly decreased in the active group during May and June both years of treatment (pG0,02). Only two light local adverse events were reported in the active group and none with placebo. Conclusion: Preseasonal treatment with a sublingual monomeric grass pollen allergoid in tablets improved rhinorrea, sneezing and conjunctivitis and reduced the steroid consumption. Tolerability was optimal. Phleum pratense alone is sufficient for allergen specific immunotherapy against allergy to Pooideae grass pollens Charlotte Hejl 1 , H Ipsen 1 , J N Larsen 2 , and N Johansen 3 . 1 ALK-Abelló, Vaccine Research, HLrsholm, Denmark; 2 ALK-Abelló, Scientific Affairs, HLrsholm, Denmark; 3 ALK-Abelló, Serology Research, HLrsholm, Denmark. Background: The grass family Poales contains more than 10.000 species which can be divided into subfamilies based on their taxonomy. One of the larger subfamilies is the Pooideae subfamily also denominated as the temperature pasture grasses. In spite of the large number of grass species, grass pollen allergic patients' IgE show a high degree of cross-reactivity between Pooideae grass species. Currently, immunotherapy for grass pollen allergy is often performed with mixtures of grass species. However, high clinical efficacy of grass pollen immunotherapy (SIT) using Phleum pratense (Phl p) alone has been demonstrated in several studies. The aim of the study was to investigate the antibody cross-reactivity towards Phl p and grass pollen extract mixes. Methods: The amount sIgE reacting with Phl p pollen extract and four different grass pollen extract mixes was determined in sera from grass pollen allergic patients. The capacity of Phl p to inhibit the reaction of grass pollen allergic patients' IgE towards the four grass pollen extract mixes was performed using excess inhibitory concentration of non-labeled Phl p extract. The four mixes each comprised between 5 and 10 Pooideae grass pollen extract species. Phl p SIT induced IgG4 was measured to Phl p pollen extract, extracts of nine individual Pooideae grass species and the four grass pollen extract mixes. All allergen specific antibody assays was performed using an ADVIA Centaur solid phase immunoassay. The correlation between the reaction of IgE towards Phl p and the four grass pollen extract mixes, irrespective of the number included in the mixes, is statistical significant (0.96Y0.98, Spearman). Phl p has the capacity to inhibit the reaction of the patients specific IgE to the four grass pollen extract mixes indicating a high degree of cross-reactivity of the patients IgE. SIT with Phleum pratense induces IgG4 which exhibit a statistical significant correlation (0.92Y0.99, Spearman) in their reaction with the individual nine Pooideae grass pollen extracts and the four grass pollen extract mixes. Thus, IgG4 induced by SIT with Phl p cross-react extensively with both single and mixed grass pollen extracts. Conclusion: Immunochemical and clinical data together support the conclusion that Phl p alone is as effective for allergen specific immunotherapy against allergy to Pooideae grass pollens as any mix or single specie fro the Pooideae family. Background: The clinical efficacy of subcutaneous immunotherapy (SIT) in the treatment of allergic respiratory diseases is well documented, but its main drawback of is the risk of systemic reactions (SRs). This 3-years prospective study evaluated the safety of SIT with standardized extracts in real life in patients with allergic rhinitis and/or asthma. Methods: Between January 2003 and June 2006 a total of 248 patients (140 male, mean age 31.4 years, range 7Y65) referred to our Allergy Unit were prescribed SIT for their respiratory diseases. 176 of the patients had seasonal/ perennial rhinitis and 72 asthma with or without rhinitis. Allergen extracts were all standardized by biological methods (ALK-Abellò and Stallergenes). 204 patients were treated according to a traditional (12 weeks) schedule, whereas 44 underwent a cluster protocol (7 weeks). Patients were treated for grass (35%), birch (13%), parietaria (31%), mite (21%). Systemic reactions (SRs) were categorized into immediate SRs (occurring within 30 minutes) and late (930 minutes). They were graded in four levels of severity according to EEACI Immunotherapy Position Paper (0= No symptoms; 1= Non specific symptoms probably non IgE-mediated; 2= rhinitis/asthma responding adequately to treatment; 3= non life-threatening SRs, e.g. urticaria, angioedema or severe asthma; 4= Anaphylactic shock). Results: A total of 9,650 injections were administered in the 248 patients. SRs registered are shown in table 1 and divided according to the two phases of the schedule (build-up and maintenance). Results. There were 18 SRs (7.3% patients and 0.20% injections). 13 occurred in the build-up phase and 5 in thye maintenance. SRs were more frequent in females (66%). Grass pollens were most frequently responsible of SRs (65%) than other allergens (parietaria 18%, tree 5%, mite 10%) SRs were Grade 1= 8 cases; Grade 2 = 7 cases; Grade 3 = 1 case, Grade 4 = 2 cases (Epinephrine administered). Both severe reactions occurred within 30 minutes after injection. Late reactions were less common (7 SRs). Conclusion: If properly used, SIT has a low and acceptable rate of side effects. In our population female gender and build-up phase were the two main risk factors. Keywords. Specific immunotherapy, injection, systemic reaction. Adjuvant effects of heat-killed wildtype Lactobacillus casei on allergen immunotherapy in an allergy mouse model Lay Hong Lim 1 , Chiung Hui Huang 1 , Hai Yan Li 1 , Yuan Kun Lee 2 , Bee Wah Lee 1 , and Kaw Yan Chua 1 . 1 National University of Singapore, Paediatrics, Singapore, Singapore; 2 National University of Singapore, Microbiology, Singapore, Singapore. Background and Aim: Probiotics, like lactobacilli have been used as a management strategy for allergy diseases. Its effects on specific immunotherapy have been previously studied in humans. The aim was to evaluate the possible adjuvant effects of probiotics on allergen-specific immunotherapy in an allergy mouse model. Methods: C57BL/6 mice were pre-sensitized by epicutaneous patching with recombinant Der p 2, and subsequently orally fed with either heat-killed wildtype L. casei (Lc) NaHCO 3 buffer for five weeks (n=6 per group). All mice then received two subcutaneous (s.c.) immunizations of Der p 2 to mimic allergen immunotherapy, followed by aerosol challenge with Der p 2 a week later. Results: The Lc fed Der p 2 sensitized mice showed significantly lower Der p 2 specific-IgE and IgG1 after s.c. immunizations and airway challenge compared to the NaHCO 3 buffer control group. Spleen T-cells and mesenteric lymph node (MLN) cells of Lc fed mice showed suppression of Th-2 (IL-5, IL-13, IL-10, IL-4) and pro-inflammatory (TNF-! cytokines), with an increase in TGF-" production in MLN cells. In addition, these Lc-fed mice but not the controls showed attenuated lung inflammation as demonstrated by the significantly reduced lung histopathology and total cell count in BALF. These results suggest that combined administration of the heat-killed L casei and allergen could effectively down-regulate the pre-existing Th-2 allergic responses. Conclusion: Probiotics such as L casei have the intrinsic adjuvancity and immunomodulatory properties that could potentially be exploited to improve the efficacy of allergen-specific immunotherapy for allergic respiratory diseases. Vaccinations with recombinant ProDer p 1 produced in E.coli prevent the development of house dust mite allergy David Walgraffe 1 , Christel Mattéotti 1 , Céline Marchand 1 , Lida Garcia 1 , Michel Vandenbranden 2 , and Alain Jacquet 1 . 1 Université Libre de Bruxelles/ IBMM, Laboratory of Experimental Allergy, Gosselies, Belgium; 2 Universite Libre de Bruxelles, SFMB, Bruxelles, Belgium. Our recent data indicated that recombinant ProDer p 1, an enzymatically inactive Der p 1 precursor form, produced in E.coli displayed very low IgE binding capacity but retained its T-cell reactivity. To confirm this hypoallergenic character, the present study investigated the prophylactic potential of ProDer p 1 produced in E.coli in a Der p 1 sensitization murine model. Gel fitration chromatography as well as FTIR spectroscopy demonstrated that ProDer p 1 is produced in E.coli as aggregates containing an higher "-sheet content than ProDer p 1 produced in wild-type P.pastoris or in wild-type CHO cells. These conformational changes could likely explain the drastic reduction of its in vitro IgE binding activity towards human allergic sera. Compared with ProDer p 1 produced in CHO, vaccination of naBve mice with ProDer p 1 coli adjuvanted with alum induced a mixed Th1-Th2 immune response characterized by the weak production of specific IgG2a, IgG1 antibodies and the absence of the specific IgE titers. Unpredictably, the drastic reduction of specific IgE titers was maintained after mice sensitizations with natural Der p 1/alum and subsequent challenges with aerosolized house dust mite extracts. Moreover, the Th1-Th2 bias was confirmed as vaccination with ProDer p 1 coli induced the secretion of IFN+ and IL-5 whereas the control allergic group secreted only IL-5. ProDer p 1 coli prevented the development of airway eosinophilia following house dust mite extract challenges of immunized mice. Furthermore, the increase in airways sensitivity to inhaled methacholine was reduced by the prophylactic vaccination. Taken together, our results indicated that ProDer p 1 coli could represent an hypoallergen suitable for the prevention of house dust mite allergy. Tolerance during dose-increase phase of specific immunotherapy with standardized house-dust mite extract in asthmatic children Li Xiang, Hong-yan Zhang, Ji-mei Zhao, Jian-xin He, and Kun-ling Shen. Bejing Children_s Hospital Affiliated to Capital University of Medical Science, Center of Asthma, Beijing, China. Background: To determine the tolerance during dose-increase phase of specific immunotherapy (SIT) with standardized house-dust mite extract in asthmatic children. Methods: Subcutaneous SIT with standardized house-dust mite extract was administrated to 24 asthmatic children with allergy to house-dust mite (Dermatophagoides pteronyssinus). We monitored the immediate and late side-effects after each allergen injection during dose-increase phase of immunotherapy treatment. Lung function test was done before and 30 minutes after each injection for every patient. Results: All patients completed a total of 370 injections during dose-increase period. 27 injections (7.30% of all injections) caused immediate side-effects within less than 30 minutes after injection in 14 patiens (58.33% of all patients), in which 26 side-effects (96.3% of all early reactions) were mild local reactions (G5cm in diameter) and 1 episode of mild asthma occurred. 21 patiens(87.50% of all patients) reported 38 late side-effects(10.27% of all injections) 30 minutes after injection, 9 late side effects occurred in 7 patiens were mild systemic reactions which included asthma, allergic rhinitis, allergic conjunctivitis and allergic dermatitis, all those systemic reactions responded well to ordinary treatment. Others late side effects were 28 mild local reactions and 1 moderate local reaction (5cm in diameter). The change rate of lung function parameters after each injection showed a wide range of variance such as FEV1(j18.9%15.8%), PEF(j35.6%33.3%), FEF 25 (j29.5%51.7%), FEF 50 (j38.648.4%), FEF 75 (j49.1%249.2%), MMEF(j38.2%101.7%), but there was no significant difference among all injections during dose-increase period. Conclusion: Subcutaneous SIT with standardized house-dust mite extract showed a good tolerance in asthmatic children. A majority of local reactions and all systemic reactions were mild and responded well to ordinary treatment and adjustment of dose-increase schedule. No significant change of lung function was observed during dose-increase phase of SIT. 45 patients of simple perenial bronchial asthma and steroiddependent bronchial asthma were studied before and after Immunotherapy with house dust mite (D.farine.pteronyssinus) alleregn vaccine. The three years course of specific Immunotherapy (SIT) in 45 patients with perenial bronchial asthma and steroid-dependant asthma showed significant reduction in amount of drug intake and attacks during the treatment. The purpose of study was to investigate the long term effect on the severity and of attacks in worst months as well as the amount of drug intake (I.C.S and oral steroids) in perenial and steroid-dependent asthma even after discontinuation of dust mite immunotherapy. The inclusion criteria during the study was patients suffering from perenial bronchial asthma sensitive to house dust mite (3+/4+) by skin prick test, eosiniphilia, and specific IgE, while 10 patients were on standard control with symtomatic medications. A total of 55 patients were followed up for three years during Immunotherapy and three years after immuntherapy, SIT treated group had significantly less intake of ICS and BD while patients without Immunotherapy were found to have attacks in perenial bronchial asthma, while in steroid dependant asthma severity of the disease was found to decrease only during immunotherapy. The result of the study revealed that specific Immunotherapy for three years with standardised allergen extracts of house dust mite results in decrease in severityand intensity of attack in both perenial; and steroid-dependent bronchial asthma, while on discontinuation of SIT, steroid-depenent astma patients reverted back. Effects of immunotherapy on symptoms, PEFR and spiromery in patients with allergic asthma to house dust mite (HDM) on inhaled steroid therapy Gobinda Das. Shaheed Suhrawardy Hospital, Allergy and Clinical Immunology, Dhaka, Bangladesh. The present study was designed to investigate the effects of immunotherapy (IT) with an extract of Dermatophagoides pteronyssinus (Alk-Abello, Spain and Credisol, India) during a 36 months period in patients with allergic asthma to house dust mites. We included 50 patients (mean age 18 years) treated with a combination of IT and inhaled corticosteroids (ICS) in comparison to another 50 (mean age 20 years) treated with ICS alone. We evaluated symptom scores, Salbutamol use, peak expiratory flow rates (PEFR) and spirometry during 24 months of therapy with ICS and in the 12 months after ICS interruption. The two kinds of treatment were efficient and comparable in relation to symptom score, salbutamol use, morning PEFR, FVC and FEV1, but patients treated with IT and ICS had a faster improvement of symptom score and PEFR variability. The interruption of ICS after 24 months of therapy was linked to an impairment of all end points which were more pronounced in patients previously treated only with ICS. These findings suggest that in selected asthmatic patients allergic to house dust mites, the association of IT and ICS is more effective than therapy with this ICS alone and to a lower rate of relapse after the interruption of therapy with ICS. pteronyssinus are the main sources of the house dust allergens and an important etiological factor in allergic respiratory diseases. Specific allergen vaccination is an important tool to reduce allergic symptoms and prevent more severe respiratory symptoms with patients. The aim of the study is to evaluate the incidence of local and systemic reactions during the HDM immunotherapy. We investigated a group of 1200 patients allergic to dermatophagoides pt. The age and sex analyses included patients aged 15-30 showing no significant difference. We divided two groups depending on degree of the skin reactivity: First group: skin reactivity of 2+ and 3+(no-1100) and second group: skin reactivity of 4+ (no=100). We have commenced with the subcutaneous specific immunotherapy in accord with the standard protocol. We analyzed the distribution if local and systemic reactions during the immunotherapy in both groups. Out of 27000 injections (97%) had not reaction. 810 injections caused local reaction (190 in the first group and 620 in the second group). We treated them with oral antihistaminic and local with ice. We can conclude that specific immunotherapy with house dust mites is safety. The majority of reactions were local and occurred within less than 30 minutes after inections. Premedication with antihistaminics is important especially in the beginning of the immunotherapy. Specific immunotherapy for house-dust mite in a patient with anisakis allergy: adverse reaction Maria Teresa Ventura, Maddalena D'Erasmo, Roberto Armando Tummolo, Augusto Arsieni, and Rosalba Buquicchio. University of Bari, Medical School, Policlinico, Department of Internal Medicine, Immunology and In, Bari, Italy. The nematode Anisakis simplex belongs to the Anisakidae family. The third-stage larvae are commonly found in the entrails and in the muscles of many fishes and if accidentally ingested can cause clinical disorders and sensitization in humans. Previous studies have shown cross reactivity between Anisakis simplex and different dust mite species. We observed gastrointestinal symptoms during specific immunotherapy for house dust mite in a patient with Anisakis sensitization. In a patient suffering from allergic asthma with sensitization to house dust mite developed gastrointestinal symptoms during Sublingual Immunotherapy (SLIT). Sensitization was evaluated by skin prick tests with common inhalant allergens, standard food extracts, commercial extracts of Anisakis simplex and Radioallergosorbent Test (RAST) to specific IgE antibodies. We obtained that Skin Prick Tests (STP) with common inhalant allergens were positive to graminacee (++j), cypress (++j) and house dust mite (+++j). SPT with standard food extract and commercial extract of Anisakis simplex demonstrated sensitization to Anisakis simplex. RAST showed specific IgE antibodies against allergens of Anisakis simplex. Moreover, it is important to underline that allergenic cross-reactivity between Anisakis simplex and other nematodes, such as arthropods, is well known. In a recent work, Johansson and his group underlined an association between sensitization to Anisakis simplex and four different dust mite species (Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae and Dermatophagoides pteronyssinus). In their study, serum samples collected from 69 subjects allergic to dust mites were analyzed for IgE to Anisakis simplex by CAP FEIA and immunoblotting inhibition. They found that 14/69 patients had detectable levels of IgE antibodies to Anisakis simplex. In the CAP FEIA inhibition, all four mite extracts inhibited, to various degrees, the IgE response to Anisakis simplex on the solid phase. In the light of these findings, it is possible to hypothesize that immunotherapy for dust-mite could pose an unintentional challenge in patients with gastroallergic anisakiasis, due to crossreaction with anisakis simplex antigens. Even if further observations of adverse reactions to sublingual immunotherapy with dust mite in patients sensitized to Anisakis are needed, our data suggest that oral treatment should be discouraged in these patients. Double-blind placebo-controlled study of treatment with a depigmented and glutaraldehyde-polymerised extract of dermatophagoides pteronyssinus and D. farinae: improvement in skin sensitivity Background: There is a high prevalence of asthma in the Canary Islands, Spain. Most asthmatic patients are sensitised to house dust mites. Exposure to mite allergens is constant and levels regularly exceed 10 mcg of Der p 1 per gram of dust. Immunotherapy using depigmented and glutaraldehyde modified allergen vaccines produces clinical benefit, as evaluated by improvement in the shock organ, symptom and medication scores, quality of life and skin sensitivity. Objective: To evaluate the evolution of skin sensitivity in a group of asthmatic patients sensitised to both mites in a double-blind and placebo-controlled study of 1 year of duration. Materials and Methods: Sixty-four patients were included, and 52 finished the study (26 in the active group and 26 in placebo). The native extract of D. pteronyssinus contained 20.35 mcg of Der p 1 and 12.30 mcg of Der p 2, and the D. farinae extract contained 19 mcg of Der f 1 per mg of freeze-dried extract. Due to the polymerisation process, Der p 1, Der p 2 and Der f 1 were not detectable in the modified extract. Patients were skin tested at baseline and at the end in duplicate, on the volar surface of the forearm using the same batch of native D. pteronyssinus extract at 0.002, 0.02, 0.2 and 2 mg/ml. The vial of maximum concentration had a potency of 100 HEP/ml and contained 40.70 mg of Der p 1/ ml and 17.22 of Der p 2. Glycerinated saline solution and histamine HCl 10 mg/ ml were used as negative and positive controls, respectively. Skin reactions were recorded after 15 minutes. The results were expressed as the 10 HEP value. The Wilcoxon test was used to compare the results. Results: Patients treated with the modified allergen extract received a total accumulated dose of 593 mcg of freeze-dried modified material of D. pteronyssinus and 453 mcg of D. farinae. The active group showed a significant difference from baseline (P=0.029, Wilcoxon), whereas the placebo group did not (P=0.900). The active group needed at the end of the study a median of 21.9 mcg more of allergen extract to achieve the value of 10 HEP, whereas in the placebo group the figure was j0.31. Conclusion: This double-blind placebo-controlled trial demonstrates that the treatment of mite allergic asthmatic patients with a mixture of depigmented and glutaraldehyde-polymerised extracts of D pteronyssinus and D. farinae induces a significant decrease in the skin sensitivity. Specific Immunotherapy with depigmented polymerized allergen extracts of domestic mites in patients with persistent rhinoconjunctivitis Angelika Sager 1 , and Hartwig Lauter 2 . 1 LETI Pharma GmbH, Medical Department, Witten, Germany; 2 Fachkrankenhaus Kloster Grafschaft, Allergology, Schmallenberg, Germany. Background: There is limited knowledge about the clinical relevance of a sensitization with domestic mites s.a. Euroglyphus maynei (Em), Lepidoglyphus destructor (Ld), Acarus siro(As) and Tyrophagus putrescentiae (Tp). The efficacy and safety of a SIT with these mites has not been described until now. Patients and Methods: We investigated 107 patients suffering from persistent rhinoconjunctivitis by i.c test, specific IgEs and placebo controlled nasal challenge test with domestic mite allergens. If the patient had a positive i.c test and/or specific IgEs and a positive nasal challenge test SIT was initiated with the respective depigmented polymerized allergen extract. 20 patients have been treated with the following allergen extracts either alone or in combination for 6 months: 9x As, 6x Ld, 6x Tp, 4x Em. Patients symptoms and medication consumption was recorded using a standardized questionnaires. Results: 17/20 patients (85%) improved during the course of SIT. The symptom and medication score was reduced in parallel. 1 patient did not change during SIT and 2 patients were lost to follow-up and therefore considered as treatment failure also. There were no systemic reactions and only few and mild local reactions reported. Conclusion: This is the first report of a successful SIT with Euroglyphus maynei, Lepidoglyphus destructor, Acarus siro and Tyrophagus putrescentiae. The treatment was efficacious and safe. Background: There is a high prevalence of asthma in the Canary Islands, Spain. Most asthmatic patients are sensitised to house dust mites. Exposure to mite allergens is constant and levels regularly exceed 10 mcg of Der p 1 per gram of dust. Immunotherapy using glutaraldehyde modified allergen vaccines demonstrated clinical benefit in several studies. The introduction of a depigmentation step before polymerisation inactivates the enzymatic activity, removes pigments and enhances the solubility of the polymer. Objective: To evaluate the clinical efficacy of a depigmented and polymerised vaccine containing 50% D. pteronyssinus and 50% D. farinae in patients sensitised to these mites in a double-blind and placebo-controlled trial. Materials and Methods: Fifty-one patients were randomly allocated to either receive active treatment (n=26), or placebo (n=25). The native extract of D. pteronyssinus contained 20.35 mcg of Der p 1 and 12.30 mcg of Der p 2, and D. farinae contained 19 mcg of Der f 1 per mg of freeze-dried extract. Due to the polymerisation process, Der p 1, Der p 2 and Der f 1 were not detectable in the modified extract. Patients recorded the symptom and medication scores on a diary card. The area under the curve (AUC) and the number of days free of symptoms and/or medication were calculated for each patient. The values of AUC were compared groupwise using the Mann-Whitney test, and the Hodges-Lehmann estimator was used to measure the effect size. Fisher's exact p value was used for the number of days free of symptoms or medication. Results: Patients treated with the modified extract received a total accumulated dose of 593.3 mcg of freeze-dried modified material of D. pteronyssinus and 452.7 mcg of D. farinae. The difference of AUC of symptom and medication scores between both groups was significant (pG0.05). The values of the Hodges-Lehmann were j1.6 (95%CL j18.9, j4.4) and j8.6 (95%CL j14.9, j2.2) for symptom and medication scores, respectively. Patients treated with the modified allergen had more days free of symptoms and medication (Fisher's exact p value G0.0001). Conclusion: This double-blind placebo-controlled trial demonstrates that the treatment of mite allergic, asthmatic patients with a mixture of depigmented and polymerised extracts of D pteronyssinus and D. farinae produces a clinical benefit in symptom and medication scores, with an important effect size (zero value was not included in the 95% confidence limits). Clinico-immunologic study on immunotherapy with Periplaneta americana extract: a double blind placebo controlled trial Deepsikha Srivastava 1 , Shailendra N. Gaur 2 , Naveen Arora 1 , and Bhanu P. Singh 1 . 1 Institute of Genomics and Integrative Biology, Allergy and Immunology, Delhi, India; 2 Vallabhbhai Patel Chest Institute, Dept. Of Respiratory Medicine, Delhi, India. Background: Cockroaches produce several potent allergens associated with sensitization and development of asthma. In highly susceptible or symptomatic patients, allergen-specific immunotherapy may be beneficial, but the data are limited. The present study is aimed to evaluate the role of immunotherapy in cockroach allergic patients using standardized whole body extract. Methods: Twenty patients belonging to allergic asthma, rhinitis or both were recruited in active group and 20 in placebo group. The patients in active group were administered with P. americana extract subcutaneously whereas the other group received placebo injections for one year. To assess the benefits of immunotherapy, the data was analyzed for symptom/drug score, airway reactivity and immunoglobulins (IgE, IgG1 and IgG4) at the baseline and after completion of one year of immunotherapy. Immunoblots of specific IgE and IgG4 were made before and after therapy. Results: Good improvement was observed in clinical status of patients with respect to in vivo and in vitro parameters. Changes in symptom score, airway reactivity and IgG4 values were highly significant in comparison to baseline values and placebo group. Skin test and IgE values showed non-significant reduction. Immunoblotting of specific IgG4 has shown development of more bands of increasing density after 1 year of immunotherapy. While IgE blot showed similar pattern of bands before and after immunotherapy. Conclusion: Cockroach immunotherapy is well tolerated in patients of allergic rhinitis and asthma. IgG4 immunoblotting has shown a correlation of clinical efficacy with the immunologic response. At the question of the cytokines therapy of the nosocomial pneumonia Anna Kostyushko, and Elena Markelova. Siberian Branch of Russian Academy of Medical Science, ?The Fareastern Scientific CentreX, Research Insti, Vladivostok, Russian Federation. Purpose: There were 74 patients with the hospital pneumonia of serious current under our observation. Patients were distributed to the way of introduction of the immunotroping drug. The first group (22 patients) received recombination IL-2 on 500 000 ÌÅ dose intravenously. The second group (24 patients) received hypodermic introduction of the same dose of the drug, and patients of the third group (28 person) received the combined introduction of recombination IL-2: 250 000 ÌÅ dose of inhalation with the help the ultrasonic inhaler simultaneously with hypodermic introduction of 250 000 Y 500 000 ÌA dose. The course of treatment consisted of 2Y3 procedures. The dynamic research of the basic clinic-laboratory indexes were given to all our patients. Results and Conclusion: It was fixed, that all three ways of recombination IL-2 introduction provided positive dynamics of indexes of immunity on the background of basic anti-inflammatory therapy of the pneumonia as the result of our research. However, the condition of the local immunoreactivity of lungs was recovered only at combined inhalation and hypodermic introduction (patients, contents of metabolites of oxide of nitrogen and !1-acidic glycoprotein in the condensate of the exhaled moisture was normalized, reaching similar indexes of healthy people). It is important to note, that complete regress of radiological indexes was only at combined application of recombination IL-2 inhalationly and subcutaneously. The analysis of frequency rate of recombination IL-2 introduction showed, that it was not enough single-pass introduction of the drug for the reduction of indexes of the local immunity, and rates of their reduction in the course of the repeated combined way of introduction were identical and depended on gravity of the condition and the expressiveness of the inflammation in bronchopulmonary system. The increase of frequency rate of recombination IL-2 introduction more than 2Y3 times did not accelerate rates of clinical convalescence, reduction rate of immune indexes and biochemical criteria of the degree of the expressiveness of the inflammation in respiratory paths. Thus, we believe that it was efficient combined recombination IL-2 application of the 250 000 ÌÅ inhalation dose and 500 000 ÌÅ dose subcutaneously 2 Y 3 times with the interval of 72 hours for treatment of patients with the serious hospital pneumonia. Background: Diagnosis of asthma or non-asthma is critical for clinical research. Using methacholine challenge testing (MCT), a well established bronchial challenge test for assessing airway responsiveness, we have identified groups of patients who have been incorrectly diagnosed as asthmatic or non-asthmatic. Methods: We screened 68 volunteers participating in clinical research in asthma after they provided informed consent. All volunteers were questioned about their general health, asthma symptoms medication and family history of asthma. Atopic status was determined with skin prick tests to a standard panel of aeroallergens. A baseline spirometry was performed prior to MCT. MCT was conducted following a recognised dosing protocol (Crapo, R.O. et al Am J Respir Crit Care Med 2000 Vol 161 pp 309Y329) using a Mefar MB3 breath activated dosimeter. Volunteers considered to be asthmatic commenced at the lowest methacholine dose of 0.03125 mg/ml and continued up the dosing protocol until a PC20 was reached. After each dose their FEV1 was obtained at 1 1/2 mins and at 2 1/2 mins post-inhalation with a subsequent 2 mins rest prior to the next dose. Those considered to be non-asthmatic started their MCT challenge from 0.5 mg/ml dose of the methacholine and continued up-dosing until the highest dose. The definition of asthma for the purpose of our studies was a PC20 below 8mg/ml at MCT. Non-asthma was defined as completion of the MCT at 16mg/ml without attaining a PC20. Results: Of the 68 subjects screened for clinical research studies, 29 claimed to have a physician diagnosis of asthma, but in 8 this was not confirmed by our MCT criteria. Of these 8 subjects, 7 had normal spirometry (FEV1 980% predicted) while 1 had FEV1 of G80% predicted.Conversely, 39 volunteers reported non-asthma. Of these 2 had symptoms at methacholine challenge testing and attained a PC20 G8mg/ml, thus fulfilling the definition of asthma according to our criteria. Conclusion: Diagnosis of asthma and non-asthma based on existing symptoms, lung function testing and physician based diagnosis is surprisingly unreliable. Asthma in pregnancy: diagnostic methods Zahra Pourpak, Katayoon Bidad, Hasan Haidarnejad, Giti Shaiestehfar, and Mostafa Moin. Immunology, Asthma and Allergy Research Institute, Immunology Department, Tehran, Islamic Republic of Iran. Introduction: Asthma is the most common respiratory disorder to complicate pregnancy and represents a significant public health issue. In many occasions, especially in the last trimester due to the pressure of unborn child, these patients are unable to perform standardized spirometry. The Impulse Oscillometry Technique (IOS) may be suitable in this application since it allows the assessment of airway obstruction by requiring minimum patient cooperation. Aim: Our aim was to identify the relationship between R5, X5, and Resonant Frequency to the 1s forced expiratory volume (FEV1) and forced vital capacity (FVC) in pregnant women. Methods: In 42 asthmatic pregnant patients aged 18Y41 yr (mean 28.5 yr) spirometry and impulse oscillometry (Erich Jaeger, Germany) was performed according to standard guidelines and manufacturers instructions. Relationships between spirometric indices and total Resistance (R5), central resistance (R20), peripheral reactance (X5) and Resonance Frequency (FRes) were obtained using correlation and regression analysis. Volumes are expressed in litres and resistance in kpa.1.s. Results: Mean T SD of FEV1 was 2.32T0.46, FVC was 2.93T0.64 , FEV1/ FVC was 80.90T15.81 , R5 was 0.49T0.14 , X5 was j0.11T0.14 and FRes was 19.71T5.00. There was significant (pG0.001) correltion between the R5 and FRes, each with FEV1.R5: FEV1 = 3.14 Y 1.34 R5, R 2 = 0.21FRes: FEV1 = 3.90 Y 3.24 fres, R 2 =0.22. Conclusion: These results suggest that Impulse Oscillometry is a valuable tool to assess airway obstruction as it is simple and requires minimal subject cooperation. This new field of oscillometry application may be particularly useful in patients who are unable to perform spirometry. Further studies are required to determine the sensitivity and specificity of this technique. The association between FeNO and acute exacerbation of asthma in children Sun-Hee Choi, and Yeong-Ho Rha. College of Medicine, Kyung Hee University, Department of Pediatrics, Seoul, Republic of Korea. Rationale: Asthma is a Th2 cell mediated chronic inflammatory airway disease. Bronchoscopic biopsy is the best method to evaluate airway inflammation but it is too invasive for children. Measurement of exhaled nitric oxide (FeNO) is a relatively simple, noninvasive, and very reproducible test for detection of endogenous inflammatory signals in childhood. Recently measurement of FeNO level is emerging as a non-invasive tool for airway inflammation. The aim of our study was to determine the relationship between FeNO and acute asthmatic exacerbation, and to decide whether measurement of FeNO could predict acute asthma exacerbation. Methods: Thirty eight children with mild to moderate persistent asthma aged from 3 to 15 years who presented to Kyunghee Medical Center were studied. FeNO levels were measured by chemiluminescence during exhalation into the NO analyzer. Patient's data were based on out-patient records. We compared FeNO levels between exacerbation group and non-exacerbation group. Results: The mean FeNO level in the asthmatic children was 16.7T13.9 ppb. In the range of abnormal FeNO level (Q10 ppb), there was significant difference between asthma exacerbation group and non-exacerbation group (p=0.004). There was also significant correlation between FeNO level and acute asthma exacerbation (p=0.003). Conclusion: Exhaled nitric oxide levels were related with acute asthmatic exacerbation in childhood asthma. Thus measurement of FeNO can be a promising clinical tool for predicting asthma exacerbation in mild to moderate persistent asthmatic children. The analysis of the level of bronchial hyperresponsiveness in astmatic children (2 -18 years) by dosimetric, oscilometric and tidal breathing methods to histamine and methacholine and analysis of polymorphisms genes ADAM33 and STAT6 Radek Vrtel 1 , Radek Vodicka 2 , Martin Zapalka 1 , and Frantisek Kopriva 1 . 1 Teaching Hospital of Palacky University Olomouc, Pediatrics, Olomouc, Czech Republic; 2 Teaching Hospital of Palacky University Olomouc, Medical Genetics, Olomouc, Czech Republic. Background of the study: The main pathogenic mechanism of bronchial asthma is chronic eosinophil-based inflammation of the bronchial mucosa. Airway hyperresponsiveness (AHR), is known to be a major risk factor for the development of asthma and to be equivalent of asthma severity. Methods: Main aim of this project, supported by IGA MZ CR No. NR8383-3/ 2005 , is to test if the level of AHR is associated with certain alleles or haplotypes of genes (ADAM 33 and STAT 6) involved in immune system activity in course of allergic illness development and airway hyperresponsiveness. 80 children with bronchial asthma had histamine and methacholine brochnoprovocation tests in a sequence with increasing levels using dosimetric method and MedicAid and DeVilbiss nebulizers. Oscilometric assessment I was followed by baseline spirometry, and with oscillometric assessment II after a decrease in FEV19 20% versus baseline. Examinations of responsible polymorphisms of ADAM 33 and STAT 6 genes were accomplished by sequence analysis in 80 children with asthma bronchiale, their sibs and parents and 50 control healthy children. Results: Examinations of responsible polymorphisms of ADAM 33 and STAT 6 genes were accomplished by sequence analysis in 80 children with asthma bronchiale, their sibs and parents and 50 control healthy children. In the evaluation of the result it could be demonstrated that changes in resistance R5 and R20 between oscillometric assessments I and II more than 40% are consistent with a decrease in FEV1 more than 20% in both dosimetric method. The statistical evaluation of the association of genes polymorphisms with asthma bronchiale occurrence and airway hyperresponsiveness level in children will be presented by poster. Conclusion: These results show that resistance R5 over R20, as measured with IOS (impulse oscillometry), significantly correlates with the-gold standard-FEV1. Description of influence of particular sequence changes could contribute to additional insight to the airway hyperresponsiveness genetical background. Differential flow analysis of exhaled nitric oxide in asthmatic patients Valeriya Nemtsova, Irina Tikhonova, and Inga Fedotova. Kharkov Allergological Center, Allergology, Kharkov, Ukraine. Background: The majority of asthmatic patients achieve control of their illness, but not all. It is therefore crucial to develop strategies that help the clinician monitor the disease, improving the response to treatment. Methods: We have quantified the inflammation in central and peripheral airways by measuring exhaled nitric oxide (NO) at multiple exhalation flows in 42 asthmatic patients at different levels of severity (mild = 11; moderate stable = 10; moderate during exacerbation = 9; severe = 12, 6 of whom were receiving oral corticosteroids) and 15 healthy control subjects. Results: Bronchial NO (bNO) in patients with mild asthma (2,283+/j346 pL/s) [mean+/jSD] was higher than in patients with moderate stable asthma (1,317+/j61 pL/s, pG 0.001), in patients with severe asthma receiving inhaled corticosteroids (ICS) [1,023+/j72 pL/s, pG 0.001], and healthy control subjects (697+/j31 pL/s, pG 0.001). There were no differences between bNO in patients with mild asthma compared to patients with severe asthma receiving ICS and oral corticosteroids (2,335+/j269 pL/s). Patients with exacerbations showed a higher bNO (3,521+/j373.2 pL/s, pG 0.01) compared to the other groups. Alveolar NO was higher in patients with severe asthma receiving oral corticosteroids (3.1+/j0.2 parts per billion [ppb], pG 0.0001) than in the other groups but was not significantly higher than in patients with moderate asthma during exacerbation (2.7+/j0.4 ppb) . No differences were seen in NO diffusion levels between the different asthma groups. All the measurements were highly reproducible and free of day-to-day and diurnal variations. Conclusion: Differential flow analysis of exhaled NO provides additional information about the site of inflammation in asthma and may be useful in assessing the response of peripheral inflammation to using therapy. Monitoring and treatment of childhood asthma in singapore: a questionnaire study in physicians Colin Tan, Belle Wong, Daniel Goh, and Hugo Van Bever. National University Singapore, Department of Pediatrics, Singapore, Singapore. Introduction: Asthma is the most common chronic disease in children and in Singapore over 25% of children have ever been diagnosed with asthma by age 15. Most children are seen in the primary care setting, thus it is of value to study the management practices of general practitioners and pediatricians in comparison with gold standards. The aims of the study were to investigate: a) methods of monitoring of childhood asthma, b) practices in managing acute asthma exacerbations, and c) choice of therapy in maintenance treatment. Methods: 2100 questionnaires of 35 questions were sent by post to general practitioners and pediatricians throughout Singapore. 173 valid responses were received and results were compared to 2006 Global Initiative for Asthma (GINA) guidelines. Results: Of respondents 76.3% were general practitioners. For monitoring childhood asthma, most physicians (89.1%) did not use symptom score cards/ diaries, while 37.6% did not use peak-flow meters/spirometry. For acute treatment, 83.8% used a short-acting â-agonist, but only 41.0% used oral corticosteroids in outpatients. In the maintenance treatment of asthma, across all age-groups (infants, preschoolers, and older children) the most commonly used first line drug was uniformly an inhaled corticosteroid. However, combinations of long acting beta-agonists (LABA) and inhaled corticosteroids were uniformly the second most popular maintenance treatment. Maintenance treatment was frequently used by most doctors in treating preschoolers (62.4%) and older children (64.7%). In contrast, most doctors seldom (45.1%) or never (21.4%) applied maintenance treatment to infants. The vast majority (158; 91.3%) of doctors never used immunotherapy in asthma management, and 2.9% of respondents believed immunotherapy was not available locally. Conclusion: Greater usage of diaries/score cards can be encouraged along with objective peak flow/spirometry measurements. Management of acute exacerbations is appropriate, but corticosteroids are under-prescribed by most doctors. LABA continues to be prescribed for maintenance despite a lack of established safety profile for infants, along with recommendations that they only be used selectively in patients that are poorly controlled by inhaled corticosteroids. Immunotherapy is not widely used and more awareness/ instruction should be generated among doctors. Risk of HBV reactivation in asthma or COPD patients treated with corticosteroids Tae Objective: To evaluate the course of HBV infection in stable HBV carriers using ICS or SC for the treatment of asthma or chronic obstructive pulmonary disease (COPD). Methods: HBsAg seropositive patients using ICS were reviewed retrospectively from Jan 1997 to Mar 2007. We reviewed data of 183 patients who met our inclusion and exclusion criteria including dose and duration of ICS and SC treatment, HBV DNA titer, HBeAg/Ab and AST/ALT serum level, and usage of antiviral drugs such as lamivudine. Results: Among the 183 patients who were using ICS, 66 patients were treated with SC at least on one event. HBV reactivation occured in 5 out of 66 (7.6%) subjects who have ever used SC and 2 out of 117 (1.7%) subjects under ICS treatment alone. Among the 5 subjects with HBV reactivation who have ever used SC, 4 (80%) subjects were treated with SC chronically due to difficult-tocontrol asthma with more than 9000mg of total cumulative dose of prednisolone. 2 subjects with HBV reaction who were using only ICS were using moderate dose ICS. Conclusion: Chronic use of SC and high total cumulative dose of SC are risk factors of HBV reactivation in patients with asthma and COPD. ICS treatment alone is not a risk factor of HBV reactivation. Prophylactic antiviral therapy should be considered in patients who needs chronic or high dose SC treatment. Background: The prevalence of asthma is increasing in the last years. Also gastro-esophageal reflux disease (GERD) is increasing in the last years. It is known that gastro-esophageal reflux (GER) may be present and also may aggravate asthma. There are no information regarding GERD and asthma in Romania. Aim: Our study was to evaluate the presence of GER in allergic asthma in Transylvania. Methods: We evaluate the patients with 56 allergic asthma for the presence of GER. The pts had moderate persistent asthma treated with glucocorticoid inhalers (and/or LABA). The pts had a 24-hours oesophageal pH-monitoring. Reflux episodes identified reflux events as the percentage of time where the pH was less than four. The pts had also an upper endoscopy. The pts were also ask about there asthmatic symptoms during nights and regarding the heart burns. Results: The mean age was 36 years (18-54 years); 32 of the pts were women. There were only 3 pts (5.7%) having heart burns. On 24-hours oesophageal pH-monitoring we found 18 pts (32%) having acid reflux during day and night. On endoscopy there were only 7 pts (12.5%) with esophagitis. There was no relationship between spirometry results on one hand, and parameters of gastroesophageal reflux on the other. The pts receive also for 4 weeks term a proton pump inhibitor (PPI) (pantoprazol 80 mg/day). The measures of symptoms (mean daily daytime and nighttime asthma symptom scores) did not change significantly during the 4 weeks treatment with PPI . There was no influence on rescue medication in the majority of the pts. Only 2 pts reported an improvement of the asthma, with lower need for rescue medication. In our Romanian patients we found no correlations between GER. Conclusions: Our results indicate that acid in oesophagus or its short term inhibition by proton pump inhibitors, has no influence on allergic asthma. The clinical implication of different pathophysiologic manifestations between asthma and pure COPD Background: Although physiologic and anatomic differences of chronic obstructive pulmonary disease (COPD) and asthma are separated into distinct categories of disease, in clinical practice a significant proportion of patients have characteristics that are classically associated with both conditions. Methods: To determine whether patients with fixed airflow obstruction have distinct cellular and biochemical characteristics depending on their asthmatic features, we characterized 30 consecutive outpatients presenting with fixed airflow obstruction by bronchodilator response, peak expiratory flow variability, serum total immunoglobulin E, allergen skin prick test, methacholine bronchial challenge test (MCT), and bronchoalveolar lavage (BAL). In addition, to determine significance of MCT on differentiating asthma from COPD, we exam MCT on 111 outpatients divided to asthma, pure COPD, and control group. Results: We found that subtypes of T lymphocytes did not show significant difference in number and ratio on both groups, but neutrophils in BAL fluid was significantly lower in group with asthmatic features. Additionally, their cytokine analysis showed significantly higher level of IL-4 in asthmatic patient compared with COPD patients, while lower level of IL-5 and INF-gamma in asthma group compared with COPD group. In MCT results, area under the receiver operating characteristic (ROC) curve of sensitivity and specificity has shown that best cut-off value of PC 20 is 15.4 mg/ml in classifying the asthma and pure COPD. Conclusion: We conclude that despite of similar fixed airflow obstruction, subjects with asthmatic features have distinct cellular and biochemical characteristics compared with pure COPD. In addition, we suggest that the MCT using the new cut-off value could be a more useful diagnostic tool for distinguishing asthma from pure COPD. Based on these findings, we expect that this distinction of asthma from pure COPD using the different pathophysiologic manifestations could provide more proper and specific therapeutic management for the patients. Airway remodeling in patients with allergic bronchopulmonary aspergillosis Hidenori Tanimoto, Yasuo Takeuchi, Masami Taniguchi, Akemi Saito, Kiyoshi Sekiya, Hiroshi Yasueda, and Kazuo Akiyama. National Hospital Organization, Sagamihara National Hospital, Clinical Research Center for Allergy and Rheumatol, Kanagawa, Japan. Background: Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity reaction to Aspergillus fumigatus (Asp.f) induced by IgE and IgG antibodies which progress to central bronchiectasis (CBE) even at an early stage. We hypothesized that the airway remodeling (=irreversible change of bronchial wall) in patients with ABPA is accompanied by not only CBE but also persistent bronchial narrowing at an early stage of ABPA. Methods: We evaluated bronchial reversibility (BR) using salbutamol and airway hyperresposiveness (AHR) using acetylcholine (Ach) in ABPA patients with CBE (=classical ABPA) and without CBE (=seropositive ABPA), and compared the BR and AHR of these patients with those of patients with allergic fungal asthma without the IgG antibody to Asp.f. Nineteen classical ABPA, 11 seropositive ABPA, and 21 allergic fungal asthmatics were participated in this study. Results: All the subjects were positive for the IgE antibody to Asp.f. All the ABPA patients with or without CBE were positive for the IgG antibody to Asp.f. In all classical ABPA patients, but not in seropositive ABPA patients, typical CBE findings were detected in HRCT. There were no differences in age, total IgE level (RIST), and %FEV1 among the three groups. However, not only classical ABPA patients, but also seropositive ABPA patients showed significantly attenuated BR after salbutamol inhalation (median increase in FEV1: 170, 210, 340 ml, pG0.05) . Moreover, about one-half of ABPA patients showed normal or very mild AHR, on the other hand, none of the allergic fungal asthmatics showed normal AHR. Conclusion: These results suggest that ABPA patients, even at an early stage, may show irreversible bronchial narrowing resistant to bronchodilators or bronchostimulator inhalation. Follicular bronchiolitis: A rare mimicker of ''severe asthma'' Belle Peralejo, Peter Benincasa, and Leonard Bielory. University of Medicine and Dentistry of New Jersey, Dept of Medicine, Division of Allergy/ Immunology, Newark, NJ, United States. Background: Atypical presentations of asthma and abnormal imaging are factors that prompt the pursuit of a lung tissue diagnosis. We present a case of follicular bronchiolitis (FB) in a patient thought to have severe asthma. Methods: A 35 year old male non-smoker developed productive cough, dyspnea and wheezing for 2 years. He was treated with inhaled steroids, long acting beta-agonists, montelukast, antibiotics and prednisone for late onset asthma. Symptoms worsened with oral steroid taper. Results: Pulmonary function tests revealed severe obstruction (FEV1 36%, FEV1/FVC of 86%) with significant bronchodilator response (30%) and mildly decreased diffusion capacity (68% predicted). Skin tests were positive to dust mite, mold, cats and dogs. Immunological evaluation ( : 1,3,8,9,14,26,51,56) . Further workup for infection (HIV, HTLV-1/2, AFB, yeast, bacteria, CMV), sarcoid (ACE levels, calcium and gallium scan), autoimmune disease (ANA, ANCA, MPO, Pr-3 Ab), Alpha-1 antitrypsin deficiency and hypersensitivity pneumonitis was negative. CT scan of the chest showed bilateral peribronchial thickening in central bronchi with few small non-specific lymph nodes in the mediastinum. Right upper lobe wedge resection showed acute and chronic bronchiolitis with lymphoid hyperplasia consistent with FB. He improved with chronic steroids and azithromycin (FEV1 72%, FEV1/FVC 92%). Conclusion: FB is a rare entity consisting of abundant lymphoid follicles limited to the peribronchiolar area. It may be idiopathic or associated with hypersensitivity phenomenon or collagen vascular disease. A few cases are reported associated with immunodeficiency such as HIV or common variable immunodeficiency (CVID). Initially, he was thought to have an immune deficiency but repeat analysis revealed normal immunologic function. His disease may, therefore, be idiopathic or related to a hypersensitivity reaction not well understood. He is controlled with steroids and azithromycin. Results: During study period, 1106 nasopharyngeal aspirates were examined from hospitalized children with respiratory symptom, of these, viruses were detected in 785(70.9%). From this population, 58 had hospitalization due to acute asthma exacerbations. Of these, infectious agents were detected in 38(65.5%) of the patients. RSV was detected in 18(31%), IFV in 10(17.2%), RV in 7(12.1%), CoV in 5(8.6%), AdV in 4(6.9%), PIV in 5(8.6%), HMPV in 2 (3.4%) . Coinfection was identified in 12(20.7%) of the samples. Exacerbation of asthma symptoms was greatest in winter(38%). The highest number of samples positive for respiratory syncytial virus was recorded in January(59%). Conclusion: In conclusion, we confirmed that acute exacerbations of asthma necessitating hospitalization was often associated with respiratory viral infection, especially RSV in young children. Also, there were no significant differences between young children and school-aged children in the prevalence of respiratory virus. We found that the causative virus in hospitalized children with asthma exacerbations follow the epidemics of respiratory virus in hospitalized children with respiratory infection. Introduction: It has been suggested that chronic chlamydia pneumoniae infection could be a cause for adult onset of asthma. There are data to suggest that infectious organisms, particulary the atypical bacteria, Clamydia pneumoniae, may be involved in asthma pathogenesis. It is not known whether this organism was allowed to persist after an infection, or was present prior to the development of asthma. The significance of atypical bacteria in the exacerbation of asthma is stil unclear. Aim of the study: The purpose of this study was to determent weather antichlamydial treatment with azitrhomycin will improve asthma symptoms and lung function in asthmatic patients. Materials and methods: 30 patients (mean age 35,5 years) with moderate to moderately severe asthma were treated a median of 6 weeks with azithromycin 1000 mg once weekly. All patients had chlamydia pneumoniae infection detected by specific IgA=1:40 and specific IgG= 1:256 against Chlamydia pneumoniae. Post treatment lung function and symptom score (caugh, wheezing, dyspnea) were compared with baseline values. Results: After 6 weeks of treatment with azithromycin there was significant reduction in symptom score (pG0,01) and significant improvement in lung function FEV1 (pG0,01), Wilcoxon matched Pairs test. Conclusion: Treatment with azithromycin significantly improved asthma symptoms and lung function indicating that Chlamydia pneumoniae may play an important role in enhancing the inflammatory processes in lower airways. Awareness of inflammation as the main pathogenetic mechanism in asthma, renewed the interest for the role of infaction in the ethiology. Influence of 8week therapy with Omeprazole on asthma symptoms, pulmonary function and quality of life in asthmatics with acid reflux symptoms Kamelija Busljetic, Tatjana Caparoska, Biserka Kaeva, Sava Pejkovska, and Kamelija Busljetic. University Hospital, Pulmo-allergology, Skopje, Macedonia, Fyrom. Acid reflux symptoms has been found to occuere in 32 to 80% of asthmatic patients. Although the association is evident, it has been difficult to establish a casual relationship between them. Acid-suppressive treatment has been inconsistent in improving asthma control. The aim of the study was to determine whether a proton-pump inhibitor improves asthma control in asthmatics patients with acid reflux symptoms. Twenty-two patients receiving usual asthma cove including an inhaled corticosteroid. Patients had acid reflux symptoms and moderate to severe persistent asthma. They received omeprazole, 20mg bid for 8 weeks. The measurements were daily asthma symptoms by diary, daily morning and evening peak expiratory flow, FEV1, FVC, asthma quality of life with standardized activities(AQLOS) questionnaire score, rescue salbutamol use. In this study we have shown that daily asthma symptoms, salbuterol use, peak expiratory flow, FEV1, FVC at 8 week did not improve significantly with omeprazole treatment. But there has been significant improvement in nighttime asthma symptoms in patients with nocturnal asthma. The AQLOS emotion function domain improved at 8 weeks(p=0,025) with omeprazole therapy. The conclusion is that daytime asthma outcome did not improve with 8-week omeprazole therapy,but there seams to be a subgroup of asthmatic patients who benefit from excessive antireflux therapy. A 9 years old male with cervical lymphangioma and asthma: a case report Horacio Del Olmo Tellez, Socorro Orozco Martinez, and Jose Guadalupe Huerta Lopez. National Institute of Pediatrics in Mexico, Allergy and Immunology, Mexico City, Mexico. Lymphangioleiomyomatosis (LAM) is rare idiopathic interstitial lung disease predominantely affecting women. Although Van Stossel first described pulmonary LAM in 1937 the natural history of LAM remains poorly understood. The true incidence and prevalence of LAM are unknown. Although current worldwide registries report LAM as a disease with no geographic preference. Most of the delay in diagnosis results from misdiagnosis and the lack of early specific clinical findings. Common misdiagnosis include asthma, emphysema, and chronic bronchitis. Tazelaar and coworkers describe nine cases of a distinct lymphatic vascular lesions that they termed diffuse pulmonary lymphangiomatosis. Six patients included wheezing or asthma (44%) and dyspnea (22%) presented for 3 months to 20 years. We presented a 9 years old boy, who began sufferind at 8 months of life with 38 grades fever, hiporexia, wheezing, dyspnea, cianosis. He received Cefuroxime and Ceftriaxone treatment without diminished symptoms. He enter to National Pediatrics Institute with sinusitis or laringothraqueitis probably diagnosis, the patient continued with dyspnea and inspiratory estridor, the cervical ultrasonography described hemangioma, biopsy reported right lateral lymphangioma without malignancy, it was partially extracted. Tha patient suffered thraqueostomy because compression in hypopharynx posterior wall with 50 percent diminution, supraglotic inflammation and edema. Moreover, our patient presented severe bronchoespasm with salbutamol and dexamethasone nebulizers treatment. Because asmatic mother background, at 1 year old, he received allergy valoration, He developed nasal constipation, pruritus, and rhinorrea in the last 3 months. He received beta 2 agonist and budesonide, transfer factor, bacterial vaccine. Diagnosis of severe moderate asthma, later he developed again, volume increase in lymphangioma zone, he received OK 432. Laboratory results: dermatophagoides farinae specific IgE 1.30 UI/ml, dermatophagoides pterinissinus 2 UI/ml, total IgE 1922 Ui/ml, 7.6% eosinophilia, Prick test positive: Chenopodium, Lolium, Quercus, Timothy, Fraxinus, ligustrum, and DPT, he started sublingual immunotherapy. This is an interesting case because lymphangioma is a rare disease, mainly in children; the association with atopia, causes a difficult control asthma. There are not references with lymphangioma and asthma in the literature, our treatment is controverted and novel. A severe acute asthma exacerbation complicated by pneumomediastinum and atelectasis of the lungs: a case report A severe exacerbation of asthma is a subacute or acute episode of asthmatic symptoms aggravation accompanied by an increased airway obstruction, respiratory insufficiency and a greater risk of complications. Pneumomediastinum and subcutaneous emphysema are rare but well-known complications of acute asthma. The generally accepted explanation for the development of pneumomedistinumis that free air tracks from ruptured alveoli along peribronchial vascular sheaths toward the hilum of the lung. From there, it extends proximally within the mediastinum. Atelectasis of the lungs is due to the formation of mucous plugs and usually responds well to the treatment of the underlying disease. This is a csse report of a 47-year old female patient L.D., who developed pneumomedistinum and atelectasis in the course of a severe asthma exacerbation. The patient has a 27-year long history of asthma and in 2004 she was vitally endangered and mechanically ventilated. In the course of this exacerbation, she developed progressive dyspnea, cough and fever. During acute coughing, an edema and crackles developed in the face, neck and chest regions, accompanied by a sudden deterioration of dyspnea, tachypnea and cyanosis. The arterial blood gas analysis at rest revealed a severe manifested partial respiratory insufficiency, and the chest X-ray established subacute emphysema. As no satisfactory regression of the disease was achieved, the chest CT scanning was performed on the eighth day, revealing atelectasis of the posterobasal left lung segment. Conclusion: pneumomediastinum and atelectasis are rare but possible complications which may accompany a severe asthma exacerbation. Dramatic in appearance, they usually have a benign course and good response to conservative treatment. Serge Chooklin, Roman Vatseba, and Ihor Bihalskyy. Medical University, Departments of Immunology, Surgery, Lviv, Ukraine. Background: The respiratory complications in patients undergoing the abdominal surgery frequently develop and aggravate course on the presence in these patients the bronchial asthma. Mechanisms of its pathogenesis and possible ways of prophylactic remain the debatable. By that, the important role plays cytokines, chemokines, adhesion molecules. Methods: The blood levels of interleukins 4, 5, 8, 13, 16, 18, eotaxin, myeloperoxidase, IgE were measured in patients with bronchial asthma, which underwent upper abdominal surgical intervention. Concentrations of some cytokines were measured by bronchoalveolar lavage. Results of investigation in patients without asthma who underwent surgery were regards as control. Results: The increasing of 8, 16, 18, IgE, eotaxin, myeloperoxidase serum levels was noted in asthmatic patients. The level of IL-13 decreased in these patients simultaneously. The high levels of IL-8 and myeloperoxidase were noted in bronchoalveolar fluid. These changes was more significant be respiratory complications (pneumonia, edema). Asthmatic patients with bacterial infection had significantly higher serum MPO and IL-8 levels in compared with asthmatic patients without infection. Conclusion: Inhibition synthesis of proinflammatory cytokines, chemokines is an important link in the prevention of respiratory complications during surgery in patients with asthma. The applying of IL-13 and interferon gamma may be a perspective in this view. Etiology of chronic cough in a population of children reffered to a pulmonologist Mohammed Herrag, and Mustapha Iraqui. IBN Sina University Hospital, Allergology-Pulmonology, Rabat, Morocco. Chronic cough, has been estimated to affect 7 to 10p.100 of children. When the diagnosis proves elusive, children with chronic cough are often referred to a paediatric pulmonologist for consultation to determine the reason for chronic cough. During the period from July 1994 to June 2005, 160 children were referred for chronic cough (longer than 3 week's duration). The mean age at presentation with chronic cough was 6 years. there were 83 girls and 77 boys. The most cause of chronic cough in this series was asthma in 48p.100. Other causes are gastro oesophageal reflux in 28p.100, tuberculosis in 8p.100, bronchiectasis in 8p.100, sinusitis in 5p.100 and psychogenic cough in 2p.100. cough resolved after evaluation and treatment in all cases Mojgan Kiani Amin, and Fatemeh Bagherabadi. Qazvin Medical University, Allergy and Immunology, Qazvin, Islamic Republic of Iran. Background: Cough is a reflex response of the lower respiratory tract to stimulation of irritant or cough receptors in the airways mucosa. The most common cause in children is reactive airways(asthma). Specific lower respiratory stimuli include excessive secretions, aspirated foreign material, inhaled dust particles or noxious gases,and a inflammatory response to infectios agents or allergic processes. Useful information that may aid in distinguishing the cough origin may include a history or family history of atopic conditions (asthma, eczema, urticaria, allergic rhinitis) a seasonal or environmental variation in frequency or intensity of cough. Case Report: We presented 3-members of a family with chronic cough (more than 6-weeks duration) which started after an upper respiratory tract infection and coninued in spite of antibiotic and bronchodilator therapy, In their past history there was not any atopic condition, The young son had difficult breathing especially in their house and got better outside, In allergy clinic we couldn t find wheezy breathing in rest and also after activity in their son, and PFC tests were in normal limits for parents, (the 4-year-old boy was incooperative) In the next step (and second visit) when they admitted in emergency ward we could find a problem in their house, they had been using a wall heater and its airway has been closed by a bird nest. They were exposed to noxious gases for a long time. Montoux test in BCG-vaccined children with asthma Soheila Alyasin. Nemazee hospital, Pediatrics-Allergy and clinical immunology, Shiraz, Islamic Republic of Iran. Introduction: Recent studies have proposed a decline in tuberculosis infection is a factor underlying the rising severity and prevalence of atopic disorders in developed countries. There are also conflicting reports on the effect of Bacillus Calmette-Guerin (BCG) vaccination on the subsequent development of atopy and asthma. BCG as the most potent inducer of TH1 is believed to suppress TH2 dependent atopic reaction. Methods: Trying to prove this hypothesis, in a case-control simple sampled 100 patients younger than 5 years old who were known case of asthma with variable severity received 5 units intradermal tuberculin test. The patients were also BCG-vaccined at birth. As the control group 100 healthy children previously vaccined at birth, who were age-adjusted with case group, were also test with tuberculin test (PPD). After 48 hours, induction area was measured in two vertical and longitudinal diameters. At the same time, the severity of asthma in the case group was determined (according to classification of National Institutes of Health, National Institutes of Heart, lung and blood), by asking mothers about the frequencies of night and day symptoms or intervals of attacks. The patients were divided to three groups: Mild, Moderate and severe groups (mild intermittent and mild persistent groups were classified as one group). Results: In our case group, mean size of PPD skin test response was 1.7mmT2.99mm but in the control group mean size was 4.42mmT3.69mm( pG0.0001) . Also in another test we divided our case group to two groups ; PPD skin test less than 5mm and more than 5 mm; again with a significant P value. Conclusion: As a conclusion, patients with definite asthma had a significant weaker response to PPD test. This is indicator of weaker TH1 response in allergic patients, can be proposed that stimulation of TH1-immune system by BCG vaccination at birth can influence subsequent development of allergic diseases dominantly mediated with TH2-Immune system. This may promise a revolution in the future of asthma. Regarding the severity of asthma, no significant evidence supported the relation between severity of asthma and PPD skin test response, it may be explained by dominancy of moderate asthma group. Relationship between coronary spasticity and airway responsiveness in patients with coronary spastic angina Inga Fedotova, Valeriya Nemtsova, and Irina Feclina. Kharkov Allergological Centre, Allergology, Kharkov, Ukraine. Background: Several reports have suggested a possible link between bronchial asthma and coronary spasm, but the possibility of a relationship in coronary spastic angina (CSA) has not been clarified. Methods: Airway responsiveness to methacholine and coronary spasticity to acetylcholine were examined in 39 patients with CSA and 28 patients with chest pain syndrome (CPS). Sixteen healthy subjects as the control group were enrolled in the study and their airway responsiveness compared with that of the CSA and CPS patients. Results: The incidence of airway hyperresponsiveness was significantly higher in the CSA group (71%) than in the CPS (21%) and control (16%) groups (pG0.001). The geometric mean of the log minimum dose (Dmin), defined as the cumulative dose at the point at which respiratory conductance began to decrease, was significantly lower in the CSA group (0.73 log units) than in the CP (1.18 log units) and control (1.41 log units) groups (pG0.01). Conclusion: Results of this study demonstrate that acetylcholine-induced coronary spasticity is significantly related to methacholine-induced airway responsiveness in patients with coronary spastic angina. A generalized hyperresponsiveness of the vascular and nonvascular smooth muscles, including that through cholinergic mechanisms, may exist in patients with coronary spastic angina. Sherif Alsayed 1 , and Khalid Elnagar 2 . 1 New Jeddah Clinic Hospital, Chest Departement, Jeddah, Saudi Arabia; 2 New Jeddah Clinic Hospital, Internal Medicine, Jeddah, Saudi Arabia. Objectives: To detect the impact of anxiety on shaping the prevalence of gastro-intestinal reflux disease (GERD) in patients with sever chronic obstructive pulmonary disease (COPD) based on FEV1 % less than 50%. Methods: We examind the prevalence of symptomatic GERD ,using the Vigneri score, cough and dyspnoea with the modified Medical Research Council questionnaire in 29 male patients with COPD. Esophageal 24 h pH monitoring was used to document the diagnosis of GERD in symptomatic group. Beck Anxiety inventory was used to figuer out the impact of anxiety on expressing GERD Symptoms. Results: Reflux disease symptoms was recorded in eighteen patients of the studied group (62%). GERD was diagnosed, based on esophogial 24 h PH monitoring, in only 11 patients of those who expressing symptoms of GERD. Mean of Anxiety score was higher among symptomatic group than non symptomatic reflecting the attribution of differnt symptoms of anxiety to the GERD syndrom. Conclusion: Patients with severe chronic obstructive pulmonary disease have a high prevalence of symptomatic gastro-oesophageal reflux. However True GERD was documented in a fewer number of them. Psychological factors, such as anxiety and somatisation may play a role, particularly in those patients without esophageal inflammation. An experimental study on effect of antioxidants on pulmonary functions of smokers Backround: Cigarette smokes contain large number of free radicals, which cause oxidative damage of airways and lower lung functions. A prospective interventional study was carried out to observe the effects of supplementation with antioxidant vitamins on lung functions of smokers. Methods: 200 healthy male smokers were divided into 4 equal groups of whom 3 groups were supplemented daily with either vitamin A 10,000IU or vitamin C 500mg or vitamin E 200mg and 4th group with combination of all three in same dose for 2 months. FVC, FEV1, FEV1/FVC and PEF were measured at beginning, after 2 months of supplementation and at the end of next 6 months without supplementation. Results: After 2 months all groups showed significant improvement of all measured parameters lung functions. The difference of improvement among various supplementation groups was not significant. However, combined vitamins yielded higher trend of improvement. All groups showed decline of lung functions at the end of six months without supplementation indicating insignificant retention effect. Conclusion: Clinicians may prescribe anti-oxidant supplementations for smokers to improve their lung functions. However, these supplementations should be given for a long period as no significant retention effect was detected. Symptom-limited stair climbing as a valuable predictor of postoperative cardiopulmonary complications after thoracic surgery Magdy Ibrahim 1 , Nancy Mahmoud Abdelaty 2 , and Mahmoud Elprince 2 . 1 Faculty of Medicine, Suez Canal University, Cardiothoracic Department, Ismailia, Egypt; 2 Faculty of Medicine, Suez Canal University, Chest Department, Ismailia, Egypt. Study objective: Thoracotomy is associated with high rate of postoperative cardio-pulmonary complications (POCs). We hypothesized that symptomlimited stair climbing predicts POCs after thoracic surgery. Methods: A prospective evaluation of 40 patients undergoing thoracotomy. The 28 men and 12 women completed symptom-limited stair climbing. A separate investigator, blinded to the number of flights of stairs climbed, assessed 30-day actual outcomes for POCs, including pneumonia, atelectasis, mechanical ventilation for 9 48 h, reintubation, myocardial infarction, congestive heart failure, arrhythmia, pulmonary embolus, and death within 30 days of surgery. The operations performed included 30 lobectomies, 1 wedge resection, 1 pneumonectomy, and 8 decortications. Results: POCs occurred in 12 of 40 patients (30%). Of those unable to climb one flight of stairs, 85.5% developed a POC. No patient able to climb the maximum of five flights of stairs had a POC. The inability to climb two flights of stairs was associated with a specificity of 92%, negative predictive value of 81% and positive predictive value of 75% for the development of a POC, while the inability to climb the maximum of five flights of stairs was associated with a sensitivity of 100% and negative predictive value of 100% for the development of a POC. The number of days in the hospital postoperatively decreased with a patient_s increased ability to climb stairs. Conclusion: Symptom-limited stair climbing offers a simple, inexpensive, valuable means to predict POCs after thoracic surgery. Hepatopulmonary syndrome among patients of cirrhosis of liver and portal hypertension vascular dilatations were classified as the positive group while others were labeled as the negative group. Results: 90 patients were included in the study, of whom 24 (26.66%) had a positive(CE); 12(13.33%) of them were found to have PaO2G70 mmHg and were qualified for the diagnosis of ''clinically significant'' HPS;and other 12(13.33%) with PaO2970 mmHg were diagnosed as ''subclinical HPS''. Using an increased alveolar-arterial difference for the partial pressure of oxygen (AaDO2) as an indication of hypoxaemia,the prevalence of HPS was considerably higher (915 mm Hg,(26.66 %); and920 mm Hg,(22.22%); than using reduced partial pressure of arterial oxygen (PaO2) as a threshold (G80 mm Hg, 20.0%; andG70 mm Hg, 13.33%). For AaDO2 as the cut off, the positive predictive value for a diagnosis of HPS was low (35.29%, and 38.46% respectively). In contrast, PaO2 as a cut off had considerably higher positive predictive values (52.94%, and 75% respectively). Introducing PaO2G60 mm Hg as the cut off, the positive predictive value increased to 100%. The Child-Pugh score correlated significantly with the severity of HPS. Cyanosis, clubbing and orthodeoxia were significantly commoner in the 12 patients of clinically significant" HPS. Conclusion: The study results showed presence of HPS and (IPVD)among patients of chronic liver disease. Expression of c-met in malignant pleural mesothelioma: an immunohistochemical study Heba M. Wagih 1 , Amira Gamal 2 , Nancy Mahmoud Abdelaty 3 , and Ayman Ekram 2 . 1 Faculty of medicine, Suez Canal University, Pathology Department, Ismailia, Egypt; 2 Faculty of medicine, Suez Canal University, Puplic Health Department, Ismailia, Egypt; 3 Faculty of medicine, Suez Canal University, Chest Department, Ismailia, Egypt. Background: Most patients with malignant pleural mesothelioma present in advanced stages of disease. Response rates and survival with currently available therapies are poor. Therefore, it is critical to identify the molecular markers of mesothelioma which would provide a way of understanding this neoplasm and targeting these markers in therapy. Objectives: To assess the immunoreactivity of c-Met in malignant pleural mesothelioma and to analyze the potential link of the c-MET expression to some clincopathological parameters such as tumor subtype, TNM stage and patients' survival. Methods: A total of 20 patients (7 females and 13 males) with pathologically confirmed MPM; age range, (35 to 63 years) were included in the study. The patient records for the clinical, radiological and laboratory investigations and the results of closed pleura biopsies were analyzed. Pleura biopsies were stained for c-Met using immunohistochemical technique applied to paraffin sections. Results: Of the studied tumors 18 (90%) were immunoreactive for c-Met. There were no significant relations between c-Met and patient age (p=0.569) or gender (0.755). Also there was no relation between c-Met expression and clinical symptoms. All tumors that showed distant metastasis were c-Met positive. While all c-Met negative tumors showed no metastasis. However, the difference was statistically insignificant. There was also no relation between c-Met and tumor subtype (p=0.40) or tumor stage (p=0.257). However, all T3 and T4 tumors were c-Met positive and the two c-Met negative tumors were of T2. The 2 c-Met negative patients showed one-year survival. Whereas (7/18) of patients with positive c-Met died. However, again the difference was statistically insignificant (p=0.755). Background: Sleep disordered breathing (SDB) is a prevalent condition in adults associated with increased cardiovascular morbidity. Circulating levels of C-reactive protein (CRP), a pro-inflammatory protein, are associated with increased risk for atherosclerosis.Therefore, CRP may be one of the links between OSAH and cardiovascular disease (CVD).We hypothesized that Patients with OSA have higher CRP values than do control subjects. Methods and Results: We studied 28 patients (20 males and 8 females ) with newly diagnosed (OSA), who were free of other diseases, had never been treated for OSA, and were taking no medications. After polysomnography, venous blood was collected at 5 AM and serum levels of CRP were investigated. We compared CRP measurements in these patients to measurements obtained in 20 control subjects (14 males and 6 females ) who were matched for age and body mass index, and in whom occult OSA was excluded. Levels of CRP were significantly higher in patients with OSAS than in obese control subjects (CRP PG0.000). We evaluated relationship between the Levels of CRP and sleep study parameters such as Apnea Hypopnea Index (AHI), Desaturation Index(DI), Average of Mean Saturation ,Average of Lowest Saturation (LSAT) and Percentage of Sleep Time with Saturation less than 90 %. The results showed significant statistical positive correlations between CRP values and AHI (pG 0.00), DI (pG 0.00) and percentage of sleep time with saturation less than 90 % (pG 0.00) in the OSAS patients. Conclusion: Levels of CRP are elevated in patients with OSAS. Therefore, OSAS is associated with increased risks for cardiovascular morbidity and mortality. The severity of OSA is proportional to the CRP level. Validity of multidimensional body mass index (B), airflow obstruction (O), dyspnea (D) and exercise capacity (E), bode index as predictor of hospitalization for COPD Nancy Mahmoud Abdelaty 1 , Mahmoud Elprince 1 , Nanees A. Ismail 2 , and Magdy Abd-Elsalam 3 . 1 Faculty of Medicine, Suez Canal University, Chest Department, Ismailia, Egypt; 2 Faculty of Medicine, Ain Shams University, Puplic Health Department, Cairo, Egypt; 3 Faculty of Medicine, Ain Shams University, Chest Department, Cairo, Egypt. Background: We hypothesized that the BODE (body mass index, airflow obstruction, dyspnea, and exercise capacity) index would better predict hospitalization for COPD than FEV1 alone. Study objectives: the purpose of this study was to test in a cohort of patients with COPD, how well a multidimensional grading system that assessed the respiratory and systemic expressions of COPD would better categorize and predict outcome in these patients. Patients: A total of 150 patients with COPD(ages 45-83 yr; 89% male) recruited from the outpatient clinic of Suez Canal University Hospital were enrolled in 30 months, prospective study and followed up for a mean period of 12 months from January 2004 YJune 2006. Measurements: The BODE index was calculated for each patient using variables obtained within 4 weeks of enrollment. The main outcome measure was the number of hospital admissions for COPD during follow-up. The following variables were assessed for each patient: age, sex, pack years of smoking, FVC%, FEV1%, the best of two 6 minute walk tests done 30 minutes apart, degree of dyspnea, body mass index (BMI). We evaluated the relationship between FEV1%, the level of dyspnea, BMI, the best of two 6 minute walk tests done 30 minutes apart and BODE scores with the number of hospital admissions. Results: After 30 months, 126 patients were available for the follow-up examination (follow-up rate, 84 %). During the follow-up period, 85 (67%) of patients required at least one hospital admission and 6(4.8%) died. In multivariate analysis a significant effect of BODE score on the number of hospital admissions was found (95% confidence interval [CI] , 0.36 to 0.61; pG 0.000). In comparison, there was a significant but smaller effect of the pack years of smoking, BMI and BMI score on the number of hospital admissions {(95% confidence interval [CI] , 0. 03 to 0.05; pG 0.000), (95% CI,-0.32 to-0.09; pG0.01) and (95% CI,-1.6 to-0.12; pG0.05) respectively}. FEV1%, the level of dyspnea, and 6 minute walk test were significant predictors of hospitalization in univariate analysis (pG0.000) but were excluded in multivariate analysis. Conclusion: The BODE staging system, which includes in addition to FEV1 other physiologic and clinical variables, is a better predictor of hospital admissions than FEV1 in COPD. Acute and chronic eosinophilic pneumonia-our own experience with diagnosis and treatment Jan Plutinsky, Daniel Magula, Stefan Petricek, Dana Laukova, Stanislav Majernik, and Ivan Marget. Specialized Hospital St.Zoerardus, 2nd Pneumology, Nitra, Slovakia. Eosinophilic pneumonia (EP) is a rare disease. Some EP may result from other diseases, while in others the cause is unclear. EP is divided into acute (AEP) and chronic (CEP) forms. The aim of the study: The authors share their own experience with the diagnosis, differential diagnosis and treatment of AEP and CEP. Methods: Since 1995, AEP or CEP have been diagnosed in 21 patients (pts). Clinical examination, radiograph, CT or HRCT scan, laboratory examinations with peripheral blood count of eosinophils (Eo), microbiology, mycobacteriology, fiberbronchoscopy or histology were used. The pts were divided into two groups: AEP and CEP. The ANOVA and Kruskal-Wallis tests were used for statistical analysis. Results: 13 female, median age 66 yrs (36Y87) and 8 male, median 67 yrs (55Y85) had one of the two forms: AEP in 7 pts and CEP in 14 pts. The main clinical symptoms were dyspnoe in 20 pts (95.2%), cough in 19 pts (90.5%), fever in 17 pts (80.9%), pain in 5 pts (23.8%). AEP or CEP was the result of concomitant disease in 3 pts with toxoplasmosis and in 1 pt. with sarcoidosis, rhinitis and allergic bronchopulmonary aspergillosis. In 15 pts no concomitant disease was identified. A statistical significance was found between the age of pts with AEP (56T12.2) and CEP (67.8T12.9) (pG0.05). There were no statistically significant differences between the absolute Eo count in male (614.8T279.3) and female (709.3T811.3) and in the groups of AEP (860.7T976.7) and CEP (579.6T429.3). 9 pts have been continually treated with corticosteroids, 8 pts temporarily and 4 pts have not received any treatment. The longest treated patient has been receiving corticosteroid for more than 9 yrs. Conclusion: EP is a very interesting disease. The clinical symptoms are similar to typical pneumonia, but treatment with antibiotics is unsuccessful. The treatment with steroids is determined on an individual basis. Dividing AEP and CEP is not so clear cut. We could consider using the results of corticosteroids treatment as a way dividing AEP and CEP since the treatment in AEP is only temporary and in CEP on-going. Recent studies have shown that osteoporosis and vertebral fractures are quite common in postmenopausal women with chronic obstructive pulmonary disease (COPD). Few data are available in correlation between bone mass density (BMD) and men with COPD.This study was designed to investigate the prevalence of osteoporosis in men with COPD. with special regards to the role of glucocorticoids (GCs) use in these patients. We aimed to determine factors that influencing bone metabolism and the clinical variables of this group of patients. We also tried to answer the arising question: should COPD patients be routinely evaluated for BMD? Methods: The study included 56 male patients with documented COPD for at least two years, their age ranged 24-66 years. Subjects were divided into 3 groups: group 1. consisted of 18 patients, who were oral GCs users, group 2. consisted of 18 patients who were inhaled GCs users and group 3. consisted of 20 patients, never GCs users (this group was considered as the control group). All subjects underwent measurement of BMD. pulmonary function tests (PFTs) and a number of biochemical markers of bone metabolism. The associations between BMD. PFTs. GCs use. biochemical markers and clinical variables were analyzed. Results: Of all 56 patients with COPD. the prevalence of osteopenia and osteoporosis. as defined by WHO criteria was 26.8% and 21.4% at the lumber spine. 30.4% and 23.2% at total hip. 35.7% and 28.5% at femoral neck and 32.1% and 28.8% for total body respectively. Patients included group 1 had the lowest BMD at any site (pG0.0001). group 2 patients had over all bone mass loss, that was indistinguishable from those who were received oral GCs. group 3 patients had less bone mass reduction than the other two groups. Of the clinical and biochemical markers measured. N-telopeptide was significantly correlated with bone mass (PG0.01), but there was no correlation with other markers. The lowest mean of FEV1 (Forced Expiratory Volume in one second) was observed in group 1 patients. BMI (Body Mass Index) was weakly correlated with bone mass in the 3 studied groups. Conclusion: Bone mass loss is a common problem in male patients with COPD. while the use of oral GCs increase the frequency of osteoporosis, inhaled GCs therapy offered no protection from bone loss. COPD patients who had never treated with GCs had also a substantial risk for osteoporosis. We advocate early screening and preventive intervention. Psychotherapy improve exercise tolerance in patients with Pink Puffer syndrome Alsayed Sherif, and Khaled Elnagar. New Jeddah Clinic Hospital, Internal Medicine, Jeddah, Saudi Arabia. Anxiety is common in the ''pink puffer'' syndrome associated with chronic obstructive pulmonary disease (COPD). The degree of anxiety correlates well with perceived dyspnoea. This study examines the effect of psychotherapy on anxiety, exercise tolerance, and dyspnoea. 10 patients with stable COPD (mean forced expiratory volume in one second (FEV1)=1.25 L) had six 90 min sessions of cognitive and behavioural psychotherapy at weekly intervals. Patients completed the Beck Anxiety Inventory (BAI), 1 day before and 1 weeks after therapy. FEV1, forced vital capacity (FVC), blood gas tensions and 6 min walking distance (6MWD) were measured. 10 control patients attended weekly for lung function and 6MWD for 6 weeks, but had no psychotherapy. There were no differences in mean baseline (BAI) score, lung function, blood gas tensions or 6MWD between groups. After treatment, the BAI score had decreased from 43.10 to 20.6 (pG0.001), in association, the mean 6MWD had also improved in the psychotherapy group only, from 333 to 559 m (pG0.001), an increase of 75%. In conclusion, six sessions of cognitive and behavioural psychotherapy produced a good improvement in exercise tolerance in anxious patients with chronic obstructive pulmonary disease (Pink puffer). Exhaled breath analysis with electronic nose in patients with lung cancer, chronic obstructive pulmonary disease, asthma and pneumonia Immanuels Taivans Background: Since developement of electronic nose (EN), it has been well recognized that there is a great potential in applying this technology to the field of medicine. Analysis of various biomarkers in exhaled breath allows completely safe, noninvasive, sensitive, and specific tool for the primary screening for different lung diseases. The aim of this study was to test whether exhaled breath analyzed with artificial nose could identify and discriminate between asthma, COPD, lung cancer and pneumonia. Methods: Exhaled air was collected in plastic bags and immediately analyzed by electronic nose instrument (9185, Nordic Sensors AB) containing 14 different odour sensors. Multifactorial logistic regression analysis and support vector machine was used to find relationships between the sensor response, on derivative, on integral and the clinical diagnoses of patients. Results: 62 individuals Y 25 asthma, 13 lung cancer, 13 pneumonia, 12 other lung disease patients and 10 healthy volunteers were tested. Given table represents p values of significant relationship between electronic nose sensors and clinical diagnosis. Some sensors (1, 6, and 13) gave specific responses to particular disease; some other sensors (3, 5, and 7) shared the response with two diseases. The highest sensitivity and specificity was found for lung cancer diagnosis compared to healthy control group It was 100% and 92,9% respectively. Artificial nose is able to discriminate among different lung diseases. Further development of this approach is necessary to create new screening and monitoring methods for different lung diseases. Proteolytic activity of Per a 10 from Periplaneta americana enhances allergenicity in a mice model Thangam sudha V, Naveen Arora, and Bhanu P Singh. Institute of Genomics and Integrative Biology, Allergy and Immunology, Delhi, India. Background: Cockroach allergen sensitization has been implicated as a major cause of allergy/asthma. Recently we have demonstrated a serine protease (Per a 10) as an immunodominant allergen from Periplaneta americana (American cockroach). So, we aimed to investigate the immune-inflammatory responses to active serine protease in mice model of airway inflammation. Methods: Balb/c mice were sensitized with proteolytically active allergen or OVA through intraperitoneal and subcutaneous routes and challenged intranasally with Per a 10 allergen (active / inactive), PBS or OVA. The cellular infiltration (total cell count and EPO activity in BALF), Th1/Th2 cytokines in BALF and spleen culture supernatant, serologic (specific IgE, IgG1) and histopathologic events in the various animal groups were compared. Results: Mice challenged with active serine protease allergen or OVA showed tissue damage in the lungs, elevated serum IgE, IgG1, eosinophils and expression of inflammatory cytokines in BALF and spleen culture supernatant, indicating the induction of a Th2 response. Administration of proteolytically inactive allergen (blocked irreversibly with AEBSF) induced significantly lower levels of cellular infiltration in lungs, systemic IgE production and Th2 cytokines. Quantitative evaluation of lung tissues showed decrease in total inflammation score in mice challenged with inactive protease. Conclusion: Serine protease allergen from P. americana induces allergic response in a mice model and its proteolytic activity enhances the airway inflammation. The inactivated allergen may have potential for therapeutic applications. Cassia siamea pollen: an important aeroallergen of tropical countries Dola Boral 1 , Kashinath Bhattacharya 2 , Sunirmal Chanda 3 , and Swati Gupta-Bhattacharya 4 . 1 Krishnath College, Department of Botany, Berhampore, India; 2 Visva Bharati University, Department of Botany, Santiniketan, India; 3 Center for Study of Man and Environment, Department of Environment, Saltlake, India; 4 Bose Institute, Department of Botany, Kolkata, India. Background: It is well known that allergy in India and other countries appear to be one of the commonest major health problems. Cassia siamea Lam. of the family Fabaceae is an important avenue tree in tropics. Its pollen grain has been known as one of the important pollinosis causing airborne bioparticles. The present study was demonstrated to identify its important allergenic components and also to study the cross reactivity between the members of the family Fabaceae. Methods: The seasonal periodicities of airborne Cassia pollen were recorded in a 2-year aerobiological survey using a seven-day Burkard volumetric sampler (Burkard manufacturing Co., Rickmansworth, Herts, England). The allergising potential of Cassia pollen antigen was investigated by in vivo skin prick tests and in vitro IgE-enzyme linked immuno sorbant assay (ELISA). The IgE specific immunoblotting confirm the IgE sensitive reaction between the pollen allergen and patient`s sera. In the present study, the whole pollen extract of C. siamea pollen was characterized by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). The presence of glycoprotein in the crude allergenic extracts of Cassia siamea pollen was detected by PAS (Periodic Acid Schif) technique. Results: An intensely positive result (+++) was observed in response to SPT with the antigenic extract of C. siamea pollen. Within 175 patients 30.85 % shows positive allergic reactions. Immunochemical studies revealed the presence of ten allergenic components in the molecular weight ranging between 8Y83 KDA. Three major allergenic protein bands (22, 17, 9 KDA) were isolated by IgE-specific immunoblotting with individual sensitive patients sera. Background: Mite allergy is a disease induced by inhalation of mite bodies and feces present in the house dust, leading to asthma, nasal inflammation, and dermatitis. The identification of allergens is needed for the specific immunotherapy. Our allergenomic analysis of Dermatophagoides farinae body extract by two-dimensional immunoblotting followed by mass spectrometry has revealed a number of new allergens. The purpose of this study is to characterize a 15-kD allergen, DFA22, with high frequency and intensity of patients' IgE binding. Results: Based on the partial amino acid sequences obtained by mass spectrometry, a full length gene encoding DFA22 was isolated by PCR. Analysis of its deduced amino acid sequence and predicted molecular conformation suggested that DFA22 is a new member of group 2 mite allergens (ML protein). It was noted that DFA22 has a higher homology with a sheep scab mite allergen, Pso o 2 (62% identity in amino acid sequences), and a storage mite allergen, Lep d 2 (46%), than a house dust mite allergen, Der f 2 (40%), indicating that there are at least two subgroups in the group 2 mite allergens. In order to characterize the immunochemical profile of the new allergen, a recombinant protein was produced using a vector pCold-TF, which can express trigger factor (TF) fusion protein at low-temperature, and Escherichia coli Rosetta-gami, which has an ability to form precise disulfide-bond. The rDFA22 was purified by His-tag affinity chromatography, removal of TF, and anion exchange chromatography. Mass spectrometric analysis demonstrated that three disulfide-bonds characteristic to the group 2 mite allergens are formed in the rDFA22. ELISA indicated that its IgE binding frequency was 51% (n = 35; RAST scores to mite 9 3). The binding of patients' IgE to rDFA22 was partially inhibited by rDer f 2 (~32% inhibition) and Pso o 2 (~56%), implying that these allergens shear common IgE epitope(s). Conclusion: DFA22 is an important novel allergen that cannot be disregarded to achieve successful specific immunotherapy against mite allergy. It is also necessary to reconsider the molecular differentiation of the group 2 mite allergens. Experiences with the setting of a new polllen trap as a member of the Hungarian airborne pollen network in southwest Hungary Tamas Major, and Marianna Rakvács. Kaposi Mór Teaching Hospital, Pulmonology, Mosdos, Hungary. The Hungarian network of aeropalynology includes traps thruout the country. In order to optimise this network, the Hungarian Institute of Aerobiology has set up a new trap in the Hospital for Chest Diseases of the Hungarian Calvinist Church In Hungary in Mosdós, Somogy county. The area has high impotance in the view of the researchers, because it is located close to the most poulated recreactional region, the Lake Balaton and this part of the country is probably the most pollen Binfected[ territory in Hungary. The equipment is a standard spore trap (Burkard), which is used all over the counry. Our aeropalinlogy team includes: three assistants, two clinicians and a technician. The team has already been reporting the pollen counts from the begining of the polens season to the National Network on a weekly basis. Objectives: To assess the situation of pollen counts and epidemiological data on allergies in the region. Second objective was study the influence of temperature, relative humidity, rainfall and wind direction on the occurrence of pollen allergens in the atmosphere. Our results showed that, the occurrence of pollen grains in the atmosphere markedly relates to meteorological factors. We evaluated a correlation between the concentration of pollen grains in the atmosphere of the region and temperature, relative humidity, rainfall and wind direction during the vegetation period. For our analysis we selected one representative of each phytoallergen group (trees, grasses, weeds). We have chosen the Betula genus of trees, the whole Poaceae family of grasses and ragweed (Ambrosia artemisiifolia) to represent weeds. The taxons mentioned represent the most significant allergens in Hungary. Our data provides predictive information on the forthcoming patient flow and the severity of their allergic symptoms. Comparison of allergen spectrum in asthma and allergic rhinitis children with 3~14 years of age Background: To compare the allergen spectrum in asthma and allergic rhinitis children with 3~14 years of age. Methods: Inhalant allergen skin prick test (SPT) were done in 527 asthma and 620 allergic rhinitis children respectively. The positive rates of different allergen and allergen spectrum in asthma and allergic rhinitis were analyzed. Results: There was no significant difference on the positive rates of SPT in asthma and allergic rhinitis (77.8% vs78.9%, P 9 0.05). The main inhalant allergens related to asthma and allergic rhinitis were Dermatophagoids pteronyssinus, Dermatophagoids farinae ,alternaria, cat dander and mugwort. The positive rates of Der.p, Der.f and molds mixture in asthma were higher than those in allergic rhinitis (64.6% vs 49.5%, 59.8% vs 47.9%, 8.8% vs 3.9%, P G 0.05); The positive rates of summer-autumn pollen and mugwort in allergic rhinitis were higher than those in asthma (25.6% vs 19.3%, 26.0% vs 19.3%, P G 0.05). 40.2% asthmatics and 46.2% allergic rhinitis children were single allergen sensitization. The multiple sensitization to mites, molds and pets were common in asthma and allergic rhinitis. Conclusion: Mites, molds, pets and summer-autumn pollen were the main inhalant allergens related to asthma and allergic rhinitis children with 3~14 years of age. The allergen spectrums were similar in asthma and allergic rhinitis. Mites and molds allergy were more popular in asthma while summerautumn pollen in allergic rhinitis. Background: Sensitization to food and airborne environmental allergens data is still rare reported. This study evaluated the epidermal skinprick test prevalence and types of pollen causing allergic symptoms have not been still well defined. Methods: We selected consecutive patients with respiratory symptoms sugestives of allergic respiratory pathologies. They were studied by means of standard prick tests with a battery of pollen extract, and seric specific IgE analysis. A Hirst spore trap runs on a continuous-basis on the roof of the BObservatorio Atmosférico de Izaña [ headquarters, since October 2004. The results corresponding to the period 2004Y2006, obtained following the counting method of the Spanish Aerobiology Network (Red Española de Aerobiología, REA), are presented. Results: The most abundant pollen types have been (by decreasing order of the annual indexes): Urticaceae, Asteraceae (99% Artemisia), Arecaceae, Myrica, Poaceae, Ericaceae, Chenopodiaceae-Amaranthaceae, Olea, Cupressaceae, Polygonaceae, Pinus, Moraceae, Plantago Mercurialis, Myrtaceae, and 43 other pollen types (G1% of the pollen spectrum). Herbs predominate in the pollen spectrum (60%) during the first half of the year. Trees (34%) predominate during the summer time. Finally shrubs (5%) pollinate mostly in spring. Most of the cited pollen types are able to cause allergy. Prevalence of pollen allergy during the period 2004Y2006 was 16% of patients with respiratory allergy studied in our medical center. The most frecuent types of pollen sensitization were: Artemisia vulgaris (64,5%), Poaceae family (Gramineae) (45,2%), Parietaria judaica (24,6%), Chenopodium album (9,4%) , Salsola kali (6,2%) y Plantago lanceolata (4%) . The prevalence of sensitized patients to molds were less than 5%. Conclusion: Pollen is a relevant cause of allergic respiratory diseases in our area. More studies are needed to improve the accuracy of diagnosis of pollen allergic patients. Distribution of allergens among allergic rhinitis patients living shiraz (Iran) region S. Hesamedin Nabavizadeh 1 , and Sohila Al-yasin 2 . 1 Yasuj University of Medicine, Pediatric Allergy and Immunology, Yasuj, Islamic Republic of Iran; 2 Shiraz University of Medicine, Pediatric Allergy and Immunology, Shiraz, Islamic Republic of Iran. Objective: Allergic rhinitis is the single most common chronic allergic disease worldwide. House dust mites, grass pollens and fungal spores play has been identified to play a major role in the pathogenesis of allergic rhinitis. To determine the distribution of allergens among allergic rhinitis patients living in the shiraz region. Methods: We investigated 56 allergic rhinitis patients (29 females, 27 males; mean age 22.3+/j14.1 years) who had symptoms. Results: The history and clinical examination revealed that 14 patients (25%) had seasonal symptoms and 42 patients (75%) had perennial symptoms. Common pattern of multiple allergens were tree mixture allergens (58.9%), grass mixture (42.8%), mite (19.6%), candida (10.7%), tomato (5.3%), fruite (3.5%), and feather mixture (3.5%) and cladosporium, oak, ash (3.5%), alternaria, maple, hen egg, aspergillus (1.7%). Skin prick test were negative in 12 cases (21.4%). According to the prick test results, sensitivity to one allergen was found in 5 patients (8.9%) and sensitivity to multiple allergens in 51 patients. Family history were positive in 29 patients. Conclusion: The most common allergens were tree mixture and grass in the shiraz region. It was concluded that the distribution of allergens was associated with the climatic, environmental and socioeconomic features of the region. Larger representative sample involving multi-centers in Shiraz should be encouraged in the near future. A research on triticum aestivum allergy in east of turkey Nejat Ozanguc. Gozde Hospital, Allergy and Immunology Center, Malatya, Turkey. : The patients who visited the Center of Allergy and Immunology in Malatya/Turkey and diagnosed with seasonal allergic rhinitis (SAR), bronchial asthma (BA) and SAR+BA. The result of the skin prick tests applied to these patients with aeroallergens showed that 120 patients had allergic reaction to only triticum aestivum (TA) and 37 patients tested positive for ta with other aeroallergens in a total of positive 293 tests. There were 134 male test participants and 159 female participants in this research. 48 male and 72 female tested positive for TA only. 18 male and 19 females tested positive for TA and other aeroallergens. All these patients are currently in our allergen immunotherapy program.These results were worth giving attention to and I was informed by the Malatya Agriculture Bureau that, Malatya and surrounding areas are the capital of growing wheat wh to make bread, which is TA. On the other hand, Malatya is exporting the 20% of the world's apricot consumption. I am working on a detailed crosssensitisation research between TA and apricot tree pollen. Since, there is no apricot tree pollen extract produced by any company, I gathered this pollen in a powder form and made connection with companies to produce the antigen to use in aeroallergen skin prick tests. Islamabad is one of the most beautiful capitals of the world with mountains and greenery all arround. But it has also highest pollen counts in asia with huge number of with Pollen allergy patients. In spring and fall numerous patients visit emergency department of different hospitals. Rapeseed mustard is the second most important edible oil in India. Allergy to Brassica seeds have been reported earlier from Austria, Finland etc, but only scant information is available from Indian subcontinent. We have investigated the allergenicity to B. juncea seed flour by in vivo and in vitro methods in Indian atopic cases. Antigen from B. juncea was extracted in 1:20 w/v by standard protocol. To assess sensitization, skin prick test was carried out with antigen extract (1:10 w/v) of B. juncea on the above cases. Total IgE and B. juncea specific IgE was estimated by Enzyme Linked Immuno Sorbent Assay. To determine the allergenically important protein, immunoblot was carried out. A total of 159 atopic cases suffering from allergic rhinitis, bronchial asthma or both were selected from general atopic population, referred to clinical units for treatment. Of these, 37 (23.27%) were suffering with allergic rhinitis, 44 (27.67%) with bronchial asthma and 77 (48.42%) with bronchial asthma as well as allergic rhinitis. The mean age was 28.5 yr. (10Y50 yr.) and sex distribution (female vs. male) was 80 vs. 79. The mean age of 50 healthy volunteers included was 25.63 yr. Of these, 15 cases (9.43 %) showed 2+ & above skin test response against seeds of B. juncea. The total IgE of 159 cases ranged from 12 IU/ml to 2208 IU/ml. Raised allergen specific IgE was also observed in the sera of skin test positive cases ranging from 0.221 to 0.848 Optical Density value. The antigen extract separated into 23 protein bands on Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis, in the mol wt range of 14Y97 kDa. The Immunoblot revealed 12 IgE binding protein fractions in the sera of patients showing raised IgE Brassica specific antibodies. Some heterogeneity in the IgE binding protein fractions amongst individual patients is observed. Thus we conclude, that sensitization against seeds of B. juncea does exists in Indian population. Malek Alaouie 1 , Rouwayda Dana 2 , and Fares Zaitoun 3 . 1 Allergy Asthma and Immunology Center, Allergology Department, Hamra-Beirut, Lebanon; 2 Essex Chemie AG, Medical Affairs Department, Beirut, Lebanon; 3 Allergy Asthma and Immunology Center, Research Department, Beirut, Lebanon. Background: Aerosolized pollen are an important cause of respiratory allergy worldwide. The availability of pollen counts in westernized countries is essential for patient education and medical treatment. Aerobiological pollen data is scarce in the Middle East and had never been previously assessed in Lebanon. In order to identify and quantify pollen prevalence in Lebanon, a national aerobiology program was launched starting in the capital city of Beirut. Methods: A Lanzoni pollen trap was placed on a roof top in Beirut 20 meters above ground level. A seven-day cycle drum with a 168-hour film coated with silicon adhesive was used to capture pollen grains. Slides were stained with gelatin fuschin & examined with a light microscope using a 40x objective. Pollen identification and counting was recorded on a weekly basis. Results: A total of 9491 pollen grains/m3 were collected for the 12 month period of 2006 (except for a brief interruption during the 2006 summer military conflict). Twelve different types of pollen were identified which were Alnus, Cupressaceae, Oleaceae, Pinaceae, Juglandaceae, Mercurialis, Chenopodiaceae, Artemesia, Parietaria, Plantago, Apiaceae, and Poaceae (table 1). The 4 most common pollen found over the study period was Oleaceae (27% of total) followed by Cupressaceae (24%), Pinaceae (18% of total), and Mercurialis (17%) (table 2). The peak pollen level recorded was 327 pollen grains/m3 on April 17th with an average daily count of 93 pollen grains/m3 throughout the month of April. The lowest total pollen count was in the month of September with an average daily level of G2 pollen/m3/day. Conclusion: This was the first aerobiologic pollen data ever recorded in the country of Lebanon. Typical elevation of pollen counts in the spring was observed however the level was surprisingly high for an urban area. The finding of tree pollen (cupressaceae and Alnus) in winter months along with identification of a particular pollen (Mercurialis) not previously suspected were important findings for this allergen information-poor country. Since pollen collection took place in only one location (Central Beirut) and for only one year, extrapolation to a nationwide impression can not be made. Nonetheless the exciting pollen data generated in 2006, a first for Lebanon, will serve a pioneering role for the study of aerobiology and allergic disease in this country and region. A ten year survey (1996Y2006) of the major pollen aeroallergens in Sydney, Australia Connie Katelaris, Therese Burke, and Pamela Burton. Campbelltown Hospital, Clinical Immunology and Allergy, Sydney, Australia. Background: Airborne pollen is a major cause of allergic symptoms during the spring months in Sydney, Australia. Establishing the predominant pollen aeroallergens and the timing of highest pollen levels greatly assists medical personnel in developing effective treatment plans for those individuals suffering from allergic symptoms related to pollen sensitivity. This survey identifies which pollen aeroallergens are most prevalent during the spring months in Sydney and suggests possible reasons for the differences observed between seasons in an attempt to determine effective predictors of severe pollen seasons. Methods: Aeroallergen monitoring was performed during the spring months 1996Y2006 in Sydney, Australia. A Burkard 7 day volumetric spore trap was used for pollen collection and slides were analysed using standard protocols. A daily pollen count was expressed for each species in grains per cubic meter (CuM). Data was collected from August to November of each year (spring in the southern hemisphere).Daily rainfall data was obtained from the Bureau of Meteorology and expressed as millimetres of rain on a daily basis for the entire ten year period of the survey. Pollen data was then collated into 3 groups Y tree, grass and weed pollen. Each group was analysed for seasonal trends and correlated with rainfall data. For each year of the survey, the season duration was identified, as well as peak day and peak count, average daily pollen count and total season pollen count for each group. Results: Tree pollens make up the majority of the total pollen count in Sydney, followed by grasses and weeds. Major tree pollen species identified in Sydney include Cypress,Pine, Plane and Casuarina. Tree pollens tend to reach peak levels in mid to late September, and grass and weeds pollens in mid to late October. The highest average daily pollen counts for all groups occurred in years with the highest autumn/spring rainfall. Average daily pollen counts ranged from 23Y147 grains/CuM (trees), 1Y25 grains/CuM (grasses) and 0Y15 grains/CuM (weeds).The period of pollen season duration also varied considerably each year. The results of this survey indicate the major pollen aeroallergens present during the spring months in Sydney, Australia. The survey also highlights considerable variability between pollen seasons and the need for ongoing aerobiological pollen monitoring in the future because of highly variable climatic conditions that clearly impact upon the pollen counts. Influence of meteorological variables on pollen counts in Sydney, Australia: longitudinal data examining effects within and between pollen seasons Therese Burke, Pamela Burton, and Connie Katelaris. Campbelltown Hospital, Clinical Immunology and Allergy, Sydney, Australia. Background: Airborne pollen is a significant cause of allergic symptoms in sensitised individuals. Various meteorological factors have an effect on pollen levels, on a day to day basis throughout the pollen season, and on a seasonal basis over time. By understanding the effects of these meteorological factors, it may be possible to develop effective indicators which may assist in predicting high pollen days within a season to assist in symptom management. Method: Airborne pollen was collected between 1996Y2006 in Sydney, Australia. A Burkard 7 day volumetric spore trap was used to collect data and slides were made up and analysed using standard protocols. Pollen counts were expressed as grains per cubic Meter (CuM) for various species and then grouped into trees, grasses and weeds. Meteorological data was obtained and included minimum temperature, maximum temperature, daily rainfall and humidity. Spearman rank correlations were used to assess the effect of the meteorological variables upon the day to day pollen counts and the seasonal pollen counts as a whole. We then examined cross correlation plots between first differenced 4 day moving averages in order to detect short term associations between fluctuations in meteorological variables and fluctuations in pollen counts. Results: Examining daily counts within a given pollen season, there was a statistically significant positive correlation between daily maximum temperature and the daily pollen count for all pollen species (p G 0.05). Daily rainfall and high humidity were negatively correlated with the daily pollen count for all species (p G 0.05). Day to day changes in maximum temperature were moderately positively correlated with a simultaneous change in the total pollen count (r = 0.35), but weakly negatively correlated with a change in the total pollen count 4 days later (r = j0.15). Examining season to season associations, mean seasonal pollen counts were negatively correlated with mean seasonal daily maximum temperature ( r = j0.782 total pollen and r = j0.600 for grasses). Conclusion: On a day to day basis, increases in daily maximum temperature lead to an increase in pollen levels and daily rainfall leads to a simultaneous decrease in the daily pollen level. Overall, cooler temperatures across a season often lead to a higher total pollen count. Ongoing pollen monitoring is needed to further examine the effects of meteorological factors on daily and seasonal pollen counts. The characterization of native thaumatin-like allergen named Cry j 3 from Japanese cedar (Cryptomeria japonica) pollen and another protein at 35kDa constituted only 7% of the total protein content. In pollutted pollens the concentration of the protein at about 45 kDa considerably diminished to 14.6 % of the total protein content but the amount of the protein fraction at 35kDa increased to 50.2%. Immunoblots, conducted to evaluate sera IgE reactivity of cypress allergic subjects to the new major protein of polluted pollen extracts, showed that the protein of 35 kDa was also the major allergen of C.arizonica pollen extracts in allergic subjects living in Tehran. Important differences observed between protein profiles of nonpolluted and polluted pollen extracts, reflected the fact that environmental pollution induce the augmentation of an allergenic protein which may have a negative repercussion on the health of urban popullations. The allergen-specific IgE reactivity pattern of Chinese house dust mite allergic patients Background: Two species of house dust mites (HDM), Dermatophagoide pteronyssinus (Der p) and Dermatophagoide farinae (Der f), have been recognized as major allergen sources in Chinese allergic patients according to epidemiological surveys using in-vivo or in-vitro assays. The specific IgE reactivity pattern of European and American HDM allergic populations are well characterized. However, few studies have addressed the allergen-specific IgE pattern of Chinese HDM patients. Methods: Serum samples from 62 patients who had allergic asthma and/or allergic rhinitis and who showed skin prick test and UniCAP positive to Der p and Der f were obtained from South and Southwest of China. Sera were analyzed by crossed radioimmunoelectrophoresis (CRIE) and immunoblotting to visualize the allergen-specific IgE reactivity patterns. Results: 23 allergens of Der p and 16 allergens of Der f were detected by CRIE. The most frequent allergens were identified to belong to HDM group 1 and 2. All sera had specific IgE against both Der p 1 and Der f 1, while 92 % of sera recognized Der p 2 and 95 % reacted against Der f 2. Antigen 16 of Der p showed IgE binding by 69%. After immunoblotting, patients' IgE mainly showed binding to proteins of around 25 and 14 kDa which were identified as HDM group 1 and 2 allergens, respectively. The reactions of the sera by immunoblotting were 53 % for Der p 1, 63 % for Der p 2, 31 % for Der f 1 and 59 % for Der f 2. Conclusion: The HDM specific IgE reactivity patterns of Chinese mite allergic patients are similar to that of other patient populations. In addition to the commonly recognized Der p 1, Der p 2, Der f 1 and Der f 2, a novel major allergen, denoted antigen 16, was identified in the Der p extract. The analysis indicates that the Der p and Der f extracts used in this study can be used worldwide both for diagnosis and treatment. Immunoglobulin E and Immunoglobulin G epitope-mapping of Blo t 5, the major allergen of Blomia tropicalis Fong Cheng Yi, Hugo P Van Bever, Bee Wah Lee, and Kaw Yan Chua. National University of Singapore, Paediatrics, Singapore, Singapore. Background: Blomia tropicalis is one of the most predominant domestic mite species in tropical and subtropical countries. Blo t 5 is a major allergen with reported IgE sensitization rates of 43Y92% in Blomia tropicalis allergic subjects. Aims: The objective of this study was to determine the IgE, IgG1 and IgG4 epitopes of Blo t 5 in mite allergic children. Methods: Five overlapping peptides, F1 (aa 1Y45), F2 (aa 41Y80), F3 (aa 70Y117), F4 (aa 1Y80), and F5 (aa 41Y117) were generated by PCR from Blo t 5 full-length cDNA. The IgE, IgG1, and IgG4 reactivities to these peptides and full length Blo t 5 were determined by ELISA assay with 20 Blomia tropicalis skin test positive children. The IgE ELISA unit was determined by a semipurified human IgE as standard curve, whereas IgG ELISA unit was determined by OD405nm X dilution factor. Results: The IgE epitope mapping experiments revealed very low levels of IgE binding to small peptides at N-terminal, F1 and F2 (p G 0.005) compared to F3, and larger peptides, F4 and F5, indicating that Blo t 5 major IgE epitopes were located at C-terminal and conformational dependent. In contrast, higher levels of IgG1 to F1 and F3 were found compared to F2 (p G 0.005) indicating that the major IgG1 epitopes are located at both the N-and C-terminal of Blo t 5. The peptides F1, F2, F3 and F4, bound lower levels of IgG4 (p G 0.005) than F5 suggesting that the IgG1 and IgG4 epitopes are unique and the major IgG4 epitopes seem to be located at C-terminal and conformational dependent. Conclusion: The data revealed both IgE and IgG4 epitopes are found mainly in the C-terminal of Blot 5 and are highly conformational dependent, whereas IgG1 epitopes are found throughout the whole Blot 5. The results suggest that recombinant hypoallergenic Blo t 5 for immunotherapy can be achieved by modifying the IgE epitopes while retaining the IgG epitopes. Mohammad fereidouni 1 , Mehdi Bakhshaei 2 , and Abdolreza Varasteh 1 . 1 Mashhad University of Medical Sciences, Immunology Research Center, Mashhad, Islamic Republic of Iran; 2 Mashhad University of Medical Sciences, Otorhinolaryngology-Head and Neck Surgery, Mashhad, Islamic Republic of Iran. Background: Allergic rhinitis is a prevalent allergic disease throughout the world. A recent study showed that allergic rhinitis is a common problem in Mashad, Iran, However, Little is known about the relation between skin test reactivity to aeroallergens and allergic rhinitis in Iran. Objective: The aim of this study was to characterize the most frequent indoor and outdoor (grass and tree) inhalant allergens involved in allergic rhinitis in Mashad, Iran. Methods: A total of 99 cases (females: 49, males: 50, aged between 3 to 68, mean age 25 years), that were diagnosed with current active allergic rhinitis based on the ISAAC rhinitis Core questionnaire and clinical presentation, underwent a standardized skin prick test with 28 (7 indoor, 13 grasses and 8 trees) aeroallergens. Results: 75% of patients were reactive to at least one of the aeroallergens and 69% of patients had positive reactions to more than 3 allergens but just 3% of patients were mono sensitized. Sensitization was more prevalent for outdoor allergens than for indoor allergens in patients (74% vs. 31%).The individual pollen allergens with the highest positive rates were pollens of Careless weed (Amaranthus palmeri) 67.4%, Russian thistle (Salsola kali) 64%, Redroot (Amaranthus retroflexus) 61%, Burning bush (Kochia scoparia) 58%, Lambsquarters (chenopodium album) 56%, Eastern cotton wood (Populus deltoides) 54%, White ash (Fraxinus americana) 53%, American sycamore (Platanus occidentalis) 43%. The most common Indoor allergens were Dermatophagoides pteronyssinus %20, cockroach mix (American and German) 20%, house dust 20%, Dermatophagoid farinae 17%. There was a significant but weak correlation between sum of all wheal sizes and serum total IgE level (PG0.001) and also between the number of positive skin test responses and the serum total IgE.The sum of mean wheal sizes was larger in men than in women (371 vs. 219 mm2) for outdoor allergens but it was not statistically significant. Redroot, Russian thistle and Careless weed had the greatest mean wheal size (60, 40 and 37 mm2, respectively). Conclusion: The present study revealed that the prevalence of the Skin Prick reactivity to outdoor allergens particularly grasses (weeds), is high among Iranian allergic rhinitis patients, although Indoor allergens do not seem to be very common among patients. Solution structure of Phl p 3, a major allergen from Timothy grass pollen Irena Matecko 1 , Kristian Schweimer 1 , Arnd Petersen 2 , Wolf-Meinhard Becker 2 , Roland Suck 3 , and Paul Rösch 1 . 1 University of Bayreuth, Department of Biopolymers, Bayreuth, Germany; 2 Molecular and Clinical Allergology, Research Center Borstel, Borstel, Germany; 3 Allergopharma Joachim Ganzer KG, Allergopharma Joachim Ganzer KG, Reinbek, Germany. : More than 20% of population in industrial countries suffer from seasonal allergic rhinitis, commonly known as Bhay fever[. Pollen from grasses, trees and weeds are some of the most frequent allergen sources. Structural similarities found among the allergens from diverse allergen sources represent the molecular and structural basis for immunologic cross reactivity and thus explain why allergic patients can exhibit clinical symptoms and IgE reactivity to various unrelated allergen sources. Binding of an allergen to the corresponding IgE is an important step in the immunoreaction. Knowledge of the structure and binding regions is therefore important to understand this process in order to improve diagnosis of disease and to prepare better allergicspecific immunotheraphy. Timothy grass (Phleum Pratense) pollen is an important allergen source in Northern and Central Europe and its set of allergens is the best characterized within different grasses. Phl p 3 was recently isolated from Timothy grass pollen extract. From the amino acid sequence and homology comparison belongs to the group 3 allergens. In this work we determined solution structure of Phl p 3 by NMR. This solution structure and comparison with the structures from other member of the group 3 (like Phl p 2 and Phl p 1) should help us to find specific IgE-reactive eppitopes and to find better cure for the disease. Investigating pollen asthma deaths in islamabad, Pakistan Ministry of Agriculture, Plant Biology, Islamabad, Pakistan; 5 Pakistan Museum of Natural History, Botany, Islamabad, Pakistan. Introduction: Severe asthma and deaths have been reported in Islamabad during the spring allergy season, which affects thousands of residents. We investigated the deaths of 3 patients due to asthma, during the last ten days of March. In order to find a possible cause of these symptoms, we investigated the siblings of the deceased. Case 1: Male, 19 years, with a history of rhinitis during spring season, developed sudden difficulty in breathing while walking in the market, just after sunset. He was shifted to the nearest hospital. He died just on reaching the hospital, approximately 15 minutes after the onset of the attack. Case 2: A female medical doctor, 27 years, mother of 2 children aged 1 and 2 years, developed asthma suddenly about 1 hour after sunset. She was rushed to the hospital but, like the previous case, expired before reaching the hospital, approximately 20 minutes after onset of symptoms. Case 3: A 30 year old female, mother of 2 children aged 2 and 4 years, expired at home just before midnight, approximately 25 minutes after the onset of the first symptoms of asthma. Commonalities in all cases: All the deceased patients had a previous history of allergic rhinitis and occasional asthma during the spring pollen season in Islamabad. None had ever suffered from severe asthma, or been hospitalised. Very prominent is that the counts of Paper mulberry pollens were in excess of 30,000 grains / cm3 that day, and humidity was 980%, due to a forthcoming thunderstorm and rain. None of the patients was taking regular preventative treatment for allergy or asthma, at the time of the episode. None had been investigated for allergy during their lifetime. Methods: We contacted the siblings of the expired patients and found a positive history of seasonal spring allergy in 2 brothers of case 1, 4 sisters and one son of case 2, and one sister of case 3. One sibling each of cases 1 and 2 never reported any symptoms of asthma ever, while case 3 did not have any other sibling. Results: Skin prick allergy testing of all the symptomatic patients revealed strong results (more than 10 mm wheal) to pollens of paper mulberry (Broussenetia papyrifera) pollens. Conclusion: It is probable that the deceased patients suffered from allergy to pollens of paper mulberry, and expired due to the thunderstorm effect, during the peak of paper mulberry pollen allergy. Incrimination of Blomia tropicalis as a potent allergen in Indian house dust Sanjay Podder, and Goutam Kumar Saha. University of Calcutta, Department of Zoology, Kolkata, India. Allergy to house dust mites particularly to the genus Dermatophagoides is a common feature in tropical climate like Kolkata, India leading to various nasobronchial allergic manifestations. Analysis of house dust samples collected from the houses of the asthmatic patients of Kolkata revealed a variety of acarine fauna of which the mites belonging to the genus Dermatophagoides predominated, comprising 60.4% of the total acarine population followed by Blomia tropicalis (16.6%) and Austroglycyphagus geniculatus (6.8%). Perusal of literature revealed that works on the role of house dust mites in allergic etiology mainly concentrated with the genus Dermatophagoides alone. No work incriminating the role of Blomia tropicalis in house dust allergy has yet been reported from India, although it contributes significantly towards the domestic acarine population. High prevalence of this mite species in the patients`environment tempted us to search for an allergic etiology towards this species of mite. Allergy skin test with house dust and house dust mites revealed an encouraging result against Blomia tropicalis. 72% allergic asthmatics of Kolkata responded positively towards this species of mites. The total serum IgE level in patients varies between 54 to 4040 KU/ ml (mean 369+487 KU/ml). Detection of allergen specific IgE antibodies against this species of mites also confirms its role in house dust allergy. 30% patients showed very high, 20% high and 40% showed moderate IgE level against this mite antigen. The present study incriminates Blomia tropicalis as an important source of allergen in house dust in Indian condition. Aeroallergen sensitivity in patients with allergic rhinitis and or asthma Manimaran Marappan. DD Hospital and Research Centre, Pulmonology and Allergy, Chennai, India. Background: Allergic rhinitis and asthma are prevalent through out the world. For the efficient diagnosis and its effective therapy it is very important to know about the prevalance and pattern of sensitisation in a located geographical area. Little is known about this in south coastal city of Chennai in India, Southern Asia. Aim: Aim of this study was to characterise the most frequent inhalant allergen involved in Allergic rhinitis and Asthma. Subjects and Methods: A total number of 104 cases [males 54 and females 50, aged from 6 to 60] who were diagnosed to have Allergic rhinitis and or asthma were subjected to undergo Standradised SKIN PRICK TEST with 22 common Allergens. Reactions was considered to be positive if the mean wheal diameter was atleast 3 mm more than that of the negative control. Each patient was evaluated for sensitisation to allergen, number of positive reaction and mean wheal size. Results: 78 % of the patients were reactive to atlest one of the allergen and 61 % of the patients had positive reactions to more than 3 allergens. The most common allergens were, D.PTERONYSSINIUS 65%, D.FARINAE 60% HOUSE DUST 52% MOSQUITO MIX 44%. Among Pollens PARTHINIUM 40% AND CHENOPODIUM 28% and Among food allergens CITRUS MIX 45%. Conclusion: The present study revealed that the prevalance of SKIN PRICK REACTIVITY to allergens particularly to HOUSE DUST MITE is high among with peoples in Chennai. Sensitization to locally prepared allergenic extracts of house dust mites in patients with respiratory allergy at the eastern of kingdome of Saudi Arabia Emad Koshak 1 , Hussein Almehdar 2 , Tarek Gheith 3 , and Magdy Abdelreheim 4 . 1 King Abdulaziz University, Internal Medicine, Jeddah, Saudi Arabia; 2 King Abdulaziz University, Biological Sciences, Jeddah, Saudi Arabia; 3 Zagazig University, Microbiology and Immunology, Zagazig, Egypt; 4 Ein Shams University, Allergy unit, Cairo, Egypt. Introduction: The prevalence of sensitization to allergens from house dust mites (HDM) in patients with respiratory allergens appears to be directly correlated with exposure. Objective: To explore sensitization to locally prepared allergenic extracts of HDM in patients with respiratory allergy at the Eastern of Kingdome of Saudi Arabia (KSA). Methods: House dust collected from different houses at KSA was put on culture media plates containing a mixture of charcoal, gypsum and water, which was then dried and wood excelsior with crushed maize for HDM nutrition was added. These were incubated in chamber with an adjustable relative humidity at 73% and temperature at 23 -C for one month with regular observation. Three types of HDM were isolated and cultured: {(Dermatophagoid Pteronyssinus (DP), Dermatophgoid Farinae (DF) and Blomia Tropicalis (BT)}. Allergenic extracts were prepared locally using the weight/volume method concentration 1/10 for skin prick test (SPT). Sensitization was based on a positive reactivity to standard SPT performed on patients with respiratory allergy. Results: 100 patients with the primary diagnoses of allergic rhinitis in 64 patients (64%) and bronchial asthma in 34 patients (34%) were studied. They aged from 8 to 62 years old (mean=32 T13SD). SPT was positive to: DP in 53 patients (53%) (44% polysensitization with other HDM and 9% monosensitization), DF in 46 patients (46%) (42 % polysensitization with other HDM and 4% monosensitization) and BT in 2 patients (2%) (2% polysensitization with other HDM). Conclusion: Prevalence of sensitization to locally prepared allergenic extracts of HDM, especially DP followed by DF, was predominant in patients with respiratory allergy seen in the Eastern of KSA. The sensitization to BT in few patients is described for the fist time and needs further evaluation in relationship to its significance in this geographical area. Properly manufactured extracts to other local allergens may help to determine new regional environmental factors for allergy. Sensitisation to house dust and house dust mites in nasobronchial allergic patients of Kolkata Metropolis, India Sanjoy Podder, Goutam Saha, and Salil Gupta. University of Calcutta, Zoology, Kolkata, India. Background: Recent studies revealed the presence of huge quantity of reported allergenic mites viz. Dermatophagoides pteronyssinus, D. farinae and Blomia tropicalis in the indoor environment of Kolkata. Immediate hypersensitivity to these dust mites along with house dust may play a role in pathogenesis of nasobronchial allergic diseases. So, The present study evaluated the sensitisation rates towards house dust and house dust mites among patients suffering from various nasobronchial allergic manifestations residing in Kolkata metropolis area, India. Methods: The Skin Prick Test was performed on a total of 1079 patients( 585 male and 494 female and 50 healthy controls) using extracts of four allergens viz. Dermatophagoides pteronyssinus, D. farinae and Blomia tropicalis and house dust allergens. Total serum IgE was measured by using EIA technique and specific IgE to these allergens measured by following Pharmacia Immuno CAP 100 System. The influence of age and gender, if any, on allergen sensitivity was also investigated. Results: The sensitisation rates among nasobronchial allergic patients to house dust and house dust mite allergen tested were as follow: house dust (96.22%), Dermatophagoides pteronyssinus (75.06%), Blomia tropicalis (72%) and D. farinae(63.72%). The frequency of positive skin response was found to be independent of age and gender. The total serum IgE level in patients varies between 54Y4040 IU/ml ( mean 369 T 26.51). Specific IgE antibody test proved that 83% patients showed sensitivity towards at least one of the allergen tested. Discussion: Nasobronchial allergic patients are highly sensitive to the house dust and other three allergenic mites. Thus, these dust mites and house dust should be considered important allergenic sources of this area. A 19 year old white male with past history of Crohn`s disease presented to ED with one-week history of pain, rash and edema involving lower extremities. Pain was localized to his lower extremities bilaterally extending from mid shin to distal toes, particularly worse on the medial aspect of left foot. He also noticed pruritis and palpable rash in the same area. His palpable pruritic rash had improved to just a petechial rash by the time he presented. He did bring pictures of the rash when it was at its worst. There were no symptoms of exacerbation of Crohn`s disease. He was compliant with his medication. The occurrence of rash was temporally related to injection of Adalimumab, his last dose was 2 weeks prior to admission. Physical exam revealed edema of both lower extremities with scattered nonblanching petechial rash. There was also tenderness and decreased range of motion of ankle. There was no sensory deficit. Laboratory results including, CBC, CMP, PT, PTT, Rheumatoid factor, Antihistone antibody, Anti-DS DNA antibody, C3 and C4 levels were within normal range. ESR was 19, CRP was 9.080. ANA, ANCA, Serum Cryoglobulin and Hepatitis screen were negative. Skin biopsy results revealed mixed cellularity vasculitis, suggestive of resolving leukocytoclastic vasculitis. After discharge, he received next dose of Adalimumab resulting in redevelopment of the same rash only more severe in presentation. Vasculitis appears to be a rare but potentially serious complication with anti-TNF therapy that likely represents a type III hypersensitivity reaction. In patients who develop this reaction, safe practice is to stop the TNF antagonist and treat with a regimen of steroids and antihistamines with or without immunosuppressive agents. This case represents a very rare hypersensitivity reaction to humanized antibody like Adalimumab. Drugs that antagonize TNF-á have demonstrated their effect controlling inflammatory activity and inhibiting the progression of structural damage on patients with chronic arthropathies. Otherwise the use of these drugs can produce quite frequently some side effects, being skin reactions one of the most frequent after infectious diseases. A wide amount of skin reactions have being described with these drugs, such as systemic erythematous lupus, necrotizing vasculitis, discoid lupus, multiform exudative erytema, Stevens-Johnson syndrome, toxic epidermal necrolysis, eczema, urticaria, etc. We introduce the case of a 57 year old patient with rheumatoid arthritis, which developed a skin reaction with a high suspicion of being a hypersensitivity drug reaction. After two years being treated with etanercept (25 mg, twice a week) and leflunomide (20 mg a day), he developed 15 minutes after administrating the drug a vesicular itchy rash on the injection sites with itchy isolated wheals all over his body. We performed some skin tests including prick-test, intradermoreaction and patch-test with etanercept, infliximab, adalimumab and leflunomide. We used a blood sample of this patient to do a basophiles activation test (BAT) with the same drugs. We performed subcutaneous provocation with etanercept and adalimumab as well. In order to rule a possible irritative reaction out, we performed prick-test and intradermoreaction with etanercept, in five healthy controls. Prick-test and intradermoreaction with etanercept were positive and negative for the other drugs. BAT was positive with etanercept and negative with leflunomide, infliximab and adalimumab. Due to these results, we carried out subcutaneous provocation with etanercept and adalimumab, being positive to etanercept and negative to adalimumab. The skin test in healthy controls was in all of the cases negative. We presented the first case of a patient with a type I hypersensitivity reaction with etanercept, demonstrated with test in vivo and in vitro, and confirmed with subcutaneous provocation. Despite the skin reactions with this type of drugs are quite frequent, there is not enough evidence published trying to clarify the immunologic mechanism implicated on it. This article opens a new expectative on the study of the side effects and skin reactions of these drugs, taking into account that they are very effective drugs in chronic arthropathies and that their use will probably be widely spread. Oral hydroxychloroquine desensitization in a 50 year old male with cutaneous lupus Cherry Ann Distor, Lara Aleta, and Madeleine Sumpaico. Philippine General Hospital, Section of Allergy-Immunology, Manila, Philippines. Background: Hydroxychloroquine is an antimalarial drug with immunomodulatory and cytotoxic properties used in the treatment of lupus erythematosus. Adverse cutaneous reactions in the form of rashes is its most common side-effect occurring in about 3Y10%. Though, a mild reaction, it is the often the reason for cessation of treatment. This is the first reported case of rapid oral desensitization to hydroxychloroquine. Methods: We present a rapid oral hydroxychloroquine desensitization protocol in a patient who developed pruritus to hydroxychloroquine. Results: This 50 year old patient diagnosed with cutaneous lupus presented with a 1-year history of malar rash, arthralgia and diagnostic tests consistent with cutaneous lupus. He was started on prednisone, calcium carbonate and aspirin. After one year, he has was started on hydroxychloroquine for better control at 200 mg per day. On the 2nd week of treatment, he developed pruritus within two hours of intake. He was then referred for hydroxychloroquine desensitization. We used a rapid desensitization protocol as opposed to a published slow desensitization protocol by Mates, et al covering 36 days. Our patient was started at 1/30th of the total dose doubling q 30 minutes reaching the cumulative dose for the day of 200 mg. There was no pruritus and other adverse reactions observed. The patient is still being maintained on hydroxychloroquine at the present time. Conclusion: This is the first reported rapid hydroxychloroquine desensitization protocol. This demonstrates that rapid desensitization may be used when a drug is essential in an allergic patient. Oral mitotane desensitization in an 18 year old female with adrenocortical carcinoma Cherry Ann Distor, Julia de Leon, and Madeleine Sumpaico. Philippine General Hospital, Section of Allergy-Immunology, Manila, Philippines. Background: Mitotane is a chemotherapeutic drug used in the treatment of unresectable adrenocortical carcinomas. It is associated with various sideeffects that may affect intake. This is the first reported case of oral desensitization to Mitotane. Methods: We present an oral mitotane desensitization protocol in a patient who developed pruritic maculopapular rash and angioedema to mitotane. Results: This 18 year old patient diagnosed with adrenocortical carcinoma presented with a 5-year history of amenorrhea, hirshutism, striae and obesity. She underwent work-ups and excision biopsy on exploratory laparotomy revealed adrenocortical carcinoma. Prednisone was started and her symptoms subsequently resolved. On the 6th month post-surgery and medical treatment, there was recurrence of the same symptoms. She was restarted on prednisone and underwent another mass excision. Prednisone was discontinued. Mitotane was initiated at 500 mg tablet three times a day. After 1 week, she developed pruritic maculopapular rashes associated with angioedema and periauricular swelling within two hours of intake. She was then referred for mitotane desensitization. We used a protocol starting at an initial dose of 1/300th of the total dose doubling q 15 minutes reaching the total cumulative dose after 3 days of 1500 mg. She tolerated the procedure well with no untoward sideeffects. Presently, she is still on mitotane. Conclusion: This is the first reported successful mitotane desensitization protocol. This demonstrates that desensitization may be an option when a good alternative drug is not available in an allergic patient. Ravishankar Sargur 1 , William Egner 2 , and R. Robinson 3 . 1 Sheffield Teaching hospitals NHS Foundation Trust, Clinical Immunology and Allergy Unit., Sheffield, United Kingdom; 2 Sheffield Teaching hospitals NHS Foundation Trust, Clinical Immunology and Allergy Unit., Sheffield, United Kingdom; 3 Chesterfield Roayl Hospital, Medicine, Chesterfield, United Kingdom. Background: Immunologic responses to insulin in the form of IgG and IgE antibodies are common. Clinical problems related to these immunologic reactions are uncommon. Gell and Coombs type I, II, III and IV reactions have been reported. The antigenicity may be the result of any of several factors: insulin as a heterologous protein, altered tertiary structure of insulin, presence of non-insulin protein contaminants, or pharmaceutical formulation additives. IgE mediated local reactions are the most common and almost always subside spontaneously. With the use of human insulin these hypersensitivity reactions have become rare. Case report: We report a Type 2 diabetic who developed an interesting combination of significant immediate cutaneous reactions and delayed Type III and Type IV reaction to all the insulins including Human and recombinant preparations. Skin tests performed with all forms of commercially available insulin and insulin analogues were strongly positive, but skin tests were negative for solvents, excipients and additives. High titre specific IgE and IgG antibodies against insulin were demonstrated against bovine, porcine and human insulin. Attempted desensitization with subcutaneous insulin-human mixtard was initially unsuccessful. An intensive desensitisation regimen with subcutaneous administration of Human actrapid followed by human mixtard over 72 hours was done with the following Modified Insulin Desensitisation Schedule Results: On day 1, 0930 dose resulted in systemic urticaria after 45 minutes, the next dose was reduced to 1/3rd of the previous dose which caused the reaction and increments done by 1 units per injection. Hypoglycemia was prevented by infusing 5 % dextrose throughout the desensitisation procedure. After 96 hours patient was tolerating a Human mixtard dose 20 units and 10 units. Conclusion: This is an unusal case of insulin allergy resluting from multiple immunologic mechanisms which responded to a intensive desensitisation regimen. Within 6 months of successful induction of tolerance there was reduction in the IgG and IgE anti Y insulin titres. Cutaneous hypersensitivity reactions to inhibitors of cyclooxygenase-2 Souhad al Faqih. National Hospital, Allergy, Khartoum, Sudan. Background: Most patients with cutaneous hypersensitivity to nonsteroidal anti-inflammatory drugs (NSAIDs) exhibit clinical tolerance to specific inhibitors (coxibs) of cyclooxyge-nase-2 (COX-2). However, a subset of such patients will develop urticaria or angioedema when exposed to coxibs. The mechanisms of urticaria and angioedema due to coxibs are presently unknown. In this paper we discuss the prevalence of skin reactions due to coxibs and possible mechanisms mediating such adverse manifestations. Methods: A total of 206 patients (female 144, male 62, mean age 31.1T13.7 years) with urticaria and/or an-gioedema due to nonselective NSAIDs were studied between September 1999 and September 2005. Drug pattern showed 167 crossreactors (81%) and 39 single-reactors (19%), with a cutaneous clinical pattern in 122 (59.2%), mixed reactions (respiratory/cutaneous) in 82 (39.8%), and systemic reactions only in 2 (0.9%). Oral provocation tests were carried out with preferential and specific COX-2 inhibitors. In addition, a PubMed search of published studies on the tolerance to specific COX-2 inhibitors in NSAID-hypersensitive subjects was performed. Conclusion: Positive oral challenges with nimesulide were observed in 31% of studied patients, with meloxicam in 20.6%, with celecoxib in 18.4%, with etoricoxib in 11.2%, with valdecoxib in 10.3%, and with rofecoxib in 9.4%. While most NSAID-intolerant patients tolerated coxibs during oral challenges, a small subgroup of patients developed urticaria. Although there are few studies for valdecoxib and etoricoxib, these results are in agreement with the majority of published studies dealing with tolerability to rofecoxib (reaction rates 0Y34.7%) and celecoxib (reaction rates 0Y33.3%). Cutaneous reaction rates correlated with in vitro inhibition of COX-1 by different COX-2 inhibitors. Two possible mechanisms are proposed to explain urticaria induced by coxibs, an IgE-mediated allergic response, and inhibition of COX-1. The importance of drug challenge in the work-up of ADR to chemotherapeutic agents Rimini hospital, Pharmacy, Rimini, Italy. The desensitization with allopurinol is an effective treatment in selected patients, with cutaneous symptoms that do not respond to therapies alternatives. Clinical case. Old patient, with gout arthropathy (uric acid:11 milligram/decilitre), allergic to allopurinol, with pruritus and maculopapular rash. Associate pathologies: renal insufficiency (creatinine: 2,1 milligram/ decilitre); Myelodysplastic syndrome. The therapy with altenative drugs (sulfinpyrazone, rasburicase) was not effective in reducing the level of uric acid. Method preparation: one tablet of 100 mg allopurinol was weighed and crushed in a mortar ;1800 mg of rice starch (F.U.Italian) were added progressively in the mortar, by standard geometric diluition technique to obtain an homogeneous allopurinol dispersion in the mixed powder (total weigth 2000 mg).To obtain unit doses containing 50 mcg, 100 mcg, 200 mcg, 500 mcg, 1mg, 5 mg, 10 mg, respectively were weighed 1 mg 2 mg 4 mg 10 mg 20 mg 100 mg 200 mg from the mixed powder. To obtain unit dose containing allopurinol 25 mg, one tablet of 100 mg allopurinol was crushed in a mortar and 1:4 of one tablet was weighed. The doses have been manifactured in capsules. The successive treatment comes carried out following the outline proposed from Fam AG. In consideration of associated pathologies, in particular of the renal insufficiency, the dosage increment has been carried out every 5 gg. Employed doses: 50, 100, 200, 500 mcg, 1 mg, 5, 10, 25, 50, 100 mg. Treatment is lasted in total 60 days and cutaneous adverse reactions have not been taken place, neither pruritus symptoms. The first dosages have been given in hospital, and the following in outpatients department, with the necessary observation. We think that this procedure is easy and sure for patients with this type of pathology. Stevens-Johnson Syndrome from hemodialysis-associated hypersensitivity reaction in a 61-year old male Leon Bautista, III, Cesar Joseph Gloria, and Madeleine Sumpaico. University of the Philippines-Philippine General Hospital, Allergy-Immunology Department, Manila, Philippines. Background: Stevens-Johnson syndrome (SJS) is often described as diffuse, severe, mucocutaneous eruption involving two or more mucosal surfaces and majority of cases are attributed to drug exposures. Hemodialysis-associated hypersensitivity reactions are well documented in literature and majority would present as flushing, urticaria, periorbital and facial edema, bronchial hypersecretion and hypotension. This is the first reported case of hemodialysis-associated SJS. Methods: To present a patient who developed SJS after hemodialysis. Results: This is a 61-year old male diagnosed with chronic kidney disease, in uremia. Less than 24 hours after the initiation hemodialysis, patient developed generalized, pruritic, erythematous macular rahes. This was thought to be an adverse reaction to present antibiotics thus, all antibiotics were discontinued. After the second hemodialysis, rashes progressed with increase in erythema and exfoliation. This was thought to be a an anaphylactoid reaction to ethylene oxide present in dialyzers. Thus, pre-medication prior to subsequent hemodialysis was done. But on subsequent hemodialysis these lesions became localized flaccid bullae, later developing epidermal detachment involving the eyes and oral mucosa. He received intravenous hydrocortisone up to 200 mg/kg/day (equivalent to 3mg/kg/day methylprednisolone). However, patient`s condition worsened requiring endotracheal intubation. He subsequently died due to multi-organ failure. The strong history surrounding each hemodialysis sessions with the progression of his lesions plus the high index of suspicion pointed to ethylene oxide hypersensitivity as the cause of the SJS. Conclusion: This is the first reported case of hemodialysis-associated SJS. This case report demonstrates the importance of early detection, referral, and management of hemodialysis-associated hypersensitivity reaction. Replacing ethylene oxide with steam or gamma radiation may help solve this problem. The impact of polypharmacy intervention on adverse drug reactions and cost of care in new orleans seniors Prem Kumar, Kareem Elshatory, E. Elshatory, Carlos Vital, Sandhya Mani, and Annette Fiorillo. Louisiana State University Health Sciences Center, Allergy/Immunology, New Orleans, United States. Background: In the United States (U.S.), more than 100,000 deaths annually are attributed to serious adverse drug reactions (including but not limited to drug allergy). The estimated total cost of these events in 1996 was 12.2 billion (U.S.) dollars. Polypharmacy, particularly in seniors, contributes to the increased likelihood of adverse drug reactions (ADRs). Polypharmacy is defined as the use of five or more prescription medications on a long-term basis. There are no reports of intervention studies from the state of Louisiana (U.S.) where fifty percent of seniors are on polypharmacy in contrast to twenty percent prevalence in the rest of the United States. The goal of this pilot clinical observational study is to demonstrate whether cessation of inappropriate medications would decrease adverse drug reactions and lower the monthly cost of medications. Methods: Nine patients, ages 66 to 83 years, that met the definition of polypharmacy were evaluated. Patients with known Human Immunodeficiency Virus (HIV/AIDS), transplant recipients, and malignancy were excluded secondary to known requirement of multiple medications. The Medication Appropriateness Index (MAI) was utilized to stop inappropriate medications. ADRs, Mini Mental Status Exam (MMSE) scores and monthly cost of medications were recorded before and after intervention. Results: The mean number of medications used daily decreased from 13.5 medications to 7.3 medications before and after intervention respectively. The ADRs decreased from 3.0 occurrences before intervention to 0.3 occurrences after intervention (pG0.001), while MMSE mean scores improved from 25.5 prior to intervention to 28. Tuberculosis continues to be a major cause of morbidity and mortality worldwide. Currently available drugs are effective for treatment of the disease or latent infection, but may cause serious allergic reactions, often difficult to manage. Once the diagnosis allergic drug reactions have been established, re-introduction of the drug, leading to tolerance can be considered, induction of tolerance is usually possible in most patients. We report the observations of 6 women, their age varies between 29 and 70 years, without immunodeficiency, treated for pulmonary tuberculosis TPM+ (2 patients), pleural tuberculosis confirmed histologically (3 patients) and a TPM-(1 patient) and having received an antituberculosis treatment containing SRHZ for the two first and RHZ for the last four ones with adequate posology. All our patients presented cutaneous reactions of gravity moderated at severe, which have occurred between 3 and 45 days after the beginning of the treatment. The treatment was stopped until disappearance of the lesions in all the cases. The reintroduction was practised successfully at five of our patients and the therapeutic protocol was changed in a patient considering the gravity of the lesions. Cutaneous adverse drug reaction induced by a generic substitute of zyloric with a residual sensitization to allopurinol Marie-Lorraine Chandeclerc, Jean-Fran0ois Cuny, Jean-Luc Schmutz, and Annick Barbaud. Hopital Fournier, Dermatologie, Nancy, France. Cutaneous adverse drug reaction (CADR) due to a generic drugs seem to be rare or have been rarely reported except with antiepileptic drugs. We report on the first case of a CADR that occurred when a drug that had been well tolerated for 15 years was replaced by a generic substitute. A 59 year old man, who had taken Zyloric* pills for 15 years developed a maculopaplar rash (MPR) 3 days after the replacement of Zyloric* pills by its generic substitute called Allopurinol Ge*. Four days after stopping the intake of Allopurinol Ge* the MPR spontaneously disappeared. Biological findings remained normal without liver involvement. Two months after, the patient took Zyloric* again and developed generalized eruption with a more severe involvement of the flexure areas, the 3 days following the re-intake of the drug. This second CADR was less severe than the previous one. Three months later, the intake of Zyloric* was disrupted and the CADR vanished in 3 days. There was no other modification in the usual treatment. Dermato-allergological investigations were performed three months after the disappearance of the CADR and after stopping Zyloric*. Drug patch and pricks tests were done. The results were negative for both tests. To our knowledge, we report on the first case of CADR due to allopurinol, appearing after replacing well tolerated Zyloric* by one of its generic substitutes. The generic substitute was responsible for inducing a sensitization to allopurinol, which unfortunately led a relapse of the CADR when Zyloric* was reintroduced, even though the clinical eatures were less severe in the second CADR. There are no common excipients between Zyloric* and its generic. In the few reported cases in literature of CADR occurring after the replacement of a drug by its generic substitute, the adverse effect does not relapse when the original drug is readministrated. Thus, to our knowledge, here is the first case in which the reintroduction of the original drug was associated with a relapse of the CADR. From our observation, new mechanisms must be evoked for this type of therapeutic accident, i.e. the role of excipients other than those listed on the pharmacological notice or a non tolerated commercialized form of the drug that induces a sensitization to the active compound. The value of skin tests in the investigation of suspected systemic corticosteroids hypersensitivity Nadège Michelet 1 , Philippe Trechot 2 , Jean-Luc Schmutz 1 , and Annick Barbaud 1 . 1 Fournier Hospital, Dermatology, Nancy, France; 2 Central Hospital, Pharmacology, Nancy, France. Background: To establish the value of patch tests (PT), prick tests (pt), intradermal tests (IDT) in suspected immediate or delayed hypersensitivity to corticosteroids (CS) systemically administered. Materials and Methods: 67 patients (52 women, 15 men, mean age 50,6 y.o) referred for immediate (13, 4%) or delayed (86,6%) cutaneous adverse reaction (CAR) possibly due to a CS systemically administered, were enrolled. All had PT with a special series of 12 CS diluted in petrolatum, read at 20 minutes, 48h, 96h and one week. When negative, pt were carried out with tixocortol, budesonide, betamethasone, betamethasone dipropionate, hydrocortisone 17 butyrate, triamcinolone acetonide and the involved CS. If PT and pt were negative, IDT were carried out at least with one CS of each group of the Goossens classification (1): hydrocortisone (group A), triamcinolone acetonide (group B), betamethasone (group C), betamethasone dipropionate (group D1), with progressively increasing dosages. If skin tests were negative, oral provocation tests with the involved CS or another CS (same group or different group) were proposed. Oral administrations were performed under hospital surveillance, versus placebo, with progressively increasing concentrations to the full dosage. Results: 12 patients (18%) had positive results either on skin testing or oral administration. PT were positive in 5 cases (7.5 %), none had positive pt, 3 patients had positive IDT, one had a positive repeated open application test. In 58 patients all tests were negative. Thirty-eight accepted one or more oral administrations (OA) of CS (50 OA were performed). Among them, 1/21 had a positive OA to the involved CS, 1/9 had a positive OA to a CS belonging to the same group; 2/20 had a positive OA with a CS belonging to another group. Conclusion: Hypersensitivity to systemic CS is supposed to be rare but do exist, as we confirmed a CS sensitization in 12 patients with an immediate or delayed mechanism. We demonstrated that skin tests are useful in the management of such patients. They have a mean value in investigating suspected CS hypersensitivity, but they are well tolerated and seem to have a good negative predictive value (92.1 % in our series). They allow to confirm a sensitization (positive tests) and to lead the OA in the initially responsible chemical group of CS or in another group with a low risk of reactivity. Stefan Cimbollek, Teresa González-Quevedo, Maria Robledo Avila, Angustias Nieto, and Manuel Díaz. Hospital U. Virgen del Rocío, Allergy Unit, Seville, Spain. Background: Proton pump inhibitors are widely used but only few cases of anaphylaxis have been reported. In some cases cutaneous tests have proved useful and in only one case cytometric cellular allergen test (FAST) was performed and confirmed the existence of specific IgE. Case report: We present a 26 year old patient whith previous history of urticaria. In one episode he associated nausea and shortness of breath. He did not correlate symptoms to food ingestions. He used ocassionally omeprazole due to acid reflux. Methods: Skin prick tests and intradermal tests were performed with omeprazole and pantoprazole (1/100 and 1/1000 dilutions) with negative results. Controlled oral challenge was programmed. The patient started with generalized flushing and pruritus, eyelid edema, abdominal pain and shortness of breath 45 minutes after last dose of omeprazole. Physical examination did not reveal wheezing nor hypotension. The patient recovered in two hours following intravenous infusion of 500 ml of saline, an iv push of one ampoule of pheniramine and 80 mg of prednisolone. FAST, also called basophil activation test, was performed with negative result. Conclusion: We present a case of anaphylaxis to omeprazole which could not be confirmed with cutaneous nor basophil activation tests as previously reported. Drug allergy in pollinosis: is the pattern changing? Background: We have presented at a previous EAACI meeting a study on the association between pollen and drug allergies. In this study $-lactams were more frequently associated with pollen allergy than non steroidal antiinflammatory drug (NSAIDS). The aim of the present study is to verify if the association pattern has changed. Methods: 952 patients allergic to pollens and 141 with hypersensitivity to drugs have been studied in the last five years. 47 patients were simultaneously allergic to pollens and drugs (4,9% of pollinosis) (33,3% of drug allergy). Clinical history, skin prick tests for pollens and prick and ID when necessary for drugs, specific IgE when possible done in all the 47 patients. Results: 32 females (68,1%) and 15 males (31,9%), previously Females (54,4%) Males (45,6%). 40 patients (85,1%) attended by pollinosis and 7 (14,9%) by drug allergy. In pollen allergy clinics was rhinitis in 45 and asthma alone in 2. Drug allergy: urticaria-angioedema in 32 (68,8%), shock in 2, bronchospasm in 2. Skin prick tests-grass pollens 32 (68%), Parietaria 19 (40,2%), olive tree 12 (25,5%). Drug allergy: $-lactams 19 (40,4%), NSAIDS 24 (51,1%) and other drugs (16,2%). There was no correlation between allergy to one specific pollen and a group of drugs. Conclusion: The association of pollinosis with drug allergy is not very frequent. Reporting to the results previously obtained in these series there is a greater number of grass pollens allergy instead of Parietaria and a greater incidence of NSAIDS hypersensitivity instead of $-lactams. These results can be explained by a decrease of antibiotics prescription to allergic patients and an increase in the use of NSAIDS in the last few years. Background: Drug hypersensitivity is a common clinical problem associated with significant morbidity and mortality. According to Volchek (2004) , adverse drug reactions are a major health issue encountered frequently in both the inpatient and the outpatient setting. Over the past few years, with the increasing number of medications that have been manufactured, there has been an increasing emphasis on pharmacotherapeutic vigilance in order to reduce the incidence of such reactions. Objective: The purpose of this investigation was to review records of patients referred to for adverse drug reactions and determine the most commonly implicated drugs, most common clinical manifestations as well as to look into the source of the referral or the consult. Methods: Chart review and frequency distribution Results: A total of 1,694 patients were referred to the Allergy Section from 2002Y2006 with a slowly increasing trend each year. Forty percent of these patients were seen at the outpatient clinic and 60% were in-patient referrals. Among those admitted, the number of adult and pediatric cases were nearly the same (54% and 46% respectively). In the out-patient clinics, adult referrals outnumbered pediatric cases by more than two-thirds. The most common manifestations in the inpatient setting were cutaneous reactions (maculopapular rashes, urticaria, angioedema), followed almost equally by jaundice, difficulty breathing, fever and chills. Rare generalized reactions such as Stevens-Johnson Syndrome (SJS) and toxic epidermal necrolysis (TEN) were also reported, averaging 10 cases per year. The most commonly implicated drugs for both in-and outpatient settings was still beta-lactam antibiotics, followed by anti-tuberculosis medications, anticonvulsants, ASA/NSAIDs and others (L-asparaginase, vitamin K and allopurinol). Over the five year period there was an increasing trend of. Conclusion: Results have shown that over the past five years, although the implicated medications are the same, there has been an increasing trend in the reporting of adverse drug reactions. This is most probably due to the fact that there is a heightened awareness regarding the issue. An increasing number of brand names and manufacturers may also lead to an increased number of adverse reactions to medications. Key words: adverse drug reaction 847 A 10-year update from the Tan Tock Seng Hospital (TTSH) prospective inpatient drug allergy reporting system (1997 Y 2006) 848 Negative predictive value of drug skin tests in investigating cutaneous adverse drug reactions Julie Waton 1 , Philipe Trechot 2 , Carole Loss-ayav 3 , Jean-Luc Schmutz 1 , and Annick Schmutz 1 . 1 Fournier Hospital, Dermatology, Nancy, France; 2 Central Hospital, Pharmacology, Nancy, France; 3 Marin Hospital, Epidemiology, Nancy, France. : Drug skin tests are useful in etiological analyses of cutaneous adverse drug reactions (CADR), in order to determine if the drug can be readministered or to avoid a cross reaction with a substitute drug. The aim of this study is to evaluate the negative predictive value (NPV) of drug skin tests. Patients and Methods: After a retrospective analysis of the files of patients referred for CADR between 1996 and 2006, we have enrolled those having CADR with clinical features, onset`s delay after the drug intake and drug imputability strictly determined, having had negative drug skin tests followed by drug administrations. There were oral provocation tests (OPT) or substitution tests (ST) with a drug of the same class of those of the drug suspected during the CADR. Methods: The files of 9 067 patients have been reviewed. Clinical presentation and suspected drugs registered. Skin tests with the group of drug incremated have been done by prick and intradermal techniques in progressive concentrations when prick were negative informed consent was obtained from all the patients. Specific IgE or leucotrienes release has been done in cases these techniques are available. Results: 448 (4,9%) out of 9 067 patients reported one or more drug adverse reactions: (females 69,7%, males 30,3%). 90% of the patients reported urticaria/angiodema and 10% rhinitis and/or asthma. NSAIDS were implied in 63,6% of the patients, $-lactams in 34,2%, local anesthetics in 4,7%. Associations of reactions to NSAIDS and $-lactams were frequent (22,3%) other drug less implied were sulfa drugs, quinolones, macrolides, ACE inhibitors, angiotensin II blockers, statins, corticosteroids, vitamines, allopurinol, tetracyclin, gentamycin, furadantine, curarisers, tramadol, ranitidin, thyocholchycosid, fibrates, fluconazol. Specific IgE and CAST (leucotrienes release) were positive only in less than 50% of the cases. Conclusion: NSAIDS and $-lactams are the more frequent cause of adverse reactions in out-patients clinical history, skin tests step by step are the more practical first diagnostic approach of drug allergy in out-patients. Drug hypersensitivity can cause asthma, allergic rhinitis or both (Fermaid Widal triad) in a non-negligible number of patients atopic or non-atopic. So-Hee Lee 1 , Sang-Min Lee 1 , Hyouk-Soo Kwon 1 , Chang-Han Park 1 , Heung-Woo Park 1 , Yoon-Seok Chang 2 , Sun-Sin Kim 1 , Sang-Heon Cho 1 , Kyung-Up Min 1 , and You-Young Kim 1 . 1 Seoul National University Hospital, Department of Internal Medicine, Seoul, Republic of Korea; 2 Seoul National University Bundang Hospital, Department of Internal Medicine, Seongnam, Republic of Korea. Background: Antibiotics skin testing (AST) is a useful procedure for identifying patients with IgE mediated hypersensitivity to antibiotics. However the procedure is not standardized and is performed in diverse and sometimes inappropriate protocols among hospitals in Korea. Objective: To examine protocols of AST in major hospitals of Korea and compare it with that in the literature. Methods: Routine protocols of AST performed in general wards of 12 general hospitals were collected via allergists working in each hospital by using questionnaire. In addition, we reviewed literatures searched from Pubmed/ Medline. Results: All of the 12 allergists responded to the questionnaire. All hospitals carried out AST on volar forearm by intradermal method without any controls. Two hospitals used saline as a negative control only if false positive result was suspected. Penicillins and cephalosporins were tested in 10 hospitals and 11 hospitals. In detail, both were tested in 9 hospitals, only penicillins in one hospital, and only cephalosporins in 2 hospitals, among which penicillin was never prescribed in one. In addition, semisynthetic penicillins was tested in 8 hospitals, carbapenems in 9 hospitals, vancomycins in 4 hospitals, aminoglycosides in 3 hospitals, and monobactams in one hospital. The testing concentration differed upto 10000 folds among the 12 hospitals. Standard of interpretation also varied between hospitals. The generalized method of AST in the literatures was intradermal testing on volar forearm. ASTwas recommended as a routine practice for penicillins and cephalosporins but not for other antibiotics in most of the literatures. Conclusion: The protocols of AST varied greatly among 12 general hospitals in Korea and differed from that commonly used in literatures. Thus standardized guidelines for AST are required for the safe usage of antibiotics. Case report: A 19-year-old healthy male took amoxicilin for a respiratory infection. After two days of treatment, some hours after the last dose, he suffered pruritus, erythema, pain, and local inflammation of penis, with vessels blisters and wounds of 1Y2 cm. He recovered after oral corticoids, antihistamines and topic antibiotic, plus permanganate potassium washing. The wounds lasted 2Y3 weeks and left skin pigmentation. On another occasion, he took paracetamol again for a sore throat. After 3Y4 days of treatment he suffered similar wounds and needed the same treatment Following this, he has tolerated other NSAIDs. Methods and Results: With written consent we made the following study: We performed a patch test with paracetamol 10% concentration that was negative at 48 and 96 hours. Prick and ID tests with paracetamol were both negative too. An oral controlled challenge test with paracetamol up to 500 mg was also negative. After this, we recommended that the patient be taken 500 mg. of paracetamol every 8 hours three more days. After two days of treatment the penis wounds appeared again and other wounds between the 1st and 2nd finger of the left hand appeared too. This wound was a 1-cm diameter, erithematous and descamative plaque. These wounds recovered after oral corticoids and antihistamines that were administered during 5Y6 days. After the challenge the patient recalled that in the first reaction he had taken paracetamol plus amoxicilin. We performed cutaneous and oral challenge tests with amoxicilin with negative results and the patient took amoxicilin for 5 more days with good tolerance. Patch test with TRUE TESTi and challenge tests with other paraamino drugs (sulfonamides and hydroclorotiazide) were all negative. Conclusion: We present an infrequent adverse reaction due to paracetamol confirmed by a challenge test. It is a skin reaction suggestive of dual fixed exanthema in both penis and betwen 1st and 2nd finger of the left hand. We have ruled out cross-reactivity with other para-amino drugs. Although paracetamol allergy is infrequent, it is important to investigate and to study all the drugs involved in an adverse reaction. Bernard Arnoux 1 , Philippe Jean Bousquet 2 , Nazima Soohun 3 , Laure Rouanet-Bousquet 3 , and Pascal Demoly 3 . 1 INSERM, U454, Montpellier, France; 2 University of N^mes, Dept de l'Information Mdicale, N^mes, 854 Adverse cutaneous reactions due to glycopeptides-induced adverse cutaneous reactions: results of immunoallergical investigations in 9 patients Amandine Perrin 1 , Philippe Trechot 2 , Nadine Petitpain 2 , J. Fran0ois Cuny 1 , J. Luc Schmutz 1 , and Annick Barbaud 1 . 1 Hopital Fournier, Dermatology, Nancy, France; 2 CHU, Pharmacovigilance, Nancy, France. Objective: With the emergence of hightly resistant staphylococcus, vancomycin (V) and teicoplanine (T) usage has considerably increased but they can induce cutaneous adverse reactions (CAR) for which the value of immunoallergical investigations has not yet been determined. Methods: Retrospective study (2006Y2006) including 9 patients (6 women, 3 men; mean age = 54.3) with CAR suspected to be due to glycopeptides, V: 4 cases, T: 4 cases and successively V then T: 1 case. Red man syndromes were not included. Imputability was estimated according to Moore's criteria. The CAR were 6 maculopapular rashes, 1 linear bullous dermatosis, 1 DRESS and 1 urticaria. Six weeks later, according to ESCD's guideline for drug skin tests, immunoallergological investigations were carried out with successively drug patch tests, prick tests and intradermal tests with all the drugs taken during the CAR. Results: Skin tests confirmed the responsibility of glycopeptides in 4/9 cases, with cross-reactivity between V and T in 2/5 cases. Following negative immuno-allergical investigations, 3 patients had well tolerated provocation tests with glycopeptides. Conclusion: In literature, T seems to be safer than V (ototoxicity, rashes, nephrotoxicity) for a similar efficacy, inducing very few hypersensitivity reactions such as red man syndrome (3 cases published). Allergic crossreactivity (V and T), already reported in literature, is not so rare (2 of our patients) but unconstant (3 of our patients had indeed well tolerated provocation tests with the other glycopeptide and we precise that all the skin tests were negatives for V and T in these patients. In that way, this study has important clinical implications, demonstrating that skin tests are useful in CAR due to glycopeptides, in determining the responsible drug and if the tests are negative, allowing the readministration of another glycopeptide with hospital surveillance, as we demonstrated that cross-reactivity between glycopeptides is not constant. A link between angioedema caused by inhibitors of angiotensin converting enzyme and eotaxin and ECP Malgorzata Basinska, Tomasz Basinski, Robert Pawlowicz, and Bernard Panaszek. Wroclaw Medical University, Department of Internal Medicine and Allergology, Wroclaw, Poland. Background: Inhibitors of angiotensin-converting enzyme (iACE) and Blockers of angiotensin II type 1 receptor (AT1 blockers) are commonly used in hypertention and chronic heart failure treatment. These therapies are believed to be safe. However, angioedema is a rare but a well recognized potentially life-threatening adverse effect of iACE therapy. The aim of the study was to estimate angioedema threat in patients treated with short and long acting iACE in comparison to patients receiving AT1 blockers. Eosinophilic cationic protein (ECP) and eotaxin serum concentrations were used as indicators of the angioedema threat, since bradykinin is involved in the activation of this chemokine and subsequently eosinophils. Methods: Patients were divided into the following groups, according to the treatment they have been receiving: AT1 blockers, short acting iACE, long acting iACE and control. Patients with a history of angioedema of etiology unrelated to the study were excluded. Eotaxin concentration in the serum was determined by standard solid phase sandwich ELISA. ECP level in the serum was assessed by the Pharmacia CAP system ECP fluoro enzyme immunoassay (FEIA) method, according to the manufacturer`s instructions. Results are shown as mean T SD. Student`s t test for unpaired samples was used for statistical analysis. Results: There were no statistically significant difference with regard to serum ECP level between the AT1 blockers and control group. The same was true for eotaxin. The levels of ECP in sera of patients taking short acting iACE (12.9T3.30 6g/mL, pG0.001) and long acting iACE (10.8T4.9 6g/mL, pG0.05) were significantly higher than in control group (5.22T2.5 6g/mL). The level of eotaxin was higher in serum of patients taking short acting iACE (177.5T57.6 pg/mL, pG0.05) and higher in serum of patient taking long acting iACE (156.1T44.8 pg/mL, pG0.05) compared to the control group (112.3T21.5 pg/ mL). Conclusion: Serum concentration of eotaxin and ECP play potential role in the pathogenesis of iACE induced angioedema and their concentration may be a predicting factor of this adverse event. Thus, patients treated with AT1 blockers are at the significantly lower risk of angioedema in comparison to patients taking iACE. Fixed drug eruption caused by traditional herbal medicine (jamu) Background: Fixed drug eruption (FDE) are so named because they recur at same site with each exposure to the medication. Jamu is the term commonly used to describe traditional herbal medicine products from Indonesia. The medicinal elements may be made from roots, bark, leaves, fruits, flowers, seeds or other parts of a plant. Jamu pegal linu indicated for the treatment of rheumatism, body ache and bone ache, muscle joint, giddiness, toothache and backache usually consumed by hard workers. Commercial products are available in the form of powders, pills, capsules, mixtures and creams. Some illegal retailers adding phenylbutazone, allopurinol and paracetamol in to jamu. FDE caused by jamu has never been reported yet. Case: A 40-year-old man presented with multiple black macules on the upper left eyebrow, left and right cheeks, lower lip and left and right hands. The lesions initially appeared small and only a few, enlarge slowly, and became obvious and of concern to the patient in the past 2 weeks. The lesions were slightly itchy but there was no pain. He was otherwise healthy. He denied taking any medication except traditional herbal medicine (jamu) for the past 1 year when feeling tired. Family history was negative. Laboratory examinations were within normal limits. Skin biopsy from one of his lesions showed some vaculolar degeneration of the basal cells, pigmentary incontinence, mild superficial perivascular limpho-histiocytic infiltrates with a few eosinophils. Discussion: The clinical presentation and histopathologic alterations are consistent with the diagnosis of fixed drug eruption. The patient was treated 857 Anaphylactic reactions during general anaesthesia: diagnostic approach and prevalence of anaesthetics involvement Mauro Cancian 1 , Muzio Meroni 2 , Raffaele Bendo 1 , Elena Ossi 1 , and Giuseppe Realdi 1 . 1 Clinica Medica 1-University of Padova, Medical and Surgical Department, Padova, Italy; 2 ISTAR-University of Padova, Pharmacology and Anesthesiology Department, Padova, Italy. Background: Although anaphylactic reactions during general anaesthesia show a very low incidence, the mortality reaches 5% or more, and additional patients may experience significant brain damage. Neuromuscular blocking drugs are usually reported as the most common cause of anaesthesia-elicited anaphylaxis, but the possible involvement of induction agents, antimicrobials, latex, plasma expanders and other drugs should be considered. Unfortunately, the diagnostic approach is limited by the lack of validated commercial kits for detection of specific antibodies, as well as by the occurrence of pseudoallergic reactions, that are not mediated by circulating IgE. Methods: We evaluated 20 consecutive cases (14 F, 6 M; age range 16Y68 yrs) of well documented anaphylaxis during general anaesthesia, by laboratory assays (WBC count; Eosinophil cationic protein; Serum Tryptase; Plasma C4 and C1-INH; IgA and total IgE levels; RAST for inhalant and food allergens, latex and beta-lactams) and skin tests (Patch and/or Prick testing; intradermal injection) for inhalant allergens, eggs, soybeans and exotic fruit, latex and drugs administered before or during anaphylaxis onset. Drug skin tests were performed with commercial formulations of beta-lactams, heparin, atropine, benzodiazepins, vecuronium, norcuronium, mivacurium, rocuromium, fentanyl, remifentanyl, propofol, droperidol and thiopental, at different dilutions. Results: Two patients showed evidence of allergy to norcuronium and mivacurium, respectively. Three subjects were allergic to amoxicillin and six to latex. Heparin skin testing elicited a local weal-and-flare response in one patient, whilst diffuse urticaria and itching were observed in one case. Finally, apparent anaphylaxis followed by cardiac arrest during anaesthesia in an asthmatic subject, seemed to be an adverse effect of a difficult intubation rather than a true allergic or pseudoallergic reaction. Unfortunately, in the remaining six patients we were not able to identify any triggering factors, also because no diagnostic tests are available to assess NSAIDs hypersensitivity. Conclusion: Latex allergy still represents, at least in our experience, the main cause of anaphylaxis during general anaesthesia. Although the reliability of skin testing with commercial drugs has not been well established till now, this procedure can help to prevent the risk of further episodes in patients with a documented anaphylactic reaction during anaesthesia. Four cases of steroid allergy with bronchial asthma Yoko Murakami, Jyunko Ikei, Hiroshi Odajima, Rumiko Shibata, and Sankei Nishima. Fukuoka National Hospital, Pediatrics, Fukuoka City, Japan. Background: Corticosteroid preparations have anti-inflammatory and immunosuppressive properties and are widely used in the treatment of asthma and allergic disorders. However, steroids themselves can rarely induce hypersensitivity reactions. The severity of the reaction can vary from a rash to anaphylaxis or death. We report four cases regarded as the steroid allergy that we experienced in our hospital recently. Case Report: Case 1 was the 9 years old girl and hospitalized with moderate asthma attack. Rash developed in several minutes when she was given aminophilin continuous intravenous infusion and hydrocortisone succinate (HC-S) 6mg/kg intravenous drip. An intradermal test by HC-S was positive and we diagnosed the drug allergy. We changed it to predonine (PSL) intravenous infusion, rash didn't develop but persistence of wheeze and decreased FEV1.0. The symptom and lung function were improved when all steroid were canceled. Case 2 was 10 years old girl with many kinds food allergy and hospitalized with moderate asthma attack. She was given aminophilin continuous intravenous infusion, hydrocortisone succinate (HC-S) 7mg/kg intravenous drip and PSL 1.0mg/kg$B!_#4(B/day intravenous injection for 8 days. She showed persistence of wheeze and decreased$B!!(B-FEV1.0. She was diagnosed steroid allergy and pertussis infection. By an intradermal test, she showed positive for PSL, HC-S, methylprednisolone succinate(MP-S), hydrocortisone sodium phosphate. Wheeze and FEV1.0 were improved by all steroid cancellation. Case 3 was a 23 years old female patient hospitalized with moderate asthma attack. It was similar to case 2, she had intractable wheeze and decreased FEV1.0 persisted. By an intradermal test, she showed positive for PSL, HC-S and she was diagnosed steroid allergy. Case 4 was a 10 years old boy with moderate asthma attack developed urticaria and hypotension after receiving HC-S intravenous drip. In this patient the intradermal tests HC-S and PSL showed positive and diagnosed steroid allergy. Conclusion: 3 in 4 cases did not show anaphylactic reaction. There were many cases which showed persistence of wheeze and decreased FEV1.0 during treatment of asthma. We thought that the steroid allergy that took the course to protract of asthma attack could be present from these cases. It is important to considerate striod allergy in treatment of acute exacerbation of intractable asthma. Angioedema after administration of methylprednisolone to treat drug allergy: a case report Enrico Compalati, Laura Guerra, Anthi Rogkakou, Antonio Scordamaglia, Giovanni Passalacqua, and Giorgio Walter Canonica. Allergy and Respiratory Diseases Clinic, Genoa University, Department of Internal Medicine, Genoa, Italy. Background: Despite the large use of corticosteroids (CS), allergic reactions to these drugs remain uncommon; Immediate allergic reactions to CS are uncommon events usually provoked by excipients. Both immunologic and nonimmunologic mechanisms are proposed. Delayed reactions are compatible with a T-cell mediated response. Cross-reactivity has been observed between different topical corticosteroids, but the classification of crossreactivity between them does not seem to be completely useful in adverse reactions to systemic corticosteroids. Case report. We signal the case of a 39year-old woman with no atopy but contact sensitization to tixocortol-21pivalate 1%. The woman, before a recent administration of mepivacaine for dental surgery, had proceeded with CS preventive administration (Prednisone 50mg 13hh, 7 hh and and clorfenamine 1h before the intervention). One hour after the last dose of premedication, followed by anahestetic administration, patient presented facial angioaedema. Further administration of metilprednisolone to treat the adverse event had the effect of increasing symptoms. Management: Patient was studied for corticosteroids allergy. Patch test, performed with Standard Series, confirmed mentioned sensitization. Skint tests were carried out with metilprednisolone succinate, betametasone fosfate, triamcinolone acetodine. Patient showed immediate local reaction after intradermoreaction with metilprednisolone; no positivity for other CS tested. Comments: Tixocortol-21-pivalate and prednisone belong to group A corticosteroids, so a possible cross reaction may be suggestive. As CS are poorly soluble in water or saline, they are coupled with esters. In this circumstance patient reacted to both the previous administration of prednisone and then to metilprednisolone; these two preparations are different only for the presence of succinate ester in the second one, so an effect of this additive may reasonably be excluded. Therefore all other excipients included in the tested preparations were commonly existing in both tolerated and untolerated drugs. Conclusion: Drug adverse reactions appearing altough preventive administration of systemic CS should be reconsidered for possible allergic response to CS. The classification of cross reactivity between CS, with patch testing and intredermal testing, may provide useful support to confirm diagnosis and to identify alternative tolerated molecules. Background: Erythema multiforme is a reaction pattern of blood vessels in the dermis with secondary epidermal changes and manifests clinically as characteristic erythematous iris-shaped papular, little or no mucous membrane involvement; vesicles but no bullae or systemic symptoms. Eruption is usually confined to extremities, face, classic target lesions. Case: A 56 year-old girl with erythema multiforme minor developed typical lesions involving most of the skin. The eruptions developed after she had taken some medicines for antihypertension. Histopathology result showed focal hyperkeratosis, acantosis, focal spongiosis in epidermis and inflammatory cell infiltrate: lymphocytes and eosinophil in dermis without vacuolar degeneration. Discussion: Mild forms have appearance of little or no mucous membrane involvement, vesicles but no bullae or systemic symptoms. In this case there were erythematous patchs and vesicles in his skin. This eruption is usually confined to extensor aspects of extremities, and classic target lesions. Clinical manifestation characteristic by classic target lesions that characterized for erythema multiforme but histopathology examination showed appearance like drug eruption without vacuolar degeneration, that mean there is discrepancy of clinical appearance and histopathology. Mild form of erythema multiforme may developed to severe erythema but in this case the lesion improved after treatment with steroid equivalent of dexamethasone 30mg orally a day in divided doses. Conclusion: There is discrepancy of clinical appearance and histopathology. Erythema multiforme in this patient well responded to corticosteroid, withdrawal of all possible offending drugs, and supportive therapy. Prompt diagnosis and treatment give good result. Keywords: erythema multiforme, mild forms, classic target lesions, vacuolar degeneration. Severe reaction due to fluoroquinolones -Toxic epidermal necrolysis case report Ioana Adriana Bujor, and Diana Dumitrascu. 3rd Medical Clinic, University of Medicine and Pharmacy BIuliu Hatieganu^, Allergy Department, Cluj Napoca, Romania. Background: Toxic epidermal necrolysis (TEN) is an uncommon fulminating mucocutaneous disease associated with high prevalence of mortality. Frequently TEN is associated to drug administration. TEN due to fluoroquinolones are rare, 7 case reports in Medline with TEN due to ciprofloxacin. Allergic reactions due to fluoroquinolones are very rare. Anaphylactoid and/or anaphylactic reactions are estimated to occur in 0.46-1.2 in 100,000 patients. Fluoroquinolones are new antibiotic drugs used mostly in treatment of urinary and respiratory infections. Allergy to fluoroquinolones may have many clinical manifestation: anaphylactic or anaphylactoid reaction, acute urticaria with or without angioedema, fixed drug eruption, serum sick-like illness, nummular eczema. The most frequent used quinolones in Romania are ciprofloxacin and norfloxacin. 2812 patients were evaluated in the department of Allergy, in Cluj Napoca between Jan 2005YDec 2006. Only 14 (0.49%) cases of allergic reaction due fluoroquinolones were reported. Case report: A 29-years-old smoking man was admitted in our hospital with malaise, with an eritematous eruption in the whole skin with small areas of denudation of epithelium, ulcers in nasal and oral mucosa. Those phenomena were associated with a severe itching and burning sensation of the skin. The patient began 10 days ago treatment with ciprofloxacin for an acute bronchitis, that was the only drug he took in the last 6 months. After a complete examination the patient presented also hepatocitolisis. After treatment with corticosteroids, antibiotics intense care of the skin the patient recover complete. Evolution in 2 months was Brestitutio at integrum^. Conclusion: Toxic epidermal necrolysis (TEN) is a rare, but the increasing rates of reports for this syndrome due to ciprofloxacin could reconsider first choice option for this drug. More studies should be done in order to find the prevalence of TEN to ciprofloxacin. Toxic epidermal necrolysis suspected due to nimesulide Background: Nimesulide is a new nonsteroidal anti-inflammatory drug with antipyretic and analgesic properties. Pruritus and rash are the most common cutaneous side effects associated with nimesulide. Toxic epidermal necrolysis induced by drugs is a rare, potentially lethal syndrome characterized by the sudden onset of widespread blistering of the skin. To the best of our knowledge there have been no previous reports of toxic epidermal necrolysis induced by the ingestion of nimesulide in the medical literature. Case: A 68 years old woman, Buginesse. Previous history she ever had TEN six years ago probable caused by paracetamol and metamphiron. The diagnosis was established based on anamnesis and clinical manifestations. The lesions consisted of generalized erythema, vesicles, bullae, Nikolsky sign was positive, erosions, and epidermolisis more than 30%. Treatment was parenteral infusion, dexamethasone intravenous 10 mg, cimetidine 1 ampule, ciprofloxacin injection per drip and topically with silver sulfadiazine and framycetin sulfate for erosion and denuded area. Discussion: This case was diagnosis as toxic epidermal necrolysis based on anamnesis and clinical manifestations. The most frequent cause is drugs, 80% of cases have strong association with spesific medication. The probable cause was nimesulide. Clinical manifestations toxic epidermal necrolysis are extensive epidermal necrosis, and exfoliation of the skin. There is almost no normal skin seen and in this form over 80% on the skin surface is involved. Day 10th of hospitalization death ensued from sepsis and multiple organ failure. Conclusion: Toxic epidermal necrolysis is presumed to be a spesific immune response, a recurrent exposure to the trigger should induce a more brisk and intense (and in this case fatal) episode Multiple organ failure are the usual Stevens Johnson Syndrome in Bandar Lampung, Indonesia M. Syafei Hamzah. Dr. Abdul Moeloek Hospital, Dermatovenereology Department, Bandar Lampung, Indonesia. Background: Stevens Johnson Syndrome (SJS) is severe form of erythema multiforme with related mucocutaneus disorder, often with severe constitutional symptoms and associated high rate of mortaliy and morbidity. Subject: To find out the incidence, etiology, treatment, lenght of stay and complication. Methods: A 3 years retrosprctive study (January 1, 2004 through December 31, 2006 of patients admitted to dermatology department of Dr. Abdul Moeloek Hospital Lampung. Results: Among 24 cases reviewed there were 11 (45,8%) male and 13 (54,2%) female. The youngest patient was a 10 years old girl and the oldest one a 54 years old female. We assume that the caused were antibiotic (penicillin derivate) 10 (41,6%), analgesic/antipyretic 8 (33,3%), anticonvulsant (carbamazepin) 3 (12,5%), and 3 (12,5%) patient unknown. There was a variety in the length of stay of hospitalization from 1 until 27 days with an average of 7,2 days. Systemic corticosteroid was the of choice against fatality, 19 (79,2%) recovered, 5 (20,8%) patient died, the cause of death were 3 (60%) broncopneumonia, 1 (20 %) septicemia and 1 (20 %) gastrointestinal bleeding. Conclusion: The incidence of Steven Johnson Syndrome in Dr. Abdul Moeloek Hospital was found female is bigger than male, the highest group of age was 24Y44 (54,1%) and the drugs most commonly involved were antibiotics (41,6 %) followed by analgesic/antipyretic (33,3%), broncopnemoni was the mostly cause of death (60%). Stevens Johnson syndrome suspected due to sulfasalazine Eny Susilowati 1 , Faridha S. Ilyas 1 , Mahmud Ghaznawie 2 , Anni Adriani 1 , and Irma Helina Amiruddin 1 . 1 Medical Faculty Hasanuddin University / Dr Wahidin Sudirohusodo General Hospital, Dermato-Venereology, Makassar, Indonesia; 2 Medical Faculty Hasanuddin University, Pathology Anatomy, Makassar, Indonesia. Background: Stevens Johnson Syndrome (SJS) is a diffuse, severe, mucocutaneous eruption involving two or more mucosal surfaces with or without visceral involvement most often induced by drugs and occasionally elicited by infections. Sulfasalazine is a sulfa drug. The metabolite of sulfasalazine may trigger an immune-response leading to cutaneous adverse drug reactions. Case: A 29 years old woman presented symptoms of skin, mucous membrane and eye lesions. She had taken sulfasalazine for several days before the eruptions. Physical examination revealed erythema, and blisters affected on the face, neck, arms, upper trunk, erosions of the lips, and conjunctivitis. Histopathology examination of skin lesion showed Toxic Epidermal Necrolysis (TEN) appearance. The patient was treated with ringer lactate, dexamethasone 5 mg per 6 hours, gentamycin 80 mg per 12 hours intravenously, and topical steroids. After being hospitalized for 14 days, the patient cured without complications. Discussion: This case is diagnosed as SJS based on anamnesis and clinical examinations, confirmed by histopathology finding subepidermal bullae with necrosis of entire epidermis and inflammatory infiltrate, according to TEN appearances. This case may be a degree of overlap as forms of SJS can evolve into TEN. Sulfasalazine is a sulfa drug, used primarily as an anti-inflammatory agent. SJS due to sulfasalazine is rare. Conclusion: SJS due to sulfasalazine is rare. SJS is a life threatening cutaneous adverse drug reaction and can evolve into TEN. Early appropriate treatment may produce an excellent prognosis in SJS. Key words: disparity SSJ-TEN, sulfonamide, corticosteroid 865 Analysis of Stevens-Johnson syndrome and toxic epidermal necrolysis in Japan Zenro Ikezawa, Yumiko Yamane, Yuko Ikezawa, Yuki Shigehira, Chieko Watanabe, and Michiko Aihara. Yokohama City University Graduate School of Medicine, Enviromental Immuno-Dermatology, Yokohama, Japan. Background: (B Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are the representative severe cutaneous adverse drug reactions with high mortality, in which the treatnent has not been established. Methods: To present the current clinical characteristics and treatments of SJS and TEN in Japan, we retrospectively analyzed the 43 cases of SJS (15 males and 28 females, aged 44.8 in average) and 54 cases of TEN (27 males and 27 females, aged 45.5 in average) from reports of SJS and TEN published in medical journals from 2000 to 2005. Results: Thirty cases of SJS (69.8%) and all cases of TEN were caused by drugs, such as nonsteroidal anti-inflammation drugs (NSAIDs), antibiotics, and anticonvulsant drugs. As the organ involvement, hepatitis was most common in both of SJS and TEN. Renal dysfunction and respiratory disorder were also involved in some cases. Major complication was sepsis. As the treatment, corticosteroid was administered systemically in most cases, and 26 cases (60.5%) of SJS and 32 cases (59.3%) of TEN were treated with corticosteroids alone. Combination of plasmapheresis and/or immunoglobulin therapy with corticosteroid therapy was performed in some cases. The mortality rate of patient with SJS was 2.3%, and with TEN was 7.4%. The mortality in TEN decreased remarkably as compared to 21.6% (58/269) in the previous 17 years from1981 to 1997. The serious patients with TEN were performed plasmapheresis and high-dose immunoglobulin therapy in combination with steroid therapy and they were alive. Discussion: These results suggest that the combination of the abovementioned 3 treatments might be useful in the serious patients. More experiences is needed to confirm the effect. Adverse effects of DPT immunization in children up to 6 years of age in Tabriz/Iran Background: Vaccines, once hailed as "magic bullets" against disease, sometimes fly astray. In fact, shots of common vaccine DPT (diphtheriapertussis-tetanus may in rare cases trigger serious health problems. In reviewing existing data, the several studies found a link between the pertussis component of the DPT vaccine and anaphylaxis, a life-threatening allergic reaction. Vaccines have been highly effective at eliminating or significantly decreasing the incidence of many once-common diseases. However, the public is now focusing on the potential adverse effects of vaccines. The aim of this study was to identify the adverse effects of DPT immunization in children up to 6 years of age in Tabriz/Iran. Methods: A descriptive cross-sectional survey was conducted in the city of Tabriz in North-West Iran. Cases were defined as those notified with Adverse effects of DPT immunization over a 6 months period (between September 23, 2006 -to March 20, 2007 in Tabriz health centers. All of the children aged 2 months up to 6 years who recived DPT vaccine in our selected health centers, were carefuly checked up to 72 hours following immunization. A questionnaire was used to elicit information related to Adverse effects of DPT immunization in children. The data was anayzed by software of SPSS/ Win. Results: The findings indicated that fever, limb swelling, unusual crying, irritability, vomiting, erythema were 47.8%, 42%, 37%, 25%, 13%, 11.8% respectively. Conclusion: As immunizations are usually given to healthy people for prevention of serious diseases, the risks for known adverse reactions to the vaccine must be weighed against the potential benefits. Nurses and health providers should be prepared to answer patients' or parents' parents questions about common as well as alleged vaccine-related adverse events and must utilize the Vaccine Information Statements for vaccines covered under the National Vaccine Injury Compensation Program. Also health care providers should continue to counsel parentss about the benefits of the vaccine and recommend vaccination for clients. Key Words: Adverse effects, DPT, Immunization, Children up to 6 years. The anticonvulsant hypersensitivity syndrome Souhad al Faqih. Sedac, Allergy, Khartoum, Sudan. Background: The anticonvulsant hypersensitivity syndrome is a potentially fatal multisystemic reaction to anticonvulsant medications. Objective: The purpose of this study was to investigate the clinical characteristics of anticonvulsant hypersensitivity syndrome. Results: A total 32 subjects, aged from 6 to 72 years, diagnosed as having anticonvulsant hypersensitivity syndrome based on clinical and histopathological findings, were included in the study. In 22 of the 32 cases, the anticonvulsants had been administered prophylactically after craniotomy and in 10 cases for epilepsy. When the cases were assessed for skin lesions, maculopapular eruption was registered in 22, Stevens-Johnson syndrome in five, and toxic epidermal necrolysis (TEN) in five. Treatment included suspension of the offending drug and then, except for the cases with toxic epidermal necrolysis, administration of corticosteroids. The 22 cases that required anticonvulsant therapy were treated with valproic acid. In all cases, we observed rapid clinical improvement corroborated by laboratory findings. Conclusion: It is essential that due importance be given to the development of an eruption in individuals to whom anticonvulsants are administered after craniotomy because anticonvulsant hypersensitivity syndrome is likely to be life-threatening. Drug allergic reactions related to the vitally important drugs -medical tactics Gulnar Batpenova 1 , Tair Nurpeissov 2 , and Gauhar Kozhakeeva 2 . 1 Kazakh State Medical Academy, Dermatovenerology Department, Astana, Kazakhstan; 2 Institute of Internal Diseases, Republican Allergological Center, Almaty, Kazakhstan. Background: Generally accepted practice of treatment of DA is a primary cancellation of Bguilty^drugs. At the same time in some cases, we face with such events, when cancellation of the given drug can bring to unavoidable consequences. Therefore, there is a dilemma between the danger of cancellation syndrome and the development of heavy manifestations of the drug allergy. Aim: To describe the running tactic of patient with a diabetes mellitus with the concomitant polyvalent DA related to a number of drugs. Methods and Results: A patient 62 years old is continuously receiving human insulins. This patient has in anamnesis such episodes of allergic reactions related to a number of drugs of different pharmaceutical groups. The patient got a heavy allergic dermatitis to an Isosorbide in a dripping form. The cancellation of this drug and the prescription of antihistamines didn't improve the situation. The patient examination with analyzer UniCap-100 brings to light an existence of specific IgE c73 substances (Insulin Human). The abolition of this drug was impossible due to the danger of decompensation of the diabetes. Tests in vivo were impossible in consequence of the activity of cutaneous process. That's why the client passed the Damage Measure of Neutrophils Test (DMNT) in the modified scheme. The hypersensibility related to a number of drugs was displayed. From such types of insulin that obtained negative results Insulin NPH and Insulin Rapid were administrated. Parallel to this, the patient was prescribed to take a intravenous Prednisolone (120 mg/day for 5 days), after that, the treatment was transferred to steroids per os in the dose of 10 mg/day for one month with the gradual dose decrease and the following cancellation. Glycemic profile had normalized and effects of heavy allergic dermatitis had regressed completely. Conclusion: In the case of the appearance of the DA on the vitally important drugs, the cancellation of the given drug is contraindicated. It's because, in the case of further necessity of emergency infusion, the risk of anaphylactic shock is increasing. The DMNT (sensitivity is about 95%, specificity-about 80%) in the modified scheme is the effective method due to the impossibility of conduction of more trustworthy tests in vitro and in vivo. Monitoring showed the effectiveness and safety of the prednisolone course for the treatment of DA. Connubial systemic cutaneous adverse drug reaction Corinne Vernassiere, J.F. Cuny, J.L. Schmutz, and Annick Barbaud. Fournier Hospital, Dermatology Department, Nancy, France. Observation: A 86 y.o. women has had for 24 hours, an itchy maculopapular exanthema (MPR) on the trunk and the arms. The daily treatment had not been modified, except concerning the switch from MOPRAL\ (omeprazole) to INIPOMP\ (pantoprazole) 48 hours before. The biopsy sample was compatible with a drug allergy. There was an eosinophilia (1296/mm3). In her history, 11 years before, cutaneous tests were performed for calcium channel blockers and penicillin sensitizations. Patch tests were positive for diltiazem. The chronology of drug intake was done with his husband. Her treatment was prepared by his husband because she had visual deficiency. A few days after the MPR`s onset a member to the family reported that 48 hours before the MPR the patient had taken by error the drugs of her husband put beside those of its wife: that were: trimetazidine, diltiazem, atenolol, lysine acetylsalicylate, domperidone and oxybutynine. The rash vanished within 3 weeks. Discussion: This observation stresses the importance of the interrogation in the assumption of a CADR in determining the chronology of drug intake. An additional difficulty is to obtain this information from an old person (lack of memory). The interrogation of the other family members, the GP and the pharmacist can be crucial. Masked drug exposure have been reported by kissing, but not by the misadministration of drug intended for a spouse. The risk could become significant because of the ageing of the population. Conclusion: 1st case masked drug allergy due to diltiazem taken by mistake (intended for a spouse). Wahjuni Husain 1 , Andi Muhammad Adam 1 , Mahmud Ghaznawie 2 , and Faridha S. Ilyas 1 . 1 Medical Faculty Hasanuddin University / Dr. Wahidin Medical Faculty Hasanuddin University, Pathology, Makassar, Indonesia. Background: Drug hypersensitivity syndrome (DHS) is an idiosyncratic and life-threatening adverse drug reaction that begins acutely in the first 2 months after initiation of drug and is characterized by fever, skin rash, facial edema, or an exfoliative dermatitis, lymphadenopathy, hematological abnormalities (eosinophilia) and multi-organ involvement. In this case, there were no other systemic organ involvement. Case: A 26-years-old woman presented with fever, maculopapular rash, facial edema and lymph nodes enlarged 2 weeks after taking cefadroxil and mefenamic acid after fine needle aspiration biopsy (FNAB) for suspect thyroid infection. After taking mebhydrolin napadysilate the rash become extended as an exfoliative dermatitis, facial edema, and lymphadenopathy. Laboratory examination including leucocytosis and eosinophilia. There were no other biologic abnormalities including normal liver function test, chest radiograph and cardiac echography. Histology examination showed hyperkeratosis, mild spongiosis, vacuolar degeneration of basal cells with infiltration of lymphocytes, neutrophil and eosinophil. Discussion: The patient fulfilled the clinical criteria with her presentation of fever, generalized maculopapular rash, facial edema, lymphadenopathy and eosinophilia. The pathogenesis of DHS is not fully understood, including genetic background, altered drug metabolism, and virus infection. The suspect drug was mebhydrolin napadysilate. The treatment was dexamethason intravenously started with 15 mg tid. Topical steroid and emollient were given also. After 3 weeks, dexamethasone intravenously was substitute with methyl prednisolone 24 mg orally a day and tapered. The rash progressively improved. Conclusion: Drug hypersensitivity syndrome is a severe skin reaction with systemic manifestations. Internal organ involvement can be asymptomatic. The suspect drug was mebhydrolin napadysilate. The treatment of dexamethasone intravenously continued with methylprednisolone oral and withdrawal of possible offending drug gave a good result. Background: Adverse reactions to pharmaceutical and diagnostic products constitute a major hazard in the practice of medicine. Multiple risk factors have been shown to modify the clinical expression of immunologic drug reactions. These can be associated with the drug itself, with the disease state in which the drug is used(chronic disease and infections), or the individual patient receiving the drug. Similarly, some allergic drug reactions are much more likely to occur if the drug is administered to patients with particular disease backgrounds, as when aminopenicillins are given to patients with atypical or abnormal lymphocytes or sulfonamides to patients with AIDS. Case Report: We represent a 63-year-old man with prolonged history (more than one year)of intermittent headache and use of NSAIDS. He had recurrent history of anaphylaxis and urticaria after getting NSAIDS at first, and with the other analgesics and also antihistamines there after. His laboratory evaluation, includind cell blood count and differentials and complement assay was in normal limits, He consulted with a neurologist, and brain CT-scan was done. It showed CNS tumor. In literature review it was found a few chronic diseses such as cyctic fibrosis or cryogobulinemia (or also immune deficiencies) can cause drug allergy but there was no association between multiple drug allergy and malignancies especially CNS tumors. Background: Dapsone Hypersensitivity Syndrome (DHS) is a rare well recognized serious adverse effect characterized by fever, skin rashes, generalized lymphadenopathy, hepatitis and hepato-splenomegaly. Hipersensitivity reaction occur during first 6 weeks, but can occur after 6 months. Objective: To find out the total cases for 2 years of dapson hypersensitivity syndrome in the wahidin sudirohusodo general hospital, Makassar, Indonesia. Methods: A retrospective study was carried out taken from medical record and registration book. (January 2005 -December 2006 . Results: Five patients with dapson hypersensitivity syndrome were reported. All patients received dapsone as a part of MDT for leprosy. Fever, jaundice, skin lesion were detected in 5 cases, but 1 case appearance like morbiliform. Lymphadenopathy (2 cases), splenomegaly and acut psychosis (1 case). Laboratory data on admission included : low hemoglobin, leucocytosis, elevated level of serum liver enzymes, erythrocyte sedimentation, raised bilirubin and hypoproteinemia. The interval between start of dapsone therapy and appearance of symptoms varied from 2Y7 weeks. But one case the clinical manifestation occur 8 months after consumption of Multi drug treatment(MDT). All cases were treated with corticosteroids after withdrawing dapsone, MDT was continued by replacing dapsone with minocyclin. Background: Lemierre's syndrome is an acute oropharyngeal infection with suppuration of the lateral pharyngeal space, bacteremia and septic thrombophlebitis of the internal jugular vein, causing metastatic infection. It is a rare disease with an estimated incidence of 1 case per million. Methods: We report a patient diagnosed with Lemierre's syndrome who presented with a rash after 2 weeks of piperacillin-tazobactam, clindamycin and phenytoin. Results: This 27 year old male with no significant past medical illness presented with a one week history of a dento-alveolar abscess of the right molar, fever, trismus, dysphagia with erythema and swelling of the neck. Work up showed a retropharyngeal extension of the dento-alveolar abscess, pneumonia, pneumomediastinum and necrotizing mediastinitis. HIV enzyme immunoassay was negative. The patient underwent cervical mediastinotomy with subxiphoid drainage and was given piperacillin-tazobactam 4.5g IV q8 and clindamycin 300mg IV q6. During the course of his hospital stay, the patient went into arrest, had post-gliottic seizures and was started on phenytoin 100mg cap q8. On the 2nd week of piperacillin-tazobactam and clindamycin and on the 8th day of phenytoin, the patient presented with erythematous, pruritic maculopapular rashes initially on the upper extremities later on becoming generalized. Antibiotics were shifted to meropenem and phenytoin was shifted to levetiracetam. The patient received diphenhydramine, ranitidine and prednisone, which resolved the rashes. He also completed 6 weeks of meropenem and was discharged improved. Conclusion: This is the first reported case of drug hypersensitivity in a patient diagnosed with Lemierre`s syndrome. The importance of discontinuing the offending drugs and shifting to structurally unrelated drugs is emphasized. Hydrochlothiazide-induced lichenoid eruption: Apropos of 2 cases Karim Aouam 1 , Hichem BelHadj Ali 2 , Amel Chaabane 1 , Montassar Amri 2 , Naceur Boughattas 1 , and Jameleddine Zili 2 . 1 Faculty of Medicine, Pharmacology Department, Monastir, Tunisia; 2 Monastir's Hospital, Dermatology Department, Monastir, Tunisia. Background: The aetiology of lichen planus is unknown. Stress, autoimmunity and infection have been suggested as possible causes. Many drugs can provoke eruptions that clinically and histologically are similar to lichen. We describe in two patients a probable association between treatment with irbesartan-hydrochlorothiazide and lichenoid eruption. Case Reports: Case 1: An 80-year-old woman presented to the dermatology clinic with a 4month history of acute onset of violaceus papules which started on her arms and spread to her trunk, back and legs. She had been taking amiodarone for auricular fibrillation for 15 months and irbesartan-hydrochlorothiazide for hypertension for 8 months. On physical exam, the patient had accurate violaceus and scaly plaques on the trunk, back and limbs. There were no ocular, buccal or genital lesions and she had no systemic symptoms. Blood count and biochemical profile were normal. Skin biopsy of involved skin showed typical features of lichen planus. Treatment with desonide and cetirizine provided no relief. Irbesrtan-hydrochlothiazide was substituted by irbesartan and the skin eruption and prutitis had resolved within two weeks. Case 2: A 56-year-old man presented with acute onset of a pruritic rash which stated on his trunk and spread to his whole body. His past medical history included hypertension and Sharp syndrome for which he was taking irbesartan-hydrochlorothiazide and methotrexate. Methotrexate and irbesartan-hydrochlorothiazide were started approximately 2 years and 8 months before skin eruption respectively. On physical exam, the patient had accurate violaceus and scaly plaques on the trunke, back and limbs. There were bilateral shallow erosions of the buccal mucosa surrounded by a network of lacy white striae. Blood count and biochemical profile were normal. Skin biopsy of involved skin showed typical features of lichen planus. Treatment with desonide and cetirizine provided no relief. Irbesrtan-hydrochlothiazide was substituted by irbesartan and the skin eruption and prutitis had resolved within two weeks. Conclusion: These observations illustrate a rare association between lichen and hydrochlorothiazide (On the basis of the Naranjo algorithm, it is probable that the systemic reaction was due to hydrochlorthiazide therapy). Clinicians should be aware of this potential adverse effect and recommend with-holding the drug intake when temporal relation is evocative. Prevalence of allergic contact sensitization among hostel students In Ajman University of United Arab Emirates (UAE) Tarik Al Zubaidy 1 , Taki Almosawi 2 , and Zeena Sameer 1 . 1 Ajman University, Faculty of Pharmacy and Health Sciences, Ajman, United Arab Emirates; 2 Al-Mafraq Hospital, United Arab Emirates. Background: It is estimated that a round 20-30% of people all over the world suffer from allergy. Contact with certain materials (allergens) is supposed to cause allergic skin sensitization culminating in development of contact dermatitis in some individuals. Some people are unaware of their sensitization as it shows no or mild symptoms. The prevalence of specific allergens varies by geographic area, population, age and sex. Objective: The present study aimed at determining the relative frequency of positive reactions to different sensitizers among hostel female students in Ajman University-UAE. Materials and Methods: Sampling approach was simple and descriptive. 46 female hostel students aged between 19Y26 years were tested using the Universal standard patch test series (TRUE test) containing 23 common allergen sensitizers and a negative control. Cross sectional questionnaire was distributed for the same students and those with chronic diseases or pregnancy were excluded. Results: The results were analyzed by SPSS. In contrast to previous European reports which suggest that nickel sensitization is the most common among females, this study shows that thimerosal is the common sensitizer among female hostel university students (32.6%), followed by nickel 15.2% and colophony 8.69%. Conclusion: Thimerosal is present in contact lens solutions and cosmetics and most positive students who were unaware of their sensitization have used contact lenses for cosmetic purposes. This study should raise the alarm in using inert materials in contact lenses industry or at least warn females from the consequence of sensitization. Background: Sesame and perilla have been a Korean traditional crop and mainly consumed as seasonings or seed oil. However, sesame food allergy is an increasingly common problem in many countries. This study aimed at comparing protein pattern of sesame with that of perilla and analyzing the cross reactivity between sesame and perilla. Methods: Seeds were placed in 1:1 ethyl ether and extracted with slow stirring at 4-C for 2days in PBS (pH 7.4). To analyze protein patterns and specific IgE to sesame or perilla, crude extracts were separated by SDS-PAGE and then performed with Western blotting using sera diagnosed as allergic to sesame or perilla. Sesame extract with various concentrations is pre-incubated with pooled sera before inhibition test, and then those sera were added to the ELISA plate pre-coated with perilla extract. Results: 1. Both sesame and perilla had the higher protein concentration and contained protein bands of 6 Y 82.2 kDa. However, after cooked, the proteins with high molecular weight disappeared and only 6 Y 14 kDa protein bands existed weakly. 2. The IgE binding bands were also detected at 6 Y 14 kDa in sesame and perilla by Western blot analysis. 3. In inhibition test, sesame showed low inhibition rate (G45%) against perilla by inhibition ELISA test. Conclusion: Sesame and perilla showed different protein pattern and low cross reactivity each other. It suggests that perilla can be the replacement of sesame in consumption. However, because they have the possibility of food allergy, the further study about their major allergens is needed. Biochemical and allergenic properties of natural and genetically modified soybeans Soybeans are an important source of protein in human`s dietary intake today. Some soybean proteins may be allergens and cause allergic reactions. In this connection, the aim of the present study was to investigate the biochemical and allergenic properties of extracts from natural and genetically modified (GM) soybeans. The protein extracts were obtained from whole natural and GM soybeans, their capsules and cores and analyzed in SDS-PAGE, western blotting and ELISA. The SDS-PAGE analysis has revealed similar results of protein composition of allergenic extracts of core and whole natural and GM soybeans. The main protein band of whole soybeans and soybean core have low a molecular weight (m.w.) from 8,5 kDa to 20 kDa. The protein band with m.w. of 20 kDa is largest and conform a protein group of Kunitz trypsin inhibitor (Hor.v1, CMb, BDR). The protein fraction of capsule of soybeans with m.w. 14 kDa conforming soybean profilin (Gly m 3) predominate natural soy, whereas that with m.w. 34 kDa conforming vacuolar protein (Gly m Bd 30 k) predominate GM soy. We investigated 220 sera of patient`s blood different age with atopic dermatitis and food allergy using ELISA. A frequency of revealed IgE-Abs to soy was increasing from 20 % in infant patients to 57,8% in adolescent patients. The increased levels of IgE-and IgG-Abs to capsule of natural soybeans were determined most often. We revealed IgG-Abs to wide protein spectrum of whole natural and GM soybeans using western blotting, where the largest protein fraction had m.w. of 30 kDa. The soybean capsule is known to contain proteins, which are homologous to the major allergens of birch pollen (Bet v1); with this in view high frequency of revealed IgE-Abs to the capsule of natural soybeans is fully explained in the context of crossreactive antigens. While analyzing the blood serum of the patients with hypersensitivity to soy we revealed increased levels of IgE-Abs to birch pollen (81 % of cases) with high level of IgE-Abs found in 25% of patients. Therefore it was revealed that extracts of natural and GM soybeans contain proteins with identified allergic properties, which may cause the development of allergic diseases related to IgE-Abs and soy enteropathy related to IgG-Abs. Moreover, due to a pronounced homology with allergens of birch pollen, foodstuff containing soybeans may strengthen sensitization and intensify allergic reactions in Pollinosis patients. Different proteomic approaches to the identification of wheat flour allergens Background: Food antigens penetrating into the gastrointestinal tract do not generally cause clinical symptoms, because most individuals acquire tolerance. However, some food components are recognized as the cause of adverse reactions in susceptible individuals. Wheat belongs among six major foods, which are responsible for approximately 90% of food allergic reactions in children. Although cereals are essential in the daily nutrition, only a few proteins have been identified as cereal allergens on the molecular level. Methods: To characterize wheat flour allergens, we prepared salt extract from seven of the most frequently used wheat cultivars and compared them by SDS PAGE. Proteins from the selected cultivar Sulamit were separated by 2dimensional gel electrophoresis , analyzed by immunoblotting and IgEreactive proteins were identified by mass spectrometry (MALDI-TOF, QTOF and LCQDECA nLC-MS/MS ion trap) and tested for the ability to activate patient`s basophils (expression of CD63 and CD203c markers was measured by flow cytometry). We developed ELISA to quantify patient`s IgE Abs reactive with defined allergens. Wheat extract was also treated with pepsin. Results: Patient sera can be divided into three groups: IgE Abs of the first one recognizes wheat components G30kDa, the second 930kDa and the third one reacts with components ranging from 6-170kDa. The pattern of wheat components recognized by IgE Abs was significantly influenced by conditions of protein separation and treatment with digestive enzymes. Using SDS PAGE, 2-DE, immunobloting and mass spectrometry we identified 19 potential wheat allergens. Employing newly developed ELISA, levels of IgE Abs against proteins from the Sulamit extract and one of the dominant wheat IgE binding component, "-amylase inhibitors type 1 and 3, were quantified and shown to be significantly elevated in sera of allergic patients compared to those of healthy controls. Furthermore, the salt extract from the cultivar Sulamit as well as "-amylase inhibitors were shown to by able induce significant activation of basophils of allergic patients. Conclusion: This study characterized the spectrum of wheat components recognized by IgE Abs of allergic patients, analyzed the impact of proteins separation and digestion on immunogenicity of wheat allergens, compared the sensitivity and specificity of newly developed specific IgE ELISA assay and in vitro basophils activation test using identified allergens. Heat-induced denaturation, aggregation and changes of immunoreactivity of milk proteins Background: Milk for liquid consumption or for processing requires an initial heat treatment to control the growth of microorganisms. However, heat treatment of milk causes the heat denaturable whey proteins to aggregate with .-casein (.-CN) via thiol-disulfide bond interchange reactions and other changes. Influence of the soluble macroaggregates presence and other heatinduced changes on immunogenicity of the commercial milks is still a controversial issue. The aim of this study was to contribute to increase knowledge in this area. Methods: The kinetics of heat-induced precipitation of the main whey proteins $-lactoglobulin ("-LG) and !-lactalbumin (!-LA) were examined in water solutions of individual proteins, in fresh milk, whey, and model mixtures of the both whey proteins with .-CN at 95-C during 30 min. At the same time, the kinetics of formation of heat-induced aggregates of the main whey proteins with .-CN was investigated in fresh milk, whey, and the model mixtures using GP-HPLC method. Dot blot assays using sera of 19 allergic patiens containing specific IgE antibodies against milk proteins were employed to assess the immunochemical reactivity of the milk proteins containing samples before or after heat treatment. Immunoreactivities of 6 commercial milk samples (pasteurized, UHT and fermented) with different content of the soluble macroaggregates were also assessed by commercial ELISA tests with monoclonal antibodies against individual whey proteins and using the sera of allergic patiens. Results: The decrease of the main whey proteins $-LG and !-LA was significantly higher in milk or whey than in solutions of the individual proteins. The experiments with the model protein mixtures showed that presence .-CN is necessary to form the aggregation complexes of the whey proteins at 95-C. The kinetics of formation of the heat-induced aggregates in milk and model protein mixtures was complex with two different maxima during 30 min time interval. Conclusion: Surprisingly, we have not observed significant changes of the immunochemical reactivities of the milk proteins samples after the heatinduced aggregation under the conditions of the dot blot assays. However, the experiments will be continued to confirm the results. Characterization of the putative allergens in nine Chinese adults with food allergy to limpet Bernard Thong 1 , Fook-Thim Chew 2 , Tan-Ching Ong 2 , Kang-Ning Wong 2 , Yew-Kuang Cheng 1 , Khai-Pang Leong 1 , Chwee-Ying Tang 1 , and Hiok-Hee Chng 1 . 1 Tan Tock Seng Hospital, Singapore, Rheumatology, Allergy and Immunology, Singapore, Singapore; 2 National University of Singapore, Biological Sciences, Singapore, Singapore. Background: Limpet, a gastropod mollusc, is the second most common cause of IgE-mediated food allergy among adults attending our Clinical Immunology/Allergy clinic. The aim of this study is to characterize the putative allergen(s) and ascertain possible cross-reactivity with common inhalant and food allergens in our patients with limpet allergy. Methods: Consecutive adults presenting from 1 July 1994 to 31 April 2002 with immediate hypersensitivity following ingestion of limpet were studied. Sera stored at j70-C were assayed for specific IgE to common inhalant and food allergens. IgE western blotting and tandem mass spectrometric analysis were performed to identify the allergens. ELISA inhibition assays were carried out to compare the allergenicity of limpets with other gastropods, as well as to recombinant tropomyosin and arginine kinase allergens from dust mites and cockroaches. Results: There were 9 patients (6 males) and all were ethnically Chinese. Their mean age was 38 T 17 (19Y60) years. Eight patients had previously ingested limpet, with allergic reactions occurring during a median of 2 occasions. Six patients had history of asthma, allergic rhinitis and/or atopic eczema. Seven patients (2 refused skin testing) tested positive on prick-prick test to canned limpet, 3 were also positive to abalone, and 2 to pacific clam. On invitro specific IgE assay, 77.8% were positive to shellfish mix, 77.8% to Dermatophagoides pteronyssinus, 55.6% to Dermatophagoides farinae, 44.4% to Blomia tropicalis, and 33.3% to cockroach mix. Western blot and protein mass spectrometric analysis showed that the putative limpet allergens were a 37kD tropomyosin and a 40kD arginine kinase. ELISA inhibition studies using recombinant dust mite and cockroach tropomyosin as well as arginine kinase showed moderate cross reactivity between these allergens and the allergens from limpet. There was no significant inhibition of binding by abalone and clam. Conclusion: The putative limpet allergens in our patients are a 37kD tropomyosin and a 40kD arginine kinase, both of which are major allergens in other gastropods (abalone and pacific clam), crustaceans, American cockroach and house dust mite. Prior sensitisation to house dust mite and cockroach in our patients could have played a role in limpet food allergy. Surprisingly there was limited cross-reactivity with abalone and pacific clams, two closely related gastropod molluscs. Cross reactivity among soy, penaut and lupine Marianne Polgár 1 , É va Gelencsér 2 , and Gyöngyi Hajós 2 . 1 Heim Pál-Madarász Children Hospital, Gastroenterology, Budapest, Hungary; 2 Central Food Research Institute, Biotechnology, Budapest, Hungary. Background: Soy protein is a common dietary protein, the intake is about 1Y2 g day in Europe among adults according a multicentre study. Soy based infant formulae are often used in infancy and early childhood in cow`s milk allergy and in healthy children instead of cows milk, too. The prevalence of allergic reactions (gastrointestinal, skin, and pulmonary) elicited by soy protein is around 6% in atopic children. Severe or fatal reactions seem to be rare, but the rate higher in children with peanut allergy. Lupine is a newly used legume in the human diet, which theoretically can give also cross reaction with soy and peanut. Aim of the study was to determine the cross reaction among soy protein, peanut and lupine. Methods: Immunoblot technique with SDS-PAGE protein separation and ELISA method. Polyclonal antibody and IgE soy positive human sera were used to detect reactions. Results: Immunological cross reactivity was demonstrated between the major allergens of studied legume proteins. The beta-conglycinin and their oligomeric subunits in soybean have shown cross reactivity with the Ara h1 in peanut and the 7S vicillin allergens in lupine. Furthermore it was observed an immunological cross reaction among the soybean glycinin allergens and the Ara h3 in the peanut and the vicillin 11S allergens in lupine, while the soybean profilin cross reacted with Ara h5 peanut allergen. The 2S allergens of prolamin superfamily in soybean as trypsin inhibitors and lectins shown cross reactive epitopes with Ara h2, Ara h6 and Ara h7 allergens in peanut. Soybean, peanut and lupine protein extracts did not differ in their IgE reactivity with soy positive human sera. Conclusion: Cross reactivity among the major allergens of soy, peanut and lupine was proved with human soy positive sera in vitro. Results draw attention to the possibility of anaphylaxis as a result of allergic cross reaction in patients with peanut and soy allergy. On this basis, it has to emphasise the need of precise labelling for food components to avoid the anaphylaxis. Jiménez Díaz, Allergy Department, Madrid, Spain; 7 Allergy Clinic, Reumannplatz, Vienna, Austria; 8 IDI-IRCCS, Center for Clinical and Experimental Allergology, Rome, Italy; 9 Hô pitaux Universitaires, Service de Pneumologie, StraQbourg, France. Background: Milk allergy is the most common IgE-mediated food allergy in early infancy. Cow`s milk contains about 25 proteins of which eight have been identified as allergens but there are varying results regarding the frequency of IgE recognition and the allergenic activity of the individual allergens. We developed a microarray for milk proteins which allows to determine indiviual patients' IgE reactivity profiles as well as the biological activity of the diseasecausing allergens using small amounts of patients' sera. Methods: We dotted different milk components including whole milk extracts, milk fractions as well as purified and recombinant milk allergens, and recombinant and synthetic fragments of milk allergens onto microarray and determined the IgE reactivity profiles of 78 milk allergic patients. In parallel, allergenic activity of the milk components was determined using rat basophil leukaemia (RBL) cells transfected with human Fcå receptor that had been loaded with the very same sera. Results: We found that 84.6% of milk allergic patients showed IgE reactivity to whole cow's milk extract. A high number of patients had IgE antibodies to sheep's and goat's milk, 82.1 % and 67.9%, respectively. But only 30.8% of the tested patients showed IgE reactivity to mare's milk. The most frequently recognized cow's milk allergens in our study were alphalactalbumin (62.8%), alpha-casein (48.7%), beta-lactoglobulin variant A and B (48.7 and 50.0%, respectively), and beta-casein (43.6%). Lactoferrin and BSA seemed to be of lesser importance since recognition frequencies were below 5 %. The most potent cell activating component in the humanized RBL assay was cow's milk which induced a relevant degranulation in 47.4% of the patients followed by goat's and sheep's milk (35.9 and 28.2%, respectively). Among the single components the caseins showed the highest biological activity (25 to 34.6% of the patients' sera). Conclusion: Using microarray technology in combination with data from basophil release assays it is possible to test simultaneously the frequency of IgE reactivity and the allergenic activity of a multitude of allergens using just a few drops of patient's serum. This test system will be important for diagnosis and therapy of cow's milk allergy. Epitope mapping on gliadins for patient sera reacting to modified gluten Background: Wheat gluten can be modified by enzymatic or chemical treatment (wheat isolate) in order to improve its functional properties, increase solubility and thus broaden its application in food industry. However, these modifications were shown to induce IgE mediated allergies in individuals usually tolerant to wheat. Leduc et al. (2003) published the first case of anaphylaxis after ingestion of a reconstituted meat containing a wheat isolate. Laurière et al. (2006) also showed that hydrolysed wheat proteins can induce severe allergic reactions by contact or ingestion. The objective of our work was to identify epitopes detected by IgE from patients with reactions to chemically deamidated gluten (product in which some glutamine residues were modified into glutamic acid). Methods: Sera from twelve patients with anaphylaxis, urticaria or WDEIA containing IgE antibodies to laboratory or industrial deamidated gluten in ELISA or RAST were analysed by pepscan with overlapping synthetic peptides. IgE-binding epitopes were searched among entire sequences of ã and w2-gliadins with identification of critical animo acid and analysis of effect of substitutions of glutamine into glutamic acid. Results: Eight sera reacted with repetitive peptides of ã or w2-gliadin containing a common octapeptide rich in proline and glutamine. Five critical positions for IgE binding have been identified on this epitope. Substitution of one of its glutamine into glutamic acid induced its recognition by two additional sera and simultaneous substitution of two glutamine into glutamic acid increased the intensity of responses obtained for the ten positive sera. Conclusion: Patients with reactions to deamidated gluten possessed IgE antibodies directed to linear sequences of the repetitive domains of ã and w2gliadins. IgE antibodies from most of the patient sera analysed in this study bound to an immunodominant epitope of 8 residues. Deamidation of two glutamines of this epitope increased significantly its recognition by patient IgE. This epitope is similar to a B epitope evidenced for celiac patients. These results show that some industrial treatments may increase the risk of allergy development. Omega-5-gliadin allergen specific IgE antibodies are clinically useful in the diagnosis of food allergy Background: The relationship between food-specific IgE titers and the outcome of food challenges is well established for egg and milk through threshold values and probability curves. For wheat or components of wheat no such relationships have been established. Objective: To evaluate the clinical use of wheat and the wheat component omega-5-gliadin specific IgE titres in relation to the outcome of food challenge. Methods: Two hundred and forty nine children with suspected wheat intolerance, from 3 clinics in Japan, were included in the study. Age ranged from 6 months to 15.4 years (mean=3.8 years, median=3.0 years). The diagnosis of food allergy was judged through confirming symptom in food challenge tests. In relation to their visit for food challenge, allergen specific IgE antibody levels for wheat and omega-5-gliadin were measured (ImmunoCAPi, Phadia AB, Sweden). Results: One hundred and eighty five children were challenged. The most frequent symptom for a failed challenge was related to the skin, then cough and lower respiratory systems, 48%, 24% and 13%. Three children had an anaphylactic reaction. Fifty four percent of the children had one symptom, 32% showed 2 symptoms, 13% showed 3 symptoms and 1 child had 4 symptoms. The specific IgE titres for both wheat and omega-5-gliadin were higher for the children who failed the food challenge, Table 1 . In a simple logistic model both wheat and omega-5-gliadin showed a significant relationship between the probability of challenge reaction to the food and the specific IgE antibody concentrations, wheat, OR 1.41, 95%CI:1.17Y1.40 and omega-5-gliadin OR 1.97, 95%CI:1.51Y2.55. In a multivariate model using both wheat and omega-5-gliadian, omega-5-gliadin showed the strongest association with a failed challenge provocation, OR= 1.95 95%CI:1.41Y2.67. Conclusion: Our results show that omega-5-gliadin specific IgE antibody titres are related to the probability for a failed oral food challenge. Omega-5gliadin IgE in combination with wheat IgE antibodies are a good predictor of wheat challenge outcome. Oral allergy syndrome due to plant-derived foods: a clinical review of 63 patients over a period of 6 years Naoko Inomata 1 , Akiko Morita 1 , and Zenro Ikezawa 2 . 1 Yokohama City University Hospital, Department of Dermatology, Yokohama, Japan; 2 Yokohama City University School of Medicine, Department of Environmental Immuno-Dermatology, Yokohama, Japan. Background: $B!'(BThe clinical features of many patients with oral allergy syndrome (OAS) due to plant-derived foods have rarely been reported in Asian countries. Objective: We aimed to determine the causative foods of OAS due to plantderived foods based on clinical features and skin prick tests (SPTs). , and there were twice as many females as male. The most frequent causative foods were found to be apple, peach, kiwi, and melon in 13, 12, 12, and 11 patients, respectively. CAP frequency was shown to be similar to that of SPT regarding apple, whereas it was less than that of SPT regarding melon, peach, and kiwi. A significant correlation between the frequencies of SPTand CAP was found regarding apple (R=0.39, pG0.05) but not peach, kiwi, and melon. Forty-one of 63 patients with OAS (66.1%) had pollinosis and/or allergic rhinitis. In patients with OAS due to apple, the positive ratio of CAP response against alder pollen was higher than that in patients with OAS due to melon. In patients with OAS due to melon, the positive ratio of CAP responses against ragweed pollen, grass pollen, and mugwort pollen was higher than that in patients with OAS due to apple. PFS is a type of Oral Allergy Syndrome (OAS), a common cause of food allergy in adults. To date, there have been no published studies on the prevalence and characteristics of PFS in a UK cohort. This study, designed to test a diagnostic questionnaire, also provided information on the foods involved and and allergy history associated with PFS. Subjects with reported springtime hay fever completed a PFS diagnostic questionnaire (PFSDQ) and were allocated to either group 1 (has PFS), group 2 (no PFS, ?food allergy) or group 3 (no PFS or food allergy). They then completed a questionnaire (PFSPQ) on their rhinitis, concomitant asthma, medication, symptom severity, reactions to foods, and severity of food symptoms. Diagnosis of PFS was made using clinical history, prick by prick tests (PPT) to fresh foods and oral food challenge. 119 subjects completed the study (mean age 36 years, 85 female); 88 subjects (Group 1 Y 58, Group 2 Y 30) reported reactions to 73 different foods. A final diagnosis of PFS was made in 60 of the 199 subjects (50%). There were no differences in age or gender between Groups 1Y3 or for final diagnosis. Most variables in the PFSPQ did not differ between Groups 1Y3 or for final diagnosis. These included family history of atopy, childhood allergy, age of onset of rhinitis, symptom severity scores for rhinitis, the presence or absence of asthma, number of years with a food allergy or onset of food symptoms before, after or at the same time as the onset of their hay fever. Subjects most commonly reported reactions to apple (39 subjects), hazelnuts (22), almonds (21), milk (18), strawberry (16), walnuts (16), peach (14), cherry (14), Kiwi (14) and wheat (13). Subjects in Group 1 reported symptoms to significantly more foods than subjects in Group 2 (pG0.02). Apple, hazelnut, peach, almond, kiwi and cherry were the commonest foods to have reported symptoms confirmed by positive PPT. Milk and wheat had the lowest concordance with PPT; only 21% of reported reactions to milk were confirmed and 0% of reactions to wheat. These data show that 50 % of a cohort of UK subjects with reported springtime hay fever had PFS. The commonest foods to cause reactions were similar to other published studies, although kiwi appeared to be a more significant precipitant of PFS symptoms than previously reported. In this study, age, gender, allergy history, asthma and time of onset of food symptoms were not associated with a diagnosis of PFS. Usefulness of skin prick test using bifurcated needle for the diagnosis of food allergy among infantile atopic dermatitis Mika Ogata 1 , Akinori Shukuya 2 , Chizuko Sugisaki 3 , Kaori Ikematsu 4 , Takashi Komata 5 , Takanori Imai 2 , Morimitsu Tomikawa 2 , Hiroshi Tachimoto 2 , and Motohiro Ebisawa 3 . 1 National Hospital Organization Kumamoto Medical Center, Pediatrics, Kumamoto-city, Japan; 2 National Hospital Organization, Sagamihara National Hospital, Pediatrics, Sagamihara-city, Japan; 3 National Hospital Organization, Sagamihara National Hospital, Clinical Research Center for Allergy, Sagamihara-city, Japan; 4 Tokyo Jikei University School of Medicine, Pediatrics, Tokyo, Japan. Objective: We investigated the usefulness of skin prick test (SPT) for the diagnosis of egg white (EW) CAPRAST-negative egg allergy infants and followed up the EW-CAPRAST in this study. Subjects and Methods: Data of negative SPT using Bifurcated needle (BF) were analyzed from the data of 202 infantile atopic dermatitis patients, who had received SPT from January in 2001 to April in 2005. From the analysis of SPT results, the average and standard deviation of negative value were obtained to set the range of positive SPT result. Among 202 cases, 89 suspected-egg allergy infants with negative IgE CAPRAST against EW at the time of first visit were recruited to this study to examine the usefulness of SPT. Positive conversion of EW-CAPRAST was checked in 78 cases (65: egg allergy+, 13: egg allergy-) who had been followed up in our outpatient clinic. Results: Range of negative SPT control value (mean+ 2SD) using BF among infants could be set as less than 2 mm for wheal and 5 mm for erythema. Among 89 suspected-egg allergy infants with negative EW-CAPRAST, 72 infants (80.9%) were diagnosed as egg allergy by the combination of elimination and provocation test, interestingly 39 infants (54.2%) showed positive SPT results in 72 egg allergies. In the follow up study of 78 negative EW-CAPRAST cases, 47 EW-CAPRAST out of 65 egg-allergy cases turned positive later infantile period (mean EW-CAPRAST: 9.6+/j16.7 Ua/ml at 9.9+/j5.6 months old). EW-CAPRAST of 7 cases in 13 non-egg allergies also turned positive in the follow up, however EW-CAPRAST titer was relatively lower compared to that of egg allergies (1.1+/j1.5 Ua/ml at 13.3+/j2.6 months old). Conclusion: We experienced fairly number of atopic infants with negative EW-CAPRAST at the first outpatient visit, who were later diagnosed as egg allergy. In about half of these cases, SPT was extremely useful for the diagnosis. In the follow up of egg-allergy infants, three quarter of EW-CAPRAST tuned positive around 10 months old. EW-CAPRAST of atopic infants without egg allergy also turned transiently and slightly positive. In the conclusions, SPT seemed to be more useful than EW-CAPRAST for the diagnosis of egg allergy in early infantile period, however provocation test may be required for the definitive diagnosis in suspected-egg allergy infants without any proof of egg-sensitization. Optical near-field immunochip biosensor for rapid allergen detection and clinical diagnosis Background: An optical immunochip biosensor has been developed as a rapid method for allergen detection and in vitro immunosensing. The optical near-field phenomenon underlying the setup design of the sensor is called resonance-enhanced absorption (REA) utilizing gold nanoparticles as signal transducers in a sensitive interferometry-setup. Combined with visual readout that eliminates complex detection instrumentation, this assay system excellently meets the requirements of a rapid method for allergen detection and provides opportunities for the development of a miniaturised multiarray-sensor with improved sensitivity for allergy testing in clinical diagnosis. Methods: REA is observed when light-absorbing metal clusters are positioned at a nanodistance from a highly reflective mirror and illuminated with white light of the visible and near-infrared spectrum from the particle side. As a direct detection method based on distance-dependent light interference between the mirror and the resonantly excited metal clusters, a REA based capture assay has the power to link affinity binding with three-dimensional structural information. Results and Discussion: The immunochip biosensor showed concentrationdependent colour development by capturing biomolecule-functionalised gold nanoparticles on allergen or immunoglobulin coated chips. Semi-quantitative immunochemical responses are directly visible to the Fnaked`eye of the analyst with sensitivity comparable to common immunoassay procedures in microtitration plate format (ELISA). Unspecific background signals were minimized to the absolute target specificity of the antibodies and antisera used in the bioassay. We have applied this method to the detection of the egg white allergens ovalbumin and ovomucoid in complex food samples. To achieve improved interpretation and cross-validation relating to the immunochemical characterization of allergenic substances of processed food at the protein and peptide level, complementary investigation (in terms of applicability, reproducibility, and reliability of results) was carried out with alternative immunological methods (performed in ELISA and immunoblot format). In conclusion, the REA-based imunochip sensor is easy to fabricate, is reproducible in its performance, has minimal technical requirements, and will enable high-throughput screening of affinity binding interactions in technological and medical application for determining allergenicity. AllergenOnline, a peer-reviewed protein sequence database for assessing the potential allergenicity of genetically modified organisms and novel food proteins Richard Goodman 1 , Motohiro Ebisawa 2 , Hugh Sampson 3 , Ronald van Ree 5 , Stefan Vieths 5 , John Wise 1 , and Steve Taylor 1 . 1 University of Nebraska, Food Science and Technology, Lincoln, NE, United States; 2 National Sagamihara Hospital, Pediatrics, Kanagawa, Japan; 3 Mount Sinai School of Medicine, Division of Pediatric Allergy and Immunology, New York, NY, United States; 4 Academic Medical Center, Experimental Immunology, Amsterdam, Netherlands; 5 Paul Ehrlich Institute, Allergy, Langen, Germany. Background: New genetically modified (GM) organisms and novel processed food proteins are being introduced into commerce. Countries require safety assessments include a bioinformatics (sequence) comparison of the new protein to determine whether the new protein is a known allergen or is so similar to an allergen that currently allergic consumers would be at risk of reactions. If a match is found, the product would have to be tested using sera from appropriately allergic subjects. The quality of the database is a primary determinant of the search efficacy. A peer review process was used to update AllergenOnline and tools for an effective bioinformatics search are provided, free to the public. Methods: A list of 1758 Ballergens[ was compiled from NCBI using keyword searches, plus entries in the Allergen Nomenclature list of the International Union of Immunological Societies (IUIS) and AllergenOnline version 6. Sequences were grouped based on taxonomy and sequence identities. Peer reviewed articles were identified for each group. Criteria were developed to evaluate the evidence based on in vivo and in vitro tests using subjects with clinically defined, relevant allergic disease. Published data for each group was compiled and evaluated by primary and secondary reviewers, then the panel to remove groups with insufficient evidence of allergenicity (no IgE binding or in vivo reactivity, with subjects having at least minimal descriptions of allergies to the source of the protein). Free full-length and sliding 80 amino acid alignment tools are provided to allow public searches of the database. Key examples from the literature are used to illustrate the appropriate use of the database. Results: AllergenOnline version 7 contains 1251 sequences with published data demonstrating at least evidence of specific IgE binding using sera from subjects having some allergic symptoms when exposed to the source of the protein. Many isoforms are listed for some proteins. Others are represented by a single sequence. Bioinformatic comparisons with transgenic proteins from approved products and the literature were evaluated. No approved product exceeded Codex Guideline criteria, but some from the literature (presumably not in commerce) did, indicating a likely need for serum IgE testing. Conclusion: The curated allergen database will markedly improve the reliability of sequence matches for assessing the potential allergenicity of these new foods. Background: The quality of in vitro IgE tests used for food allergy diagnosis mainly depends on the analytes used. Usually total extracts of the suspected foods are used. However, in the recent past a rising number of studies about component resolved diagnosis have attracted rising interest. In many cases the use of purified single allergens either of natural origin or applied as a recombinant product has shown a superior performance when compared to total extracts. Within the EC funded IP Europrevall a library of 31 highly purified allergens from 10 foods listed in the new Labelling directive of the European Commission was established for developing novel diagnostics. Methods: Highly purified allergens either natural or recombinant from 10 foods (apple, peach, hazelnut, peanut, celery, cow`s milk, goat`s milk, hen`s egg, fish and shrimp) were characterized regarding their structural integrity, biological and allergenic activity. Protein mass was verified by mass spectrometry and sequence integrity by MALDI-TOF and N-terminal sequencing. Presence of isoforms in natural allergen preparations were identified by 2D Gel electrophoresis and secondary structure was evaluated by NMR maps of low molecular weight proteins. Allergenic activity was characterized using a panel of sera from food allergic patients. Results: Purification protocols for natural and recombinant allergens were established and quality assessment of the purified batches performed. Depending on either natural or recombinant origin different quality criteria were defined and met for inclusion into the allergen library. Structural integrity of the purified proteins was verified and IgE binding activity was assessed. Conclusion: This panel of well characterized allergens will be used to prove the concept of component resolved diagnosis and presents the necessary tools for setting up novel diagnostics. Using a significant number of allergic patients`sera across Europe it will contribute to identify relevant sensitization patterns and to improve dietary recommendations for the food allergic patient in the future. Dominant epitopes and allergic cross-reactivity of the thaumatinlike protein act d 2 Background: Some important food allergens have been identified as pathogenesis-related proteins (PRs). Thaumatin-like proteins (TLPs) belong to the family 5 of PRs. Several structures of plant TLPs have been solved in our group, including the TLPs from sweet cherry (Pru av 2), kiwifruit (Act d 2) and apple (Mal d 2). Comparing different structures belonging to the same protein family can lead to the identification of conformational epitopes. Methods: ELISA inhibition analysis using the sera of kiwi allergic patients was used for determination of immunological cross-reactivity between Act d 2 and other TLPs (thaumatin, zeamatin, Pru av 2 and Mal d 2). In order to identify putative cross-reactive epitopes virtual epitope mapping was applied. The similarities and differences in surface topology were compared. Results: ELISA inhibition assays for Act d 2 showed a high cross-reactivity with zeamatin, medium cross-reactivity with Pru av 2 and Mal d 2, and low cross-reactivity with thaumatin. On the other hand, the sequence comparison between Act d 2 and other TLPs yielded a different order of similarity: zeamatin9 thaumatin9Pru av 29 Mal d 2. Combining the immunological data and virtuale epitope mapping three putative epitope regions could be determined on the surface of Act d 2. Conclusion: High sequence homology/identity between two allergens is not necessarily the indicator for high cross-reactivity. We used the comparison of surface features of cross-reactive and non cross-reactive allergens to derive conformational epitope regions. Bet v 1 homologous proteins in kiwi fruit-relevant allergens? However, till now nothing is known about Bet v 1 homologous proteins present in kiwi. Preliminary efforts to purify natural Bet v1 homologues in sufficient quantities from kiwi failed so far. Methods: Two individual sequences (from green and gold kiwi fruit) homologous to Bet v 1 were identified from an EST cDNA library, subcloned into expression systems pETBlue-2 and pMW 175, respectively and expressed in E. coli. The resulting recombinant proteins were purified and characterized according to established methods. IgE binding activity of the purified recombinant Bet v 1 homologous proteins from kiwi fruit was tested by IgE-ELISA, IgE immunoblots, and inhibition assays. Results: Alignment of recombinant kiwi Bet v 1 homologues and recombinant Bet v 1a revealed an amino acid sequence identity of 50%. Well characterized purified recombinant Bet v 1 homologues from green (Actinidia deliciosa) and gold kiwi fruit (Actinidia chinensis) were recognized by birch pollen and kiwi sensitized patients in IgE immunoblots and ELISA experiments. The presence of a natural Bet v 1 homologous protein in green and gold kiwi fruit was confirmed by IgE immunoblot inhibition experiments using natural kiwi fruit extracts and immuno tissue prints. Cross-reactivity of the two recombinant proteins with varying intensities to natural Bet v 1 related protein from green kiwi and to rBet v 1a was demonstrated by ELISA and immunoblot inhibition assay. Conclusion: This is the first evidence of cross reactive Bet v 1 homologous allergens present in gold and green kiwi fruits. The present data point out that Bet v 1 homologues are relevant allergens in kiwi, especially for patients sensitized to tree pollen and kiwi fruits and might have been neglected so far due to low abundancy in the conventional extracts used for diagnosis. The evolutionary biology of allergens of the cupin and the Bet v 1 superfamilies Heimo Breiteneder, Merima Bublin, Stefan Wagner, and Christian Radauer. Medical University of Vienna, Department of Pathophysiology, Vienna, Austria. Background: The massive accumulation of sequence data of plant proteins in recent years has made the classification of allergens into protein families possible. By comparing sequences and structures, related proteins can be grouped together into families, and related families can be grouped together into superfamilies. This much broader picture now allows us to view allergenic proteins in their evolutionary context. Methods: Representative protein sequences were compiled by manually searching the Pfam database of protein families. Multiple sequence alignments and generation of neighbor-joining trees was performed using ClustalX. Phylogenetic trees were drawn with TreeView. Results: The cupin superfamily comprises numerous functionally highly diverse protein families from all groups of organisms including archaea. However, allergenicity within the cupins is confined to the vicilin and legumin plant seed storage proteins. The Bet v 1 superfamily contains the PR-10 family, the family of major latex proteins and ripening-related proteins, the norcoclaurine synthases, and the cytokinin-binding proteins. The PR-10 proteins from certain taxa such as the Fagales and Rosaceae are the only allergenic members of this superfamily. The divergence of the allergenic members of the Bet v 1 superfamily from the related major latex proteins, ripening related proteins and norcoclaurine synthases represents an ancient evolutionary event. Apparently the ancestors of modern day allergens were present long before any type of immune system had evolved. Conclusion: This study of the distribution of allergenic and non-allergenic members of protein families which was funded by the Austrian Science Fund grant SFB F01802 will provide new insights into the evolution of allergenicity and the factors that make proteins allergenic. Optimizing allergen components in litchi and peach extracts for in vitro specific IgE assays Anita Kober 1 , Hans Ö man 2 , Maria Lundberg 2 , and Maryam Poorafshar 1 . 1 Phadia AB, R and D, Uppsala, Sweden; 2 MIAB, R and D, Uppsala, Sweden. Background: Allergy to fruits is often associated with pollinosis and is then often related to milder symptoms such as oral allergy syndrom (OAS). However, especially in warmer regions more severe symptoms are also common. Fruits are here more prevalent foods and are also often eaten in more sun-ripe forms. Allergy to fruits of the Rosaceae family is strongly associated with Bet v 1 in northern regions but in warmer regions allergic reactions to these fruits are more often related to lipid transfer proteins (LTPs) and sometimes also profilins. The aim with this study was to explore new extraction buffers and different parts of fruits trying to optimize extraction of labile Bet v1 analogues as well as more stable profilins, LTPs and other allergen components. Methods: Peach was selected as a model for Rosaceae fruits where Bet v 1 homologues, profilin and LTPs are important allergen components. Litchi was selected as a fruit where the exposure to the population in Europe is increasing with changing feeding habits and where profilin is suggested to be an important allergen. Extracts were produced using different buffers and procedures and different parts of the fruits (pulp/peel) were used. Samples from patients from different regions with different symptoms (from mild to severe reactions) were analysed using ImmunoCAP coupled with improved extracts as well as single components. IgE antibodies to components were also assayed with a microarray using only 30 6l of sample. Results: Optimal extraction performance was achieved by a cold extraction procedure containing polyvinylpyrrolidone. Both pulp and peel were needed for optimal performance. For selected samples, mainly reacting to Bet v1, increased results as measured with ImmunoCAP was seen in 76% (62/81) for Peach f95. In litchi profilin was the dominating allergen component in several patient samples both measured by ImmunoCAP and using the microarray assay. The results were confirmed by inhibition studies. Conclusion: Improved extraction methods can enhance the content of both Bet v 1 homologues and profilin in fruit extracts used for in vitro assays for specific IgE. Microarrays are useful tools to analyse component profiles in samples using very small volumes of sample. Profilin (Lit c1) is an important allergen in litchi and may cause severe reactions. In peach LTP (Pru p3) and the Bet v 1 homologue Pru p1 were the more important components in the tested patient population. Characterization of saffron pollen allergens Sepehr Salehi, Majid Baseri Salehi, and Mehrdad Shariati. Islamic Azad University, Kazeroun Branch, Biology, Kazeroun, Islamic Republic of Iran. Grass pollen is one of the most important causes of IgE-mediated allergic reactions. Hypersensitivity to saffron pollen is an occupational allergy and is a serious problem especially among people dealing with saffron (farmers). Saffron is a native plant of Iran and this study was performed on saffron pollen in Khorasan province. Saffron pollen allergenicity was proven using skin prick test. The aim of this study was to characterize IgE-binding components of saffron pollen. At first saffron pollen was extracted with phosphate-buffered saline (PBS 125mM, pH 7.2Y7.4) at 4-C for 5 hours with gentle shaking. After centrifugation and delipidation, the extract was concentrated by polyethyleneglycol (PEG20000) at 4-C. Concentrated saffron pollen extract was fractionated by SDS-PAGE and transferred to nitrocellulose paper by western blotting technique. After western blotting, using sensitive individual sera and anti-human IgE-HRP immunoblot analysis was performed. Finally one specific IgE-binding band was showed with molecular weight of 20-27 KD. This study indicated that sensitive sera contain specific antibodies directed against allergens present in saffron pollen extract. Also this component can be bind specifically to IgG. The role of Interleukin 4 and 5 (IL4, 5) in development of chronic inflammations of stomach in patients with food allergy Zbigniew Bartuzi, and Magdalena Bikowska. Collegium Medicum Bydgoszcz Nicolaus Copernicus University, Department Allergy, Clinical Immunology and Intern, Bydgoszcz, Poland. The chronic gastritis can cause protracted iatrogenic effect of certain medicaments, for example non-steroid anti-inflammatory drugs also. Still, to little is known about the extent and role of hypersensitivity in patients with atopic diathesis in the development of chronic gastritis. The aim of the study was to determine serum concentrations of IL-4, IL-5 in patients with chronic gastritis and food allergy, who had been infected with H. pylori. The study was conducted on patients with atopic diathesis, suffering from dyspepsia and abdominal pain, who were monitored in our Outpatient Clinic for Food Allergy. The study group consisted of 71 patients, including 42 females aged 16=V54 years (mean age 35.5 years) and 29 males aged 18=V60 years (mean age 36.2 years). The control group was formed of 40 patients with dyspepsia and abdominal pain related to chronic gastritis and confirmed in endoscopy and histopathological investigation. specimens. The patients underwent endoscopy The samples were stained with eosin and hematoxylin (H and E) and with modified Giemza stain. Selected serum cytokines: IL-4, IL-5, nwere determined with ENDOGEN kits(Cambridge ). Serum levels of interleukin-4 in patients with food allergy ranged between 0.2 to 291 pg/ml, with mean value of 27.85=Ó 34.89 pg/ml. In the group of non-atopic patients with chronic gastritis, the range of determined values was 1.5 =V60.1 pg/ml =V mean concentration was 13.26=Ó 7.0 pg/ml. In control patients with functional dyspepsia, the concentration of IL-4 obtained ranged between 0 and 13.2 pg/ ml =V mean IL-4 concentration was4.31=Ó 2.8 pg/ml. IL-4 concentrations in various groups are presented in table 1. The concentrations of interleukin-5 in patients with food allergy ranged between 0 and 111.3 pg/ml (mean value 7.43=Ó 3.20 pg/ml). In control group of non-atopic subjects with chronic gastritis, IL-5 levels were 0=V20.6 pg/ml (mean level 4.03=Ó 1.60 pg/ml). The comparative analysis of IL-5 and IL5 levels between patients with food allergy and control group with chronic gastritis showed statistically significant differences (pG0.05). Conclusions: 1. Chronic exposure to the sensitising food in patients with food allergy results in the significant elevation of IL-4 and IL-5 levels. 2. Patients with food allergy and chronic gastritis, who had been infected with H. pylori manifest simultaneous increase in IL-4, IL-5, which suggests the participation of allergic factors in the pathogenesis of inflammation. Summary of blinded-food challenges against hen's egg and cow's milk allergies in the past 11 years Takatsugu Komata 1 , Takanori Imai 1 , Mika Ogata 1 , Sakura Sato 2 , Morimitsu Tomikawa 3 , Hiroshi Tachimoto 3 , Akinori Shukuya 3 , and Motohiro Ebisawa 2 . 1 Sagamihara National Hospital, Department of Pediatrics, Sagamihara-city, Kanagawa, Japan; 2 Sagamihara National Hospital, Clinical Research Center, Department of Allergy, Sagamihara-city, Kanagawa, Japan; 3 Sagamihara National Hospital, Pediatrics, Sagamihara-city, Kanagawa, Japan. Objective: The aim of this study was to analyze the results of 175 singleblinded food challenges (SBFC) against hen's egg(HE) and cow's milk(CM) allergy patients. Methods: We analyzed the data of the patients who had received the SBFC in Sagamihara National Hospital from 1995 to 2005. The total numbers of examination was as follows; HE: 92 challenges (male: 65, female: 27, mean age: 72.6 months), CM; 83 challenges (male: 62, female: 21, mean age: 71.0 months). SBFC was performed using dried powder (4g of whole egg powder or 6g of CM powder) with 120g strawberry puree as vehicle and the test was carried out along the AAAAI office procedure manual (JACI 82: 986, 1988) . Results: Most of the subjects received the SBFC prior to the entrance of primary school whether or not they could take school lunch. The positive rate of challenge test was as follows; HE: 70/92 (76%), CM: 37/83 (45%). The most common symptom induced by HE challenge was GI reaction (83%), followed by skin reaction (69%) and lower respiratory symptoms (17%). We further confirmed challenges with heated hen's whole egg against 62 subjects in 70 diagnosed raw egg allergy patients. Interestingly a discrepancy of the result between raw egg challenge and heated egg challenge was seen. Twenty nine percentage (18/62) of raw egg allergy patients could take heated whole egg. Egg white specific IgE level of patients who could not pass the challenge (18.0UA/ml) was higher than that of those who passed (8.0UA/ml) (pG0.05). For CM challenge, skin reactions were the most common (89%), followed by lower respiratory symptom (65%) and GI reaction (19%). There was no statistical difference of CM specific IgE level between failed challenges (19.5UA/ml) and passed challenges (12.1UA/ml). Higher SPT positive reaction to CM and higher complication rate of bronchial asthma were recognized in failed cow's milk challenge group. Conclusion: SBFC for HE and CM was useful for the diagnosis at the time of entrance to primary school. To improve the quality of life of raw egg allergy patients, it is recommended for the patients to receive heated egg challenge. It seems that HE challenge test tended to induce GI reactions more than CM challenge and that persistent CM allergy may have something to do with the complication of bronchial asthma in school age children. Cow`s milk protein allergy presenting as necrotizing enterocolitis: a case report Sarayutdh Tonglor, Pantipa Chatchatee, and Jarungchit Ngamphaiboon. King Memorial Chulalongkorn Hospital, Pediatric Allergy and Immunology, Bangkok, Thailand. Background: Gastrointestinal symptoms are common manifestation of cow`s milk protein allergy. How ever necrotizing enterocolitis resulting from cow`s milk protein allergy is rare. Case report: A 3,300 gm. full term female infant had been fed with cow`s milk formula since birth for ten days then switched to breast feeding alone. At twelfth day of life, she gradually developed abdominal distension and vomiting 2-3 times a day and was brought to see the pediatrician. Her physical examination had no other abnormal finding exept abdominal distension. So, she was advised to got some simethicone for abdominal discomfort and continued breast feeding. She still had the same symptoms that was vomiting and progressive abdominal distension. At twenty fourth day of life, she had developed more frequent vomiting, mucous bloody diarrhea, severe abdominal distension, hematochezia and pneumatosis intestinalis was seen on her X-ray finding. Laboratory finding included normal complete blood count, negative blood and stool culture. Necrotizing enterocolitis was diagnosed and successfully tolerated with parenteral antibiotics. She was placed on extensive hydrolysate formula which was well tolerated. The prick skin testing, atopy patch testing and radioallergosorbent testing were all negative for cow`s milk protein. Three months after, she grew very well with normal weight and height. She was challenged with soy-based formula at four month of age and well tolerated. At seventh month of age, she was challenged with cow`s milk protein and developed vomiting, abdominal distension within four days. The diagnosis of cow`s milk protein allergy was made based on positive symptoms after challenge. Conclusion: Cow`s milk protein allergy should be evaluated in patient presents with necrotizing enterocolitis without obvious risk factors. Calcium dependence of IgE reactivity of recombinant alphalactalbumin, a major cow`s milk allergen compared the results of RAST and open food challenge tests (OFC) among children who were under elimination therapy because of suspected allergy to some foods. Patients and Methods: The objects were 218 children, 139 male and 79 female, aged between 3mo and 12yr, suspected of food allergy. From January 2002 to May 2007, they were challenged with foods which seemed to have provoked allergic symptoms previously or which doctors advised not to take because of high scores of RAST tests and eliminated before the challenge. The clinical symptoms after ingestion of foods were observed for 3 days, documented and evaluated thereafter. Symptoms were classified into three categories; none, mild and moderate. Results: Foods challenged were egg white (61), egg yolk (58), cow milk (27), noodle (13), buckwheat (9), chicken (6), peanuts (5), soybean (4), tofu (3), mayonnaise (2) and others (30)(Figures in the parentheses are numbers of challenges). Positive and negative family history of allergic diseases was 84% and 16%, respectively. Symptoms after ingestion of foods were none in 78%, mild in 14% and moderate in 8%. No life-threatening symptom was noted. Specific IgE titers (RAST scores) against suspected foods were measured in 109 children. There was significant correlation between the symptoms and RAST scores (P$B!a(B0.017). On the other hand, 73% of high score cases (RAST score 5 and 6) were symptom free. These results show that there is discrepancy between Symptoms and RAST score titers. Conclusion: OFC is a simple and reliable method for clinical diagnosis of food allergy in children. The effect of soy-based formula on clinical symptoms and the fatty acid compositions in Infants with atopic dermatitis Jae-Won Oh 1 , Eun-Young Lee 2 , and Ha-Baik Lee 1 . 1 Hanyang University College of Medicine, Pediatrics, Seoul, Republic of Korea; 2 Gyunghee University, Nutrition, Seoul, Republic of Korea. Atopic dermatitis and food allergy commonly occur in early childhood. Especially infants with cow's milk allergy were reported to have the reduced growth and special nutritional needs. Formulations of soy-based infant formula have changed over the years to improve digestibility, the stability and availability of minerals, and protein quality. soy-based formulas are a rich source of essential fatty acids, linoleic acid and "-linolenic acid. This study was intended to investigate whether soy based formula has an effect on clinical symptom and the fatty acid composition in erythrocyte in infants with atopic dermatitis. The infants were recruited from pediatric allergic clinic in Hanyang University Guri hospital from September 2004 to August 2005. One group was supplemented soy based formula (SBF, n=21) and the other was supplemented hydrolyzed cow's milk as controls (HCM, n=20) for 12 weeks. The clinical severity was measured by using SCORAD index. To evaluate nutrition status of the subjects, anthropometric indices compared with Standard Value of Growth and Development for Korean children. Blood samples were collected to analyze fatty acid composition and immune parameters. Specific IgE measurement and Skin prick test was performed. The mean age of the SBF group was 9.0 T 2.1 month, and the HCM group was 8.8 T 1.9 month, and their SCORAD scores were mild or moderate states that significantly improved after experimental period (before SBF 40.9 T 2.4, after SBF 17.2 T 0.6, before HCM 45.7 T 24.6 after HCM 13.3 T 21.7). Although SBF grows consumed more "-linolenic acid (18:3, n-3) intake, eicosapentaenoic acid and docosahexaenoic acid were decreased after SBF. In the other hand, arachidonic acid (C20:4, n-6) was increased after SBF feeding (before SBF 8.3 T 2.8, after SBF 8.8 T 0.6), however, decreased after HCM (before HCM 8.6 T 1.0, after HCM 8.2 T 0.8). In conclusion, SBF supported to improve symptoms with normal growth and nutritional status in infants with atopic dermatitis but there were no evidences to improve fatty acid compositions although giving a number of lá-linolenic acid intakes. It might be suggested that %-6-desaturase was reported to decrease in atopic dermatitis, SBF could not show to recover this enzyme deficient condition in this study. Egg white-specific IgE levels as predictors of clinical reactivity in the follow-up of egg allergy E Montesinos, R Felix, JC Cerda, A Martorell, J Garde, and M Pont. University General Hospital, Allergy Unit, Valencia, Spain. Background: Our objectives were to describe the egg white proteins allergy survival time in our population and analyze the correlation between egg white IgE levels and oral contolled challenge test (OCCT) results. Methods: We performed a retrospective study including 42 patients diagnosed to egg allergy in the first 2 years of life who were controlled at 3,4 and 5 years of age. In the annual controls skin-prick tests, determination of egg white-specific IgE levels and OCCT were carried out until tolerance developed. In different moments of the follow-up we determined cut-off points of egg white-specific IgE levels that provided a probability Q 95 of egg white tolerance, analyzed by Receiver-Operating Caharacteristic (ROC) curve. Results: The median survival time of egg allergy was 48.7 months. The specific IgE levels cut-off points correlated with OCCT results differ in the different periods of follow-up. The egg white-specific IgE levels which were predictors of the clinical reactivity (positive predictive value (PPV) Q 95%) in the different periods were: between 25 and 36 months 1.52 kU/L, between 37 and 48 months 1.35 kU/L and between 49 and 60 months 2.59 kU/L. For these levels the negative predictive value (NPV) was 40%. Conclusion: The monitorization of egg white-specific IgE levels in allergic children to egg white allows us to predict the clinical reactivity along the egg allergy follow-up. Egg white-specific IgE levels greater than those referred would not be allow the OCCT in our population, while with lower levels we would find tolerant patients. Conclusion: Children with allergy to EHF demonstrated more severe symptoms of AEDS than those without allergy to EHF. The atopic background of clinical symptoms was more expressed in children with allergy to EHF. Amino-acid based formula should be recommended in therapy of children sensitized to EHF. Hypersensitivity to cow`s milk protein in a premature infant manifested with feeding intolerance and hypereosinophilia Khanitha Siripool, Wasu Kamchaisatian, Suwat Benjaponpitak, Surat Tungchaisin, and Chalerat Direkwattanachai. Faculty of Medicine Ramathibodi Hospital, Mahidol University, Division of Pediatric Allergy/Immunology, Bangkok, Thailand. Background: Cow`s milk protein hypersensitivity is the most common food hypersensitivity in infants. The current diagnostic approaches include a thorough history taking, skin testing and measuring of serum specific IgE to suspected food, elimination of suspected food and food challenge test. Eosinophilia occurs frequently in premature infants and has been associated with total parenteral nutrition and with cow`s milk protein allergy, a response to foreign antigen presented to body through gastrointestinal tract, airway or intravenous routes. Methods: We reported a premature male infant, with gestational age of 33 weeks and birth-weight of 980 gram, who developed feeding intolerance to premature infant formula since 3 days old. He was treated as necrotizing enterocolitis and was stopped feeding for a few days which showed improving of the symptoms. Then he was started feeding again on 7 days old and had feeding intolerance with increasing of peripheral blood eosinophilia from 440 / mcL to 9,640 /mcL at the age of 26 days. Results: The allergy consultation was appreciated for evaluating the cause of hypereosinophilia. After thoroughly excluding other causes of eosinophilia, the patient was suspected to be allergic to cow`s milk protein. He was changed the feeding to extensively casein-hydrolysated formula. The specific IgE to cow`s milk was done and revealed a value of 7.77 kUA/L with total IgE of 114.3 IU/mL. His symptom of feeding intolerance was improved with a good weight-gaining. The peripheral blood eosinophilia was gradually decreased to 2,220 /mcL within 5 days. One week later, the infant was reintroduced feeding with cow`s milk protein formula for 3 days and developed vomiting, abdominal distension and retention of gastric contents with a striking peripheral blood eosinophilia to 16,592 /mcL. The feeding was changed back to extensively casein-hydrolysated formula and the symptoms of feeding intolerance were improved with decreasing of eosinophil count to 900 /mcL on the age of 60 days and body weight of 2,000 gram. Conclusion: Herein, we report a case of premature infant with feeding intolerance and hypereosinophilia resulted from cow`s milk protein hypersensitivity. Eosinophilia in premature infant may be one response to foreign antigen mediated through gastrointestinal tract resulted in food hypersensitivity. A case of cow's milk allergy, which was succesfully treated with an amino acid-based formula but not with breast feeding Yoshiyasu Ogata 1 , Masafumi Zaitsu 1 , Shuichi Yamamoto 1 , Eriko Muro 1 , Natsuko Nishi 1 , Katsuya Tashiro 1 , and Rahmawati Minhajat 2 . 1 Saga university, Pediatrics, Saga, Japan; 2 Saga university, Pathology, Saga, Japan. Background: Hematochezia in infancy should be considered as one of the symptoms of cow's milk allergy (CMA). The most effective treatment of CMA is complete avoidance of cow's milk protein. Objective: We have experienced a male infant with CMA, who tolerated an amino acid-based formula but not breast feeding. Case: The patient was a full-term baby boy initially raised with breast milk and small amount of CM-based formula for about one month. From one month of age, the patient was exclusively breast-fed. At 3 month of age, a supplement of CM-based formula was started on a pediatrician's recommendation, because of slight poor weight gain. After ingesting a CM-based formula, he developed visible rectal bleeding immediately. A presumed diagnosis of CM allergy was made and mother was instructed to stop CM-based formula and to avoid drinking CM. Because mucus blood mixed stool continued, he was admitted to our hospital for diagnosis and treatment. Physical examination showed he was no apparent abnormal findings. Stool culture was negative for routine pathogens. The number of eosinophil in stool smear was elevated. Serum IgE was 10 IU/ml and specific IgE for milk was positive. Lymphocyte proliferative responses to CM-based formula and breast milk were positive, but to an amino acid-based formula was negative. A skin prick test was positive for CM. Rectocolonoscopy up to the sigmoid colon was performed, showing macroscopic evidence of nodular lymphoid hyperplasia with small erosions in rectum. Histological examination of biopsy specimen revealed colitis with predominant eosinophil-infiltration in the lamina propria. Through these findings we considered that this is a case of CMA and that breast milk contained a small amount of CM. We asked mother to stop breast feeding and CM-based formula and started to an amino acid-based formula. After avoidance of breast feeding, resolution of hematochezia took place with in two days. At the age of 14 months, the patient grows well on an amino acid-based formula and food without CM protein. Conclusion: It is difficult for mothers to avoid completely milk protein, because a lot of food contain CM as ingredient, and therefore breast milk may contain small amount of CM protein often. In this case, we successfully treated the patient with an amino acid-based formula without breast feeding. And he has no problem for his growth. An amino acid-based formula is a powerful tool of treatment of CMA in infancy. A prebiotic oligosaccharide mixture inhibits pathophysiological changes in mice orally sensitized against casein or whey proteins Bastiaan Schouten 1 , Betty van Esch 1 , Gerard Hofman 1 , Linette Willemsen 1 , and Johan Garssen 2 . 1 Utrecht Institute for Pharmaceutical Sciences, Pharmacology and Pathophysiology, Utrecht, Netherlands; 2 Numico-Research BV, Immunology, Wageningen, Netherlands. Background: Human breast milk is thought to protect the neonate from development of allergies. In particular, oligosaccharides in human milk may be beneficial. These prebiotics may have direct immunomodulatory capacities or exert their effects by enhancing growth of lactobacilli and bifidobacteria. To test the effects of prebiotic oligosaccharides on allergic sensitization, a diet containing a 2% oligosaccharide mixture of fructo-, galacto-and acidic oligosaccharides (GOS:FOS (9:1, 1,8%) and AOS (0,2%)) was tested in mice orally sensitized against casein and whey proteins. The prebiotic mixture mimicks the oligosaccharide composition in human milk. Methods: For a period of 6 weeks, five week old female C3H-HeOuJ mice were sensitized weekly by gavage (i.g.) with casein or whey (20 mg), using cholera toxin (CT) as an adjuvant. Mice were fed with the control or prebiotic (GFA) diet, starting two weeks prior to the first sensitization. At week 7 the mice were challenged subcutaneously (ear) (10 6g) and orally (100 mg). Serum levels of total IgE and allergen specific IgE, IgG1 and IgG2a were measured. The acute allergic skin reaction was determined by measuring ear swelling. Furthermore, isometric contraction of the colon was determined to get insight into motility changes of the intestine. Results: In the whey and casein sensitized animals the allergic skin reaction was significantly reduced in the prebiotic group. This was most obvious in the casein group (control vs GFA diet, 202 +/-23 vs 73 +/-11 6m; pG0.01). Casein and/or whey specific IgG1 and IgE levels were unaltered, while in whey sensitized animals IgG2a levels were enhanced by the diet (control vs GFA diet; 0.11+/-0.003 vs 0.40 +/-0.13 A450; pG0.05). The hypocontractility of the colon of casein sensitized mice was restored to normal levels by the prebiotic diet (control vs GFA diet, 1342 +/-404 to 3515 +/-1106 mg; pG0.05). Conclusion: A mixture of GOS/FOS/AOS oligosaccharides reduces systemic and local allergic symptoms in mice orally sensitized to cow`s milk proteins, when provided during the sensitization phase. Balb/c mouse model of allergy to wheat gliadins: biochemical and clinical manifestations Marie Bodinier 1 , Michaël Leroy 1 , sandrine Ah-Leung 2 , Fany Blanc 2 , Olivier Tranquet 1 , Sandra Denery-Papini 1 , Jean-Michel Wal 2 , and Karine Adel-Patient 2 . 1 INRA, BIA unit, allergy team, Nantes, France; 2 INRA, Unit of Immunology and Food Allergy, CEA Saclay, Gif-sur-Yvette, France. Background: Animal models demonstrating IgE of the same specificity as human IgE and clinical symptoms upon challenge close to those observed in allergic patients are of great interest for the understanding of mechanisms involved in the induction and regulation of allergy. Food allergy to wheat (FAW) affects 0.1 to 0.4% of the global population. Major allergens have been characterized in this pathology. Water/salt soluble albumin/globulin fraction contains major allergens for children with the atopic eczema/dermatitis syndrome (AEDS) and the insoluble w5-gliadins are involved in adults with wheat-dependent exercise-induced anaphylaxis (WDEIA). Aim: The aim of this study consisted of developing a mouse model of food allergy to a gliadin extract, a wheat fraction containing major allergens by testing different protocols. Methods: Three strains of mice, Balb/c, C3H/HeJ and B10.A, were immunized by 4 successive intra-peritoneal injections of gliadin extract. The influence of the dose of gliadins (10 and 20 6g per mouse) was evaluated on allergen specific IgE and IgG1 production, on the cytokines production by activated T lymphocytes and on the elicitation of an allergic reaction. The development of the allergic reaction was compared by testing the Th2 type cytokine production and the eosinophil influx in the bronchoalveolar lavage (BAL) fluids 24h after a single intra-nasal stimulation with a total gliadin extract. Results: Balb/c mice demonstrated higher gliadin specific IgE and IgG1 productions than the other mice, whatever the dose administered (i.e. 10 or 20 6g). This was confirmed by the Th2 type cytokine profile produced by activated splenocytes. The significant eosinophil influx and the Th2 type cytokine produced in the BALs also showed the elicitation of the allergic reaction in Balb/c mice, whatever the sensitisation dose. Low to no elicitation was evidenced in B10.A and C3H/HeJ mice at the 10 6g dose. Conclusion: This study demonstrates for the first time the interest of Balb/c strain as a mouse model of food allergy to wheat. Our project is to further characterise the fine specificity of induced IgE and to compare the responses obtained in humans and mice. The number of IgE-binding cells in stomach mucosa in patients with peptic ulcer disease and food allergy versus without food allergy Zbigniew Bartuzi. Collegium Medicum Bydgoszcz N. Copernicus University, Department Allergy, Clinical Immunology and Intern, Bydgoszcz, Poland. Immunoglobulin E is the main antibody responsible for immediate type reactions. It can bind with FceRI receptors on mast cells and basophiles including those in the alimentary tract. Aim: The aim of this study was to compare a quantity of cells connected with IgE antibody at stomach mucosa in patients with peptic ulcer disease and food allergy, versus group of patients with peptic ulcer disease and without food allergy The study group consisted of 33 patients with ulcer disease and food allergy. The control group was formed of 21 non-atopic patients with ulcer disease. We evaluate a quantity of cells connected with IgE antibody in gastric mucosa by method of direct immunofluorescence. Results: The number of IgE-binding cells in stomach in patients with peptic ulcer disease and food allergy was average 15,55 instead in patients with peptic ulcer disease 8,52. We found statistical significant difference of quantity of cells connected with IgE between patients with peptic ulcer disease and food allergy, versus patients with peptic ulcer diseases and without food allergy (pG0,02). Conclusion: This results confirm the important role allergic mechanisms in pathogenesis of inflammatory changes in stomach mucosa observed in patients with peptic ulcer disease and food allergy. Relationship between ulcerative colitis, Irritable Bowel Syndrom (IBS) and food allergy Xueyan Wang, Xuefei Yang, Ruiqin Ma, Ming Zhang, and Huan He. Bejing Shijitan Hospital, Allergy, Beijing, China. Background: Allergic diseases are common diseases that involve many clinical disciplines. Food allergy, the offending food and food additives are concerned problems for doctors. Objective: to explore the relationship between ulcerative colitis, irritable bowel syndrome(IBS) and food allergy. [Method] During the periods of 2005 to 2007,30 patients with ulcerative colitis or IBS were tested food skin test, serum food specific immunoglobulin G( sIgG), ANA. CIC, C3, C4, CH50, ADNs. Results: 13 patients have positive result to egg, 8 patients to wheat, 4 patients to tomatoes, 3 patients to beans, 3 patients to fish, 2 patients to shrimp, 2 patients to peanut, 2 patients to corn, 2 patients to beef. Most of them combined with more than two kinds of food. All of them were visited 3 months and 6 months after examination. After avoiding positive food for 3 months, 15 patients with colitis have improvement, abdominal pain relieved, After 6 months, 22 patients recovered. Conclusion: It is valuable to test food sIgG. According to the elevated sIgG, these patients were asked to change food habits. 73.3% patients have improvement. 26.3% patients have no effect. The study helps us to know the close relationship between digestive disease and food allergy. The study of the Chinese herbal medicine-Lonicera extract from water solution on the ovalbumin-sensitized BALB/c mice Fei Li, and Haiqi Li. Children's Hospital, Department of Primary Child Care, Chongqing, China. Background: Few studies are available for treating food allergy by used TCMs. Objective: To investigate the effects of the Chinese herbal medicine Lonicera on the ovalbumin-induced hypersensitivity BALB/c mice. Methods: BALB/c mice sensitized with OVA were treated 4 hours after the first challenge and then twice daily for 10 days with different Lonicera concentrations extracted from water solution, i.e. 1g/ml as high dosage , 0.5g/ ml as middle, and 0.25g/ml as low. Results: Part I: The conditions of hypersensitivity mice were markedly improved by the high or middle Lonicera doses, such as reduced levels of serum OVA-specific IgE , inhibited the abnormal monocular cell proliferations, alleviated the degranulation and aggregation of intestinal mast cells, increased the percentage of intact mast cells and significantly reduction of histamine release, as well as downregulated the ratio of IL-4/IFN-, and promoted the mRNA expression of IL-10 and TGF-$ in PLNMC. Part II: The counts of Bifidobacterium and lactobacillus of the stool samples of the sensitized mice were significantly increased by the high or middle Lonicera doses, and meanwill the counts of Enterobacteriaceae and Staphylococcus were markedly declined. Conclusion: Our data first showed the immunological regulation effects to food allergy from the high or middle Lonicera concentrations extracted from water solution, such as anti-inflammation and immunoregulation of Th1/Th2 responses as well as activation of T regulatory cells. And the inbalance of intestinal microflora in OVA-induced allergic mice were improved by the Lonicera ext, which were helpful to improve the clinical manifestations of food allergy. Utility of fast in food allergy: a case report Paola Minale 1 , Federica Fumagalli 2 , Annalisa Kunkle 3 , Matteo Caiti 4 , Costantino Troise 1 , and Giorgio Walter Canonica 2 . 1 San Martino Hospital, Genoa, Allergy Unit, Genoa, Italy; 2 University of Genoa, Allergy and Respiratory Diseases DIMI, Genoa, Italy; 3 San Martino Hospital, Genoa, Diagnostica Citofluorimetrica, Genoa, Italy; 4 University of Genoa, Dept of Internal Medicine -DIMI, Genoa, Italy. We describe the case report of a 29 years old man who experienced an episode of urticaria and angioedema 30 minutes after having taken 100 mg of nimesulide and having drunk a beer. He was known to suffer from allergic rhinitis and food allergy (skin prick test were positive to: Dermatophagoides Pteronissinus, Dermatophagoides farinae, Grass pollen, olive tree pollen, birch pollen, artemisia, hazel tree pollen, peanuts, nuts). He had had urticaria and angioedema after the ingestion of peanuts and nuts. As regards the last episode, both the drug and the food were suspected to be the cause of the reaction, therefore we performed skin prick test to beer and malt (previously tolerated), which resulted positive; the FastImmune Basophil Activation Assay (Flow-Fast) for beer confirmed the skin prick test positivity; Fast for nimesulide was negative; to confirm this data the patient has been challenged to nimesulide that was tolerated. According to our clinical and laboratory findings, we concluded that the diagnosis was urticaria due to beer allergy. Fast reveals the presence of a very specific and sensitive cellular activation marker that permits to diagnose IgE-mediate reactions and pseudoallergies. In our case report has been helpful to confirm the positivity to beer allergen skin prick test and exclude a drug reaction. Therapeutic indications to amino acid-based formula in infants with atopic eczema dermatitis syndrome Beata Cudowska, Maciej Kaczmarski, and Ma 3 gorzata Sawicka-ukowska. Medical University of Bialystok, III Department of Paediatrics, Bia 3 ystok, Poland. Background: Allergy to cow`s milk protein is one of the casual factor of atopic eczema dermatitis syndrome (AEDS) in infants. Treatment is based on diet, generally using an extensively hydrolyzed formula (EHF) or amino-acid based formula (AABF). The aim of study was to establish the therapeutic indications to amino acid-based formula in infants with atopic eczema. Materials and Methods: 30 children aged 1-36 months with AEDS were qualified to the study. Allergy to cow`s milk proteins was established by the measurement of total and specific IgE and verified by oral food challenge following standards. According to the obtained results infants were divided in two groups: 16 children with allergy to EHF and 14 children without allergy to EHF. Results: In all of children allergy to cow`s milk formula was confirmed. In the group of children with allergy to EHF, hypersensitivity to casein hydrolysates was confirmed in all of them, to whey hydrolysates in 9/16 of infants. Severe atopic dermatitis was recognized more often in infants with sensitization to EHF (SCORAD index: mean 60,01 T25,14 vs 52,01 T24,43 in the control group; p=0,03). Total IgE was elevated significantly in the group of infants with severe AEDS and allergy to EHF (mean 623,1 T735,2 vs 88,21 T124,2; p=0,0001). Conclusion: Amino acid-based formula should be recommended in children with severe atopic dermatitis and allergy to extensively hydrolyzed cow`s milk formula. Allergy to casein is the most common in infants with hypersensitivity to EHF. Assessment of birch pollen immunotherapy effect on pollen-food allergy syndrome to apple with in vitro passive sensitization assay Ramon Bencharitiwong, and Anna Nowak-Wegrzyn. Mount Sinai School of Medicine, Allergy and Immunology, New York, United States. Rationele: Polllen-apple allergy syndrome affects approximately 50% of birch tree pollen-allergic adults. Allergy to apple results from cross-reactivity between apple and birch pollen proteins. Subcutaneous immunotherapy with birch pollen was reported to improve apple allergy in a subset of adult subjects. Skin tests and serum allergen-specific IgE antibody levels correlate poorly with clinical expression of apple allergy in birch-allergic individuals. Oral food challenge to apple remains the most accurate diagnostic test, however, standardized protocol is not easily available and the procedure is labourintense. We sought to determine whether RBL passive sensitization assay might be used to evaluate effects of birch pollen immunotherapy in subjects with pollen-food allergy syndrome to apple. Methods: Birch-allergic individuals had apple allergy confirmed with doubleblind placebo-controlled oral food challenge and were randomized to birch pollen immunotherapy arm or to control arm (no immunotherapy, continued standard medical management). RBL-2H3 cells transfected with human Fc? receptor were passively sensitized with sera from birch-allergic individuals with oral allergy to apple at 1:40 dilution and following overnight incubation were stimulated with serial dilutions (10-2-10-8) of in-house freshly prepared raw Granny Smith apple extract. N-hexosaminidase content in the supernatant was used as a marker of RBL degranulation and was expressed as a percentage of total degranulation caused by Triton X. Results: Eight subjects completed birch immunotherapy course. Median baseline release was 2.6% (range; 1.2-19.5), not different from median postimmunotherapy release 4.7% (range; 1.3-20.5), p=0.9. There was a trend for decreased skin test responses to birch pollen extract (mean 13.1 mm preversus 10.3 mm post-immunotherapy; p= 0.21) and fresh apple (mean 4.4 mm pre-versus 3.3 mm post-immunotherapy; p=0.08). There was no significant difference between pre-and post immunotherapy birch IgE and Bet v 1 -IgE levels. Conclusion: RBL cells passively sensitized with sera from birch-and appleallergic individuals undergo degranulation upon stimulation with fresh Granny Smith apple extract. Mediator release to fresh apple extract is not different before and after birch pollen subcutaneous immunotherapy. The performance of the in vitro passive sensitization assay is affected by low concentration of major apple allergens in the extract. Development of the first multidisciplinary psycho-educational intervention for parents and children with food allergy Audrey DunnGalvin, and Jonathan Hourihane. University College Cork, Paediatrics and Child Health, Cork, Ireland. Aim: Development of an evidenced based intervention programme to help children and parents to manage the biopsychosocial impact of food allergy on their everyday lives. Methods: 6 focus groups with parents of confirmed food allergic children and 10 focus groups with food allergic children aged 5-13 years. Qualitative analysis using grounded theory was used to analyse transcripts. The transcripts gave rise to categories relating to risk perception, illness cognitions, coping strategies, understanding of health and well-being, meanings given to diet and food. Transcripts also revealed developmental, parent/child, and sex differences vital to creating a targeted and food allergy sensitive intervention. A psycho-education intervention was then developed and is currently being validated with strong initial findings. Conclusion: Middle childhood is a period when problems with anxiety, low self-esteem, peer comparison, and malconstructive coping strategies emerge, leading to higher risk in adolescence. Our data suggests that an intervention at this critical point in the food allergy developmental perspective is vital to ensuring successful biopsychosocial adjustment in day-to-day life. A new questionnaire for food allergy Federica Fumagalli 1 , Ilaria Baiardini 1 , Sara Pellegrini 1 , Micaela Bacic 1 , Paola Minale 2 , Fulvio Braido 1 , Costantino Troise 2 , and Giorgio Walter Canonica 1 . 1 University of Genoa, Allergy and Respiratory Diseases DIMI, Genoa, Italy; 2 San Martino Hospital, Genoa, Allergy Unit, Genoa, Italy. Despite clinical expertise suggest that food allergy has a substantial burden on patients' life, little data is now available on its impact and treatment on quality of life. The aim of our study is to evaluate patients? answers to the preliminary version of a QoL questionnaire specifically addressed to food allergy. During the development phase of the new tool, 32 items were administered to 27 patients (9 M, 18 F, mean age 40, SD 7,2) suffering from food allergy. Patients had to indicate, on a Likert scale with multiple options (1: not at all; 5: very much), how much they were been troubled by each problem. Results: A relevant percentage of patients reported a important burden in many aspects of their life: Patients live in a state of alert: they are watchful to new symptoms? appearance (21 out of 27) and to what they eat (18/27), they are afraid to become sick when they eat something at risk (20/27) or a new foods (15/27). Patients refer to feel uneasy when they go out for dinner (12/27) and they are troubled because they have to read labels on every product (13/ 27). Another relevant problem is the diet that, due to avoiding the food, could be unbalanced (10/27). So food is a cause of concern for patients (14/27), and they can't enjoy food (10/27). Conclusion: on the basis of our preliminary data, food allergy appears as a problem that deeply interfere with patients life. The increased prevalence of latex allergy is associated with the increased usage of the latex materials and with the cross reactivity to latex and food allergens, due to the common determinants of the plants. Goal: to reveal the cross sensitization to the latex allergen in the patients with the food allergy using the test of allergic alteration of leucocytes (TAAL). Materials and Methods: 37 patients with the food allergy to fruit and 18 health care workers with glove-related skin symptoms (the itching, redness and dryness, sometimes an oedema, of the hand skin) were included in the study. The diagnosis of latex allergy in the patients of the second group was confirmed by the positive use test (in 10 from 10 ones), the negative results of Contact Patch Test with rubber chemicals for type IV hypersensitivity and by revealing of IgE antibodies to latex in 6 from 7 sera with ELISA method. TAAL and skin testing with latex and fruit allergens (carrot, potato, apple, tomato, kiwi, mango, melon, avocado, apricote, banana, perch -BStallergenes[, France) were performed in all patients. TAAL: 100 mkl of the each blood sample were incubated at 37oC during 30 min with 10 mkl of the standard solution of the ammoniated latex (Stallergenes), then micropreparates were prepared, stained with acrydine orange for luminescent microscopy and a percent of the alterated granulocytes was counted. The result was considered to be positive if the number of the alterated cells was more 10%. A sample of the same blood, incubated with 0.9% NaCl, was examined as a negative control. Results: the positive results of TAAL and skin test with a battery of fruit allergens were observed in the both groups of patients. In the latex allergy patients the positive results of the TAAL and skin test with latex allergen were observed in 77.8 and 83.3%, consequently. The positive results of the TAAL with the same allergen were observed in 70.5% of patients with the food allergy, and 76.4% of these patients had positive skin tests to the latex. The high percentage of the sensitization to latex in the food allergy patients without clinical symptoms to latex, established by two methods, may be the evidence of the cross reactivity and the risk for development of latex allergy. Latex recombinant hevein b IgE antibody in atopic children sensitized to fruit, latex and pollen Rumiko Shibata, Jyunko Ikei, Youko Murakami, and Sankei Nishima. Fukuoka National Hospital, Pediatrics, Fukuoka, Japan. Rationale: The associations of latex and fruit oral allergy syndrome (OAS) with pollinosis have been reported in adult allergic patients. In childhood fruit allergy, however, no significant data had been collected on clinical features and frequency of cross-reacting IgE antibodies to latex. Latex antigen is clarified to hevein b1 to 13, and primary allergen to be related to clinical manifestation has been elucidated by IgE antibody determination for these recombinant hevein (rHev) b proteins. Objective: The aim of this study was to identify the specific IgE antibodies to latex rHev b protein in fruit OAS children and latex allergy. Methods: Twenty-six atopic children (aged 2-11 years, 5.62.6 y.o.) with fruit OAS sensitized to latex and 3 latex allergy children (3-8 y.o.) with having operation history from early stage of age were enrolled in this study. Specific IgE Antibodies to food, pollen, latex, rHev b 5, 6.01, 6.02, 8, 11 were measured by CAP-FEIA (Phadia KK). Results: The clinical manifestations of fruit OAS were due to banana (67%), melon (63%) and kiwi (20%). With a fruit specific IgE antibody of 26 cases, banana 80%, melon 78%, kiwi 54% were positive. All patients had IgE antibodies to latex (0.6~31 UA/ml) and Japanese cedar pollen. In 6 fruit OAS with clear latex elicitation history, latex IgE antibody levels were high and IgE antibody to rHev b6.01 were positive in 3 cases and rHev b6.02 in 2 cases. In 14 of 20 cases without latex allergic provocative medical history, IgE antibody to rHev b 8 (profiln) increased remarkably. Specific IgE to rHev b 5, 6.02 and 6.01 were clearly positive in 3 cases of latex allergy with operation history. Conclusion: The results provide evidence that food allergy infants with fruit OAS have the risk of early sensitization to latex and pollen. Measurement of specific IgE to latex rHev b6.01 and 6.02 was useful as a prediction of latex anaphylaxis in fruits OAS children with latex IgE antibody. Turnip/oilseed rape and mustard allergy in Finnish and French children with atopic dermatitis against food allergy are on going in collaboration with the Ministry of Health, Labour, and Welfare in Japan in order to improve quality of life of patients with food allergy. Investigation of peanut processing and its effect on allergenecity in Asian and North American populations Elisabeth Berger, Birgitta Backhans, Håkan Larsson, Maryam Poorafshar, and Ingrid Holmquist. Phadia, R&D, Uppsala, Sweden. Aim: Peanuts are cultivated over almost the whole world. Despite the widespread consumption of peanut products, the prevalence of peanut allergy is varying between different areas. Peanut is, prior to consumption, subjected to different kind of processing in different parts of the world. In this study the effect of different treatments, on a variety of peanut cultivated in North and South America, trying to mimic different eating habits was investigated. Finally the IgE-binding patterns to untreated and roasted peanuts were studied in two different populations from US and Asia. Methods: Peanut source materials, Jumbo Runner, from US and Argentina were used. The peanuts were treated in three different ways; heated (90C, 20 min), boiled (20 min), or roasted (180C, 20 min) followed by grinding and extraction. Untreated peanut was used as control. The proteins were separated on both conventional SDS PAGE and 2D gel electrophoresis. Some spots from the 2D electrophoresis gel were excised and analyzed by MALDI-TOF MS. Immunoblotting analyzes were performed with sera from US and Asia from individuals with peanut sensitization. Results: Differences in protein patterns between the two source materials were detected on SDS-PAGE electrophoresis. One prominent protein band of approx 40 kD, in material from US was missing in the peanuts from Argentina. As expected a decrease in protein amount dependent on treatment and temperature was detected. Distinct spots in 2D electrophoresis at approx 60 kD from untreated materials, were not visible after roasting. These spots were identified as major peanut allergen Ara h 1 by MALDI-TOF MS. No obvious differences in IgE-binding in Immunoblotting between the two populations could be observed. For both populations there was a decrease in IgE-binding to roasted compared to untreated peanuts, especially at approx 60 kD (Ara h 1) and at 30-40 kD. Other differences in IgE-binding patterns were due to individual differences. Conclusion: Peanuts cultivated in different parts of the world may contain different proteins and allergens. Different treatment and cooking habits can influence the allergenicity. In this study no obvious differences in IgE-binding patterns between the two populations were observed even though the populations were chosen to reflect the different eating habits of peanuts. The clinical relevance of the missing protein in peanut source materials from Argentina need to be further investigated. The outcome of food allergy among infants and children in Israel Darit Aronov, Eli Somekh, and Ilan Dalal. E. Wolfson Medical Center, Pediatrics Allergy Unit, Department of Pediatrics, Holon, Israel. Background: Food allergy is common among infants and children. Milk, eggs and soy allergy tends to resolve early in life while allergy to peanuts, tree nuts and fish usually stay for life. The knowledge about the natural history of sesame allergy is scarce. We did a survey among the children that were treated in the allergy clinic in E. Wolfson medical center. Methods: The survey was done based on the clinical records of children that were diagnosed with food allergy. A phone survey and a detailed questioner were filled. The emphasis was on the outcome of the child's food allergy. Results: We found 234 children that were suspected to have food allergy. 54 (23%) of them were found not to have food allergy. The 180 (77%) patients with food allergy were found to be allergic to several foods: Milk-125, eggs-71, sesame-30, soy-23, peanuts-14, tree nuts-9, fish-3, beef and chicken-2, other foods-6. The children were followed up for an average period of 4.7 years. In 139 cases the allergy had resolved, in 93 it was still existing, and in 51 cases it was not clear whether the allergy had been resolved or not. Among those with milk allergy, 86/125 (69%) were IgE-mediated (group I) and 39/125 (31%) were non IgE-mediated (group II). In group I there was clearly a more atopic background then in group II (P=0.0032). The allergy had been resolved in 35/39 (90%) among group II patients, as compared to only 32/86 (37%) in group I (PG0.0001). Allergy to soy was found in 23 children, 21 of them were also allergic to milk and only 2 were allergic just to soy. The number of children with soy allergy among group II patients was higher than in group I. In 19/23 (83%) the allergy resolved. 30 children had allergy to sesame. In third of them the allergy presented as anaphylaxis. Most of the children with sesame allergy were found to have atopic background. 5 of them have a first degree relative with sesame allergy. In 9/30 (30%) the allergy had resolved in an average of 2.8 years. Conclusion: The distribution of food allergens in Israel differs from other countries. We found that the atopic status and the outcome of IgE-mediated milk allergy.is significantly different compared to non IgE-mediated patients. This survey may help to understand many aspects of food allergy among children in Israel, and may help us provide appropriate guidance for the children and their families. Background: Teenagers are a high-risk group for food-hypersensitivity (FHS) fatalities and engage in risk-taking in managing their condition. Existing research has identified anxiety and impaired quality of life in children with FHS and their parents. To date, however, research has not addressed what the experience of living with FHS is like for teenagers. This study therefore aimed to describe the lived experiences of teenagers with FHS. Methods: Individual semi-structured in-depth interviews were conducted with 21 UK teenagers (aged 13-18 years) with a pre-existing diagnosis of FHS. Participants were hypersensitive to a range of foods and experienced a variety of reactions. Participants were excluded if they had a concomitant non-allergic disease. Interviews explored the lives of the teenagers generally with particular attention paid to the experience of living with FHS. Interviews were audiotaped, transcribed verbatim and were then analysed using a phenomenological approach. Results: Four main themes were identified: FHS as a way of life, managing FHS as an assessment of acceptable risk versus acceptable burden, living with FHS as coping with necessary burden, and alleviation/exacerbation of the burden of living with FHS. For participants the process of managing FHS was not described as problematic but as a way of life. Managing FHS was described as negotiable rather than prescriptive and was based on an assessment of acceptable risk versus acceptable burden. Negotiating management did not however completely relieve the burden felt and participants felt that a necessary part of living with FHS was coping with necessary burden and a variety of coping strategies were employed to this effect. Participants also described ways in which the burden of living with FHS is alleviated or exacerbated both by them personally and by others whose behaviour affects their life with FHS. Conclusion: The themes provide some explanations for why teenagers with FHS engage in risky behaviours and describe how teenagers cope with managing FHS and what factors make living with FHS easier or more difficult. The themes indicate ways in which the lives of teenagers with FHS can be improved and therefore provide new information about living with FHS from the perspective of teenagers that will be of use to healthcare practitioners working with teenagers with FHS. Cow milk allergy fo associated to rhinitis atopic Maria Alul. CIPEA: Centro de Inmunologia pediatrica, asmay alergia, Alergia pediatrica, San Miguel de Tucuman, Argentina. Background: The Allergy to the cow milk (CM)is one of the causes of manifestations allergic but common in the childhood. It has an excellent one I foretell since most of the cases they are solved from the 4 years of age. The associated clinical disorders to the allergy to cow milk in these children include atoepics conditions such as atoepics dermatitis, allergic rhinitis, asthma and other manifestations corresponding to foods allergies. Objective: To determine the association of allergy to cow milk and persistent allergic rhinitis in smaller children of 4 years. Material and Method: 5 smaller children of 4 years with diagnosis of allergic rhinitis and allergy to cow milk through Clinical History evaluated themselves. They were made Prick Test with cow milk standardized protein extracts (Casein, A-lactoglobulin and Blactoglobulin); Concentrated of Histamina 1mg/ml considering as positive control the one presence papula of 5 mm, and saline solution as negative control without presence of pápula. One quantified Ig and EspecÚ fic to cow milk in International Units mililiter by means of enzimoinmunoensayo. To the clinical aims, the specific concentrations of IgE were evaluated according to the following data: 5 UI/ml:nonsignificant ;0.51 -1.0 Ul/ml:Low;1,1 -5.0 Ul/ ml: Moderate ;5,1 -25.0 Ul/ml:High;25,1 -75.0 Ul/ml: Very elevated;9 75.0 Ul/ml:Highest. Results: Of the 5 studied children, the totality I present/display direct atopicos familiar antecedents; and manifestation of rhinitic symptoms from the incorporation of the cow milk like dietario supplement (Three children from birth and two as of the 2 months of life). Were levels of moderate specific IgE in 3 of the 5 studied children (two with supplement of CM from birth and one as of both months of life) presenting/displaying these prick positive for alpha and beta lactoglobulin and negative test for casein; with serum of IgE a superior to 100 UI/m. In other two patients of the five were levels of specific IgE to nonsignificant CM with low serum values of IgE to 50 UI/ml and negative prick test for cow milk proteins. Conclusion: According to these results it is possible to be determined that the cow milk is present in the allergic rinitis of smaller children to the four years of age, and that his replacement could improve the clinical sintomatology of the patients with medication saving for such. Prevalence of peanut and tree-nut allergy in Singapore teenagersestimates from a questionnaire survey, allergy testing and food challenges Irvin Francis Gerez 1 , Shu E Soh 2 , Jian Yi Soh 2 , Pao Zhing Ng 2 , Elizabeth Morales 1 , Stefan Ma 3 , Bee Wah Lee 2 , and Pei Chi Lynette Shek 2 . 1 National University Hospital, Paediatrics, Singapore, Singapore; 2 National University of Singapore, Paediatrics, Singapore, Singapore; 3 Ministry of Health, Statistics, Singapore, Singapore. Introduction: Peanut and tree nut allergy can potentially be life threatening. Hence, awareness of its prevalence in the population would allow physicians to better address the issues of diagnosis, management and education. Objective: To estimate the true prevalence of peanut and tree-nut allergy in Singaporean teenagers. The prevalence of food allergy in 14 to 15 year old Singaporean children was evaluated using a standardized questionnaire which was distributed to students of randomly selected schools. Students who responded positively to these food allergies were administered an extended questionnaire to determine presence of convincing food allergy. A portion of those with convincing allergy symptoms were recalled for skin prick testing (SPT) as well as underwent a graded food challenge for those who consented. Results: From a cohort of 8072 students, 6765 (83.78%) responded. The crude prevalence of self-reported allergy to peanuts was 0.90% and treenuts 0.63%. However, using the criteria for convincing food allergy, the prevalence fell to 0.33% for peanuts and 0.20% for tree-nuts. A proportion of subjects with convincing allergy symptoms agreed to skin prick testing, which showed low frequency of skin test positivity peanut 4.17% (1/24) and tree-nuts 13.3% (2/15). Finally, all food challenges (5 for peanuts and 2 for tree-nuts) carried out on 7 subjects with either a convincing history plus negative SPT or convincing history plus positive SPT, resulted in a negative outcome. Conclusion: Our results indicate that the prevalence of peanut and tree-nut allergy in Singapore adolescents is low. Evaluation based on questionnaire survey alone overestimated the prevalence of peanut and tree-nut allergy in our population. Giovanni Ferrari, and Peter A Eng. Kantonspital Aarau AG, Children's clinic, Aarau, Switzerland. Background: IgE-mediated food allergy is the most common type of adverse reaction to food in infants and young children. It presents with a wide spectrum of clinical reactions including life-threatening allergic symptoms. The study has been performed to determine the most prevalent food allergens inducing immediate clinical reactions in different age groups of Swiss infants and children. Furthermore we investigated if there are specific patterns of allergic reactions to individual food allergens. Methods: Prospective analysis of patients referred from 2004-2006 for assessment of immediate type allergic reaction to food. The study has been carried out at the Food Allergy Clinic of the Children's Hospitals Aarau and Lucerne. Individuals presenting with atopic dermatitis and evidence of delayed reaction to food were not included. Diagnosis of food allergy was based on clinical history, skin Prick tests (with commercial extracts), Prick-to-Prick testing using native food and determination of specific IgE antibodies to food proteins by CAP-FEIA system. Food challenges were performed in patients without history of life-threatening anaphylaxis when the causative antigen could not be conclusively determined. Results: 278 food allergies were identified in 151 children (67% male, 33% female) with a median age of 1.9 years (range 3 months -15 years). Overall, the most prevalent food allergen is hen's egg (23.7%), followed by cow's milk (20.1%), peanut (14.0%), hazelnut (10.4%), wheat (6.1%), fish (4.3%), kiwi and soy (2.2% each) . In infancy, the most common food allergens are cow's milk, hen's egg and wheat. In the second and third year of life, hen's egg, cow's milk and peanut are predominant whereas above the age of 3 years, peanut, hen's egg and fish are the most prevalent. Urticaria is the most frequent reaction to food allergens. However, beyond urticaria, there are different patterns of clinical reactions to individual food. Cow's milk and hen's egg allergies are mainly presented with gastrointestinal symptoms. Peanut and hazelnut with angioedema. Fish with both angioedema and anaphylaxis and wheat with both angioedema and asthma. Conclusion: 8 allergens account for 83% of IgE-mediated food allergies in Swiss infants and children. In each age group there is a specific order of most prevalent offending food proteins. Furthermore, there is a specific spectrum of allergic symptoms to individual food allergens. Immediate hypersensitivity to common food allergensY YAn investigation on respiratory allergic patients in Kolkata (Calcutta), India University of Zimbabwe College of Health Sciences, Immunology, Harare, Zimbabwe. Background: Peanuts, Arachis hypogea, are the prominent sources of Immunoglobulin E (IgE) mediated allergic reactions to food items. They are highly recognised for their nutritional value of proteins and oils. The major allergens are Ara h 1 and Ara h 2 , which result in allergenic manifestations such as eczema, urticaria, angiodema, asthma and sometimes fatal anaphylactic reactions. Objective: The aim of the pilot study was to establish whether Zimbabweans had nut allergy in general and peanut allergy in particular as well as to identify clinical entities associated with sensitization. Methods: Patients for the study were selected upon review of sensitization to peanut allergens. Sensitization was diagosed using the Skin Prick Test (SPT) and the serological evidence of allergen specific IgE antibodies with the Euroimmune assay. Most patients who came to the allergy clinic had eczema, dermatitis and urticaria. Results: 63 patients had nut allergies, peanut and hazelnut.16 patients reacted to peanut only with 6 children of 2-8 years and 10 adults of 21-72 years of age.22 patients reacted to hazelnut only, with 4 children of 1-7 years and 18 adults of 18-58 years. Cross-reactivity of peanut and hazelnut was noted in 25 patients with 10 children of 3-13 years and 15 adults of 17-68 years of age. Conclusion: Allergen specific antibodies to peanut, hazelnut ,or both peanuts and hazelnuts were demonstrated. We observed overlap in reactivity of biologically different nuts one being a groundnut (peanut, Arachis hypogea) and the other one being a tree nut (hazelnut ,Coryllus avellana) in 40% of the patients, hence our interest in further investigations to establish whether it was cross-reactivity or that it was a mere co-sensitization between the two nuts. Peculiarities of a food sensitization of children with atopic dermatitis in Ukraine Viktoriia Klymenko. Kharkiv State Medical University, Propedeutic Pediatria Department N2, Kharkiv, Ukraine. Background: A food allergy affects the considerable quantity of children because of their gastrointestinal tract immaturity and low immunologic tolerance. According to the recent data of investigations from 1/3 to 1/2 children with an atopic dermatitis have a food sensitization. But the food sensitization is different in different countries because of the national food tradition and genetic factors. For example, there are not allergy reactions for peanuts and shellfishes in Ukraine but they are characteristic for American countries. From other hand, some food has changed its allergic properties. For example, all over the world it is marked increasing allergy to soy. Objective: To find out peculiarities of a food sensitization for early aged and late school aged children with atopic dermatitis in Ukraine. Methods: Sixty seven patients with atopic dermatitis were observed. The patients were divided into two groups: the first one (28 patients) aged from 3 month to 3 year and the second one (39 children) aged 12-18 year. The food sensitization was determined by the enzyme immunoassay method for the measurements of the level of allergen specific human IgEantibodies in the blood for the first group and by a skin prick testing for the second group. Results: The food sensitization was revealed in 92,9% and 71,8% patients of the first and the second group accordingly, including the high and very high reactions for 92,9% and 21,4%, monoallergy for 7,1% and 5,1% children accordingly. The high and very high levels of specific IgE were detected for antigens of a corn (42,3 %), a chicken egg protein (39,3 %), a fish (30,8 %), a rye (28,6 %), a chicken egg (26,9 %), oats (25 %), a cow's milk (23,1 %), a chicken egg yolk (21,4 %), rice (21,4 %), a tangerine (21,4 %), a lemon (21,4 %), and an orange (21,4 %) in children of the early age. The food tolerances for a corn, a rye, a cow's milk, oats, rice, a chicken egg protein, a tangerine are formed to late childhood. Certain types of food are more likely than others to be allergen for children of late school period: a chocolate (70 %), a tomato (26,3 %), a pear (23,3 %), a carrot (20,6 %), a potato (20,5 %). Nobody from 24 examined patients has sensitization to peanuts. Conclusion: The established peculiarities of a food sensitization should be taken into account at initial preventive maintenance of atopi, and also they should be used for prescription of the empirical hypoallergic diet in Ukraine and the East Europe. Sensitization to parvalbumin and collagen in 6 Japanese patients with fish allergy Michiko Hirokado 1 , Naoko Inomata 1 , Jyunko Yamaguti 1 , Katsura Yasujima 1 , Mio Kirino 1 , Megumi Kondo 2 , Kuniyoshi Shimakura 3 , Kazuo Shiomi 3 , and Zenro Ikezawa 1 . 1 Yokohama City University School of Medicine, Depertment of Enviromental Immuno-Dermatorogy, Yokohama, Japan; 2 Kndo clinic, dermatorogy, Yokohama, Japan; 3 Tokyo University of Marine Science and Technology, Depertment of Food Science and Technology, Shinagawa, Japan. Background: Fish has been reported to be one of the most important allergenic foods causing severe allergic reactions. Furthermore, parvalbumin, a small calcium-binding protein, and collagen have been identified as important cross-reactive allergens in fish. Objective: To clarify the clinical and laboratory features of allergy to fish and to detect the cross-reactive allergens by ELISA in Japanese patients. Methods: Six patients with suspected hypersensitivity to fish were subjected to skin prick tests and measurement of specific IgE antibody to fish. Additionally, specific IgE against purified parvalbumin and collagen were investigated by ELISA. Results: The patients'(1 male, 5 females) age ranged from 8 months to 22 years. All of them also had other atopic diseases, such as atopic dermatitis, bronchial asthma and pollinosis. The diagnosis based on symptoms after fish ingestion and results of skin prick tests, was anaphylaxis and oral allergy syndrome (OAS) accompanied with symptoms other than oropharyngeal symptoms in 2 and 4 patients, respectively. The causative foods were saury, yellowtail, and flatfish in 3 patients; and horse mackerel, mackerel, tuna, salmon, Atka mackerel and flatfish in 2 patients. The SPT was positive for more than three kinds of fish in all patients. The immuno CAP-RAST test showed that 5 of 6 patients were positive for cod; 3 for flatfish; 4, for horse mackerel; 5, for salmon; 4, mackerel. Furthermore, specific IgE against the purified parvalbumin and collagen was detected in 4 patients each. Discussion: The results indicated that our patients with fish allergy tended to develop severe symptoms after ingestion of different kinds of fish due to sensitization to cross-reactive allergens. Hiroshi Narita 1 , Kumiko Kidu 1 , Tsutomu Honjoh 2 , Akihiro Kimura 3 , and Junko Hirose 4 . 1 Kyoto Women's University, Food and Nutrition, Kyoto, Japan; 2 Morinaga Institute for Biological Science, Research and Development, Yokohama, Japan; 3 Itayado Clinic, Allergy, Kobe, Japan; 4 The University of Shiga Prefecture, Department of Life Style Studies, Hikone, Japan. Background: We found that significant amounts of food proteins were secreted into human and rat breast milk as immune complexes with sIgAs. To clarify a physiological consequence of this phenomenon, animal experiments have been carried out in view of allergy prevention. Results: SD rats were divided into two groups, one group (E-group) fed only egg white proteins and the other (M-group) fed only cow's milk proteins as a dietary protein source. When infant rats fed their own mother's milk were immunized with egg white proteins by using Freund's complete adjuvant, serum concentration of anti-ovomuciod IgG was lower in Einfants than in M-infants. When infant rats were immunized with cow's milk proteins in turn, serum concentration of anti-casein IgG was lower in Minfants than in E-infants. Thus, the immune tolerance against food proteins was acquired via breastfeeding from mothers fed the food containing the relevant proteins. The acquired tolerance against ovomucoid diminished spontaneously after weaning while the infants did not feed egg white proteins. Similar immune tolerance via breastfeeding were also observed in BALB/c mice. We propose the hypothesis that breastfeeding, probably through the immune complexes of food proteins and sIgAs, serves as the natural drinkable vaccine against food allergy. Oral desensitisation in children with immunoglobulin e-mediated hen's egg allergy Paolo Meglio 1 , Paolo Gianni Giampietro 1 , Simona Gianni 1 , Nadia GM Mora 1 , Paola Palazzo 2 , Enrico Scala 2 , Adriano Mari 2 , and Elena Galli 1 . 1 S.Pietro Hospital-Fatebenefratelli, Research Center, Roma, Italy; 2 IDI-IRCCS, Center Clinical Experimental Allergology, Roma, Italy. Background: Basic treatment for food allergy has been to avoid the offending item, to date. This approach is difficult in the case of common foods, and where there is a risk of severe reaction, due to child's and child's family concern of consuming the offending food, even inadvertently. In this study preliminary data regarding 3 children allergic to hen's egg (HE) orally desensitised with increasing doses of this food, are shown. Objective: To desensitise children with severe IgE-mediated HE allergy in a period of 6 months by introducing increasing daily doses of HE in order to enable the child to assume 25 ml of HE daily (corresponding to 3.2 protein grams), or to induce tolerance of the highest possible HE dose. Study Design: These are preliminary data of a larger study concerning children at least 6 years old with severe IgE-mediated HE allergy. A positive DBPCFC with HE confirmed the diagnosis. Oral desensitisation was done with increasing doses starting from 0.06 mg of HE proteins. Specific IgE to HE were evaluated through allergenic molecule-based approach with a proteomic technology (VBC Genomics microArray). Results: The 3 children achieved the daily intake of 25 ml of HE during a 7 month mean period. During this period a reduction of IgE values (kUA/L) for Gal d 1 (Ovomucoid) was observed in all cases (in one case with negative result after desensitization). No significative differences were seen comparing the result for Gal d 2 (Ovalbumin), Gal d 3 (Conalbumin) and Gal d 4 (Lysozim)(see table) . Conclusion: We successfully desensitised 3/3 children with IgE-mediated HE allergy in a mean period of 7 months. Moreover, we dramatically reduced the risk of severe reactions after accidental or unnoticed introduction of low quantities of HE. We observed a parallel reduction of specific HE IgE against Gal d 1. We do not propose generalizing this method beyond trained staff. Systemic anaphylaxis induced by lupin ingestion: two case reports Enrico Compalati, Laura Guerra, Anthi Rogkakou, Pierangela Massacane, Antonio Scordamaglia, and Giorgio Walter Canonica. University of Genoa, Allergy and Respiratory Diseases, Genoa, Italy. Background: IgE-mediated food allergy is an important cause of anaphylaxis, approximately 6% of young children and 3.7% of adults have a food allergy. The most frequent finding in food allergic patients are: peanuts, tree nuts, milk, egg, crustaceans and fish. Case Reports: We report the cases of two young women referred to our department from emergency room for food induced anaphylaxis. The first case occurred in a 42 year-old woman after few minutes from eating lupins, with the development of cyanosis, inspiratory dyspnoea and sudden lack of consciousness. She was immediately treated with epinephrine and systemic steroids with remission of symptoms. The second case occurred to a 30 year-old woman showing anxiety, general itching, urticaria, vomit, facial angioedema, hypotension and syncope after ten minutes from assumption of gluten free cookies (patient was not celiac). The sudden administration of systemic steroids and antihistamines induced patient's recovery. Both patients underwent prick test for inhalant allergens showing positivity only in the second patient for dermatophagoides, birch, parietaria and grass pollens. No positivity was observed for common food allergens mentioned above. The two patients showed marked positivity to the prick-prick test with lupin. A deep analysis of cookies-label showed the presence of lupin flour. Previous reports described the sensitivity to lupin most of all in patients with professional exposition to this legume flour. In our case reports no specific risk factors to lupin allergy nor particular predisposition to food allergens sensitisation were evident. Discussion: Lupin products are gradually being introduced into more and more foods including beverages, bread, biscuits and pasta as an additive to wheat flour substitutive of soy flour often present in food for people affected by celiac disease. In this contest allergy to lupin may be considered as an emerging condition, related to the hidden presence of this allergen or contamination, potentially dangerous for celiac patients. A case of food protein-induced enteropathy with a polypoid lesion Food protein-induced enteropathy (FPIE) is a cell-mediated hypersensitivity disorder commonly seen in the first several months of life, presenting with diarrhea and failure to thrive caused by ingestion of a certain food. A case of FPIE with a polypoid lesion has not been reported yet. We examined a case of FPIE by allergy-related blood and skin tests and endoscopy. A two month old Japanese female patient showed repeated blood streaks in her stool. She was exclusively breast-fed and did not show any other symptoms. Food protein-induced proctocolitis was considered and the symptom was resolved temporarily by eliminating cow's milk from her mother's diet. However, she developed diarrhea at 3 months of age and began failing to thrive. At 4 months of age, gross bleeding in the stool was observed 16 hours after her mother ate cake and breast-fed. The stimulation index (SI) of the lymphocyte stimulation test (LST) for cow's milk was extremely high (783 %) and her mother stopped all dairy product intake. The patient experienced gross bleeding in her stool 24 hours after eating tofu at 6 months old. Her body weight did not increase for two months. The SI of the LST for soy beans was also high at 407%. She showed positive results in patch tests for cow's milk, soy beans, wheat and pacific cod, but did not show any positive results in skin prick tests. Also, antigen specific serum IgE levels did not rise. Fecal examinations were repeated but eosinophils and neutrophils were not found and microbiological results were all negative. For further investigation, we performed a colon endoscopy under general anesthesia. Lymphoid nodules and mucosal edema were found in the colon. A 7 mm polypoid lesion was found on the ileocecal valve and juvenile polyposis was considered. However, the biopsy specimens showed non-specific inflammation with massive lymphocyte infiltration and did not indicate a solid tumor. Food protein-induced enteropathy was diagnosed. She stopped breast-feeding and started to take highly hydrated cow's milk (MA-1, Morinaga Japan) and rice and vegetables as solid foods. She has not shown gross bloody stool since, and her body weight has increased dramatically. The endoscopic findings improved after 2 months of dietary treatment. In infants with intestinal bleeding and failure to thrive, the influence of food allergies should be considered and the LST, patch test and an endoscopic examination should be performed. The investigation of cases which performed provocation test on Food-dependent exercise induced anaphylaxis for the last 8 years Junko Ikei, Yoko Murakami, Rumiko Shibata, and Sankei Nishima. Fukuoka National Hospital, Pediatrics, Fukuoka City, Japan. Background: Food-dependent exercise induced anaphylaxis (FDEIAn) is a severe systemic syndrome induced by physical exercise after food ingestion. In recent years there has been an increasing number of reports of FDEIAn, but the actual situation is not known well. Objective: To investigate the symptom of FDEIAn and grasp the actual situation of provocation test we investigated about 19 cases that was doubted FDEIAn, and were admitted for the purpose of detailed examination for the last eight years. Methods: Nineteen children (16 boys and 3 girls) 19 cases were enrolled in this study. Age at onset was from 8 to 16 years old (average age 11 years old). IgE was measured by CAP-FEIA (Pharmacia) and skin prick test was examined in need. Provocation test by combination of food and exercise test was performed for 18 patients and 4 of them with preadministration of aspirin. Results: As for the symptom at the onset, facialangioedema or blepharedema was present in 12 cases and urticaria in 10 cases, respiratory symptom in 13 cases ,hypotension in 5 cases, unconsciousness in 4 cases. Intramuscular epinephrine injection was medicated for 4 cases. The causative food allergen was as followed, wheat in 8 cases, crustacea in 7 cases, vegetables / cone / milk / cheese for each 1 case, unknown in 4 cases. We could be able to estimate food antigen by medical history in 17 cases. Related food specific IgE was positive in 5 of them. Twelve cases were negative CAP-RAST ,and 8 of them showed positive SPT. Two cases were not be able to estimate antigen by medical history and negative CAP-RAST. And one of them showed positive SPT. Four cases in 15 were positive by provocation tests with combined foods and exercise. We performed administration of aspirin before ingestion of food and exercise for 4 examples, symptom was provocated in only one case. Conclusion: Wheat or Crustacea was a major food allergen of FDEIAn, but few case were caused by vegetables or fruit. Combination of CAP-RAST and skin prick test was useful to specify the allergen. In our cases, a positive rate was 50% by combination of food and exercise test or with preadministration of aspirin. It is need to establish about laboratory methods on children with FDEIAn such as food loading dose, exercise methods, and premedication of aspirin in future. IgE mediated anaphylaxis to sesame Emmanuella Guenova 1 , Sonia Genova 2 , Bogomil Voykov 3 , and Silvia Novakova 2 . 1 Eberhard Karl University, Department of Dermatology, Tuebingen, Germany; 2 University Hospital, Department of Allergology, Pleven, Bulgaria; 3 Eberhard Karl University, Department of Ophtalmology, Tuebingen, Germany. A 70-year-old Caucasian man presented with suspected food allergy. In the past emergent medical help was needed, because of an acute systemic anaphylactic reactions, timely associated with sesame ingestion. 15 minutes after consumption of sesame-poppy-raisin sweets suffered the patient from generalised urticaria, facial and laryngeal oedema, severe hypotension and unconsciousness. Emergency treatment with intramuscular administration of epinephrine and high doses corticosteroids and antihistamines intravenously was needed. Previous allergic reactions were denied. The patient stated occasional contact to sesame throughout his life span and non-contributory further own and familial anamnesis. Skin prick tests to common inhalant allergens, preservatives and food allergens were negative. Prick-to-prick tests with poppy seed, raisins and wheat flour remained negative as well. Surprisingly, skin prick-to-prick test with sesame seed and sesame oil revealed a pronounced positive reaction. The specific IgE for sesame was highly positive (8,37 kUA/l) whereas total serum IgE (93,5 kUA/l) and basal serum tryptase level (3,1 2g/l) remained in the normal range. A double blind, placebo-controlled food challenge with incremental doses of sesame seeds was conducted. 15 minutes after distributing 100 mg backed sesame seed, acute anaphylaxis occurred. The IgE mediated mast cell activation was proven by increase of the serum tryptase level (17,52g/l) two hours after the food challenge. Prevalence of allergy to sesame (Sesamum indicum) with its major allergens Ses i 1-7 becomes more significant due to its increasing industrial importance. Sesame represents an insidious Bhidden allergen[, found not only in alimentary products, but also in cosmetics and medical products, especially sun-protection creams and solvents for intramuscular injections. The incidence of food allergy is age-dependent, particularly affecting children and young patients. In elderly, anaphylactic reactions tend to be less severe and a primary manifestation is unusual. Nevertheless we were able to diagnose the rare case of classic IgE mediated anaphylaxis to sesame with primary manifestation in a 70-year-old patient. This shows that the potential life-threatening severity of food anaphylaxis, especially to foods with pronounced allergenic properties like sesame, should not be underestimated, irrespective of the patient's age. Assessment of quality of life in children with food allergy Noriko Minamitani 1 , Takanori Imai 2 , Takatsugu Komata 2 , Mika Ogata 3 , Chizuko Sugizaki 1 , Morimitsu Tomikawa 2 , Hiroshi Tachimoto 2 , and Motohiro Ebisawa 1 . 1 Sagamihara National Hospital, Clinical Research Center, Department of Allergy, Sagamihara-city, Kanagawa, Japan; 2 Sagamihara National Hospital, Department of Pediatrics, Sagamihara-city, Kanagawa, Japan; 3 National Hospital Organization Kumamoto Medical Center, Department of Pediatrics, Kumamoto, Japan. Objective: The dietary life of children with food allergy (FA) is restricted very much, and their quality of life (QOL) of diet would be affected. Therefore we have to perform dietary management for such children, but we have not yet known how they are restricted in their dietary life. We conducted current study to clarify the problems of dietary life in children with FA in our outpatient clinic. Methods: By questionnaire we examined QOL of diet (burden of the dietary life at home and outside, problems of nourishment, and parents' psychological burden) against the parents of children with FA or without FA (control group). In addition, only to parents of children with FA, we interviewed to qualify the troubles they had in the dietary life just after the diagnosis of FA and at the some time after starting elimination diet. Results: A total of 544 patients was surveyed. In the QOL survey, 40.1% of the children with FA had different menu from the family's menu, whereas only 21.7% of the control subjects did. In addition, 69.0% of parents of children with FA had difficulty in making menu, and 96.8% of the parents paid attention to food labeling, and 40.1% of those felt an economic burden for purchasing foods for FA children. Furthermore, 54.1% of the children with FA were not satisfied with eating-out, whereas only 35.4% of control was not satisfied. Children with FA were satisfied with the quality of school lunch by 71.9% but 95.4% for control. The most prominent problem just after the diagnosis of FA was about Bmaking menu / cooking[ in all age group. It was also a problem for the mothers to wean FA babies during infancy. However, a tendency to diversify troubles by age was recognized after starting elimination diet, such as problems concerning Bpurchase foods and eating-out[, or Bschool lunch[. Conclusion: The study revealed that children with FA were restricted in terms of the dietary life, and that their QOL was deteriorated compared with control group. Since we could obtain information of the dietetics of which children with FA and their parents desired, it is important to pursue appropriate dietary management for the children with FA. Imatinib decreases duodenal mast cell density and degranulation in epicutaneously-sensitized food allergy model of mouse Kirsi Vaali 1 , Aung Htun Lin 1 , Lizhi Sun 1 , Petri Kovanen 2 , Arnold Berstad 1 , and Kari Eklund 3 . 1 University of Bergen, Institute of Medicine, Section of Gastroenterology, Bergen, Norway; 2 Wihuri Research Institute, Helsinki, Finland; 3 Joint Authority for the Hospital District of Helsinki, Division of Rheumatology, Helsinki, Finland. Background: Balb/c mice, when sensitized epicutaneously with ovalbumin, had increased serum ovalbumin-specific IgE as well as IgG1. After repeated intragastric challenges (ig) with ovalbumin, mice developed acute diarrhea within 60 min of the 6. ig dose, whereas there was no diarrhea nor increased IgE or IgG1 in the control groups. Since the role of small intestinal mast cells was undisputable in this model, we wanted to evaluate the effect of imatinib on these cells. Imatinib (GlivecR) is a potent inhibitor of the most important growth factor receptor of mast cells, the c-kit. Methods: Imatinib (100 mg/mouse) was given orally to Balb/c female mice (20-25 g) as a pre-treatment during each 4 sensitizations and every second day from the day 39 at the same time with the 10 ig ovalbumin (50 mg/mouse) challenges. Ovalbumin-specific IgE, IgG2A, and mucosal mast cell protease 1 (MMCP-1) were measured from the serum and histamine from the plasma with standard ELISA methods. Rectal temperatures were measured and diarrhea evaluated before and after the 6.-10. ig challenge of ovalbumin. Chloroacetate esterase (CAE) was used for general mast cell staining, for immunohistochemical staining MMCP-1 antibody was used. For double staining fluorescent method with MMCP-1 antibody and TUNEL methods were used. The total amount of stained cells was calculated from the whole crosssectional specimens of duodenum. Results: There were no significant differences in the levels of ovalbuminspecific IgE or IgG1 between the imatinib-treated and the vehicle treated ovalbumin-sensitized mice indicating that the treatment did not prevent the sensitization of the mice. The serum MMCP-1 levels at the sacrification were lower in the imatinib-treated mice than in the vehicle treated group (10100 T 10967 and 14173 T 11246 ng/ml, respectively). Imatinib decreased the number of CAE+ cells (59 T 32 and 126 T 53 cells/mm 2 , p G 0.05) and MMCP-1+ cells (13 T 15 and 65 T 47 cells/mm 2 ), respectively. The combined MMCP-1 and TUNEL staining indicated that the duodenal mast cells did not go into apoptosis during the treatment. Conclusion: Imatinib treatment indicates the important role of the intestinal mast cell population, CAE+ and MMCP-1+ cells, of which MMCP-1+ cells seems to represent the majority of intestinal mast cells in the mouse food allergy model. Identification and immunochemical characterization of a common allergenic pollen Cycas circinalis Kashinath Bhattacharya. Visva Bharati University, Santiniketan, Botany, Santiniketan, India. Objective: Cycas circinalis is a common garden plant. The pollen grains of Cycas circinalis are airborne and remain suspended in the air from May to June with a peak concentration at the end of June. The allergenicity of the pollen grains is very common among the local inhabitants, but has not been reported so far. Materials and Methods: The pollen extract showed high (25.6%) skin reaction on 172 respiratory allergic patients. Sera from the subjects were tested directly by ELISA for estimating the allergen specific IgE. The results were expressed as P to N value (ratio of patient's sera in respect to normal one). The allergic fraction was separated in 12% SDS-PAGE. Results: The results showed the presence of 14 components in the range between 14-100 kDa. Sera of eight patients showing P to N value greater than 3.5 were used for immunoblotting to identify the specific IgE binding protein. The proteins from the gel were transferred to PVDF membrane. Immunoblotting was performed using eight patients sera, where anti human IgE-AP conjugate was used as second antibody and NBT and BCIP as substrate. The results showed two components of 20 kDa and 39kDa. Thus, the Cycas circinalis pollen grain is an important aeroallergen in India. The 20 kDa and 39kDa components are the major allergens present in the pollen extract. The correlation between SNPs of GATA-3 and Chinese with allergic rhinitis XiangDong Wang, Luo Zhang, and Demin Han. Head and Neck Surgery of Beijing Tongren Hospital affiliated to Capital Medical University, Beijing Institute of Otorhinolaryngology; Departme, Beijing, China. Objective: To discuss the correlation between SNPs of GATA-3 and Chinese with allergic rhinitis. Methods: 109 patients and 112 normal control people were enrolled in this study. We detect SNPs of two sites of rs1269486 and rs2229360 with RFLP. Results: We detected SNPs in both of the two sites. The genotypes of rs1269486 are G/G, G/A and A/A, and the genotypes of rs2229360 are C/C, C/ T and T/T. The distribution of G/G in patients was significantly higher than that in control group (Pgƒ0.01). The haplotype frequency of GC in patient group was significantly higher than that in control group (Pgƒ0.05), and the haplotype frequency of AC in patient group was significantly lower than that in control group (Pgƒ0.01). Conclusion: This is the first study to evaluate the association between SNPs of GATA-3 and allergic rhinitis, and we found that the SNPs of rs1269486 was significantly different between Chinese with allergic rhinitis and normal control people. The SNPs of GATA-3 may influence the phenotype of allergic rhinitis in Chinese. A novel approach to IgE epitope mapping Y Y virtual epitope mapping Fabio Dall'Antonia, and Walter Keller. University Graz, Inst. of Chemistry, Graz, Austria. Background: The determination of cross-reactive IgE epitopes on allergens is a major challenge. Mapping of discontinuous (=structural) IgE epitopes by experimental methods is extremely time consuming and possible for only few allergens and epitopes. Most structural methods for epitope determination (e.g., co-crystallization of Allergen-Fab complexes, NMR-based hydrogen exchange or MS-based methods) work only with monoclonal antibodies and therefore epitopes can only be determined one at the time and never represent the polyclonal situation found in reality. The mapping of allergen specific sequence motives onto the surface has been used or prediction of putative epitopes. However, sequence homology (alone) may be a weak predictor for cross-reactivity, as structural conservation is bound to be a major determinant of cross-reactivity. In order to improve the accuracy of the prediction of structural epitopes, we have developed a new approach combining the 3D-structural information and surface topology with the information on sequence conservation. Methods and Results: The computational methods for structure based epitope mapping are integrated in a suite of programs which combine public domain programs for sequence and structure alignment with topological comparison algorithms and evaluation of chemical properties of the solvent exposed surface. A program suite consisting of several Perl scripts, embedded into a graphical user interface written in Tcl / Tk, provides automated surface comparison. The following steps are executed by the program: &Superposition of two or more user selected atomic models &Calculation of solvent-accessible surfaces of the aligned structures &Surface comparison (calculation of differences) &Assign similarity scores to surface elements (including topology, chemical similarity, H-bonding capacity, etc.) &Averaging of similarity scores for each amino acid residue &Graphical display of results The program was tested with two families of cross-reactive allergens, where the structures of three or more members have been determined. Conclusion: We developed a novel method of IgE epitope mapping based on the structural information of two ore more cross-reactive allergens. The method uses sequence conservation as well as topological similarity for the delineation of putative IgE epitopes. The graphical user interface allows for the interactive visualization of the epitopes. Exosomes in human blood plasma contain RNA Y Y a genetic signal between cells at a distance? V. Seyed Alikhani, H. Valadi, K. Ekström, A. Bossios, M. Sjöstrand, and J. Lötvall. Göteborg University, Department of Internal Medicine, Respiratory Medicine and Allergology, Göteborg, Sweden. Background: Exosomes are vesicles of endocytic origin produced by many cell types and released in the extracellular environment as 30?100 nm large membrane vesicles. Exosomes are involved in different signalling events including antigen presentation to T-cells and development of tolerance. It has recently been shown that mast cell exosomes contain a substantial amount of functional RNA (Valdi et al., Nat Cell Biol 2007) . The aim of this study was to investigate weather exosomes in human blood plasma contain RNA and/or DNA. Methods: Peripheral blood plasma was derived from healthy blood donors and exosomes were isolated by a series of ultracentrifugations and filtrations. Purified exosomes were attached to anti-MCH class II coated dynabeads and analysed using FACS. The tetraspanins CD 63, CD 9 and CD81, known to be enriched in exosomes, were used as markers for exosome detection. We also performed electron microscopy (EM) to detect exosomes in the sample. RNA was precipitated using TrizolÛ according to the manufacturer?s protocol. To magnify the signal, the RNA was then converted to DNA using a random primer, and separated by agarose gel. We also confirmed the presence of mRNA in exosomes by using oligo d(t) primers for conversion of mRNA to cDNA. Results: The isolated vesicles from plasma were identified as exosomes by both EM and flow cytometric analysis (FACS). The results from the EM showed exosomal structures with a size of 30?100 nm in diameter. The FACS analysis showed the presence of MHC II, CD63, CD9 and CD81, all markers of exosomes. The exosomes also proved to contain significant amounts of RNA, but no DNA. Some of the exosomal RNA was identified as mRNA. Conclusion: Our results show the presence of RNA-containing exosomes in human plasma. Since exosomes have the ability to fuse with cells and deliver their RNA to that cell, it is possible that the RNA in plasma exosomes constitute a genetic signal between cells at a distance. Background: Allergic airway diseases are characterized by inflammation and hyperresponsiveness. There is evidence that, in the upper airways, hyperresponsiveness can be attributed to increased neural activity. Nerve growth factor (NGF) is a neurotrophin that has been shown to be an important mediator of neural responsiveness in the airways of bronchial asthma patients. Recent studies suggest that NGF may also be an important inflammatory mediator. Therefore, NGF could be a link between allergic inflammation and neural responsiveness However, expression of NGF has not been reported in the nasal mucosa of allergic rhinitis patients. This study was performed to investigate the expression of nerve growth factor (NGF) messenger RNA and protein and to localize the NGF protein in the nasal mucosa of patients with perennial allergic rhinitis and control subjects. Methods: Inferior turbinate mucosa samples were obtained from 20 patients with perennial allergic rhinitis and 20 matched non-allergic patients. NGF messenger RNA was extracted from the inferior turbinate mucosae, and then reverse transcription-polymerase chain reaction was performed. Western blotting was used to analyze differences in expression levels of NGF protein between patients with perennial allergic rhinitis and normal controls, and the NGF protein was localized immunohistochemically. Results: The expression levels of NGF messenger RNA and protein in the nasal mucosa were significantly increased in patients with perennial allergic rhinitis compared with those in controls. NGF protein was expressed in the epithelium, infiltrating inflammatory cells, and submucosal glands. Conclusion: This study provides evidence of NGF expression in the human nasal mucosa and its increased expression in perennial allergic rhinitis. These results suggest that NGF may contribute to the pathogenesis of perennial allergic rhinitis. Heung-Man Lee 1 , Hee Joon Kang 1 , Hyo-Hyun Park 2 , and Yong-Dae Kim 3 . Background: The exact etiologic mechanisms leading to the formation of nasal polyps have remained largely obscure. However, the formation of new blood vessels (angiogenesis) may play an important role in the development of nasal polyp. Angiogenin, a heparin-binding 14 kDa plasma protein with angiogenic activity, plays an important role in inducing angiogenesis. The aim of the present study was to investigate the expression and distribution of angiogenin in normal nasal mucosa and in nasal polyps and to evaluate the implication of such expression as regards the development of nasal polyps. Methods: Normal inferior turbinate mucosa was obtained from 20 patients undergoing surgery for augmentation rhinoplasty. Nasal polyps were obtained from 20 patients undergoing endoscopic sinus surgery for chronic polypoid sinusitis. Semi-quantitative reverse transcriptase-polymerase chain reaction was performed for angiogenin mRNA. Sections were immunostained by using specific antibodies for localization of angiogenin and quantitatively analyzed using computer-based image analysis. Western blot analysis was done. Results: Using immunohistochemistry, moderate to high levels of angiogenin were mainly localized in the infiltrating inflammatory cells of nasal polyps, and faint staining was found in normal turbinate mucosa. Semiquantitative reverse-transcriptase polymerase chain reaction and Western blot analysis showed that angiogenin expression was increased in nasal polyps compared with that in normal turbinate mucosa. Conclusion: The markedly increased expression of angiogenin in nasal polyp compared with normal nasal mucosa suggests that angiogenin may play a significant role in the formation of nasal polyp. Proteomic identification of known and novel allergens from house dust mite Dermatophagoides pteronyssinus Pattama Ekpo 1 , Sirichit Wongkamchai 2 , Sunthorn Puangyoy 1 , Nhujohn Jansong 1 , Supachok Sinchaikul 3 , Shui-Tein Chen 3 , Chaweewan Bunnag 4 , and Visith Thongboonkerd 5 . 1 Faculty of Medicine Siriraj Hospital, Mahidol University, Immunology, Bangkok, Thailand; 2 Faculty of Medicine Siriraj Hospital, Mahidol University, Parasitology, Bangkok, Thailand; 3 Institute of Biological Chemistry and Genomic Research Center, Academia Sinica, Taipei, Taiwan; 4 Faculty of Medicine Siriraj Hospital, Mahidol University, Otorhinolaryngology, Bangkok, Thailand; 5 Faculty of Medicine Siriraj Hospital, Mahidol University, Office for Research and Development, Bangkok, Thailand. Background: Dermatophagoides pteronyssinus is the most predominant house dust mite (HDM) species and the most important source of indoor allergens in Thailand. Currently, there is no subunit vaccine available for successful prevention of the allergy to HDM. Characterizations of all potential HDM allergens that cause IgE production in the allergic patients are needed for development of a proper vaccine. The present study was therefore conducted to globally identify potential allergens from D. pteronyssinus using a proteomics approach. Methods: Proteins derived from D. pteronyssinus cells were resolved by twodimensional gel electrophoresis , transferred onto a nitrocellulose membrane, and subjected to immunoblot analysis using pooled HDMsensitive patients' sera (n = 39) or pooled normal control sera (n = 5) as the source of primary antibody. The membrane was then probed with mouse antihuman IgE ("-chain specific) conjugated with horseradish peroxidase and immunoreactive proteins were then visualized using a chemiluminescence substrate. The corresponding spots in another parallel 2-DE gel stained with Coomassie Brilliant Blue that matched to these immunoreactive spots were excised and identified by quadrupole time-of-flight (Q-TOF) mass spectrometry (MS) and/or tandem MS (MS/MS). Results: A total of 33 immunoreactive spots were observed using the pooled patients'sera, whereas none was detected with the pooled control sera. Among these, 23 were successfully identified by Q-TOF MS and/or MS/MS analyses, including Der p 1, Der p 2, Der p 8, Der p 14 and other proteins that had not previously been recognized as the HDM allergens. Quantification of Fc(RI-bindable human IgE using chimeric EGF receptor Ryosuke Nakamura, Reiko Teshima, and Jun-ichi Sawada. Natl. Inst. Health Sci., Div. Biochem. Immunochem., Tokyo, Japan. Background: Quantification of human IgE (hIgE) in the patients' sera is important for the diagnosis of allergy. Most of conventional assay methods of hIgE depend on the binding of anti-hIgE antibody and hIgE in the sera. However, the hIgE concentration in the sera quantified by such methods is not necessarily correlated to the activation of mast cells. One possible reason is anti-hIgE autoantibody in the sera, which may interfere binding of hIgE to high-affinity IgE receptor (Fc(RI). Therefore, it is important to develop a new method which can detect hIgE actually contributed to the sensitization of mast cells. In this study, we have tried to establish new quantification method of surface-bound hIgE using chimeric receptor bearing the extracellular region of Fc(RI " subunit and intracellular region of EGF receptor (EGFR). Methods: The cDNAs encoding chimeric receptors with or without intracellular EGFR region were made by fusing cDNAs of Fc(RI" and EGFR based on PCR. The transmembrane domains of the chimeric receptors were from those of both receptors. The cDNAs were inserted to a mammalian expression vector, and the plasmids were transfected to HLR-Elk1 cells, which are HeLa cell-derived cell line containing Elk1-transactivable luciferase gene. The expression of the chimeric receptors were determined by confocal microscopy and Western blotting. The phosphorylation of intracellular signaling molecules induced by hIgE sensitization and crosslinking with anti-hIgE was investigated by Western blotting. The crosslinking of the chimeric receptors were measured by luciferase activity. Results: Confocal microscopic analysis revealed that the chimeric receptors with transmenbrane domain of EGFR were efficiently expressed on the plasma membrane of HLR-Elk1 cells, however, those of Fc(RI" were less expressed on the cell. Addition of hIgE to the chimeric receptor-expressing HLR-Elk1 cells caused significant phosphorylation of the chimeric receptor and MAPK, and the phosphorylation was increased when anti-hIgE was applied. However, crosslinking of hIgE with anti-hIgE did not augment luciferase expression induced by hIgE addition. Conclusion: The cells expressing Fc(RI-EGFR chimeric receptors could be a useful diagnostic tool that detects binding of hIgE to the cell surface Fc(RI. However, a new method to detect crosslinking of Fc(RI should be developed. AllFam Y the database of protein families of allergens Christian Radauer 1 , Merima Bublin 1 , Stefan Wagner 1 , Adriano Mari 2 , and Heimo Breiteneder 1 . 1 Medical University of Vienna, Department of Pathophysiology, Vienna, Austria; 2 Allergy Data Laboratories, Instituto Dermopatico dell´Immacolata, Latina, Italy. Background: Along with the identification of a large number of allergenic proteins from diverse sources, it has become clear that most allergens belong to a limited set of protein families. Most currently available allergen databases such as the official list of allergens (www.allergen.org) group allergens mainly by source organisms and do not take into account evolutionary relationships among allergens. Hence, we established AllFam, a database of protein families of allergens. Methods: Allergen data were downloaded from the Allergome (www.allergome. org) database. Protein sequences from the Uniprot (www.expasy.uniprot.org) database were compared to protein family definitions from the Pfam (www.sanger.ac.uk/Software/Pfam). The AllFam database combines these protein family data with exposure and source data from the Allergome. AllFam contains a fact sheet for each allergen family with information on biochemical properties and allergological significance. Results: The Allergome database contained 831 allergen sequences not including isoallergens. The sequences were classified into 182 protein families. However, only 94 families contained more than one allergen. The most abundant allergen families were the prolamin superfamily that comprised several subfamilies, and the profilins, a ubiquitous group of cross-reactive plant allergens. The family of EF hand containing calcium-binding proteins included polcalcins from pollen and parvalbumins from fish and amphibians. Allergenic tropomyosins were found as food allergens in crustaceans and molluscs and as inhalative allergens in mites and cockroaches. A classification of allergens by source revealed a similar number of protein families among plant and animal allergens (72 and 79 families) and a considerably lower number of fungal allergen families (50). Grouping of allergens by route of exposure showed that the distribution of food allergens is biased towards a few abundant protein families. The database contained 64 families of food allergens with the seven most abundant families accounting for 55% of all allergens. In contrast, there are 125 families of inhalative allergens with the first seven families making up only 27% of all allergens. Background: Aspergillus species are ubiquitous saprophytic fungi that cause a variety of diseases, ranging from hypersensitivity reactions to flu-like pneumonia and life-threatening invasive aspergillosis. As the lung is the primary site of initial infection with airborne conidia, we investigated the innate immune responses of bronchial epithelial cells against different forms of Aspergillus. Methods: Human bronchial epithelial cells were treated with equal numbers of killed spores or germ tubes of either Aspergillus fumigatus or Aspergillus flavus. Transcriptional activation of the cytokine genes in the presence of germ tubes or spores was determined using real-time PCR analysis. Secreted cytokines following treatment with the mentioned fungi were also measured by enzyme linked immunosorbant assay. Results: Analysis by real-time PCR showed that inflammatory cytokines such as IL-8 and IL-6 as well as the proinflammatory protease, caspase-5 were strongly upregulated by both treatments in a dose-dependent manner. Consistently, germ tubes induced a stronger response than spores. TNF-" and $-2-defensin were induced by high a concentration of germ tubes, but not by spores. Furthermore, IL1 pretreatment highly induced the expression of $-2-defensin, IL8 and IL12. Conclusion: Taken together, our results show that germ tubes of Aspergillus fumigatus and flavus are potent inducers of innate immune responses in human airway cells. Considering the presence of Aspergillus spores in the air, differentiation between transient spore contact and invasion, as represented by germ tube formation, is important in order to determine proper immunological response. Moreover, these results can also provide additional data in understanding pathophysiology of hypersensitivity reactions due to the aspergilli. Background: The importance of indoor bioaerosols has been emphasized in the last decades. It is the fact that hospital environment is often contaminated with fungal spores and control of the air quality in hospital wards has assumed great importance. Exposure to the spore floating on the air in hospital wards leading to appearance of several diseases that ranges from allergies to sever fungal infections in hospitalized patients. The aim of this study was to identify and evaluation of fungal air contamination rates in high risk wards of a teaching hospital in Shiraz, southern Iran. Materials and Methods: A settle plate assay was used for air sampling. Petri dishes containing Sabouraud and Malt extract agar were opened and exposed in high risk wards of a teaching hospital and incubated for five to ten days. The colonies are then identified, enumerated, and reported as the number depositing in one square meter of surface rooms. Results: A total of 528 microbial colonies were detected which 223 (42.23%) colonies yielded fungi. Fungal isolates belongs to 12 genera as follows: Cladosporium, Aspergillus, Penicillium, Alternaria, Trichosporon, Aureobasidium, Mucor, Rhizopus, Chrysosporium, Curvularia, Trichothecium, Stachybotrys, Stemphylium, Acremonium, Bipolaris and Yeast. 4.25 percent of rooms were as clean room and Cladosporium was the predominant fungal isolates. The highest and lowest fungal spore concentration were 1015 and zero colony forming unite in emergency and transplantation rooms, respectively. Conclusion: Cladosporium, Aspergillus and many other fungi are the major aeroallergens and causative agent of nosocomial infections. Detection of these fungi from hospital area has a potential risk of infection to these diseases. TH1, Treg and TH2 immunoregulation through TLR2 and TLR4 by Lactobacillus plantarum IS-10506 probiotics in reducing allergic reaction. Experimental study in mice subjects Anang Endaryanto 1 , Subijanto Marto Sudarmo 1 , Suhartono Taat Putra 2 , Ingrid S Surono 3 , S Suwarno 4 , and K Kusnoto 5 . 1 School of Medicine, University of Airlangga, Child Health Department, Surabaya, Indonesia; 2 School of Medicine, University of Airlangga, Pathobiology Department, Surabaya, Indonesia; 3 University of Indonesia, Seameo-Tropmed RCCN, Jakarta, Indonesia; 4 School of Veterinary Medicine, University of Airlangga, Veterinary Microbiology Department, Surabaya, Indonesia; 5 School of Veterinary Medicine, University of Airlangga, Veterinary Parasitology Department, Surabaya, Indonesia. The purpose of this study is to clarify the mechanisms and involvement of Toll-like Receptor (TLR) 2 and 4 in reducing allergic reaction by Lactobacillus plantarum IS-10506 (LIS) probiotics. In the factorial design study, Balb/c mice subjects were randomized into 6 experimental groups and were sensitized with OVA. Probiotics (LIS and placebo) and TLR inhibitors (PD98059, LY294002 and placebo) are independent variable. The expression of innate immune response (TLR2 and 4, NF.Bp65, NF.Bp105/p50); the activation of adaptive immune response (IFN,, IL-2, IL-4, IL-5, TGF$ and IL-10, sera OVA-specific IgG2a, IgA, IgE, ileum OVA-specific IgA and total IgE); and the allergic reaction (histamine and symptoms score) are dependent variable. The TLR2 and 4, NF.Bp65, NF.Bp105/p50, IFN,, IL-2, IL-4, IL-5, TGF$ and IL-10-positive cells were measured on histological slices from the small intestine of mice in all groups. Levels of sera OVA-specific IgA, IgE, IgG2a and ileum OVA-specific IgA were determined with an indirect ELISA. Measurement of total IgE antibodies in sera was done with Sandwich ELISA. Histamine levels were determined with a competitive ELISA. After final OVA sensitization, the LIS groups did not experience the severe and fatal allergic reaction, however most of mice in control group did experience. There were significant modulation of the innate and adaptive immune response in the LIS group compared with the control group, except IL-4 and total IgE. In control group, the low level of innate immune response (TLR2 and 4, NF.Bp65, NF.Bp105/p60) was in conformity with TH2skewed (TH1 G Treg G TH2) and IgE-mediated mast cell degranulation which produce histamine and clinical symptoms. LIS probiotics enhanced the innate immune response (TLR2 and 4, NF.Bp65, NF.Bp105/p60), formed the TH1skewed (TH1 9 Treg 9 TH2), and reduced the mast cell degranulation (decreasing the level of histamine and symptoms score). The involvement of TLR2 and 4 in the effect of the LIS probiotics on decreasing of allergic reaction was proved by the interaction effect between those probiotics and inhibitor TLR in reducing allergic reaction. In conclusion, LIS probiotics is involving TLR2 and 4 in reducing allergic reaction through enhancement of the TH1 and Treg immune responses and forming a new TH1-TH2 equilibrium without reducing the level of TH2. Identification of interferon-inducible gene expression in fibroblasts derived from allergic and nonallergic patients M Yoshikawa 1 , K Wada 1 , T Yoshimura 1 , N Okada 1 , H Saito 2 , and H Moriyama 1 . 1 Jikei University School of Medicine, Department of Otorhinolaryngology, Tokyo, Japan; 2 National Research Institute for Child Health and Development, Department of Allergy and Immunology, Tokyo, Japan. Rationale: Recently, asthmatic tissue-derived cells are reported to retain their characteristic features such as steroid insensitivity even when cultured for a long time. In our previous study, we identified many differences in gene expression patterns containing interferon-inducible CXC chemokine genes in fibroblasts derived from allergic (AF) and nonallergic (NF) patients, although the fibroblasts were cultured and stimulated under the same conditions. Therefore, we determined differences in gene expression of IP-10/CXCL10 and I-TAC/CXCL11 between AF and NF. Methods: Nasal fibroblasts were isolated from inferior turbinate mucosa from allergic (AF) and nonallergic (NF) patients. Cells were stimulated with IL-4, TNF-", IFN-,, IFN-$, poly I:C or their combination. After mRNA was extracted, the expression of IP-10/CXCL10 and I-TAC/CXCL11 was examined using real-time PCR. Results: IP-10/CXCL10 and I-TAC/CXCL11 were more down-regulated when stimulated with IL-4 and TNF-" in AF compared to NF, whereas their expression levels were increased when treated with TNF-" alone in AF. In addition, we found mRNA expression of these genes was significantly more strongly up-regulated by TNF-" in AF than in NF. Conclusion: We identified specific gene expression profiles containing interferon-inducible CXC chemokine genes in AF and NF. The expression of IP-10/CXCL10 and I-TAC/CXCL11 was up-regulated in AF compared to NF even after cultured for several weeks in the same culture condition, suggesting genetic or epigenetic modification may be involved in its mechanism. Additional studies are needed to clarify the exact mechanism underlying this finding. Novel two series of benzo[b]furan derivatives having potent and selective LTB4 (BLT1and/or BLT2) antagonist Yoshitaka Ohishi. Mukogawa Women's University, Faculty of Pharmaceutical Sciences, Hyogo, Japan. Leukotriene B4 is a dihydroxy fatty acid produced mainly by macrophages and neutrophils. LTB4 plays important physiological roles on leukocytes trafficking to the site of infection and clearance of invaded microorganisms. However, overproduction of LTB4 is reported to cause inflammatory diseases including bronchial asthma and inflammatory bowel diseases. Thus, much work has been done to prepare LTB4 antagonists for clinical use as anti-inflammatory drugs. Unfortunately, no antagonist has yet been developed for clinical medicinal application. Recently, a novel cell surface receptor (BLT2) for LTB4 has been isolated and its molecular cloning has also been established. Current studies on LTB4 antagonists and its receptors (BLT1, BLT2) suggest the possibility of the development of new clinical drugs for treating arteriosclerosis, immunosuppression of allograft rejection in organ transplantation, psoriasis, cancer, and rheumatoid arthritis. Stable compounds with definite conformation should be favorable as antagonists. Thus, we prepared two series of benzo[b]furan derivatives, 2alkylcarbamoyl-1-methylvinylbenzo[b]furans (1) and 2-(4-thiazolyl)-3-phenylbenzo[b]furans (2) , and evaluated their inhibitory activity for BLT1 and /or BLT2. Some of them showed more potent and more BLT2 selective inhibitory activity than positive standard compound ZK158252. Especially, 2-(2-N,Ndiethylcarbamoyl-1-methylvinyl)-7-(1-phenylethyl)benzo[b]furan ((E)-1c) and 2-[2-[(dimethylamino)methyleneamino]-5-formylthiazol-4-yl]benzo[b]furans (2a) showed potent and BLT2 selective inhibitory activities. These compounds will be assay for treatment ability concerning immflamatory and asthma in vivo screening. Relation of IgE antibody to antigen 5 of echinococcus granulosus Farideh Bagheri, Mohammad Reza Siavashi, and Alireza Khabiri. Pasteur institute of Iran, Parasitology, TEHRAN, Islamic Republic of Iran. Anaphylactic reactions, such as urticaria, edema, respiratory symptoms, and anaphylactic shock often complicate the course of Cystic Echinococcosis (CE). To investigate the role of the IgE immunoreactive Antigen 5 (Ag 5) in the sero-positive patients with CE, we determined Nterminal of 57 kDa subunit of Ag5 responsible for IgE and C-terminal of this active antigen related to induction of IgG specifically. Immunoblotting analysis showed that specific IgE to 57-kDa subunit related to inter-chain disulphide band of two 22 kDa and 38-kDa component of Ag5 and conformational epitope on this subunits. In addition, IgE specifically recognized N-terminal of 22 kDa subunit which remain bounds to the other component, whereas IgG reacted with C-terminal of 38 kDa component of Ag5. Recognation of the specific binding site on the 57 kDa subunit of Ag5 could leads to understanding the mechanism regulating IgE/IgG production in some immune circumstances that IgE tends to some dominate, whereas in other IgG predominates. Effect of the complex of beta-glucan and Zataria multiflora on oxidative burst of neutrophils in BALB/c mice Hojjatollah Shokri. University of Tehran, Microbiology and Immunology, Tehran, Islamic Republic of Iran. Background: Natural products, beta-glucan and herbal essences, have been found to enhance immune functions. The aims of this study were to purify beta-glucan from the cell wall of Saccharomyces cerevisiae and to investigate its conjugate effect with Zataria multiflora essence on oxidative burst in BALB/c mice. Methods: After preparing the cell walls from the yeast, beta-glucan was extracted from wall by alkaline-acid treatment (glucan-S1). This preparation contained protein and mannan, which were removed by DEAE sephacel chromatography (glucan-S2) and con-A sepharose chromatography (glucan-S3), respectively. Glucan-S3, Zataria multiflora essence, and their conjugate were injected into BALB/c mice intraperitoneally. Blood collected at days 4 and 7 after injection and oxidative burst was assayed by chemiluminescence method. Results: The findings showed that intraperitoneally administration of glucan-S3, Zataria multiflora essence and their conjugate significantly increased oxidative burst of neutrophils both at days 4 and 7 when compared to control group (PG0.05). Conclusion: These results suggest that the complex of à-glucan and Zataria multiflora essence can be used as adjuvant agents to stimulate immune functions in immunocompromised subjects. Effect of staphylococcal enterotoxin on inducible nitric oxide synthase in the mice with allergic rhinitis Si Whan Kim, Tae-Bin Won, Dae Woo Kim, Dong-Young Kim, Chae-Seo Rhee, Sang-Wook Kim, and Yang-Gi Min. Seoul National University College of Medicine, Departments of Otorhinolaryngology, Seoul, Republic of Korea. Background: According to recent reports, inducible form (type II) of nitric oxide synthase is expressed at higher levels in the nasal mucosa of allergic rhinitis patients, and these patients show a higher carriage rate of Staphylococcus aureus than normal adults. Aim: Are to investigate the role of staphylococcal enterotoxin B in the development of allergic rhinitis and to determine its relevance with inducible nitric oxide synthase overexpression. Methods: The nasal mucosa and serum were obtained from the sensitized and negative control groups, and the frequencies of allergic symptoms, such as sneezing, were compared. Eosinophil counts and inducible nitric oxide synthase expression in the nasal mucosa were examined. The serum levels of ovalbumin-specific IgE were measured by ELISA. Differences between the sensitized and control groups were statistically analyzed using the Kruskal-Wallis test and the Mann-Whitney U test. Results: The frequencies of sneezing, serum levels of ovalbumin-specific IgE, and degrees of inducible nitric oxide synthase expression were significantly higher in the groups locally sensitized with a mixture of ovalbumin and staphylococcal enterotoxin B or ovalbumin alone than in the negative control group or in the group locally sensitized with staphylococcal enterotoxin B alone. Conclusion: Inducible nitric oxide synthase may play a crucial role in the development of allergic rhinitis whereas staphylococcal enterotoxin B may not participate in this process. The anti-allergic effect of lactobacillus probiotics in mite-allergen sensitized murine model Kung-Chih Yeh 1 , Ren-Long Jan 2 , Shin-Hong Chen 3 , and JIu-Yao Wang 4 . 1 College of Medicine, National Cheng Kung University, Department of Microbiology and Immunology, Tainan, Taiwan; 2 Chi-Mei Medical Center, Liou-Yin Campus, Pediatrics, Tainan, Taiwan; 3 Madou Shin-Lau Hospital, Pediatrics, Tainan, Taiwan; 4 College of Medicine, National Cheng KUng University, Pediatrics, Tainan, Taiwan. Background: Clinical trails have demonstrated that probiotics can improve the allergic symptoms but the mechanisms responsible for these benefits are still unclear. We sought to investigate the effects of probiotic organisms in a mouse model of allergic airway inflammation. Methods: First, Balb/c mice were treated with two probiotic organisms Lactobacillus rhamnosus and Lactobacillus paracasei via gavaging needle for two weeks daily before Dermatophoides pteronyssinus (Der p) allergen sensitization and for another two weeks during sensitization and allergen challenge. Second, to know the dose effect of Lactobacillus paracasei, we further investigated the ability of three doses of Lactobacillus paracasei, 5 Â 107CFU/ml, 107CFU/ml and 3 Â 106CFU/ml, in the prevention of allergic airway inflammation. Results: First, we found that treatment with Lactobacillus paracasei significantly elevated Th1 cytokine, IL-12 in bronchoaveolar lavage fluid (BAL) and decreased Der p-specific IgG1 in serum but there were no changes in airway hypersensitivity, influx of inflammatory cells to the lung and serum IgE level. Treatment with Lactobacillus rhamnosus did not have any significant effects. In the second part, treatment with medium dose (107CFU/ml) of Lactobacillus paracasei significantly attenuated the influx of total leukocytes, monocytes and lymphocytes to the airway and elevated IL-12 in BAL, Der p-specific IgG2a in serum. Medium dose of Lactobacillus paracasei also decreased airway hyperresponsiveness and increased IFN-, production in Der p stimulated splenocytes. The high dose of Lactobacillus paracasei reduced the influx of total leukocytes, monocytes in BAL and IL-4 production in Der p stimulated splenocytes, increased Der p-specific IgG2a in serum and IFN-, production in Der p stimulated splenocytes. The low dose of Lactobacillus paracasei only elevated Der p-specific IgG2a in serum, IFN-, production in Der p stimulated splenocytes and reduced the splenocyte proliferation activity. Conclusion: Our results show that oral treatment with probiotics can attenuate allergic airway inflammation by increasing Th1 immune response and these effects are strain specific and dose dependent. Aerobiological and Immunochemical studies in Domjur, Howrah District, West Bengal, India Indrani Choudhuri. Bose Institute, Kolkata, Botany, Kolkata, India. Allergic diseases, like bronchial asthma, allergic rhinitis, conjunctivitis due to exposure of various airborne biological particles are very common. Pollen grains among them are known as a causative agent which cause respiratory allergy in susceptible individuals. In this program, a rural area at Howrah district, Domjur Block near Kolkata was selected for our aerobiological and clinico-immunological studies. The area was selected as it is rich in vegetation and a high density of population suffering from respiratory allergy. Aerobiological Study was done using Burkard sampler. Preparation of allergenic extracts was done using pure bulk of pollen grains using 0.1 M phosphate buffered saline (PBS). Skin tests (SPT) with different pollen extracts were performed following the method of Stytis (1982 ) . Sensitive sera were collected from the patients visiting the outpatients' Department of the Allergy Clinic of the Institute of Child Health, Calcutta. Normal sera were obtained from healthy donors with no history or symptoms of atopy. Electrophoresis and Western blotting was analyzed by SDS-PAGE according to the method of Laemmli (1970) . In order to prepare a pollen calendar, at Domjur, rich in vegetation with wide agricultural farming aerobiological survey was done. 47 air borne pollen types were recorded and identified from the air of Domjur. The dominant types were, Areca catechu, Cocos nucifera, Phoenix sylvesrtis, Riccinus communis, Lantana camara, Perthenium hysterophorus, Trema orientalis, Moringa olifera, Carica pa paya, Catharanthus roseus, Casuarina equisetifolia, Albizzia lebbeck, Cyperus rotundus etc. Among the dominant pollen types, some were subjected to Skin Prick Test. Areca catechu, Carica papaya, Catharanthus roseus, Cocos nucifera, Amaranthus spinosus, Borassus flabellifer, Lantana Camara were found to be highly positive. Carica papaya pollen was selected for the biochemical and clinical investigations. SPT reaction was studied with the relevant pollen extract on 320 respiratory allergic patients and 30% of them showed a positive response. The native polyacrylamide gel electrophoresis of Carica pollen and fruit showed the presence of a number of protein bands . In IgE specific dot immunoblotting of Carica pollen showed remarkable cross reaction with papain, an enzyme present in the latex, fruit and other plant parts. Nasal mucosal infiltration of eosinophil after nasal provocation Sung-Wan Kim, Joong Saeng Cho, Kun Hee Lee, and Seung Yeop Shin. Kyunghee University Hospital, Dept. of ORL-HNS, Seoul, Republic of Korea. The Background of the study: Eosinophils are found in the nasal lavage fluid and nasal biopsies of patients with allergic rhinitis after nasal antigen challenge. Numerous studies have been carried out to clarify the paticipation of eosinophils in a late-phase allergic reaction or airway hyperresponsiveness. However, there has been a few report describing in detail the role of eosnophils during an early-phase allergic reaction. The aim of this study was to investigate the degree of nasal eosinophil infiltration before and after nasal provocation test and to evaluate the changes of degree of nasal eosinophil infiltration according to the duration of allergic rhinitis. Methods: One hundreds and forty-five allergic rhinitis patients who were diagnosed by allergic work-up were included. The patients were divided into three groups by duration of symptoms. Group 1 was the patients whose duration of symptoms was less than 2 years (n = 40), group 2, 2 to 9 years (n = 58), and group 3, more than 10 years(n = 47). Symptom scores of the patients were checked for nasal obstruction, watery rhinorrhea, sneezing, and itching with visual analogue scale. Nasal mucosal scraping and smear was performed with curette at before and 30 min after nasal allergen provocation. Degree of eosinophil infiltration was evaluated in each slide by 6-point scale. And degree of eosinophil infiltration was compared between before and after provocation. The degree of eosinophil infiltration was evaluated and compared according to the duration of symptoms. Results: Prominent nasal symptoms of all 3 groups was nasal obstruction and it was increased by duration of symptoms. But other symptoms were more severe in group 2 than in other group. Group 2 and 3 showed higher degree of eosinophil infiltration compared with Group 1. And degree of eosinophil infiltration was increased after provocation in group 2 and 3. Conclusions: The eosinophil infiltration is increased even in early stage of allergen provocation and increase of eosinphil infiltration in the nasal mucosa after provocation is more significantly in chronic state. Eosinophil may be involved in early reaction of allergy, especially in chronic state. Association of cytochrome P450 (2C9, 2C19) gene polymorphism with susceptibility to Ankylosing Spondylitis in a population group from Algeria Sofiane Samir SALAH 1 , Habiba AMROUN 1 , Rachida ALLAT 2 , Marc BUSSON 3 , Antoine TOUBERT 3 , Rajagopal KRISHNAMOORTHY 4 , Dominique CHARRON 3 , Mohamed Cherif ABBADI 5 , Hachemi DJOUDI 2 , and Ryad TAMOUZA 3 . 1 Institute Pasteur of Algeria, Immunology, Algiers, Algeria; 2 Douera Hospital, Rhumatlogy, Algiers, Algeria; 3 Saint Louis Hospital, Immunology and histocompatibility, Paris, France; 4 Robert Debré Hospital, Immunology, Paris, France; 5 Pasteur Institute of Algeria, Immunology, Algiers, Algeria. Background: Cytochrome P450 (CYP) superfamily members CYP2D6, CYP2C9 and CYP2C19 are polymorphically expressed enzymes that are involved in the metabolic inactivation of several drugs, including, antiepileptics, NSAIDs, oral hypoglycemics, and anticoagulants. Many of these drugs have a narrow therapeutic index, and several studies indicates a prominent role of these enzymes polymorphisms in the therapeutic efficacy and in the development of adverse effects among patients treated with drugs that are CYP2D6, CYP2C9 or CYP2C9 substrates. Previous linkage and association studies has suggested the presence of a susceptibility gene for ankylosing spondylitis (AS) close to, or within, the cytochrome P450 2D6 gene which could play an key role in pathophysiology of AS. In our study, we examined whether common mutations of cytochrome P450 (CYP) [CYP2C9*2,*3, CYP2C19*2, and CYP2D6*3,*4,*6,*7 and *8] are involved in outcome of AS in Algerian patients and, find out and determine genotype frequency in Algerian population. Methods: CYP genotype was determined in 160 AS patients [fulfilling the modified Naw York criteria] and 204 healthy non-related subjects. DNA isolated from blood samples was used for the analysis of CYP2D6, CYP2C9 and CYP2C19 allelic variants by allele-specific real-time PCR. Thus, with respect to allelic variant frequencies, genotype distributions or predicted phenotypes were deduced from genotype combinations. Results: i) no significant difference was found in CYP2D6 allelic variants between AS patients and control subjects; ii) the CYP2C9*3 allelic variant is more frequent in AS patients than healthy subjects (pc=0,015,OR=2,98); iii) CYP2C19*2 allelic variant is more frequent in AS patients than healthy subjects (pc=0,009, OR=2,06) ; iiii) and the frequency of the Bintermediate metabolizers^genotype and/or phenotype for substrates of CYP2C19 is 2-fold more important in AS patients than controls (pc=0.0009). Conclusion: It would exist a possible role of CYP2C9 and C19 polymorphisms in the risk for development of AS and probably on the metabolism of some drugs used in treating AS patients. Exosomes from bronchoalveolar fluid of tolerized mice prevent allergen-specific allergic reaction Noela Prado 1 , Eva G Marazuela 1 , Elodie Segura 2 , Héctor Fernández-García 3 , Mayte Villalba 1 , Clotilde Théry 2 , Rosalía Rodríguez 1 , and Eva Batanero 1 . 1 Universidad Complutense, Bioquímica y Biología Molecular, Madrid, Spain; 2 Institut Curie, Pavillon Pasteur, Paris, France; 3 Universidad Rey Juan Carlos, Área de Histología y Anatomía Patológica, Madrid, Spain. Background: Exosomes are nanovesicles originating from multivesicular bodies that are secreted by a variety of cell types. The dual capability of exosomes to promote immunity or to induce tolerance has prompted their clinical use as vehicles for vaccination against different human diseases. In the present study, the effect of allergen-specific exosomes from tolerized mice on the development of allergen-induced allergic response was determined using a mouse model. Methods: Mice were tolerized by respiratory exposure to the olive pollen allergen Ole e 1. Exosome-like vesicles were isolated from bronchoalveolar lavage fluid of the animals by the well-established filtration and ultracentrifugation procedure, characterized by electron microscopy, western blot and FACS analyses, and assessed in a prophylactic protocol. To this end BALB/c mice were intranasally treated with tolerogenic exosomes or naBve exosomes as a control, one week prior to sensitization/challenge to Ole e 1. Blood, lungs and spleen were collected and analyzed for immune responses. Results: Intranasal pretreatment with tolerogenic exosomes inhibited IgE response along with a marked reduction of lymphoproliferative response and Th2 cytokine production. Moreover, the pathologic alterations of the lung associated with airway inflammation, such as eosinofilia, were significantly suppressed. Conclusion: These results demonstrate that exosomes can induce tolerance and protection against allergic sensitization in mice. Thus, exosome-based vaccines could represent an alternative to conventional therapy for allergic diseases in humans. Primary and secondary infection with Respiratory Syncytial Virus (RSV) before allergen challenge enhanced airway hyperresponsiveness in murine model Objective: To determine whether neonatal infection with RSV plays a critical role in the development of AHR and pathophysiological response to subsequent allergen sensitization in adult mice. Methods: Mice were infected initially at 5-7 days of age with RSV and were re-infected 5 weeks later. RSV-infected mice were exposed to ovalbumin (OVA) at 1 or 3 weeks after infection via airway followed by assessment of airway function by invasive method in changes of lung resistance to inhaled methacholine (Mch), cytokine levels in bronchoalveolar lavage fluid (BALF), and lung histopathology. Results: RSV infection and subsequent allergen sensitization resulted in development of AHR and inflammation, which is associated with increased eosinophils influx into the lungs, increased IL-5, and mucus production. Although enhanced AHR to Mch and inflammations are observed in both RSV-infected mice at neonate and subsequently re-infected with RSV after weanling, these response were more exaggerated in mice with early exposed to ova than later after RSV infection. Conclusion: Neonatal RSV infections predispose the adult to develop enhanced airway responses and inflammation upon allergen exposure. Background: Food allergy is an early and potentially life-threatening manifestation of Fatopy march'. There is limited published epidemiological data of food allergy in Chinese children. To plan for possible intervention to prevent the development of food allergy, it is important to identify its disease spectrum and risk factors in our population. This study aims to determine the prevalence and clinical features for self-reported food allergy in Hong Kong preschool children. Methods: Children aged 2-6 years living in Hong Kong were recruited through randomly selected nurseries and kindergartens for this study to ascertain the presence of self-reported symptoms of food allergy and other atopic disorders. The parental questionnaire was developed from the Chinese version of International Study of Asthma and Allergies in Childhood questionnaire, with addition of validated items on details of food allergy. Results: 3827 children from 21 local nurseries and kindergartens returned our study questionnaires, of which 3677 (96.1%) children were evaluable for food allergy. Two hundred and fifty-three (6.9%) children were born in mainland China and subsequently migrated to Hong Kong. Four hundred and twentytwo (11.5%) children were currently avoiding any food. The prevalence rates of self-reported food allergy and self-reported, doctor-diagnosed food allergy were 8.1% and 4.6%, respectively. The figures for ever wheeze, doctordiagnosed asthma and current flexural eczema were 14.0%, 5.0% and 31.9%. The six leading causes of self-reported food allergy in our Chinese children were shellfish (15.8%), egg (9.1%), peanut (8.1%), beef (6.4%), cow's milk (5.7%) and nuts (5.0%). When compared with children born and raised in Hong Kong, children born in mainland China had significantly lower prevalence of self-reported food allergy (4.0% versus 6.7%; P = 0.016), doctor-diagnosed asthma (1.7% versus 5.4%; P = 0.006) and current rhinoconjunctivitis (13.9% versus 25.4%; P G 0.001) but not current flexural eczema (P = 0.257). The occurrence of food allergy was strongly associated with current wheeze, rhinoconjunctivitis and flexural eczema (P G 0.001 for all). Conclusion: Food allergy is a significant atopic disorder in Hong Kong preschool children, and coexisting airway and skin allergies are also commonly seen in these children. Further studies to identify early life feeding and environmental factors for food allergy are needed. A patient with tracheobronchial amyloidosis and complete lung collapse however, if taken for a very long time, can have cardiac side effects. Etoricoxib is less cardio-toxic. Methods: We have tested the drug on 65 patients with previous adverse reactions to NSAIDs: 13 to salycilates, 18 to arylpropionics, 10 to arylacetic acid, 12 to oxicam and derivates, 8 to pyrazolones and 4 to acetaminophen (paracetamol-P). The reported symptomatology was urticaria and/or angioedema in 69%, rhinitis reactions in 3% and one case of anaphylactic shock (1,5%). Peroral challenges were performed at random controlled with placebo, starting with a 10 mg dosage up to a total dosage of 90 mg. in the third day. The challenge has been done in Day Hospital and the patients were cardiorespiratory monitored for 4-6 hours. Results: The tolerance test was fully carried out and well tolerated in 97% of the patients. Only two systemic reactions and a slight tiredness along with stomach-ache were reported during the first day. Conclusion: Etoricoxib can be considered a safe molecule for those patients with previous adverse reactions to NSAIDs. Background: Mashhad locates in North-East of Iran and so close to the borders of Afghanistan and Turkmenistan, also considered as a migrated city due to religious reasons. Injecting Drug Users are increasing due to its lower cost and other interests especially in marginalized regions. Sharing the contaminated syringes and injecting tools is the most important and common way of HIV transmission in the country and city. Peer education with PLHIVs among their peers and most at risk groups such as IDUs is a strategy of positive prevention which also helps as psychosocial support for PLHIVs. Mashhad Positive Club established to empower PLHIVs based on the national AIDS program and positive prevention through UNAIDS, MOH CDC and Family Planning Association support. Method: After preparing PE Protocol and call for action, we trained 7 PLHAs including 3 of them were IDU. They were going to the ruins, places for group injection (blood sharing) and some in Drop In Centers in Daravey and Nodeh, two marginalized regions of Mashhad. They have trained their peers, IDUs and other positives, by face to face talking and distributing harm reduction packages including syringes and condoms for safer sex came out through FGD. The process monitoring was being done by writing a daily and weekly report, going with another peer educator and officer direct visit. The other training program is peer group sessions including movie display titled BAIDS Borne^then a discussion between trained educators and beneficiaries. Results and Conclusion: Based on one month evaluation (1-31 July 2007) 455 IDUs (431 Male and 24 Female),%88 of them were 20-30 years old, have been trained by PLHAs and referring to DIC to exchange syringe program and getting condoms or start MMT increased about 42 percent. An indirect finding is health care workers in DIC have changed their attitude to PLHAs and see them not as a problem also as key of problem solving which would be decreasing social stigma. complications. Cold urticaria may be potentially life threatening, in any case it has a profound impact on patient quality-of-life. Second-generation antihistamines are recommended as the first-line treatment, but to date only a few have been scientifically tested for this condition. To assess the safety and efficacy of ebastine in preventing ACU symptoms, we examined 22 adult patients suffering from cold urticaria within the scope of a double-blind crossover trial administering a single dose of 20 mg ebastine or placebo before local cold challenge. The safety of ebastine was sensitively assessed with a psychometric battery testing cognitive performance and mood. After cold challenge, wheal and erythema were assessed by the investigator and the intensities of pruritus and burning were rated by the subject. We could show that Ebastine had no negative impact on any of the parameters of cognitive performance or mood. Most importantly, cold urticaria symptoms were markedly reduced after challenge in the verum group, indicating that ebastine can effectively protect patients from cold urticaria symptoms. These results demonstrate the safety of ebastine and reveal its efficacy in treating patients with cold urticaria. Dermographic urticaria symptoms can be safely prevented by ebastine Markus Magerl, Jan Schmolke, Frank Siebenhaar, Torsten Zuberbier, Martin Metz, and Marcus Maurer. Charité -Universitätsmedizin Berlin, Dermatology, Berlin, Germany. Physical urticaria includes a heterogeneous group of disorders characterized by the development of urticarial lesions and/or angioedema after exposure to certain physical stimuli. The most frequent physical urticaria is the dermographic urticaria (DU), in which shearing forces on the skin result in wheals and itching. Subsequent scratching aggravates the symptoms. Second-generation antihistamines are recommended as the first-line treatment, but to date only one has ever been tested for this condition. The aim of this study was to assess the safety and efficacy of ebastine in preventing DU symptoms. We administered 20 mg ebastine to seven adult DU patients in the scope of a double-blind cross-over trial. The safety of ebastine was sensitively assessed with a psychometric battery testing cognitive performance and mood. After challenge by shearing forces, wheal and erythema were assessed by the investigator and the intensities of pruritus and burning were rated by the subject. We could show that Ebastine had no negative impact on any of the parameters of cognitive performance or mood. Wheals, pruritus, and burning were greatly reduced for the majority of subjects. These results underline the safety of ebastine and reveal its efficacy in cold urticaria. and should be tested on a larger scale. Sensitization to environmental allergens among the inhabitants of Calcutta city and suburbs with special emphasis to inhalant fungal allergy: an aerobiological and allergological study Paediatrics, London, United Kingdom; 2 University of Southampton, School of Psychology, Southampton, United Kingdom; 3 University of Southampton, School of Medicine, Southampton, United Kingdom. Background: The relationship between artificial food colouring and benzoate perservative (AFCP) intake and behaviour has hitherto been contentious. We have previously shown in a population based DBPCFC study, an adverse effect of AFCP on parentally-rated behaviour of 3 year olds and have now confirmed the observations in 144 8Y9 year olds and 153 3 year olds using objective measures of observed behaviours in a school setting. One potential mechanism is IgE independent histamine release from circulating basophils. Methods: From the second challenge study we have genotyped the children using buccal cells and related findings to the magnitude of the behavioural response to challenge. Results: Polymorphisms of catecholamine genes, COMT Val108Met and ADRA2A C1291G, previously associated with ADHD, had no impact on responses but T939C and Thr105Ile polymorphisms of the histamine Nmethyltransferase gene (HNMT) significantly (p=0.02 and 0.04 respectively) adversely affected responses to AFCP challenge. Conclusion: HNMT polymorphisms impair histamine clearance and AFCP cause histamine release. The presence of Histamine 3 receptors in the brain provides a potential mechanism (and therapeutic target) to explain the effect we observed. Many environmental factors increase histamine including infections and many foods. This would explain the frequent claim that food intolerance and infections adversely affect behaviour in some children. This gene by environment interaction should be investigated in relation to AFCP induced urticaria, asthma and other atopic conditions. Serum specific IgE -is result enough? Joana Caiado 1 , Susana Lopes da Silva 1 , and José Costa Trindade 2 . 1 Hospital de Santa Maria, Serviço de Imunoalergologia, Lisboa, Portugal; 2 Hospital de Santa Maria, Unidade de Imunoalergologia Pediátrica, Lisboa, Portugal. Background: In vitro determination of serum specific IgE (sIgE) is an essential diagnostic tool in allergy. Clinical analysis laboratories offer different methods / technologies to determine sIgE, in some cases with highly variable and poorly interchangeable results, concerning the different allergens. Rotation between different methods may compromise allergic patients follow-up, namely in those situations that may benefit from sequential sIgE determinations, as food allergy and specific immunotherapy efficacy evaluation. Aim: To verify the availability of written information concerning the method used in sIgE determination in lab result reports from clinical analysis laboratories in Lisbon District. Methods: During 2 months (April and May 2007), 3 Immunoallergologists collected serum sIgE result reports from the patients they observed. Reports from 50 different clinical analysis laboratories within Lisbon district were obtained and analysed. Labs that did not mention the method that had been used to sIgE determination were further inquired by phone, in order to get that information. Results: Among the 50 reports analysed, only 13/50 (26%) specified the method that had been used, namely the immunoenzimatic (IE) method in 9 and the chemiluminescent (QL) method in 4. Among the remaining 37 labs, when later contacted by phone, we verified that 17/37 (46%) were using IE method, 14/37 (38%) QL and 6/37 (16%) were using both methods, according to the different allergens. In 27/50 laboratories (54%), BRAST^was used as synonym of sIgE. Conclusion: We intend to alert to the generalized lack of written information concerning the methods used in serum sIgE determinations in the lab reports from Lisbon district laboratories. This lack of written information, the persistence of the incorrect use of the term BRAST^and the simultaneous use maintenance. Treated patients and placebo were compared by Pearson test. P values less than .05 were considered statistically significant. Results: 30 adult patients were enrolled and 26 completed the study, with 1 dropout in the active and 3 dropouts in the placebo group. In SLIT patients the peak diameter of LLR was reduced 9 50% in 8/14 (57%). No change in LLR was seen in 11 of 12 placebo patients, and a systemic reaction (generalized urticaria) occurred in a patient on re-sting. The difference was significant (P = .004) between the 2 groups. No adverse event was reported and no discontinuation of SLIT was required. Conclusion: HB-SLIT significantly reduced the extent of LLR in HB allergic subjects, and its safety profile was satisfactory. This pilot study envisages the possible use of SLIT also in hymenoptera allergy, and trials in patients with systemic reactions should be the next step to confirm our observations. Mometasone furoate nasal spray effectively treats the ocular symptoms of seasonal allergic rhinitis in patients with all degrees of severity over 24 hours L Bielory 1 , MR Danzig 2 , and D Gates 2 . 1 UMDNJ-New Jersey Medical School, Asthma and Allergy Research Center, Newark, New Jersey, United States; 2 Schering-Plough Research Institute, Kenilworth, New Jersey, United States. Background: Mometasone furoate nasal spray (MFNS) has been shown to be effective in controlling the nasal symptoms of seasonal allergic rhinitis (SAR). An analysis of subject-reported symptom data in MFNS trials was performed to determine the impact of MFNS on ocular symptoms. This analysis evaluated the effects of MFNS on SAR patients with varying degrees of symptom severity and differences in symptom reduction in the morning (AM) and evening (PM). Methods: A retrospective analysis was conducted of data pooled from four randomized, double-blind, placebo-controlled clinical studies (three 2-week; one 4-week) with similar protocols that compared the effectiveness of MFNS 200 mcg QD and placebo in controlling ocular symptoms in subjects aged Q12 years with a Q2-year history of SAR. The analysis reviewed change from baseline over a 15-day period in symptom scores for eye tearing, itching, and redness on a 4-point scale (0=none; 3=severe), and calculated total ocular symptom score (TOSS). The effect size of the Days 1Y15 average for subjects with mild, moderate, or severe symptoms at baseline was compared, and TOSS was evaluated approximately 12 hours postdose (PM) and on arising, approximately 24 hours postdose (prior to dosing on next day, AM). Results: Subjects in the MFNS (n = 491) and placebo (n = 492) groups had similar baseline clinical and demographic characteristics. The mean baseline TOSS was 4.35 (PM 4.39; AM 4.32) for the MFNS group and 4.5 (PM 4.60; AM 4.43) for the placebo group. The decrease in TOSS with MFNS for the Day 1Y15 interval was -19.8% vs -5.6% for placebo (PG0.001); the difference reached statistical significance at Day 3. For the same time period, scores for tearing decreased by -22.2% with MFNS vs -13.9% with placebo (PG0.001), for eye itching by -17.3% with MFNS vs -12.3% with placebo (P=0.002), and for redness by -15.7% with MFNS vs -8.3% with placebo (P=0.002). For the mild, moderate, and severe symptom groups, the improvements in TOSS with MFNS over placebo were 0.30, 0.42, and 0.50, respectively. Reductions in individual symptom scores were significantly greater with MFNS than with placebo in both the PM (Pe0.006) and AM (Pe0.001). Importantly, differences in TOSS between MFNS and placebo in the PM (0.36) and AM (0.43) were similar. Conclusion: MFNS reduced ocular symptoms of SAR over 24 hours regardless of subjects' baseline symptom severity. Shrimp allergy: effect of vinegar soaking on allergenicity Background: Previous studies demonstrated the muscle protein tropomyosin as the major shrimp allergen. This allergen, Sa-II, was shown to be a 34-kDa protein -rich in aspartic and glutamic acids. Heat-stability of this allergen has been documented. However, no studies have shown yet the effect of vinegar on this allergen. This study aimed to demonstrate if vinegar soaking prior to cooking would have an effect on shrimp allergenicity, based on skin test results. Methods: Eighteen shrimp-allergic and eighteen non-shrimp-allergic pediatric patients were subjected to skin prick test using conventionally-prepared shrimp extract. Mean wheal diameters obtained were compared to mean wheal diameters obtained using 3 experimental shrimp extracts prepared with preliminary vinegar soaking. For the adult group, twenty-six shrimp-allergic and twenty-six non-shrimp-allergic parients were included. Results: Mean wheal diameters obtained using shrimp extract prepared with preliminary vinegar soaking were significantly smaller than mean wheal diameters obtained using conventionally prepared extract. Wheal diameters obtained for shrimp-allergic patients were significantly bigger than wheal diameters obtained fo non-shrimp-allergic individuals. Results were similar for pediatric and adult patients. Conclusion: Results indicate that vinegar soaking prior to cooking can reduce shrimp allergenicity. With resources, further validation of results using doubleblind placebo-controlled food challenges woul be ideal. Mometasone furoate nasal spray improves symptoms of perennial allergic rhinitis in children and adults in Peru Percy Ruiz. Clinica Angloamericana, ENT Department, Lima, Peru. Background: Perennial allergic rhinitis (PAR) is a highly prevalent disease globally and is the most common chronic disease in children. Nasal obstruction and other bothersome symptoms of PAR need to be treated year round. Previous studies have shown the intranasal corticosteroid mometasone furoate nasal spray (MFNS) to be well tolerated and clinically effective in relieving the symptoms of PAR. A study was conducted to assess effectiveness of MFNS and patient satisfaction with treatment in pediatric and adult subjects. Methods: In this prospective, open-label, multicenter study, 33 Peruvian subjects (age 8 to 69 years) with clinically diagnosed PAR recorded nasal symptom data in a diary. Total nasal symptom score (TNSS) was evaluated at 15 and 30 days. Subjects also rated satisfaction with treatment, and the investigators evaluated adverse events. Results: Thirty-three subjects (61% female, 39% male) received MFNS 100 or 200 mcg QD (pediatric or adult dose). Their mean TNSS was reduced from 10.7 (T 2.9 SD) at baseline to 4.1 (T 2.9 SD) at 15 days and 1.6 (T 2.2 SD) at 30 days (PG0.001 for both reductions). At days 15 and 30, 25 subjects (76%) and 31 subjects (94%), respectively, rated themselves as satisfied or very satisfied with treatment. Mild adverse events (nasal irritation, sneezing, epistaxis, and cough) occurred in 7 (21%); one subject stopped treatment due to nasal irritation. Conclusion: Mometasone furoate nasal spray 100 or 200 mcg QD safely and effectively improved symptoms of PAR in children and adults. Effect of postoperative mometasone furoate nasal spray on wound healing following endoscopic sinus surgery Mark Jorissen 1 , and Claus Bachert 2 . 1 University of Leuven, ENT Department, HNS, Leuven, Belgium; 2 University Hospital Ghent, ENT Clinic, Ghent, Belgium. Background: Chronic sinusitis and nasal polyposis are diseases of the nasal and sinus mucosa that affect approximately 15% and 4% of the population, respectively. Functional endoscopic sinus surgery (FESS) is standard surgery for both diseases after failure of medical treatment. Approximately 20% of surgery patients experience impaired wound healing following FESS, leading to recurrences of sinusitis or complications requiring further surgery. A doubleblind multicenter study found that 6 months' treatment with the intranasal corticosteroid mometasone furoate nasal spray (MFNS) 200 mcg QD after surgery for bilateral nasal polyps significantly prolonged the time to recurrence compared with placebo. This study was undertaken to investigate whether use of MFNS following FESS improves wound healing over 6 months. Methods: In this double-blind, randomized, placebo-controlled study, subjects with a diagnosis of chronic sinusitis or nasal polyps were randomized to receive 6 months' treatment with MFNS 200 mcg BID or placebo beginning approximately 2 weeks after FESS. Endoscopic scores were assessed at 1 and 2 weeks, and at 1, 2, 4, and 6 months. The primary efficacy endpoint was the investigator`s assessment of treatment outcome at 6 months. A combination score was derived from scores for inflammation, edema, and polyps. Postoperative symptom scores were recorded at 1, 2, 4, and 6 months. Adverse events (AEs) were recorded throughout the study. Results: Of the 99 randomized subjects, 91 (46 MFNS, 45 placebo) were included in the intent-to-treat analysis. Subjects in the MFNS group received oral steroids for the first 10 days postsurgery. For 24 subjects (11 MFNS, 13 placebo) who discontinued, results from the last study visit were used. Preoperative symptom scores in the two groups were similar. At 6 months, investigators`assessment of treatment outcome and reductions in total endoscopic scores in all subjects were superior, but did not reach statistical significance, with MFNS versus placebo. Combination endoscopic scores at 6 months showed significantly improved healing with MFNS versus placebo for all subjects (median scores: 0.0 vs 3.0, P=0.02) and those with nasal polyps (median scores: 2.0 vs 4.0, PG0.03). MFNS was well tolerated. Conclusion: Treatment with MFNS following sinus surgery improved healing, particularly in subjects with an initial diagnosis of nasal polyps, and was well tolerated. Mometasone furoate nasal spray reduces symptoms and improves quality of life in chinese children with allergic rhinitis Yamei Zhang. Beijing Children`s Hospital, Department of ORL-HNS, Beijing, China. Background: Approximately 40% of children and adolescents in China suffer from symptoms of allergic rhinitis (AR). Effective treatment of AR in children is important in minimizing or preventing behavioral problems and poor academic performance and may help prevent the development of comorbid disorders such as asthma, adenoidal hypertrophy, and otitis media with effusion. This study was undertaken to evaluate the effects of mometasone furoate nasal spray (MFNS) on clinical symptoms and quality of life (QoL) in Chinese children with AR. Methods: A total of 273 symptomatic children aged Q6 years and e12 years with documented seasonal or perennial AR were treated in an open-label study of MFNS 100 mcg QD for 4 weeks. Eligible subjects had a nasal stuffiness/ congestion score Q2 (on a scale of 0=no symptoms and 3=worst symptoms) and a total nasal symptom score (TNSS) Q8 (total of the individual symptom scores for rhinorrhea, congestion, nasal itching, and sneezing) at the start of treatment. The primary efficacy variable was a change in the TNSS, and the coprimary variable was a change in the individual symptom scores, while the secondary efficacy variables included overall efficacy and Pediatric Rhinoconjunctivitis QoL Questionnaire (PRQLQ) scores (assessing nasal and eye symptoms, behavior, and daily life). Results: MFNS significantly (PG0.05) reduced TNSS from baseline (mean 9.16) after 1, 2, and 4 weeks of treatment (mean scores 5.41, 3.47, and 2.10, respectively). Significant improvements from baseline were observed at each week for all individual nasal symptoms. The percentages of patients achieving a 25% improvement (indicating at least moderate symptom reduction) from baseline in TNSS at weeks 1, 2, and 4 were 74%, 95%, and 96%, respectively. Moderate-to-complete symptom relief was reported by 71% of subjects at week 1 and 99% at week 4. Significant reductions (improvements) from baseline in total and individual PRQLQ scores were seen at weeks 2 and 4. Mean behavior PRQLQ scores decreased by a statistically significant amount from 13.99 at baseline to 5.70 at week 2 and 2.71 at week 4. Mean PRQLR scores decreased from 8.30 at baseline to 3.12 at week 2 and 1.72 at week 4. Less than 2% of subjects reported adverse events. Conclusion: MFNS effectively relieved symptoms of AR, improved QoL, and was well tolerated in Chinese children with AR. Mometasone furoate nasal spray improves quality of sleep and reduces symptoms of allergic rhinitis in chinese patients Demin Han. Beijing Tongren Hospital, Department of Otolaryngology Head and Neck Surgery, Beijing, China. Background: Between 20% and 40% of urban-dwelling adolescents and adults in China report symptoms of allergic rhinitis (AR). Unlike other regions where pollen allergy is common, in China dust mites and/or silk are the more prevalent allergens. Consequently, many Chinese people with AR suffer from symptoms throughout the year. One recent study found that once-daily mometasone furoate nasal spray (MFNS) was an effective treatment for AR in Chinese people in Hong Kong. An open-label study was undertaken to evaluate further the effect of MFNS on clinical symptoms and quality of life in Chinese patients with AR. Methods: A total of 500 subjects aged Q12 years to e70 years with a physician diagnosis and documented medical history of seasonal or perennial AR were treated with MFNS 100 mcg QD for 28 days at 10 centers in China. At the screening visit, eligible subjects had a total nasal symptom score (TNSS) Q8 (maximum of 12) and a nasal stuffiness/congestion score Q2 (maximum of 3). The primary efficacy variables were change from baseline in TNSS and individual symptom severity scores for congestion, rhinorrhea, sneezing, and itching. Secondary efficacy variables were change from baseline in scores for interference with sleep and daytime activities on the Nocturnal Rhinoconjunctivitis QoL Questionnaire (NRQLQ) and the Mini-Rhinoconjunctivitis QoL Questionnaire (Mini-RQLQ), respectively. Results: MFNS significantly (PG0.05) reduced TNSS and individual nasal symptom scores from baseline at each evaluation. Mean TNSS at baseline and weeks 1, 2, and 4 were 9.24, 5.78, 3.99, and 2.98, respectively. Corresponding mean scores for nasal congestion were 2.37, 1.57, 1.14, and 0.84; similar improvement was seen with the other nasal symptoms. At week 4, 94.4% of subjects reported moderate-to-compete relief of AR symptoms. Total and individual NRQLQ scores and scores for sleep problems and for symptoms during sleep and on awakening were significantly reduced after 1 week of treatment. There were significant reductions from baseline in total Mini-RQLQ scores and scores for activity problems and symptoms during activity after 1 week. MFNS was well tolerated. Conclusion: MFNS 100 mcg QD effectively reduced symptoms of AR, improved the quality of sleep and daytime activities, and was well tolerated in Chinese patients with AR. Frequency of Dermatophagoides mite allergens in the homes of allergic patients in Mashhad and Tabriz Mashhad Hospital, Immunology, Mashhad, Islamic Republic of Iran; 6 Children's Hospital, Pediatrics, Tabriz, Islamic Republic of Iran. Introduction: Allergic diseases are common disorders with increasing rate during the last 30 years. almost 30% of people have a kind of allergic disorders during their life. Mites in house dust are the most important allergens in allergic disorders. The optimal growth conditions for mites are temperature and humidity and the most important sites are inside bedding, mattresses and carpets. The aim of this study is the survey of frequency of Dermatophagoides and their allergens in the homes of allergic patients. Methods: This descriptive, cross-sectional study has been performed in Tabriz and Mashhad cities, two major industrial centers in north western and north eastern Iran, respectively, from 2005 to 2006. Twenty three allergic patients with positive skin prick test to mites from each city were selected and all of the methods for evaluation of mites and their allergens (specific antibodies of Derp and Derf, density of mites and concentrations of mite allergens) were measured. Because of high density of mites in carpets and bedding, sampling was obtained from these two sites. Results: The frequency of mite density in the homes of allergic patients was 32.6%, 95%CI, 31.25 to 33.95; specific antibody to mite allergens was 23.9%, 95%CI: 23.78 to 24; and concentration of HDM allergens which is performed by ELISA, was not detectable. Conclusion: Mites are common in worldwide, but the incidence of them is various in different parts of world. The frequency of mites and their allergens in two large provinces of Iran are lower than other studies, performed in other parts of world. The most important factor in frequency and growth of them is environmental humidity, and the other factors such as carpets and bedding have less importance. The differences of clinical characteristics between childhood and adult allergic rhinitis patients in Asians: according to ARIA classification Hyun Jong Lee 1 , Dong Wook Kim 2 , Jeong-Whun Kim 1 , and Chul Hee Lee 2 . Background: The prevalence of allergic rhinitis varied across centres from 0.8% to 14.9% in the 6Y7-year-olds and from 1.4% to 39.7% in the 13Y14year-olds. The recent guidelines of Allergic Rhinitis and its Impact on Asthma (ARIA) group, classified it as the frequency and severity may not be appropriate for the children age group. The present study evaluates the childhood and adult patients with the ARIA classification and analyzed the clinical features of each age group. Methods: 1,375 subjects with allergic rhinitis diagnosed by skin-prick test at our hospital from December 2005 to July 2007 were included. These patients were classifed according to age; babyhood(1Y4 years), preschoolers (5Y7 years), elementary schoolers (8Y13 years), adolescents (14Y19 years), young adults (20Y35 years), adults (36Y55 years), middle age (55Y64 years), old age (65 years).Clinical features including nasal obstruction, rhinorrhea, nasal itching, sneezing, postnasal drip(PND), ocular symptoms and sleep disturbance and the allergen was analyzed in each age group according to the ARIA classification. Results: A similar distribution of the ARIA classification groups were shown among all ages; Mild & intermittent group 27.5%, moderate to severe & intermittent group 17.8%, mild & persistent 18.5%, moderat to severe & persistent 34.9%. There was a higher proportion of nasal obstruction(p=0.04) and PND(p=0.042) in babyhood & preschoolers compared to adults, but ocular symptoms(p=o.01) and sneezing(p=0.01) were shown more frequently in adults. PND, sleep disturbance, nasal itching showed no distributional difference between children and adults. For allergens, housedust mite(p=0.07) showed a higher proportion in children whereas grass(p=0.01) and weed(p=0.021) was more frequent in adults. Clinical features, IgE and serum eosinophile count showed no significant relationship in each age ARIA classification group. Conclusion: Classification of the ARIA group into different ages helded to identify the different clinical manifestations among children and adults. A new treatment strategy taking children's clinical features into account is deemed necessary. Chronic idiopathic urticaria and autoimmune thyroid disease Vincenzo Patella, Diomira Magliacane, and Giovanni Florio. U.O. of Allergy and Clinical Immunology, Agropoli Comunity Hospital, Department of Medicine of ASL SA/3, Salerno, Italy. Chronic idiopathic urticaria (CIU) persists for longer than 6 weeks, with a not determined cause. One of the interesting clinical associations is between chronic urticaria with positive autologous serum skin test (ASST) and thyroid disease. CIU and chronic tyroiditiis has not been completely investigated. Methods: We studied 94 patients (65 female and 39 male; mean age 44.7 years; range 15Y78 years) who had CIU and a history of thyroid dysfunction. Thyroid function was investigated by means of thyroid-stimulating hormone (TSH), fT3 and fT4 measurement and thyroid ultrasound. Thyroid antibodies (anti-thyroid peroxidase and anti-thyroglobulin), total serum IgE levels and ASST were performed in all patients. Results: Antibodies to thyroglobulin and to peroxidase were positive in 26/94 patients (27.7%) with CIU and morphological or volumetric abnormalities of the thyroid gland by ultrasonography. All the CIU patients with autoimmune thyroiditis had a normal thyroid function, a negative ASST and a normal total serum IgE levels (74.3 +/j 14.9 IU/ml; normal value G100 IU/ml). Antibodies anti-thyroglobulin and anti-thyroid peroxidase were negative in 68/94 (72.3%) CIU patients, of which 44/68 (64.7%) had high total serum IgE levels (395.2 +/j 180.6 IU/ml) and a negative ASST. Four of the thirty four CIU patients (11.8%) with negative thyroid autoantibodies had a positive ASST and a normal serum IgE levels, while 16/68 (23.5%) patients had a multinodular goiter, ASST negative and normal value of total serum IgE levels. Conclusion: We found a correlation between autoimmune thyroiditis and normal total serum IgE levels in CIU patients with ASST negative. High total serum IgE levels were found in CIU patients without thyroid disease. Thus, in CIU patients the tests to detect thyroid autoantibodies are relevant in particular in patients with normal value of total serum IgE levels. Further investigation of mast cell activation by thyroid antigens is incoming. Perioperative allergic reactions: a case of intraoperative anaphylactic shock Aguiar Gonzalez, Gracia Maria, Arias Franco, Maria Jesus, Belen Carnero Pinto, and Mejuto Castedo. Hospital Xeral Calde Lugo, Spain and, Rosa Hospital Keral Calde Lugo, Spain. The incidence published on perioperative allergic reactions is highly variable, fluctuating between 1:3.500 and 1:20.000 anaesthesias, with a mortality rate of 3 to 6%. During surgical procedures it is difficult to recognise an allergic reaction.Under anaesthesia , bronchospasm and hypotension are common and lacking in a set of symptoms typical of an allergic reaction;especially when they occur separately;if the response to treatment is effective, the symptoms are often not investigated and they are attributed to another cause. Furthermore, 10Y14% of the reactions,primarily the most severe, affect just one system;and that,too, leads to many cases not being diagnosed. Therefore, epidemiological monitoring of these reactions is required. This is a case of intraoperative anaphylactic shock: A 59-years old female scheduled for an abdominal hysterectomy due to uterine sarcoma; her Tsuda, T, 183 Tuckova, L, 879 Tummolo, R, 757 Tungchaisin, S, 906 Tungtrongchitr, A, 675 Tungtrongchitrc, A, 297 Tupkina, O,