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CORD-19:4ca5d854d3f99440b507fd7dfae73c8ad6fc45d5 JSONTXT 9 Projects

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Id Subject Object Predicate Lexical cue
T1 0-935 Sentence denotes DVP01 "Inactivated mycobacteria bead preparation" method for identification of mycobacteria by MALDI-TOF MS DVP02 MALDI-TOF MS for functional detection of ß-lactam resistance of positive blood cultures DVP03 Molecular typing of Stenotrophomonas maltophilia from patients with cystic fibrosis by repetitive sequence-based PCR DVP04 Photochemical inactivation allows papid diagnostics of Alpha- and Poxviruses DVP05 Rapid detection and molecular differentiation of toxigenic Corynebacterium diphtheriae and Corynebacterium ulcerans strains by LightCycler PCR DVP06 Use of recombinant proteins for optimized serodiagnosis of M. pneumoniae infections DVP07 MALDI-TOF-MS analysis of bacteria difficult to identify by standard procedures DVP08 Carbapenemase detection by phenotypic methods DVP09 Direct identification of uropathogens by MALDI-TOF-MS and interfering factors DVP10 Comparison of different methods for the isolation of tick DNA
T2 937-945 Sentence denotes Abstract
T3 946-959 Sentence denotes Introduction:
T4 960-1131 Sentence denotes Next to the clinical important Mycobacterium tuberculosis complex (MTC) Nontuberculosis mycobacteria (NTM) are pathogens especially in immunocompromised and senior people.
T5 1132-1204 Sentence denotes Species identification is important for diagnosis and optimal treatment.
T6 1205-1358 Sentence denotes MALDI-TOF identification of some Mycobacterium strains is hampered by low quality of acquired mass spectra from samples processed by standard procedures.
T7 1359-1506 Sentence denotes Therefore, a new preparation method for Mycobacteria was established and influence of an additional preceding heat inactivation step was evaluated.
T8 1507-1515 Sentence denotes Methods:
T9 1516-1611 Sentence denotes 14 Mycobacterium strains were prepared using standard ethanol/formic acid extraction procedure.
T10 1612-1722 Sentence denotes The new established Mycobacteria bead preparation protocol was tested with these 14 and additional 22 strains.
T11 1723-1826 Sentence denotes An extended protocol named inactivated Mycobacteria bead preparation method was tested with 10 strains.
T12 1827-1992 Sentence denotes Mass spectra were recorded in a microflex LT MALDI-TOF mass spectrometer and identification was done applying MALDI Biotyper 3.0 software (Bruker Daltonik, Germany).
T13 1993-2001 Sentence denotes Results:
T14 2002-2146 Sentence denotes Out of 83 Mycobacterium spp. strains tested (data not shown) 14 strains were hard to analyse by MALDI-TOF and therefore selected for this study.
T15 2147-2258 Sentence denotes Consequently, standard ethanol/formic acid extraction procedure failed to acquire mass spectra from 12 strains.
T16 2259-2423 Sentence denotes A significant improvement of quality of mass spectra was achieved by using Mycobacteria bead preparation method with glass beads for cell suspension and disruption.
T17 2424-2536 Sentence denotes Now, 11 out of 14 Mycobacterium strains resulted in good quality mass spectra and three ones in average quality.
T18 2537-2670 Sentence denotes In addition, 22 further Mycobacterium strains were processed according to this new protocol and all resulted in good quality spectra.
T19 2671-2742 Sentence denotes An additional heat inactivation step was introduced for safety reasons.
T20 2743-2893 Sentence denotes Mass spectra of 10 Mycobacterium spp. strains processed by Mycobacteria bead preparation method with and without heat inactivation step were compared.
T21 2894-3031 Sentence denotes Mass spectra revealed only minor differences, just higher peak intensities were obtained in general with the heat inactivation procedure.
T22 3032-3043 Sentence denotes Conclusion:
T23 3044-3137 Sentence denotes A fast, secure and easy to perform method was developed for mycobacterial protein extraction.
T24 3138-3299 Sentence denotes These extracts are highly accessible to MALDI-TOF analysis and good quality mass spectra are a precondition for identification of Mycobacteria by this technique.
T25 3300-3400 Sentence denotes The new preparation method was successfully tested for 36 different strains representing 30 species.
T26 3401-3506 Sentence denotes A supplemental heat inactivation step was introduced for minimizing hazards by handling living MTC cells.
T27 3507-3698 Sentence denotes The main mechanism of bacterial resistance against ß-lactam antibiotics is the expression/overexpression of ß-lactamases which destroy the ß-lactam ring of ß-lactam antibiotics by hydrolysis.
T28 3699-3790 Sentence denotes This reaction results in a mass shift of plus 18 Da easily detectable by mass spectrometry.
T29 3791-3897 Sentence denotes A rapid MALDI-TOF MS based assay was set up to detect ß-lactam resistance of bacteria from blood cultures.
T30 3898-3906 Sentence denotes Methods:
T31 3907-4061 Sentence denotes Blood culture bottles were inoculated with different E. coli and Klebsiella pneumonia strains and incubated at 37°C until they were indicated as positive.
T32 4062-4194 Sentence denotes Subsequently, 1 ml fresh positive blood culture was processed according to a slightly modified protocol of the MALDI Sepsityper kit.
T33 4195-4347 Sentence denotes The washing step was repeated once and subsequently bacteria were incubated with the corresponding antibiotics solution for 3 h at 37°C under agitation.
T34 4348-4395 Sentence denotes Ampicillin was tested with the E. coli strains.
T35 4396-4455 Sentence denotes Ertapenem was tested with the Klebsiella pneumonia strains.
T36 4456-4618 Sentence denotes After incubation of the bacteria with the corresponding antibiotics and subsequent centrifugation, the supernatant was directly applied to a MALDI sample carrier.
T37 4619-4670 Sentence denotes Dried spots were overlaid with MALDI matrix (HCCA).
T38 4671-4751 Sentence denotes MALDI-TOF MS spectra were acquired on a microflex LT benchtop mass spectrometer.
T39 4752-4841 Sentence denotes Inhibition of hydrolysis was performed in the presence respective ß-lactamase inhibitors.
T40 4842-4850 Sentence denotes Results:
T41 4851-4978 Sentence denotes The MS spectrum corresponding to DH5alpha revealed the molecular peak of Ampicillin (350 Da) and the respective sodium adducts.
T42 4979-5202 Sentence denotes The spectra derived from the ß-lactamase producing strains show peaks at 368, 394, 412 and 324 Da corresponding to the hydrolyzed form, its sodium adducts and the hydrolyzed, decarboxylated form of ampicillin, respectively.
T43 5203-5280 Sentence denotes A slight spontaneous hydrolysis was also observed in the spectra of DH5alpha.
T44 5281-5394 Sentence denotes Successful inhibition of Ampicillin hydrolysis was demonstrated by incubation in the presence of calvulanic acid.
T45 5395-5556 Sentence denotes Incubation of Ertapenem with Klebsiella pneumonia strains revealed a clear difference between the carbapenem sensitive strain and carbapenamase positive strains.
T46 5557-5666 Sentence denotes For the carbapenem sensitive strain the molecular peak of Ertapenem [M+H] + at 476 Da was clearly detectable.
T47 5667-5796 Sentence denotes Incubation of Ertapenem with the carbapenemase positive strains resulted in the disappearance of the molecular peak of Ertapenem.
T48 5797-5910 Sentence denotes In contrast to Ampicillin, the hydrolyzed form of Ertapenem was very instable and immediately broke down further.
T49 5911-6053 Sentence denotes Therefore, only the decrease of the molecular peak of Ertapenem can be employed as indicator for resistance against carbapenems at the moment.
T50 6054-6240 Sentence denotes Additional peaks appear in the spectra of the carbapenemase positive strains but these do not directly correspond to the hydrolyzed form of Ertapenem and have not been identified so far.
T51 6241-6252 Sentence denotes Discussion:
T52 6253-6407 Sentence denotes The developed approach provides a rapid method for the detection of beta-lactamase activity of bacteria within less than 4 h from positive blood cultures.
T53 6408-6418 Sentence denotes Objective:
T54 6419-6624 Sentence denotes The purpose of this study was to investigate the genotype distribution of S. maltophilia isolates from patients with cystic fibrosis (CF) by repetitive sequence-based PCR (rep-PCR) using DiversiLab System.
T55 6625-6633 Sentence denotes Methods:
T56 6634-6786 Sentence denotes Sputum samples (n=623) from 165 CF patients were inoculated onto a selective medium for improved isolation of S. maltophilia over a period of two years.
T57 6787-6885 Sentence denotes Suspected isolates were identified by conventional standardized methods, as well as by sequencing.
T58 6886-6958 Sentence denotes A total of 65 S. maltophilia isolates from 33 CF patients were detected.
T59 6959-7140 Sentence denotes For genotyping, DNA extraction was performed with the UltraCleanTM microbial DNA isolation kit (Mo Bio Laboratories, Carlsbad, CA, USA), according to the manufacture´s instructions.
T60 7141-7285 Sentence denotes DiversiLab bacterial DNA fingerprinting kit (bioMérieux, Marcy I´Etoile, France) was used for typing following the protocol of the manufacturer.
T61 7286-7377 Sentence denotes The relatedness was determined by cluster analysis using Pearson´s Correlation coefficient.
T62 7378-7522 Sentence denotes Isolates with a similarity of 95% were considered as clinical related, and isolates with a similarity >98% were considered as indistinguishable.
T63 7523-7531 Sentence denotes Results:
T64 7532-7642 Sentence denotes Rep-PCR demonstrated that S. maltophilia isolates from CF patients exhibited a high rate of genetic diversity.
T65 7643-7772 Sentence denotes A small proportion of isolates were clonally related (12%), suggesting either cross-transmissions or a common source of exposure.
T66 7773-7868 Sentence denotes Conclusions: S. maltophilia from CF patients showed a wide variation of the molecular profiles.
T67 7869-8002 Sentence denotes Further epidemiological studies such as source tracking are required to elucidate the route of colonization/infection in CF patients.
T68 8003-8289 Sentence denotes The objective of this study was to determine whether photochemical inactivation of viruses could be accomplished with high efficiency while preserving the molecular integrity of viral targets allowing subsequent diagnostic tests to be performed at a lower level of containment and cost.
T69 8290-8463 Sentence denotes We studied the effect of either 5-Iodonaphthyl 1-azide (INA) and amotosalen (AMO, also known as S-59), that are known to target either viral proteins or viral nucleic acids.
T70 8464-8689 Sentence denotes We found that Vaccinia virus (VacV), an orthopox virus with a DNA genome, and Pixuna virus (PIXV), an alphavirus with an RNA genome, were stable when irradiated with UVA alone or when exposed to either INA or AMO in the dark.
T71 8690-8831 Sentence denotes AMO followed by UVA exposure was at least one-thousand-fold more virucidal than INA/UVA on VacV and on PIXV treated under similar conditions.
T72 8832-8992 Sentence denotes Photoinactivation with either INA or AMO at conditions that abolished viral infectivity resulted only in minimal impairment of subsequent ELISA and PCR testing.
T73 8993-9277 Sentence denotes The presented data could assist in developing methods to inactivate in the field environmental and forensic samples suspected of contamination with high virulence virus, thus limiting the need for costly security and safety operations after an accidental or intentional viral release.
T74 9278-9417 Sentence denotes The systemic symptoms of diphtheria are caused by the tox-encoded diphtheria toxin (DT) which is produced by toxigenic Corynebacterium spp.
T75 9418-9584 Sentence denotes In recent years, the emerging zoonotic pathogen C. ulcerans has replaced C. diphtheriae as the major agent for diphtheria in several industrialized Western countries.
T76 9585-9785 Sentence denotes The reliable detection of toxigenic Corynebacterium spp. is substantial for both diphtheria surveillance in the public health sector and the clinical work-up of a patient with diphtherialike symptoms.
T77 9786-9975 Sentence denotes Since the respective tox genes of C. diphtheriae and C. ulcerans differ in both DNA and amino acid sequence from each other, both tox genes should be covered by novel real-time PCR methods.
T78 9976-10262 Sentence denotes We describe the development and validation of a LightCycler PCR assay which reliably recognizes tox genes from both C. diphtheriae and C. ulcerans and differentiates the respective target genes by fluorescence resonance energy transfer (FRET) hybridization probe melting curve analysis.
T79 10263-10274 Sentence denotes Background:
T80 10275-10539 Sentence denotes Detection of Enterobacteriaceae carrying carbapenemases is challenging since the MIC can be in the susceptible range and because differentiation between carbapenem resistance due to carbapenemases or other mechanisms is not possible by routine susceptible testing.
T81 10540-10548 Sentence denotes Methods:
T82 10549-10698 Sentence denotes Enterobacteriaceae with elevated carbapenem MICs were sent to the German National Reference Laboratory for multidrug-resistant gramnegative bacteria.
T83 10699-10844 Sentence denotes For this study 80 strains were used, for which carbapenemases were excluded by a bioassay based on cell-free extracts and up to 15 specific PCRs.
T84 10845-10930 Sentence denotes In addition, 80 strains with carbapenemases characterized by sequencing were studied.
T85 10931-11124 Sentence denotes In all strains the MIC for carbapenems was determined by Etest, the modified Hodge-Test (MHT) was performed on different media and combined disk tests with EDTA and boronic acid were performed.
T86 11125-11133 Sentence denotes Results:
T87 11134-11279 Sentence denotes The collection comprised K. pneumoniae (n = 82), E. coli (n = 24), E. cloacae (n = 23), E. aerogenes (n =12) and other Enterobacteriacea species.
T88 11280-11637 Sentence denotes Strains with OXA-48 (n = 24), OXA-162 (n = 4), KPC-2 (n = 12), KPC-3 (n = 11), VIM-1 (n = 15), VIM-2 (n = 2), VIM-4 (n = 4), NDM-1 (n = 4) and GIM-1 (n = 3) were included.The sensitivity of the MHT on Mueller-Hinton agar (Oxoid) was 98.8%, 88.8% and 93.4% for imipenem,meropenem and ertapenem, respectively, whereas the specifity was 71.3%, 91.3% and 86.3%.
T89 11638-11833 Sentence denotes For the subgroup of metallo-betalactamases (n = 27) the MHT was performed on four different agar media with the lowest sensitivity for the MHT with meropenem performed on MH agar by Merck (7.4%).
T90 11834-11913 Sentence denotes Addition of 20 μl 50 mM zinc to the disk increased the sensitivity only to 30%.
T91 11914-12026 Sentence denotes The sensitivity of the MBL Etest was 74.1%, whereas the combined disk test with EDTA had a sensitivity of 96.3%.
T92 12027-12039 Sentence denotes Conclusions:
T93 12040-12125 Sentence denotes In contrast to recommendations by CLSI the MHT should also be performed for imipenem.
T94 12126-12225 Sentence denotes Despite a lower specifity the importance of carbapenemase detection justifies a higher sensitivity.
T95 12226-12342 Sentence denotes Caution should be used in choosing the test media since huge differences in metallobetalactamase detection are seen.
T96 12343-12458 Sentence denotes Due to a better sensitivity the combined disk test with EDTA should substitute the MBL Etest in Enterobacteriaceae.
T97 12459-12472 Sentence denotes Introduction:
T98 12473-12556 Sentence denotes Urinary tract infections (UTI) are the most common nosocomial infections worldwide.
T99 12557-12846 Sentence denotes Standard procedures for the identification of uropathogens are based on the cultivation on agar plates and the subsequent biochemical differentiation (e.g. with VITEK™, API™ etc.) and often require more than one day so that the initial antibiotic treatment is non-specific for the species.
T100 12847-13061 Sentence denotes We developed a protocol for a fast (< 30 min) direct identification of the species of pathogens in urine samples based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).
T101 13062-13070 Sentence denotes Methods:
T102 13071-13183 Sentence denotes We analysed urine samples from inpatients sent to the Institute for Medical Microbiology for routine diagnostic.
T103 13184-13351 Sentence denotes We separated the uropathogens from the sediment consisting of leucocytes, erythrocytes, cell detritus and protein cylinders by centrifugation and filtration protocols.
T104 13352-13435 Sentence denotes The filter residue was then eluated and transferred onto the spot of a metal plate.
T105 13436-13608 Sentence denotes After inactivation with 2.5-dihydroxybenzoic acid as organic matrix the protein spectra were acquired by a MALDI-TOF-mass spectrometer (Voyager-DE™ STR, AppliedBiosystems).
T106 13609-13709 Sentence denotes For identification the spectra were compared with a database (SARAMIS™, AnagnosTec, now BioMérieux).
T107 13710-13760 Sentence denotes The whole process was completed within 30 minutes.
T108 13761-13769 Sentence denotes Results:
T109 13770-13840 Sentence denotes Direct identification of uropathogens is already successful in > 60 %.
T110 13841-14023 Sentence denotes In 90 % of the unidentifiable samples a very intense peak triplet occurs corresponding to the human alpha-defensins 1, 2, and 3 as interfering factor at m/z values of around 3440 Da.
T111 14024-14191 Sentence denotes These proteins are produced naturally in the body and cannot be removed simply by washing the filter residue because they closely interact with the bacterial membrane.
T112 14192-14277 Sentence denotes Therefore we have to improve our protocol for purification of the separated bacteria.
T113 14278-14385 Sentence denotes In ticks different pathogens can be found that may cause serious infections in humans and domestic animals.
T114 14386-14553 Sentence denotes For the reliable detection of these pathogens by PCR, isolation of nucleic acids is one of the most important prerequisites influencing the whole diagnostic procedure.
T115 14554-14627 Sentence denotes Therefore an optimized protocol for DNA isolation from ticks is required.
T116 14628-14801 Sentence denotes Since the question regarding optimal DNA extraction from ticks is only poorly addressed so far, the present study compared different methods for isolation of DNA from ticks.
T117 14802-14823 Sentence denotes Material and Methods:
T118 14824-14918 Sentence denotes Four different methods for the DNA extraction from field-collected, unfed ticks were compared.
T119 14919-15049 Sentence denotes DNA-quantification in all experiments was done by 1) NanoDrop-spectrometer and 2) quantitative PCR measurement of tick beta-actin.
T120 15050-15366 Sentence denotes In experiment 1 DNA was isolated from each 15 single homogenized nymphs per method: a) high-salt DNA isolation protocol (PeqLab, Germany) b) MagBeads DNA extraction protocol (Carl-Roth, Germany) c) column-based DNA extraction protocol (PeqLab, Germany) and d) a modified TriPure-isolation protocol (PeqLab, Germany).
T121 15367-15438 Sentence denotes Experiments a-c were done according to the manufacturer's instructions.
T122 15439-15547 Sentence denotes In experiment 2 the best DNA extraction method of experiment 1 was used for an additional enrichment of DNA.
T123 15548-15685 Sentence denotes Again 15 single nymphs per treatment were used, each pre-treated e) with liquid nitrogen or f) without pre-treatment prior DNA isolation.
T124 15686-15858 Sentence denotes These experimental runs were combined with different additional steps of proteinase K treatment prior to experiments e and f based on differences in the treatment duration.
T125 15859-15980 Sentence denotes Finally the best method of experiment 2 was used for the DNA-isolation from adult ticks in comparison to the methods a-c.
T126 15981-15989 Sentence denotes Results:
T127 15990-16137 Sentence denotes In experiment 1 the highest DNA-yields were obtained by the modified TriPure DNA isolation protocol with up to 15 times higher DNAamount (p<0.001).
T128 16138-16295 Sentence denotes In experiment 2 the pre-treatment with proteinase k overnight combined with an additional liquid nitrogen treatment showed the highest DNA recovery (p<0,03).
T129 16296-16399 Sentence denotes Finally these findings were transferable to the DNA isolation results from adult ticks (p 0,001) ( fig.
T130 16400-16404 Sentence denotes 1 ).
T131 16405-16416 Sentence denotes Discussion:
T132 16417-16551 Sentence denotes For the detection of different pathogens in ticks a reliable and repeatable method for the DNA-isolation is an important prerequisite.
T133 16552-16673 Sentence denotes Our experiments clearly show a significant different performance of various DNA extraction methods when applied to ticks.
T134 16674-16747 Sentence denotes Furthermore, modifications clearly enhanced the quantity of isolated DNA.
T135 16748-16905 Sentence denotes Using this optimized method, studies with a new qualitative and quantitative PCR are on the way to measure the amount of borrelia in single ticks in Germany.
T136 16906-17053 Sentence denotes -lactamases-producing (ESBL) Enterobacteriacea show resistance against various -lactam-antibiotics like penicillins, cephalosporins, and aztreonam.
T137 17054-17211 Sentence denotes As minimal inhibitory concentration (MIC) of these strains can be increased but below the standard CLSI-breakpoints, further confirmatory tests are required.
T138 17212-17359 Sentence denotes This study compared ESBL-screenig results of MIC-determination, disk diffusion confirmatory test and growth onto a chromogenic ESBL-detection agar.
T139 17360-17466 Sentence denotes Program 1378 E. coli from diseased food-delivering and companion animals were collected from 2006 to 2007.
T140 17467-17818 Sentence denotes MIC determination by broth microdilution to -lactam-antibiotics (penicillin G, cefotaxime, cefoperazone, cefquinome, ceftiofur), and disk diffusion confirmatory test for ESBL (ceftazidine (30μg)/ ceftazidine-clavulanic acid (30μg/10μg) and cefotaxime (30μg)/ cefotaxime-clavulanic acid (30μg/10μg) were performed in accordance to CLSI document M31-A3.
T141 17819-17917 Sentence denotes Brilliance ESBL ® chromogenic agar (Oxoid) was applied corresponding manufacturer's constructions.
T142 17918-18018 Sentence denotes ESBL-suspicious strains were verified by detection of blaCTX-M-, blaOXA-, blaTEM-, and blaSHV-genes.
T143 18019-18027 Sentence denotes Results:
T144 18028-18162 Sentence denotes At broth microdilution testing, 49 E. coli (3.55%) exhibited MIC values of 2 mg/L for cefotaxime (=CLSI ESBL-screening concentration).
T145 18163-18319 Sentence denotes Simultaneously performed disk diffusion test and screening onto chromogenic agar resulted in 27 (1.96%) and 26 (1.87%) ESBL-suspicious strains respectively.
T146 18320-18371 Sentence denotes By PCR 26 ESBL-positive E. coli could be confirmed.
T147 18372-18574 Sentence denotes All these strains were characterized by MIC values of 16 mg/L for cefotaxime, a 5mm increased zone diameter in disk diffusion ESBL confirmatory test and blue growth onto chromogenic ESBL-detection agar.
T148 18575-18587 Sentence denotes Conclusions:
T149 18588-18724 Sentence denotes Overall, disk diffusion ESBL confirmatory test as well as chromogenic ESBL-detection agar were suitable to detect ESBL-positive E. coli.
T150 18725-18765 Sentence denotes Both tests showed a sensitivity of 100%.
T151 18766-18897 Sentence denotes The chromogenic agar further resulted in a specifity of 100%, whereas by disk diffusion test one falsepositive strain was detected.
T152 18898-18974 Sentence denotes However, use of disk diffusion tests is often more staff-and time-intensive.
T153 18975-18986 Sentence denotes Background:
T154 18987-19100 Sentence denotes Enterobacteriaceae resistant to carbapenems are a growing problem in an increasing number of countries worldwide.
T155 19101-19230 Sentence denotes In Germany, carbapenemase resistance is still rare, but appears to be rising, and several hospital outbreaks have been described.
T156 19231-19385 Sentence denotes As many infections with carbapenem resistant strains are acquired abroad, special precautions have to be undertaken to control infection and local spread.
T157 19386-19446 Sentence denotes Rapid isolate typing is a cornerstone for infection control.
T158 19447-19480 Sentence denotes Outbreak description and results:
T159 19481-19577 Sentence denotes In our institution, no carbapenemase producing enterobacteriaceae have been isolated previously.
T160 19578-19778 Sentence denotes In January, 2011, a Russian patient was admitted, and screening for multiresistant microorganisms revealed colonization with a Klebsiella pneumoniae strain of increased MIC for imipenem and meropenem.
T161 19779-19902 Sentence denotes Carbapenemase production was demonstrated by modified Hodge Test and confirmed by genotyping for OXA-48 type carbapenemase.
T162 19903-20059 Sentence denotes Subsequently, four additional patients were found to be colonized with K. pneumoniae strains of identical resistance patterns and later as OXA-48 producers.
T163 20060-20208 Sentence denotes It is suspected that transmission occurred during the time span between admission and initiation of infection control measures on the index patient.
T164 20209-20360 Sentence denotes Random PCR (RAPD) was attempted to prove clonal origin of the isolates, yet, in case of these K. pneumoniae isolates the method was not discriminative.
T165 20361-20689 Sentence denotes As an alternative for rapid typing, MALDI-TOF spectra of isolates were compared yielding an excellent spectrum identity between outbreak strains and a good discrimination towards other unrelated isolates, yet, automated calculation of the dendrogram by the biotyper software was not applicable for intra-species differentiation.
T166 20690-20769 Sentence denotes Results were subsequently confirmed by pulsed-field gel electrophoresis (PFGE).
T167 20770-20781 Sentence denotes Conclusion:
T168 20782-20989 Sentence denotes In conjunction with or instead of other rapid typing methods (e.g. RAPD), the diagnostic use of MALDI-TOF spectra may be helpful for preliminary epidemiologic analysis of clinical outbreaks within few hours.
T169 20990-21268 Sentence denotes The values and limitations of the method need to be further validated for different microorganisms by a standard procedure (e.g. PFGE, MLST), yet, ease of applicability and low running costs make mass spectra analysis a promising tool for first line clinical outbreaks analysis.
T170 21270-21386 Sentence denotes Mycoplasma pneumoniae (M.p.) is the third most frequent pathogen responsible for community-acquired pneumonia (CAP).
T171 21387-21491 Sentence denotes Infections are most prevalent in children and in young adults showing epidemic peaks every 3 to 7 years.
T172 21492-21591 Sentence denotes M.p. infections are often diagnosed by PCR but mainly by serology in clinical routine laboratories.
T173 21592-21717 Sentence denotes In this study, we evaluated more than 35 recombinant M.p.-specific proteins to develop a serological line-immunoassay (Line).
T174 21718-21824 Sentence denotes Out of these, seven proteins proved to be useful in the detection of specific IgG, IgA and IgM antibodies.
T175 21825-21989 Sentence denotes In addition, the I-protein of human erythrocytes, an autoantigen, which induces cold agglutinins of the IgM-type in 50 to 70% of acute M.p. infections was included.
T176 21990-22227 Sentence denotes Furthermore, the integration of genotype-specific and conserved regions of the main P1 adhesin enables us to differentiate between infections with the two most common M.p. subtypes 1 and 2 and to estimate the prevalence for specific IgG.
T177 22228-22578 Sentence denotes To establish the Line, we tested 200 sera (46 sera with possible M.p. infection, 52 blood donors, 38 Bordetella pertussis or Legionella pneumophila positive sera, 37 sera of patients with acute non-respiratory infections and 27 sera of children; all sera were pretested by ELISA and Western Blot using the whole M.p. antigen preparation) in the Line.
T178 22579-22698 Sentence denotes When compared to a consensus result of ELISA and Western Blot the following sensitivities and specificities were found:
T179 22699-22747 Sentence denotes 93%/98% (IgG), 100%/93% (IgA) and 98%/94% (IgM).
T180 22748-22831 Sentence denotes In addition, we determined a M.p.-specific IgGprevalence of 53% for our sera panel.
T181 22832-22927 Sentence denotes A positive PCR result as reference test for confirmation of M.p. infection is well established.
T182 22928-23036 Sentence denotes Therefore, we analysed 70 sera from the network for community-acquired pneumonia (CAPNET) with positive M.p.
T183 23037-23090 Sentence denotes PCR result in the corresponding respiratory specimen.
T184 23091-23119 Sentence denotes Furthermore, 30 sera of M.p.
T185 23120-23273 Sentence denotes PCR-negative healthy persons and 30 sera from patients suffering from CAP caused by other agents than M.p. were analysed for sensitivity and specificity.
T186 23274-23391 Sentence denotes In reference to the PCR results the Line achieved sensitivities for IgG, IgA and IgM of 53, 65 and 83%, respectively.
T187 23392-23506 Sentence denotes When accumulating the results for IgA and IgM and for IgG, IgA and IgM, sensitivities of 91 and 92% were obtained.
T188 23507-23679 Sentence denotes The results demonstrate the practical usefulness of a new line blot based on selected proteins as a highly specific and sensitive tool for serodiagnosis of M.p. infections.
T189 23680-23682 Sentence denotes A.
T190 23683-23701 Sentence denotes Bittner* 1 , K.-D.
T191 23702-23715 Sentence denotes Müller 2 , J.
T192 23716-23729 Sentence denotes Buer 2 , A.W.
T193 23730-23748 Sentence denotes Rettenmeier 1 , F.
T194 23749-23948 Sentence denotes Mosel 1 1 Universitätsklinikum Essen, Institut für Hygiene und Arbeitsmedizin, Essen, Deutschland 2 Universitätsklinikum Essen, Institut für Medizinische Mikrobiologie, Essen, Deutschland Background:
T195 23949-24120 Sentence denotes The standard procedures for the identification of bacteria are based on the detection of distinct metabolic properties (biochemical test systems such as API ® , VITEK ® ).
T196 24121-24252 Sentence denotes In addition, gas chromatographic determination of species-specific patterns of long-chain fatty acids is used in some laboratories.
T197 24253-24571 Sentence denotes Bacteria isolated from tissues and fluids of patients under antibiotic treatment and, specifically, from the viscous bronchial mucus in the respiratory tract of patients with cystic fibrosis often have changed their characteristic metabolic properties and therefore elude the identification by conventional procedures.
T198 24572-24786 Sentence denotes We examined whether the analysis of bacterial protein patterns by MALDI-TOF-MS (Matrix-assisted laser desorption/ionization time of flight mass spectrometry) results in an unequivocal identification in those cases.
T199 24787-24795 Sentence denotes Methods:
T200 24796-25008 Sentence denotes Isolates obtained from tissues and fluids of patients in the clinical routine which could not be unambiguously identified by API ® or VITEK ® or by gas chromatography, respectively, were analyzed by MALDI-TOF-MS.
T201 25009-25109 Sentence denotes A small number of bacterial cells from the culture plate was directly deposited onto a sample plate.
T202 25110-25271 Sentence denotes The bacteria were inactivated and intracellular, mostly ribosomal proteins were liberated by exposure to 0.3 μl 2.5-dihydroxybenzoic acid used as organic matrix.
T203 25272-25381 Sentence denotes The spectra of these proteins were compared with the entries of a database (SARAMIS ® , AnagnosTec, Germany).
T204 25382-25482 Sentence denotes The whole process of sample preparation, measurement and database matching takes only a few minutes.
T205 25483-25491 Sentence denotes Results:
T206 25492-25668 Sentence denotes By using MALDI-TOF-MS analysis we could identify nearly 80 % of all isolates which previously had not given a clear result when standard identification procedures were applied.
T207 25669-25776 Sentence denotes In most cases the bacteria could be identified at the species level, in some cases only at the genus level.
T208 25777-26025 Sentence denotes DNA from adult ticks (n=15 per treatment) isolated by different extraction protocols: (a) high-salt DNA isolation protocol, (b) MagBeads DNA extraction protocol, (c) column-based DNA extraction protocol, and (d) modified TriPure-isolation protocol.
T209 26026-26113 Sentence denotes Rapid detection of toxin producing Clostridium difficile is essential in CDAD patients.
T210 26114-26334 Sentence denotes This study compares the performance of a new PCR assay (BD GenOHM Cdiff Assay) and a conventional ELISA (Ridascreen C. difficile Toxin A/B) for rapid detection of toxin-producing C. difficile strains directly from stool.
T211 26335-26462 Sentence denotes 531 stool samples obtained from 411 patients with suspected CDAD from a single centre were analysed by real-time PCR and ELISA.
T212 26463-26572 Sentence denotes Only samples in accordance with the criteria for diarrheal stool (Bristol Score) were eligible for the study.
T213 26573-26628 Sentence denotes Anaerobic cultures were also performed (George´s agar).
T214 26629-26686 Sentence denotes The study was approved by the institutional review board.
T215 26687-26830 Sentence denotes Using the ELISA as an analytical gold standard PCR yielded 106 concordant positive results (93,8%) and 352 concordant negative results (84,2%).
T216 26831-26889 Sentence denotes In 73 stool samples both assays showed discrepant results.
T217 26890-27020 Sentence denotes Including culture results into the discrepant analysis, PCR resulted in 165 (31,1%) true positives and 359 (67,1%) true negatives.
T218 27021-27203 Sentence denotes One false positive and 7 false negative PCR tests were observed yielding an analytical sensitivity and specificity of 99,4 respective 98,1% compared to 64,5% and 98,4% for the ELISA.
T219 27204-27295 Sentence denotes Positive and negative predictive values for PCR testing were 95,9% and 99,7%, respectively.
T220 27296-27564 Sentence denotes Although, studies dealing with cost effectiveness of PCR in the management of CDAD patients are sparse, the BD GenOhm Cdiff assay (PCR) has greater analytical sensitivity than a conventional toxin ELISA maintaining comparable specificity in samples from CADA patients.
T221 27565-27567 Sentence denotes A.
T222 27568-27584 Sentence denotes Halfmann* 1 , U.
T223 27585-27616 Sentence denotes Geipel 1 , L. von Müller 1 , M.
T224 27617-27743 Sentence denotes Herrmann 1 1 University Hospital of Saarland, Institute for Medical Microbiology and Hygiene, Homburg, Deutschland Background:
T225 27744-28021 Sentence denotes Universal screening for methicillin-resistant Staphylococcus aureus (MRSA) is a valuable tool to estimate MRSA prevalence, yet, for a single institution, the laboratory challenges of a large campaign requiring quality-assured handling and processing of samples can be enormous.
T226 28022-28131 Sentence denotes For a two-month universal MRSA admission prevalence screening project, >20.000 samples needed to be analyzed.
T227 28132-28410 Sentence denotes To reduce the manual burden for processing 500-1000 additional specimens workdaily, the Walk Away Specimen Processor (WASP, Copan, Italy), a fully automated, robot-based instrument managing all steps of tube handling, vortexing, medium inoculation, and plate labelling was used.
T228 28411-28528 Sentence denotes Here we report on our experience using WASP in this setting and its parallel use in processing other routine samples.
T229 28529-28537 Sentence denotes Methods:
T230 28538-28585 Sentence denotes All samples were collected with eSwabs (Copan).
T231 28586-28771 Sentence denotes For the MRSA project, a modified eSwab system consisting of two flocked collection swabs (for throat, and both nares, respectively) and one liquid Amies medium-containing tube was used.
T232 28772-28915 Sentence denotes Samples were inoculated on chromogenic S. aureus / MRSA biplates (Mast, Germany), which were incubated at 37°C for 48h and read at 24h and 48h.
T233 28916-29051 Sentence denotes Other routine samples like urines were processed according to standard procedures using different universal and selective media plates.
T234 29052-29060 Sentence denotes Results:
T235 29061-29169 Sentence denotes During an initial pilot phase, 800 MRSA screening samples were plated in parallel both by WASP and manually.
T236 29170-29379 Sentence denotes Initially, important differences in recovery rates between both methods appeared, with manually inoculated plates yielding about ten times elevated colony counts compared to parallel plates inoculated by WASP.
T237 29380-29634 Sentence denotes Subsequently, several modifications on the WASP specimen processing protocol (various vortexing times, quantities of medium plated or streaking patterns) were carried out with support of the readily available WASP technical team both on-line and on-site.
T238 29635-29724 Sentence denotes A specific optimized streaking pattern was found to be crucial for optimal colony counts.
T239 29725-29937 Sentence denotes After optimizing the WASP protocol, additional testing of specimen processed either by WASP or manually now revealed that even at very low colony numbers, the sensitivity of WASP was comparable to manual plating.
T240 29938-30097 Sentence denotes The readily achieved WASP connection to our LIS MLab (Dorner, Germany) further allowed for processing of a variety of different samples types at the same time.
T241 30098-30222 Sentence denotes As a basis for fast and reliable work flow, streaking quality and yield of single colonies was consistently of high quality.
T242 30223-30412 Sentence denotes Especially for urines, use of a one quadrant streaking pattern allowed an easy and fast semiquantitative estimation of colony count by showing a fir tree like growth pattern on agar plates.
T243 30413-30424 Sentence denotes Conclusion:
T244 30425-30650 Sentence denotes Integration of WASP in the laboratory procedures both for a large universal screening program and for processing of routine samples allowed for considerable reduction of manual workload and qualityconform specimen processing.
T245 30651-30789 Sentence denotes False positive candida and cryptococcal antigen detection in serum as a result of cross reactivity with different therapeutic iv solutions
T246 30790-30901 Sentence denotes The Epstein -Barr virus (EBV) is a virus of the herpes family, and is one of the most common viruses in humans.
T247 30902-31023 Sentence denotes Most people become infected with EBV at an early age, which is often asymptomatic but may cause infectious mononucleosis.
T248 31024-31110 Sentence denotes Symptoms of infectious mononucleosis are fever, sore throat, and swollen lymph glands.
T249 31111-31265 Sentence denotes Although the symptoms of infectious mononucleosis usually resolve in 1 or 2 months, EBV dormant or latent in a few cells in the throat and blood for life.
T250 31266-31365 Sentence denotes Periodically, the virus can reactivate and is commonly found in the saliva of infected individuals.
T251 31366-31427 Sentence denotes This reactivation usually occurs without symptoms of illness.
T252 31428-31593 Sentence denotes The novel recomWell EBV -test system combined with the confirmatory assay recomLine EBV IgG (Avidity), IgM is an effective tool for comprehensive EBV serodiagnostic.
T253 31594-31764 Sentence denotes The system is based on four separate ELISA (enzyme linked immunosorbent assay); recomWell EBV EBNA IgG, recomWell EBV VCA IgG, recomWell EBV EA IgG and recomWell EBV IgM.
T254 31765-31922 Sentence denotes The standardized conception design of the EIA test kits enables implementation of all assays by using a single sample dilution and standard incubation times.
T255 31923-32099 Sentence denotes The system performance was evaluated in four sample collections; healthy blood donors, EBV seronegative samples und samples representative of acute and subsided EBV infections.
T256 32100-32166 Sentence denotes The assays were characterized by high specificity and sensitivity.
T257 32167-32503 Sentence denotes As compared with other commercially available EBV EIA test systems, the recomWell EBV tests showed considerable advantages; compared to the other assays tested, the primary EBV infections were detected in very early phase of an infection due to the high sensitivity of the recomWell EBV VCA IgG, recomWell EBV EA IgG and EBV IgM assays.
T258 32504-32665 Sentence denotes Additionally, due to the high specificity of the recomWell EBNA IgG assay false positive anti-EBNA IgG findings among the acute infections were utterly excluded.
T259 32666-32981 Sentence denotes The high specificity of Anti -EBNA IgG assays is an important aspect of EBV diagnostic, as the differentation of a primary recent infection from a subsided past infection is primarily achieved through detection of anti -EBNA IgG antibodies (Screening) or measurement of the avidity of IgG antibodies (confirmation).
T260 32982-33306 Sentence denotes Finally, the results obtained here were compared with the recently published evaluation study of four commercial systems for the diagnostics of EBV primary infections (de Ory et al., 03/2011, Clinical and Vaccine Immunology, Evaluation of Four Commercial Systems for the Diagnosis of Epstein -Barr Virus Primary Infections).
T261 33308-33507 Sentence denotes Several studies have been published that claimed to identify oxacillin resistance of Staphylococcus aureus by whole cell matrix assisted laser desorption time of flight (MALDI-TOF) mass spectrometry.
T262 33508-33651 Sentence denotes In these studies selected methicillin susceptible S. aureus isolates were compared to selected methicillin resistant S. aureus (MRSA) isolates.
T263 33652-33875 Sentence denotes Nevertheless, the study design in all of those studies was not suitable to achieve this goal, because only mecA/SCCmec isogenic strains could discriminate between strain specific-and mecA / SCCmec-dependent peak variations.
T264 33876-33971 Sentence denotes It have been also described that strains specific peaks exist within the species of S. aureus..
T265 33972-34090 Sentence denotes We identified in our study mecA/SCCmec isogenic strains of S. aureus and analysed their peak profiles by MALDI-TOF MS.
T266 34091-34191 Sentence denotes A large MRSA strain collection containing 120 strains with molecular divergent backgrounds was used.
T267 34192-34294 Sentence denotes The mecA gene was found in only 104 out of 120 of these isolates after culturing the freezing culture.
T268 34295-34375 Sentence denotes The loss of the mecA gene during freezing culture has been previously described.
T269 34376-34485 Sentence denotes Single colonies of those isolates were identified by the cefoxitin resistance phenotype and mecA PCR results.
T270 34486-34655 Sentence denotes The bacteria from freezing culture were submitted to LB media containing increasing oxacillin concentrations in order to select the corresponding mecA containing strain.
T271 34656-34780 Sentence denotes In two out of 16 samples we could select a mecA containing strain and a corresponding isogenic strain lacking the mecA gene.
T272 34781-34877 Sentence denotes These two pairs of molecularly characterised strains were submitted for whole cell MALDI-TOF MS.
T273 34878-35020 Sentence denotes In order to increase the expression of the PBP2a, the oxacillin resistant strain was also cultured in LB medium containing NaCl and oxacillin.
T274 35021-35099 Sentence denotes The corresponding oxacillin susceptible strain was cultured in LB medium only.
T275 35100-35291 Sentence denotes We do not found evidence for a difference in the peak profiles in a mass-tocharge-ratio (Da) of 2000 to 15000 Da in two mecA-isogenic pairs of S. aureus strains cultured on blood agar plates.
T276 35292-35471 Sentence denotes The protein peak profiles of the mecA positive strains, cultured on oxacillin containing media, were also virtually identical compared to the mecA negative corresponding isolates.
T277 35472-35724 Sentence denotes In contrast to several reports in literature, we could show for the first time that whole cell MALDI-TOF MS-derived peak profiles are not suitable for the discrimination of mecA positive S. aureus strains in the mass-to-chargeratio of 2000 to 15000 Da.
T278 35725-35893 Sentence denotes Rapid identification of mixed-and single bacterial and fungal infections with PCR coupled to Electrospray-Ionization Timeof-Flight Mass Spectrometry (PCR-ESI/TOF MS) C.
T279 35894-35909 Sentence denotes Tiemann* 1 , A.
T280 35910-35925 Sentence denotes Hartmann 1 , D.
T281 35926-35955 Sentence denotes Knoop 1 1 Labor Krone, Molek.
T282 35956-36119 Sentence denotes Diagnostik, Bad Salzuflen, Deutschland The identification of bacteria and yeast from blood culture is a time consuming step, shortening the benefit of the patient.
T283 36120-36292 Sentence denotes Several technologies were developed to replace the second incubation step on agar plates and the biochemical characterization, most prominent analysis by mass spectrometry.
T284 36293-36403 Sentence denotes However, the resolution of complex mixtures, detection of antibiotic resistance markers are still unmet needs.
T285 36404-36662 Sentence denotes In order to overcome these limitations, a new molecular technology, PCR coupled to Mass Spectrometry, was developed, allowing detection and identification of multiple pathogens and selected antibiotic resistance markers or virulence factors in a single test.
T286 36663-36956 Sentence denotes After PCR with 18 different broad range and specific primers covering bacteria, Candida and key antibiotic resistance markers, the base composition of the amplicons was determined by mass spectrometry with the Abbott PLEX-ID analyzer, followed by bioinformatical identification and genotyping.
T287 36957-37118 Sentence denotes Clinical samples (positive and negative blood cultures) as well as spiked blood cultures were analyzed with the PLEX-ID instrument and the BAC Spectrum BC assay.
T288 37119-37260 Sentence denotes The results were compared to traditional microbiology techniques, discordant results were additionally confirmed by 16s rRNA gene sequencing.
T289 37261-37359 Sentence denotes The PLEX-ID technology displayed a high specificity, superior to traditional molecular techniques.
T290 37360-37534 Sentence denotes We were also able to demonstrate the resolution of mixtures in several clinical samples and the resolution of up to 5 different bacteria and yeasts with a single primer pair.
T291 37535-37732 Sentence denotes Evaluation of S. pseudintermedius differentiation using molecular marker analysis and MALDI TOF MS EKP01 Functional analysis of the two type II NADH dehydrogenase isoforms in Toxoplasma gondii S.S.
T292 37733-37743 Sentence denotes Lin 1 , U.
T293 37744-37755 Sentence denotes Groß 1 , W.
T294 37756-38008 Sentence denotes Bohne* 1 1 Universitätsmedizin Göttingen, Institut für Medizinische Mikrobiologie, Göttingen, Deutschland NADH dehydrogenases play an important role in the energy metabolism by transferring electrons from NADH to the mobile electron carrier ubiquinone.
T295 38009-38245 Sentence denotes Several bacteria, fungi and protozoa possess so called type II NADH dehydrogenases (NDH2s), which are structurally and functionally different from the common complex I of mammalian cells and are thus considered as putative drug targets.
T296 38246-38330 Sentence denotes NDH2s are composed of a single subunit and are not involved in proton translocation.
T297 38331-38473 Sentence denotes The apicomplexan parasite Toxoplasma gondii possesses two NDH2 isoforms, TgNDH2-I and TgNDH2-II, which are both targeted to the mitochondrion.
T298 38474-38627 Sentence denotes Type II NADH dehydrogenases can occur either as internal enzymes, which utilize mitochondrial NADH, or as external enzymes, which utilize cytosolic NADH.
T299 38628-38849 Sentence denotes We determined the orientation of both isoforms by applying a split GFP complementation approach and could demonstrate that both isoforms are internal enzymes, which face with their active site to the mitochondrial matrix.
T300 38850-38978 Sentence denotes This internal orientation was confirmed by a protease protection assay after selective membrane permeabilization with digitonin.
T301 38979-39152 Sentence denotes To obtain functional data for the contribution of the individual isoforms in the T. gondii energy metabolism, we generated single knock-out mutants for each of the isoforms.
T302 39153-39306 Sentence denotes Both mutants were viable, but displayed a decreased replication rate, which was more severe in the Tgndh2-II-deleted than in the Tgndh2-I-deleted mutant.
T303 39307-39524 Sentence denotes A functional complementation was possible by ectopic expression of the deleted gene, but not by expression of the heterologous isoform, suggesting that the two enzymes possess nonredundant, isotype-specific functions.
T304 39525-39631 Sentence denotes Development of Toxoplasma gondii in skeletal muscle cells depends on differentiation of the host cell I.J.
T305 39632-39651 Sentence denotes Swierzy* 1 , C.G.K.
T306 39652-39870 Sentence denotes Lüder Institute for Medical Microbiology, Göttingen, Deutschland Toxoplasma gondii, an apicomplexan parasitic protozoan, infects almost every nucleated mammalian and avian cell including skeletal muscle cells (SkMCs) .
T307 39871-40062 Sentence denotes Replication of the parasite and its differentiation to the dormant bradyzoite stage in SkMCs are considered as crucial for the food-borne transmission to humans via undercooked or cured meat.
T308 40063-40228 Sentence denotes Previous results from our laboratory indicated that SkMCs promote spontaneous differentiation from tachyzoites to bradyzoites in vitro more readily than fibroblasts.
T309 40229-40359 Sentence denotes Therefore, the identification of cell-type specific factors which regulate the parasite development in SkMCs is of major interest.
T310 40360-40522 Sentence denotes In this study, cell type-specific interactions were investigated using permanent mouse-derived skeletal muscle cells (C2C12) and fibroblasts as controls (NIH3T3).
T311 40523-40607 Sentence denotes C2C12 myoblasts were differentiated to polynucleated, contractile myotubes in vitro.
T312 40608-40784 Sentence denotes Immunoblotting, immunofluorescence microscopy and quantitative RT-PCR validated the differentiation efficiency of C2C12 cells by measuring the expression of myosin heavy chain.
T313 40785-40974 Sentence denotes Intriguingly, after infection with T. gondii, parasite replication was clearly lower in differentiated C2C12 myotubes than in undifferentiated C2C12 myoblasts or NIH3T3 control fibroblasts.
T314 40975-41189 Sentence denotes After 2 days of infection, parasitophorous vacuoles (PV) contained an average of 6 and 7 parasites in C2C12 myoblasts or NIH3T3, respectively, whereas only 3 parasites were found in the PVs in differentiated C2C12.
T315 41190-41326 Sentence denotes Additionally, no parasite-induced cell lysis could be observed in differentiated C2C12 in contrast to nondifferentiated C2C12 or NIH3T3.
T316 41327-41501 Sentence denotes Quantification of Toxoplasma by realtime PCR confirmed a reduced replication of the parasite in differentiated C2C12 myotubes as compared to undifferentiated C2C12 myoblasts.
T317 41502-41619 Sentence denotes Mitogen Activated Protein Kinases (MAPK) such as p38 or ERK as well as Akt/PKB regulate the differentiation of SkMCs.
T318 41620-41752 Sentence denotes Therefore, we also investigated whether MAPK might control T. gondii development in differentiated and undifferentiated C2C12 SkMCs.
T319 41753-41886 Sentence denotes First results using immunoblotting showed a considerable increase of p38, ERK and Akt/PKB phoshorylation within differentiated C2C12.
T320 41887-42062 Sentence denotes Furthermore, the phosphorylation of p38 and Akt/PKB proteins was enhanced after 24h postinfection with T. gondii in differentiated C2C12 as compared to undifferentiated C2C12.
T321 42063-42186 Sentence denotes Further experiments on the impact of MAP Kinases on C2C12 differentiation as well as the infection process are in progress.
T322 42187-42316 Sentence denotes Together, these results suggest that the host cell type and the differentiation of SkMCs regulate T. gondii replication in vitro.
T323 42317-42430 Sentence denotes This might also explain the different efficiencies of T. gondii to persist in differentiated tissues of the host.
T324 42431-42662 Sentence denotes The Candida albicans factor H binding molecule Hgt1p also functions as complement receptor 3 (CR3) and HIV binding molecule EKP07 Caenorhabditis elegans as a host model system to screen novel antifungal benzimidazole derivatives A.
T325 42663-42676 Sentence denotes Singh* 1 , W.
T326 42677-42694 Sentence denotes Abu Rayyan 1 , T.
T327 42695-42710 Sentence denotes Dietsche 1 , H.
T328 42711-42729 Sentence denotes Eickhoff 2 , K.-H.
T329 42730-42747 Sentence denotes Wiesmüller 2 , K.
T330 42748-43054 Sentence denotes Schröppel 1 1 Universitätsklinikum Tübingen, Interfakultäres Institut für Mikrobiologie und Infektionsmedizin, Tübingen, Deutschland 2 EMC microcollections GmbH , Tübingen, Deutschland Candida albicans, a commensal microorganism, is a causative agent of opportunistic oral and genital infections in humans.
T331 43055-43129 Sentence denotes Infections with C. albicans account for 70 to 90% of all invasive mycoses.
T332 43130-43350 Sentence denotes Treatment of invasive fungal infections is problematic due to several persisting therapeutic drawbacks, in particular development of resistance against the currently available drugs or toxic side effects for the patient.
T333 43351-43440 Sentence denotes Various approaches have been developed to screen new antifungal drugs for their activity.
T334 43441-43596 Sentence denotes Previously, the nematode Caenorhabditis elegans has been successfully used as a model host for human pathogens and to screen for novel antimicrobial drugs.
T335 43597-43782 Sentence denotes It has been shown that C. albicans colonizes the intestinal tract of C. elegans and establishes a persistent lethal infection accompanied by deep tissue invasion of the entire organism.
T336 43783-44013 Sentence denotes We screened a series of novel (S)-2-aminoalkyl benzimidazole derivatives for their antifungal activity against various Candida isolates using a standard EUCAST (European Committee on Antimicrobial Susceptibility Testing) protocol.
T337 44014-44370 Sentence denotes One compound ((S)-2-(1-aminoisobutyl)-1- (3-chlorobenzyl) benzimidazole (EMC120B12)), showed very auspicious results and therefore was studied in more detail to analyze its antifungal activity against various Candida species, its ability to inhibit Biofilm formation in C. albicans and to determine the activity in vivo using C. elegans as an animal model.
T338 44371-44533 Sentence denotes EMC120B12 showed the highest antifungal activity against a set of various clinical isolates that included C. albicans, C. glabrata, C. parapsilosis and C. krusei.
T339 44534-44647 Sentence denotes Furthermore, EMC120B12 showed a significant reduction in C. albicans biofilm formation under in vitro conditions.
T340 44648-44759 Sentence denotes Additionally, we found that EMC120B12 had no toxic effects to C. elegans at effective inhibitory concentration.
T341 44760-44930 Sentence denotes During in vivo infection studies, we could demonstrate that EMC120B12 can inhibit filamentation of C. albicans and prolong survival of the C. albicans infected nematodes.
T342 44931-45181 Sentence denotes EMC120B12 identified in this screen can potentially be used as an effective antifungal agent and will be further investigated for its activity against other fungal species and also in a murine model to study its activity against systemic candidiasis.
T343 45182-45272 Sentence denotes The HexA protein of Aspergillus fumigatus and its role in stress resistance and virulence.
T344 45273-45275 Sentence denotes J.
T345 45276-45288 Sentence denotes Beck* 1 , B.
T346 45289-45307 Sentence denotes Echtenacher 2 , F.
T347 45308-45518 Sentence denotes Ebel Bakteriologie, München, Deutschland 2 Institut für Immunologie, Universität Regenburg, Regensburg, Deutschland The mycelium of filamentous fungi consists of an interwoven network of multi-nucleated hyphae.
T348 45519-45580 Sentence denotes Individual cells within these hyphae are separated by septae.
T349 45581-45785 Sentence denotes However, these internal walls contain central pores allowing the exchange of molecules, nutrients and even organelles enabling the mycelium to respond as a functional entity to changes in the environment.
T350 45786-45907 Sentence denotes The draw-back of this organization is that a local damage of the cell wall is a potential threat for the whole syncytium.
T351 45908-46059 Sentence denotes To prevent uncontrolled bleeding of cytoplasm, filamentous fungi developed a unique organelle, the Woronin body, which is able to seal the septal pore.
T352 46060-46294 Sentence denotes In several fungi the Hex1 or HexA protein has been identified a major and essential component of the Woronin body and mutants generated in Neurospora crassa or Aspergillus oryzae show uncontrolled loss of cytoplasm after local damage.
T353 46295-46484 Sentence denotes Loss of Hex1 in Magnaporthe grisea impairs the virulence of this plant pathogen, but no data are yet available on the role of Hex proteins in filamentous fungi causing infections in humans.
T354 46485-46679 Sentence denotes In this study, we have analyzed the importance of Woronin bodies and in particular of the HexA protein for the stress resistance and virulence of the human pathogenic mold Aspergillus fumigatus.
T355 46680-46787 Sentence denotes The phenotype of the A. fumigatus delta hexA mutant is less dramatic compared to the N. crassa hex1 mutant.
T356 46788-46997 Sentence denotes Under standard conditions growth of the A. fumigatus delta hexA mutant is comparable to that of the wild type, but in the presence of stressors, like Congo red or SDS, growth of the mutant is clearly impaired.
T357 46998-47079 Sentence denotes We have also analyzed the localization of HexA using fluorescent fusion proteins.
T358 47080-47254 Sentence denotes Whereas a GFP-HexA fusion is targeted to organelles showing the typical characteristics of Woronin bodies, a HexA-GFP fusion shows a striking localization at the septal wall.
T359 47255-47482 Sentence denotes To further investigate the consequences of a loss of HexA, we analyzed the virulence of the delta hexA mutant in a murine model of infection and studied its phenotype in confrontation assays using human and murine immune cells.
T360 47483-47636 Sentence denotes Functional analysis of C. albicans infection-associated genes with unknown function during interaction with host cells and in complex infection models I.
T361 47637-47653 Sentence denotes Jacobsen* 1 , D.
T362 47654-47667 Sentence denotes Wilson 1 , F.
T363 47668-47680 Sentence denotes Mayer 1 , B.
T364 47681-47755 Sentence denotes Hube Mikrobielle Pathogenitätsmechanismen, Jena, Deutschland Introduction:
T365 47756-47862 Sentence denotes Candida albicans is a common fungal pathogen in humans and able to cause a variety of clinical infections.
T366 47863-48050 Sentence denotes We hypothesized that C. albicans unknown function genes, which are transcriptionally upregulated during infection ("infection associated genes") might contribute to the infection process.
T367 48051-48071 Sentence denotes Aim and Methodology:
T368 48072-48165 Sentence denotes We aimed to determine whether C. albicans infection associated genes contribute to virulence.
T369 48166-48374 Sentence denotes Therefore, based on transcriptional profiling from oral infections, liver invasion and incubation in human blood, we selected a subset of infection-associated genes with unknown function for further analysis.
T370 48375-48555 Sentence denotes Isogenic deletion mutants were constructed and subsequently tested in extensive in vitro screens, interaction with epithelial and endothelial cells and in complex infection models.
T371 48556-48564 Sentence denotes Results:
T372 48565-48708 Sentence denotes Twentyfour isogenic deletion mutants were constructed and subsequently analyzed both in vitro and in infection models of increasing complexity.
T373 48709-48822 Sentence denotes Sixteen mutants were impaired in their ability to damage monolayers of human endothelial and/or epithelial cells.
T374 48823-48917 Sentence denotes Within this subset, seven mutants additionally displayed decreased stress resistance in vitro.
T1 48918-48964 Sentence denotes Only two mutants showed filamentation defects.
T2 48965-49178 Sentence denotes To determine whether the deleted genes influenced virulence in more complex infections models, all 24 mutants were tested in ovo for their ability to kill chicken embryos infected on the chorio-allantoic membrane.
T3 49179-49359 Sentence denotes We recently showed that mortality in this model depends on the fungal ability to invade the membrane and that the pro-inflammatory host response likely contributes to pathogenesis.
T4 49360-49497 Sentence denotes Surprisingly, only seven mutants were attenuated in this model, of which five were also attenuated in damaging epithelial cells in vitro.
T5 49498-49724 Sentence denotes Three of the mutants, which were attenuated in ovo, and one mutant, which was attenuated in damaging endothelial and epithelial cells but fully virulent in chicken embryos, were subsequently analyzed in a systemic mouse model.
T6 49725-49780 Sentence denotes Virulence in mice mirrored the results obtained in ovo.
T7 49781-49804 Sentence denotes Discussion and Outlook:
T8 49805-49993 Sentence denotes Our results demonstrate that genes essential for causing full damage of epithelial or endothelial cells in vitro are not necessarily crucial for full virulence in complex infection models.
T9 49994-50257 Sentence denotes We conclude that either such damage potential is not critical for full virulence in the complex host setting or that the greater complexity of the environment in animal models stimulates additional fungal regulatory networks leading to compensation of the defect.
T10 50258-50367 Sentence denotes Thiol-dependent redox networks: analysing the effects of the anti-malarial agent Methylene Blue Introduction:
T11 50368-50539 Sentence denotes The development and spread of resistance against standard antimalarial drugs such as chloroquine (CQ) and pyrimethamine/sulfadoxin (PSD) has been a challenge for 25 years.
T12 50540-50680 Sentence denotes Thus standard drugs -standard implies that they are affordable for and available to those who need them the most -are continuously required.
T13 50681-50908 Sentence denotes A number of drugs and drug candidates, for instance methylene blue (MB), are directed against the antioxidative redox-protein network of Plasmodium falciparum which contains a total of 25 redoxins and disulfide reductases [1] .
T14 50909-51030 Sentence denotes Application of YANAsquare [2] allowed dynamic modelling of enzyme inhibition by different drugs including methylene blue.
T15 51031-51150 Sentence denotes We analysed affected pathway modes, regulatory effects on affected enzymes, and combination effects of different drugs.
T16 51151-51173 Sentence denotes Materials and Methods:
T17 51174-51304 Sentence denotes The enzyme activity of key redox enzymes with and without the influence of methylene blue was measured (in vivo) by real time PCR.
T18 51305-51410 Sentence denotes Enzyme activity of enzymes affected by CQ and PSD was collected from literature and databases (PlasmoDB).
T19 51411-51654 Sentence denotes The software YANAsquare was used for extreme pathway (EP) calculation in order to evaluate the strain-specific redox-network, folate synthesis, haemoglobin degradation as well as porphyrine and carbon-pool networks with all contained pathways.
T20 51655-51663 Sentence denotes Results:
T21 51664-51867 Sentence denotes Modelling shows that metabolic fluxes through gluthation reductase as well as a number of redox-specific enzymes change in their activity (partial inhibition, MB was applied in sublethal concentrations).
T22 51868-51962 Sentence denotes Monitoring changes of enzyme mRNA expression, these predictions were validated by RT-PCR data.
T23 51963-52099 Sentence denotes The strongest reductions (more than 10%) caused by MB are calculated for protein protection modes using thioredoxin and/or glutaredoxin.
T24 52100-52229 Sentence denotes Furthermore, some enzymes of carbohydrate and nucleotide metabolism reacted to this inhibition by specific up or down regulation.
T25 52230-52344 Sentence denotes These pleiotropic effects were considered next in a flux model where different resistance mutations were modelled.
T26 52345-52492 Sentence denotes Resistance mutations (MDR, plasmodial transporter, CQ/PSD resistance) were considered by their mode of resistance on the enzymes targeted by drugs.
T27 52493-52642 Sentence denotes The effects of the latter drugs are complementary to the fluxes affected by MB and there is favourable synergism, for instance against CQ resistance.
T28 52643-52655 Sentence denotes Conclusions:
T29 52656-52759 Sentence denotes Taken together, our data suggest that MB could become a key factor in drug combination therapy efforts.
T30 52760-52834 Sentence denotes This should be tested more as a promising local eradication strategy [3] .
T31 52835-52983 Sentence denotes A C-terminal hyphal activating motif (CHAM) of the TEA transcription factor Tec1p is essential for the activation of morphological development of C.
T32 52984-52995 Sentence denotes Albicans W.
T33 52996-53014 Sentence denotes Abu Rayyan* 1 , A.
T34 53015-53027 Sentence denotes Singh 1 , K.
T35 53028-53042 Sentence denotes Fischer 1 , K.
T36 53043-53232 Sentence denotes Schröppel 1 1 Interfakultäres Institut für Mikrobiologie und Infektionsmedizin, Medical Mycology, Tübingen, Deutschland Candida albicans is a diploid fungal commensal of humans and mammals.
T37 53233-53326 Sentence denotes The transition between yeast and hyphal forms is one of the virulence traits for C. albicans.
T38 53327-53470 Sentence denotes C. albicans has developed numerous signaling pathways to activate the expression of the stage specific genes in response to environmental cues.
T39 53471-53623 Sentence denotes The transcription factor Tec1p, a member of the TEA/ATTS transcription factor family, harbors an evolutionarily conserved N-terminal DNA-binding domain.
T40 53624-53758 Sentence denotes The deletion of TEC1 leads to defects in hyphal formation in vitro and attenuates virulence in a systemic model of murine candidiasis.
T41 53759-54162 Sentence denotes In order to elucidate the putative DNA binding and gene activating domains spanned over the C. albicans Tec1p protein, we integrated modified open reading frames (ORFs) of the wild type TEC1 under the control of the C. albicans -adapted reverse Tet-dependent transactivator (rtTA) into the tec1/tec1 mutant CaAS12 and used a simple test for Tet-induced hyphal formation as a read-out for Tec1p-activity.
T42 54163-54364 Sentence denotes Transformation of CaAS12 with wild type TEC1 ORF was not only able to restore the hyphal growth ability in liquid medium but also it showed invasive growth on solid agar after adding Tet to the medium.
T43 54365-54649 Sentence denotes The ORF constructs either with a deletion of the C-terminus from 637 to the stop codon at 744 aa or a deletion of the TEA domain from 216 to 244 aa showed abrogation in restoring the hyphal growth, invasive growth and to grow under different stress condition in the tec1/tec1 mutants.
T44 54650-54797 Sentence denotes Interestingly, the tec1/tec1 phenotype was rescued in a mutant containing an ORF with an additional 72 aa that spanned the region from 1 to 709 aa.
T45 54798-54939 Sentence denotes After expression of the modified ORFs of TEC1 in E. coli, we tested for binding of recombinant rTec1p to the promoter consensus sequence TCS.
T46 54940-55038 Sentence denotes The deletion of the proposed C-terminal CHAM of Tec1p did not affect DNA binding to the TCS probe.
T47 55039-55192 Sentence denotes In conclusion, we show that CHAM of Tec1p is not required for DNA binding but it is essential for the biological activation of morphological development.
T48 55193-55254 Sentence denotes The RNA-binding protein Khd4 of the pathogen Ustilago maydis:
T49 55255-55441 Sentence denotes Identification of target mRNAs using in vivo UVcrosslink immunoprecipitation (iCLIP) C Co-and post-transcriptional control is an essential component of the regulation of gene expression.
T50 55442-55554 Sentence denotes RNA-binding proteins are the key mediators determining the spatio-temporal expression of proteins within a cell.
T51 55555-55741 Sentence denotes Accumulating evidence indicates an involvement of RNA-binding proteins in regulating expression programmes underlying pathogenic development of the phytopathogenic fungi Ustilago maydis.
T52 55742-55814 Sentence denotes We are currently investigating the role of the RNA-binding protein Khd4.
T53 55815-56002 Sentence denotes Khd4 is the founding member of a novel class of fungal multi hnRNP K-homology domain proteins with homologues in human pathogens such as Cryptococcus neoformans and Aspergillus fumigatus.
T54 56003-56155 Sentence denotes The deletion of khd4 results in a disturbed cell morphology and severe reduction in virulence, implying an important role for Khd4 during morphogenesis.
T55 56156-56313 Sentence denotes Khd4 recognizes a distinct hexanucleotide motif which is present in almost 25% of all annotated transcripts, suggesting a large regulatory potential of Khd4.
T56 56314-56414 Sentence denotes Failure to bind this RNA element results in an aberrant phenotype similar to the deletion phenotype.
T57 56415-56478 Sentence denotes The precise role and function of Khd4 however is still unclear.
T58 56479-56624 Sentence denotes To address this question, we are currently applying in vivo UV-crosslinking and immunoprecipitation experiments to identify target mRNAs of Khd4.
T59 56625-56754 Sentence denotes The iCLIP-technique enables the identification of target mRNAs by coupling in vivo UV-crosslinking to high throughput sequencing.
T60 56755-56902 Sentence denotes Furthermore, it offers the advantage of resolving crosslink sites at individual nucleotide resolution, enabling detailed analysis of binding sites.
T61 56903-57007 Sentence denotes Presented here are initial steps of establishing iCLIP in U. maydis using a control RNA-binding protein.
T62 57008-57010 Sentence denotes A.
T63 57011-57028 Sentence denotes McCormick* 1 , I.
T64 57029-57044 Sentence denotes Jacobsen 2 , J.
T65 57045-57059 Sentence denotes Wagener 1 , F.
T66 57060-57255 Sentence denotes Ebel Bakteriologie, München, Abt. für Mikrobielle Pathogenitätsmechanismen, Jena, Deutschland Aspergillus fumigatus is a mold causing severe and systemic infections in immunocompromised patients.
T67 57256-57345 Sentence denotes The high mortality of these infections is largely due to the limited therapeutic options.
T68 57346-57465 Sentence denotes Since A. fumigatus seems to lack sophisticated virulence traits, alternative therapeutic targets have to be identified.
T69 57466-57554 Sentence denotes The ability to respond to environmental changes is a vital requirement for all microbes.
T70 57555-57676 Sentence denotes Interference with signaling pathways helping the fungus to adapt to situations of stress is therefore a promising option.
T71 57677-57798 Sentence denotes However, this approach is hampered by the fact that many signaling pathways and molecules are conserved in fungi and men.
T72 57799-57906 Sentence denotes Two-component systems (TCS) comprising a sensor histidine kinase and a response regulator are an exception.
T73 57907-57971 Sentence denotes They are found in plants and microorganisms, but not in mammals.
T74 57972-58065 Sentence denotes In this study, we have analyzed the putative group III histidine kinase TcsC of A. fumigatus.
T75 58066-58140 Sentence denotes TcsC is the only A. fumigatus protein that comprises several HAMP domains.
T76 58141-58236 Sentence denotes A similar protein architecture has recently been proposed to constitute an osmo-sensing module.
T77 58237-58448 Sentence denotes Deletion of tcsC has no severe impact on growth under standard conditions, but growth and sporulation of the delta tcsC mutant is clearly impaired in the presence of various stressors, including high osmolarity.
T78 58449-58538 Sentence denotes This demonstrates that TcsC signaling is required to cope with different forms of stress.
T79 58539-58698 Sentence denotes In vitro, the delta tcsC mutant turned out to be highly resistant to several fungicides, including fludioxonil, which are supposed to activate the HOG pathway.
T80 58699-58797 Sentence denotes Consistently, we observed a dramatic swelling of fludioxoniltreated A. fumigatus wild type hyphae.
T81 58798-59059 Sentence denotes Mutants defective in genes of the cell wall integrity (CWI) pathway are significantly more sensitive to fludioxonil suggesting either a cross-talk between the HOG and the CWI pathway and/or a requirement for cell wall dynamics to bear fludioxonilinduced stress.
T82 59060-59119 Sentence denotes The role of TcsC during infection is currently under study.
T83 59120-59122 Sentence denotes E.
T84 59123-59137 Sentence denotes Hiller* 1 , M.
T85 59138-59155 Sentence denotes Dörflinger 1 , G.
T86 59156-59175 Sentence denotes Eigenstätter 1 , S.
T87 59176-59189 Sentence denotes Brunke 2 , I.
T88 59190-59205 Sentence denotes Jabobsen 2 , M.
T89 59206-59226 Sentence denotes Marcet-Houben 3 , T.
T90 59227-59242 Sentence denotes Gabaldon 3 , T.
T91 59243-59415 Sentence denotes Schwarzmüller 4 Although Candida glabrata has become the second most important pathogenic Candida species, only few of its virulence mechanisms have been identified so far.
T92 59416-59717 Sentence denotes Within the ERA-Net consortium FunPath the genes of about 800 potential virulence factors coding for predicted proteins of the cell wall, known signalling pathways, membrane-bound receptors, transporters and transcription factors were identified by comparative genome analysis and subsequently deleted.
T93 59718-59934 Sentence denotes This library is screened with biological assays, e.g. for strains with altered cell wall stability, stress tolerance, or adhesion behaviour to get a more comprehensive idea of the virulence mechanisms of C. glabrata.
T94 59935-60016 Sentence denotes Furthermore, fitness tests in mice with pools of different mutants are performed.
T95 60017-60240 Sentence denotes A microarray based tool to read out the amount of cells from different deletion mutant strains after the tests was established which utilises the amount of genetic bar-codes (specific for the different strains) in a sample.
T96 60241-60308 Sentence denotes Up to now, several strains were found in survival assays on plates.
T97 60309-60441 Sentence denotes According to their homologues in S. cerevisiae, some of the genes deleted in mutants are presumably involved in cell wall integrity.
T98 60442-60631 Sentence denotes Since cell wall integrity is crucial for stress resistance and virulence, further analysis of deletion mutant strains which are predicted to be part of one signalling network was performed.
T99 60632-60860 Sentence denotes Based on research done in the homologue S. cerevisiae, SLG1 and MID2 are presumed to be putative cell wall sensors, whereas SLT2 is supposed to be a transcription factors involved in the activation of cell wall biogenesis genes.
T100 60861-61017 Sentence denotes Regarding adhesion on different surfaces, mid2 and slt2 showed comparative behaviour to the wild type, whereas the mutant strain slg1 was stronger adherent.
T101 61018-61223 Sentence denotes Similar behaviour could be observed while assaying cell wall stability and composition, with slg1 having considerably changed properties in comparison to the mutant strains mid2 and slt2 and the wild type.
T102 61224-61493 Sentence denotes Comparing the trancriptomes of the deletion strains with the wild type in the presence or absence of congo red showed only very slight changes in the expression pattern of mid2 and slt2, whereas in slg1 a significantly changed transcriptional profile could be detected.
T103 61494-61608 Sentence denotes The results obtained suggest that SLG1 has a different role in C. glabrata than in S. cerevisiae compared to MID2.
T104 61609-61741 Sentence denotes The function of the inspected genes of the cell wall integrity pathway in S. cerevisiae might therefore be different in C. glabrata.
T105 61742-61842 Sentence denotes African trypanosomes transmigrate across an in vitro blood brain barrier model in both directions C.
T106 61843-61858 Sentence denotes Untucht* 1 , J.
T107 61859-61871 Sentence denotes Rasch 1 , M.
T108 61872-61884 Sentence denotes Rohde 2 , S.
T109 61885-61900 Sentence denotes Bergmann 1 , M.
T110 61901-62059 Sentence denotes Steinert 1 1 TU Braunschweig, Institut für Mikrobiologie, Braunschweig, Deutschland 2 HZI, Medizinische Mikrobiologie, Braunschweig, Deutschland Introduction:
T111 62060-62205 Sentence denotes The African Sleeping Sicknesss is a vector borne parasitic disease caused by human pathogenic Trypanosoma brucei gambiense and T. b. rhodesiense.
T112 62206-62277 Sentence denotes The disease threatens over 60 million people of 36 sub-Saharan nations.
T113 62278-62358 Sentence denotes Without treatment the lethality is 100 % and 10.000 deaths occur each year [1] .
T114 62359-62482 Sentence denotes The parasites are transmitted during a blood meal of tsetse flies from Glossina genus and multiply within the blood stream.
T115 62483-62665 Sentence denotes Depending on the subspecies, the trypanosomes invade the central nervous system (CNS) within weeks, several month or even years, which leads to the fatal outcome of the disease [2] .
T116 62666-62750 Sentence denotes This project aims on the cellular mechanisms of Blood Brain Barrier (BBB) traversal.
T117 62751-62920 Sentence denotes Therefore a transwell based in vitro BBB model was established and optimized using endothelial like ECV304 [3] , glial C6 cells and a semi-synthetical basement membrane.
T118 62921-63131 Sentence denotes The cell layers were analysed by electron microscopy (EM) and barrier integrity was evaluated by bacterial transmigration comparing a non invasive E. coli HB101 strain with meningitis causing E. coli K1 strain.
T119 63132-63361 Sentence denotes J774 macrophages and blood stream cultures of different trypanosome strains, namely T. b. brucei TC221, T. b. gambiense STIB930 and T. b. rhodesiense STIB900, were used to analyse the transmigration across the in vitro BBB model.
T120 63362-63561 Sentence denotes Results: EM microscopy revealed a confluent layer of ECV304 cells with partly overlapping neighbour cells inside the transwell inserts and a loose meshwork of C6 cells underneath the insert membrane.
T121 63562-63735 Sentence denotes The optimized ECV304-C6 model was permeable for pathogenic E. coli K1 transmigration whereas apathogenic E. coli HB101 did not transmigrate even at 100fold higher inoculums.
T122 63736-63857 Sentence denotes African trypanosomes transmigrated efficiently and T. b. gambiense STIB930 strain showed the highest transmigration rate.
T123 63858-64048 Sentence denotes The analyses of macrophage transmigration across the optimized ECV304-C6 BBB model indicated that paracellular extravasation of J774 macrophages does not enhance trypanosomal transmigration.
T124 64049-64194 Sentence denotes An upside-down array of the BBB cells, representing an inverse BBB model, revealed brain to blood traversal of trypanosomes with similar amounts.
T125 64195-64206 Sentence denotes Discussion:
T126 64207-64332 Sentence denotes The optimized ECV304-C6 BBB is a reproducible in vitro BBB model and selectively permeable for meningitis causing E. coli K1.
T127 64333-64537 Sentence denotes Trypanosomal transmigration across the optimized ECV304-C6 model occurs in both directions underlining the possibility of a relapsing parasitemia after treatment with drugs that have no access to the CNS.
T128 64538-64711 Sentence denotes This model promotes the analysis of the cellular and molecular mechanisms underlying the CNS invasion, which is urgently needed for therapy improvement and drug development.
T129 64712-64842 Sentence denotes Candida albicans affects barrier function and expression of tight and adherens junction proteins of intestinal epithelial cells M.
T130 64843-64860 Sentence denotes Böhringer* 1 , O.
T131 64861-64926 Sentence denotes Kurzai Septomics Research Centre, Jena, Deutschland Introduction:
T132 64927-65044 Sentence denotes Intestinal epithelial cells (IEC) establish a tight monolayer, forming a barrier for microorganisms in the gut lumen.
T133 65045-65225 Sentence denotes Tight junctional transmembrane proteins, notably claudins, regulate paracellular permeability of the intestinal barrier, constricting diffusion of small solutes and macromolecules.
T134 65226-65314 Sentence denotes Barrier function can be affected by pathogenic microorganisms, toxins and other stimuli.
T135 65315-65502 Sentence denotes The polymorphic fungus and common commensal of the human gastrointestinal tract, Candida albicans is capable of establishing systemic disease within the host after traversing the barrier.
T136 65503-65597 Sentence denotes Here we investigated the role of active barrier disruption for invasion of IEC by C. albicans.
T137 65598-65606 Sentence denotes Methods:
T138 65607-65736 Sentence denotes Caco-2 derived C2BBe1 IEC, grown on permeable cell culture inserts served as in vitro model of the intestinal mucosal epithelium.
T139 65737-65843 Sentence denotes Barrier function of C2BBe1 monolayer was examined by measuring transepithelial electric resistance (TEER).
T140 65844-65917 Sentence denotes Cytotoxicity was determined by LDH release assay on culture supernatants.
T141 65918-65997 Sentence denotes Fixed inserts with Candida-infected IEC were imaged with a confocal microscope.
T142 65998-66085 Sentence denotes Results: IEC were grown on membrane inserts for 12 days to establish stable monolayers.
T143 66086-66165 Sentence denotes Addition of C. albicans with a MOI of 0.3 led to an increase of TEER of approx.
T144 66166-66171 Sentence denotes 50 %.
T145 66172-66312 Sentence denotes TEER peaked around 8 h after inoculation and then started to decrease until reaching background levels (i.e. insert without IEC) after 21 h.
T146 66313-66387 Sentence denotes Decrease of electric resistance came along with an increasing LDH release.
T147 66388-66518 Sentence denotes 21 h after infection cytotoxicity levelled at 30-40 %, while microscopic imaging revealed a profound disruption of the monolayers.
T148 66519-66668 Sentence denotes To examine the role of filamentation and active penetration, we infected the C2BBe1 monolayer with the non-filamentous C. albicans mutant efg1/ cph1.
T149 66669-66854 Sentence denotes In contrast to wild-type inoculated IEC monolayer, TEER of mutant inoculated monolayer increased until reaching a two-fold level, which continued until the end of the experiment (22 h).
T150 66855-66930 Sentence denotes Cytotoxicity of the latter remained at the level of non-inoculated control.
T151 66931-67058 Sentence denotes We examined the expression levels of junctional proteins to define more precisely the impact of C. albicans inoculation on IEC.
T152 67059-67211 Sentence denotes Adherens junction (E-cadherin) and tight junction (JAM-A, occludin, and claudin -1, -2 and -3) protein levels decreased markedly in infected monolayers.
T153 67212-67439 Sentence denotes Confocal imaging of Candida-infected C2BBe1 cells showed some localized Ecadherin accumulation at fungal filaments penetrating through cell-cell boundaries, whereas overall E-cadherin staining around the invasion site was lost.
T154 67440-67448 Sentence denotes Outlook:
T155 67449-67541 Sentence denotes The results indicate an active participation of C. albicans in intestinal barrier breakdown.
T156 67542-67656 Sentence denotes However, the relative impacts of host and fungus on barrier disturbance will have to be clarified in ongoing work.
T157 67657-67805 Sentence denotes Future analyses are intended to clarify localized vs. general effects of junctional protein distribution and/or degradation in C. albicans invasion.
T158 67806-67878 Sentence denotes In vivo imaging of inflammation during invasive aspergillosis in mice K.
T159 67879-67892 Sentence denotes Große* 1 , T.
T160 67893-67909 Sentence denotes Opfermann 2 , P.
T161 67910-67927 Sentence denotes Gebhardt 2 , H.P.
T162 67928-67940 Sentence denotes Saluz 2 , B.
T163 67941-67952 Sentence denotes Hube 1 , I.
T164 67953-68075 Sentence denotes Jacobsen 1 1 HKI Jena, Mikrobielle Pathogenitätsmechanismen, Jena, Deutschland 2 HKI Jena, Jena, Deutschland Introduction:
T165 68076-68158 Sentence denotes Invasive aspergillosis (IA) is an important disease in immunocompromised patients.
T166 68159-68229 Sentence denotes Immunosuppressed mice are the gold standard to investigate IA in vivo.
T167 68230-68338 Sentence denotes In mice immunosuppressed with cortisone acetate, infection is accompanied by a strong inflammatory response.
T168 68339-68457 Sentence denotes Immigrating immune cells are characterized by an increased glucose uptake, which can be used for non-invasive imaging:
T169 68458-68604 Sentence denotes Fluorine labeled glucose (FDG -2-[ 18 F]fluoro-2-deoxy-D-glucose) is used as tracer for positron emission tomography/computer tomography (PET/CT).
T170 68605-68748 Sentence denotes Alternatively, a fluorescence labeled glucose derivative (RediJect 2-DG-750 Probe, Caliper) is detectable by the In-Vivo-Imaging-System (IVIS).
T171 68749-69064 Sentence denotes Since non invasive imaging tools to monitor inflammation have the potential to reduce the number of laboratory animals needed for the investigation of IA, we aimed to determine whether the above mentioned in vivo imaging systems are suitable for the quantification of the host's inflammatory response to IA in mice.
T172 69065-69087 Sentence denotes Materials and methods:
T173 69088-69312 Sentence denotes After immunosuppression with cortisone acetate (25 mg/mouse intraperitonialy on day -3 and day 0) mice were infected intranasally with a dose of 1 x 10 5 conidia of a bioluminescent Aspergillus fumigatus strain (Brock et al.
T174 69313-69320 Sentence denotes 2008) .
T175 69321-69365 Sentence denotes PBS mock-infected animals served as control.
T176 69366-69536 Sentence denotes PET/CT imaging was performed on days -5/-4 and +3/+4 under isoflurane anaesthesia immediately after application of 100-150μl with 7-12 MBq FDG into the lateral tail vein.
T177 69537-69691 Sentence denotes IVIS measurements were done with isoflurane anaesthesia immediately after PET/CT, which was 3 hours after injection either of a control dye or 2-DG-probe.
T178 69692-69762 Sentence denotes The distribution of tracers was quantified in the right and left lung.
T179 69763-69881 Sentence denotes After the last measurement, mice were euthanized, the lung was removed during necropsy and measured alone in the IVIS.
T180 69882-69938 Sentence denotes The left and right lungs were homogenized independently.
T181 69939-70048 Sentence denotes Myeloperoxidase and cytokine levels were quantified in lung homogenates as molecular markers of inflammation.
T182 70049-70057 Sentence denotes Results:
T183 70058-70186 Sentence denotes The FDG and 2-DG-probe distribution in immunocompetent mice and immunocompromised PBS mock-infected mice were indistinguishable.
T184 70187-70288 Sentence denotes In contrast, both the FDG and 2-DG-probe accumulated in the lungs of mice infected with A. fumigatus.
T185 70289-70401 Sentence denotes Measurements of mice injected with the control dye revealed no accumulation, neither before nor after infection.
T186 70402-70413 Sentence denotes Discussion:
T187 70414-70541 Sentence denotes We demonstrate that inflammation during IA can be monitored non-invasively by PET/CT and IVIS using labelled glucose as tracer.
T188 70542-70616 Sentence denotes Both methods allow quantification of immune cell accumulation in the lung.
T189 70617-70647 Sentence denotes References Brock et al., 2008.
T190 70648-70672 Sentence denotes Appl Environm Microbiol.
T191 70673-70682 Sentence denotes 74 (22) :
T192 70683-70693 Sentence denotes 7023-7035.
T193 70694-70844 Sentence denotes Caliper: http://www.caliperls.com/products/reagents/in-vivo-imaging-reagents/xenolight-bioluminescentsubstrates/xenolight-rediject-2-dg-750-probe.htm.
T194 70845-70952 Sentence denotes Malaria parasites hijack human factor H to protect from complement-mediated lysis in the mosquito midgut N.
T195 70953-70966 Sentence denotes Simon* 1 , M.
T196 70967-70986 Sentence denotes Scheuermayer 1 , L.
T197 70987-71003 Sentence denotes Sologub 1 , P.F.
T198 71004-71017 Sentence denotes Zipfel 2 , C.
T199 71018-71031 Sentence denotes Skerka 2 , G.
T200 71032-71310 Sentence denotes Pradel 1 1 Research Center for Infectious Diseases, Würzburg, Deutschland 2 Leibniz Institute for Natural Product Research and Infection Biology, Jena, Deutschland As part of the innate immune system, the human complement is a first line of defence against microbial infections.
T201 71311-71427 Sentence denotes A number of blood-borne pathogens have evolved strategies for complement evasion, including mimicking host surfaces.
T202 71428-71762 Sentence denotes While the intraerythrocytic blood stages of the malaria parasites are protected from any attack by the human complement, the parasite gametes have to exit the enveloping erythrocyte prior to fertilization in the mosquito midgut and are then vulnerable to factors of the blood meal, including human immune cells and complement factors.
T203 71763-72013 Sentence denotes We here show that the human malaria parasite Plasmodium falciparum binds the regulatory protein factor H (FH) during sexual reproduction in the mosquito midgut, which protects the extracellular gametes from complement-mediated lysis in the bloodmeal.
T204 72014-72167 Sentence denotes The alternative pathway of the human complement is active for approximately 1 h post feeding, while FH becomes degraded in the mosquito midgut after 8 h.
T205 72168-72238 Sentence denotes Macrogametes and young zygotes bind FH, resulting in C3b inactivation.
T206 72239-72376 Sentence denotes FH comprises 20 complement control protein modules, and binding of FH to malaria gametes appears to be primarily mediated by modules 1-7.
T207 72377-72586 Sentence denotes Loss of FH-mediated protection results in impaired gametogenesis und reduced zygote numbers, and transmission blocking assays with antibodies against factor H significantly reduce transmission to the mosquito.
T208 72587-72719 Sentence denotes This is the first time that a protozoan parasite is demonstrated to camouflage using FH in order to avoid complement-mediated lysis.
T209 72720-72812 Sentence denotes Our findings might lead to novel approaches in the design of transmission blocking vaccines.
T210 72813-72908 Sentence denotes Up to now there is only little data on the interaction of human NKC with the pathogenic fungus.
T211 72909-73013 Sentence denotes However, after NKC depletion mice have been shown to be more susceptible to systemic Candida infections.
T212 73014-73148 Sentence denotes On the basis of these observations we investigated if there are direct or indirect effects of human NK Cells against Candida albicans.
T213 73149-73157 Sentence denotes Methods:
T214 73158-73268 Sentence denotes Primary human PBMCs were isolated from buffy coat of healthy donors by standard ficoll gradient (Biochrom AG).
T215 73269-73398 Sentence denotes Untouched NKC were separated by MACS using the NK cell isolation kit (Miltenyi Biotec), according to manufactorer's instructions.
T216 73399-73490 Sentence denotes The isolated cells were primed with a cytokine cocktail (IL-2, IL-15, IFN-alpha, IFN-beta).
T217 73491-73632 Sentence denotes Viable or heatinactivated Candida albicans SC5314 was used for confrontation and receptor surface expression was quantified by FACS-analysis.
T218 73633-73694 Sentence denotes Reduction of fungal viability was determined by an XTT assay.
T219 73695-73906 Sentence denotes To analyze the mechanism of NKC mediated fungal killing, cells were treated with blocking antibodies (anti-FasL, anti-TRAIL), strontium chloride (inducing granula release) or concanamycin A (perforin inhibitor).
T220 73907-73974 Sentence denotes Bioplex or ELISAs were used for quantification of secreted markers.
T221 73975-73983 Sentence denotes Results:
T222 73984-74326 Sentence denotes Co-incubation of NKC and C. albicans resulted in downregulation of Toll-like receptors (TLR2, TLR4) , cellular adhesins (CD11a, CD54) and activation markers/ ITAM-Bearing Receptors (CD25, CD314, CD335) on NKC but in parallel a marked increase of the degranulation marker CD107a as well as secretion of NK cell specific cytokines was detected.
T223 74327-74390 Sentence denotes Activation of NKC was most prominent for C. albicans filaments.
T224 74391-74557 Sentence denotes Degranulation was significantly reduced for inactivated yeast forms, whereas heat-inactivated filaments resulted in levels comparable to viable wild-type C. albicans.
T225 74558-74676 Sentence denotes The cytokine mediated priming of primary human NKC led to an enrichment of cytotoxic granules Perforin and Granzyme B.
T226 74677-74815 Sentence denotes Using these activated NKC, XTT-based killing assays showed a significant decrease of fungal viability after a coincubation for four hours.
T227 74816-74945 Sentence denotes Blocking experiments suggest that the cytotoxic effect is perforin dependent while FasL/TRAIL receptor pathways are not involved.
T228 74946-75104 Sentence denotes In parallel to these direct effects, NKC may interact with other immune cells since cytokine release shows an upregulation of neutrophil activating cytokines.
T229 75105-75117 Sentence denotes Conclusions:
T230 75118-75231 Sentence denotes Our observations suggest that NKC may play an important role in the first line defense of C. albicans infections.
T231 75232-75330 Sentence denotes In this context, direct and indirect antifungal effects of NKC are currently further investigated.
T232 75331-75456 Sentence denotes Characterisation of the innate immune activation in response to Candida albicans in a human whole blood model of infection K.
T233 75457-75473 Sentence denotes Hünniger* 1 , O.
T234 75474-75539 Sentence denotes Kurzai Septomics Research Centre, Jena, Deutschland Introduction:
T235 75540-75614 Sentence denotes Fungal sepsis is an increasing clinical problem with high mortality rates.
T236 75615-75792 Sentence denotes Although the incidence of fungal sepsis has increased recently, diagnosis is still difficult and mainly relies on the detection of the pathogen (Candida spp.) in blood cultures.
T237 75793-75954 Sentence denotes Using a human whole blood model of infection we are investigating the innate immune activation in response to Candida albicans in a situation similar to in vivo.
T238 75955-76066 Sentence denotes These analyses will allow identifying immune markers which might be potentially useful in clinical diagnostics.
T239 76067-76075 Sentence denotes Methods:
T240 76076-76215 Sentence denotes Pathogen is added to human whole blood in different concentrations and patterns of immune activation are monitored in time course analyses.
T241 76216-76324 Sentence denotes Initial assays included differential FACS staining to define activation patterns on immune cell populations.
T242 76325-76388 Sentence denotes In parallel we quantified humoral markers of immune activation.
T243 76389-76501 Sentence denotes Further investigation included the characterisation of the whole blood transcriptome in response to C. albicans.
T244 76502-76653 Sentence denotes The obtained data will be integrated using bioinformatic methods to generate a network of innate immune activation during early steps of fungal sepsis.
T245 76654-76662 Sentence denotes Results:
T246 76663-76875 Sentence denotes Fungal infection resulted in a strong and predominant activation of neutrophils detectable by an increased surface expression of markers for early activation (CD69, HLA-DR) and degranulation (CD11b, CD63, CD66b).
T247 76876-77017 Sentence denotes As neutrophil granules contain important antifungal effector molecules, the release of these granules in response to C. albicans was studied.
T248 77018-77128 Sentence denotes As expected, myeloperoxidase from primary and lactoferrin from secondary granules could be detected in plasma.
T249 77129-77304 Sentence denotes NK cells also showed increased surface levels of activation marker CD69 whereas several receptors (Fc RIII, NKG2D, NKp46) decreased markedly over time during fungal infection.
T250 77305-77480 Sentence denotes Furthermore, C. albicans blood infection induced a predominant increase in total number of platelets aggregated with granulocytes as well as PAC1 and CD62P surface expression.
T251 77481-77716 Sentence denotes Multiplex assays revealed a secretion of proinflammatory and regulatory cytokines (IFN-, TNF-, IL-1 , IL-6) as well as chemokines (MCP-1, MIP-1 / , Rantes, IL-8, IP-10) corresponding to cellular activation as detected by FACS analyses.
T252 77717-77881 Sentence denotes We also measured a strong increase in plasma concentration of anaphylatoxins (C3a, C4a, C5a) that are produced as a part of the activation of the complement system.
T253 77882-77893 Sentence denotes Conclusion:
T254 77894-78045 Sentence denotes The human whole blood model of infection enables the analysis of innate immune activation in response to C. albicans in a situation similar to in vivo.
T255 78046-78181 Sentence denotes Advantages over isolated primary cells are the physiological conditions and the possible communication between immune cell populations.
T256 78182-78410 Sentence denotes Furthermore we are currently performing comparative analyses with Gram-positive and Gram-negative bacteria to identify pathogen-specific activation patterns and get a global view on innate immunity of microbial blood infections.
T257 78411-78460 Sentence denotes Protozoans like Leishmania can undergo apoptosis.
T258 78461-78546 Sentence denotes In this process phosphatidylserine (PS) is externalized and DNA fragmentation occurs.
T259 78547-78646 Sentence denotes During leishmanial apoptosis an endonuclease (Lm Endo G) is activated, involved in DNA degradation.
T260 78647-78760 Sentence denotes In addition, an anti apoptotic peroxidase (Lm APX) has been found that inactivates reactive oxygen species (ROS).
T261 78761-78914 Sentence denotes However, the protein machinery mediating apoptosis in mammalian cells like caspases or members of the bcl-2 families are completely absent in Leishmania.
T262 78915-79033 Sentence denotes In this study we want to identify the mechanism and the proteins that are involved in the protozoan apoptotic program.
T263 79034-79147 Sentence denotes We used axenic amastigote cultures and promastigote cultures and induced apoptosis using staurosporine treatment.
T264 79148-79358 Sentence denotes First we performed quantative FACS and microscopical analyses, subsequently we used mass-spectrometry and characterized the most abundant proteins that were either up or down regulated upon apoptosis induction.
T265 79359-79568 Sentence denotes Using timelapse imaging we observed that induction of apoptosis in eGFP expressing promastigotes resulted first in rounding of the promastigotes, then in loss of eGFP and subsequently in externalization of PS.
T266 79569-79731 Sentence denotes A more detailed investigation of apoptosis revealed that reactive oxygen species (ROS) are generated in a parasite stage and apoptosis induction dependent manner.
T267 79732-79804 Sentence denotes Focusing on Lm Endo G we found an upregulation upon apoptosis induction.
T268 79805-79880 Sentence denotes In the same context, we observed a downregulation of anti apoptotic Lm APX.
T269 79881-79946 Sentence denotes Mass spectrometry quantified a total of 707 leishmanial proteins:
T270 79947-80098 Sentence denotes In promastigotes apoptosis induced the upregulation of 3 proteins and the downregulation of 7 proteins, for amastigotes this was 6 and 22 respectively.
T271 80099-80173 Sentence denotes We have selected 3 proteins for which specific inhibitory compounds exist.
T272 80174-80256 Sentence denotes Currently we are investigating these proteins for a role in leishmanial apoptosis.
T273 80257-80364 Sentence denotes Programmed cell death (PCD) is a common feature of multicellular organisms and has been well characterized.
T274 80365-80561 Sentence denotes In recent years, there has been increasing evidence for the occurrence of PCD in distinct unicellular organisms as well, however, there is only little information about putative PCD in Toxoplasma.
T275 80562-80690 Sentence denotes Therefore, identification of apoptosis-like markers and molecules that might play a role in T. gondii PCD need to be elucidated.
T276 80691-80871 Sentence denotes TUNEL assays followed by flow cytometry were used to determine DNA strand breaks after treatment of the parasite with several proapoptotic inducers (staurosporine and miltefosine).
T277 80872-81046 Sentence denotes The results showed that staurosporine and miltefosine increased the number of TUNEL-positive cells to various extents and time-dependently from 24 to 72 hours post treatment.
T278 81047-81139 Sentence denotes Treated parasites showed a slight reduction in cell size as compared to untreated parasites.
T279 81140-81290 Sentence denotes Such reduction in cell size was most obvious in miltefosine-treated parasites which also displayed the highest percentage of TUNEL-positive parasites.
T280 81291-81489 Sentence denotes Furthermore, treatment with staurosporine and miltefosine led to a higher percentage of Annexin Vpositive parasites exposing phosphatidylserine on the cell surface as compared to untreated controls.
T281 81490-81630 Sentence denotes Some proteins that are involved in mammalian apoptosis pathways have been identified by in silico analysis of the T. gondii genome database.
T282 81631-81698 Sentence denotes In this project, we have been focusing on T. gondii Bax inhibitors.
T283 81699-81818 Sentence denotes In silico analysis of the T. gondii genome demonstrated the presence of putative Bax inhibitor genes, named T. gondii .
T284 81819-81916 Sentence denotes Amino acid sequence analysis of TgBI-1, TgBI-2 and TgBI-3 showed 33-38% identity with human BI-1.
T285 81917-82066 Sentence denotes Putative TgBI-1, TgBI-2 and TgBI-3 contain 7-9 transmembrane domains and have a molecular weight of 37,974 Da, 29,322 Da and 37,345 Da, respectively.
T286 82067-82207 Sentence denotes In order to unravel possible functions of T. gondii BI proteins in regulation of apoptosis, we heterologously expressed TgBIs in HeLa cells.
T287 82208-82364 Sentence denotes Stable transfection of the genes encoding TgBI-2 and TgBI-3 into HeLa cells was confirmed by immunofluorescence microscopy as well as Western Blot analysis.
T288 82365-82540 Sentence denotes Hoechst staining showed lower levels of chromatin condensation in TgBI-2-expressing HeLa cells than in HeLa wild type cells after treatment with brefeldin A and staurosporine.
T289 82541-82719 Sentence denotes Measurement of caspase 3/7 activity demonstrated an inhibitory effect of TgBI-2 in HeLa cells after treatment with either staurosporine, brefeldin A, tunicamycin or thapsigargin.
T290 82720-82865 Sentence denotes Moreover, TgBI-2-and TgBI-3-expressing HeLa cells showed lower induction of hypoploid DNA (subpeak G0/G1) after treatment with inducers as above.
T291 82866-83012 Sentence denotes These findings suggest the existence of apoptotic-like cell death in T. gondii and protective effects of TgBI proteins in regulation of apoptosis.
T292 83013-83134 Sentence denotes Histone deacetylase inhibitors rescue macrophages from the Toxoplasma gondii-mediated inhibition of IFN-responsiveness F.
T293 83135-83147 Sentence denotes Brand 1 , C.
T294 83148-83159 Sentence denotes Lang 1 , A.
T295 83160-83182 Sentence denotes Hildebrandt 1 , C.G.K.
T296 83183-83409 Sentence denotes Lüder* 1 1 University Medical Center, Institute for Medical Microbiology, Göttingen, Deutschland Toxoplasma gondii is an eukaryotic parasite that is highly prevalent in warm-blooded hosts including 25 -30% of humans worldwide.
T297 83410-83637 Sentence denotes Infection of individuals with premature or suppressed immunity can lead to lifethreatening toxoplasmosis whereas in immune competent hosts, infection is mostly asymptomatic but leads to parasite persistence for the host's life.
T298 83638-83758 Sentence denotes Cell-mediated immunity including the production of IFN-is critical for control of the obligatory intracellular parasite.
T299 83759-83968 Sentence denotes However, by using genomewide transcriptome analysis, we have shown that T. gondii leads to global unresponsiveness of murine macrophages to IFN-and this may be essential for establishing a long-term infection.
T300 83969-84188 Sentence denotes Analysis of the underlying mechanisms revealed normal activation and nuclear translocation of STAT1 in Toxoplasma-infected macrophages though a defect in the activation of STAT1-responsive promoters in response to IFN-.
T301 84189-84463 Sentence denotes Isolation and characterization of proteins that bound to oligonucleotides containing a STAT1-responsive consensus sequence indicated a defective recruitment of non-muscle actin and Brg-1 in Toxoplasma-infected macrophages, i.e. components of chromatin remodelling complexes.
T302 84464-84653 Sentence denotes In addition, chromatin immunoprecipitation showed that T. gondii inhibited the acetylation of histone H4 in vivo, i.e. a major prerequisite for chromatin remodelling prior to transcription.
T303 84654-84803 Sentence denotes Histone acetyl transferase (HAT) activity towards histone H4 and H3 was not generally decreased in nuclear extracts of T. gondiiinfected macrophages.
T304 84804-84887 Sentence denotes Furthermore, histone deacetylase (HDAC) activity was not increased after infection.
T305 84888-85112 Sentence denotes Nonetheless, treatment of macrophages with HDAC inhibitors MS-275 or sodium butyrate prior to stimulation with IFN-largely abrogated the inhibition of IFN--induced MHC class II and CIITA gene expression exerted by T. gondii.
T306 85113-85306 Sentence denotes Together, these data unravel novel insights into the mechanisms by which T. gondii evades IFN-regulated immunity and open up novel strategies for therapeutic intervention against toxoplasmosis.
T307 85307-85309 Sentence denotes T.
T308 85310-85325 Sentence denotes Dietsche 1 , A.
T309 85326-85338 Sentence denotes Singh 1 , W.
T310 85339-85356 Sentence denotes Abu Rayyan 1 , K.
T311 85357-85371 Sentence denotes Fischer 1 , K.
T312 85372-85684 Sentence denotes Schröppel* 1 1 Universitätsklinikum Tübingen, Interfakultäres Institut für Mikrobiologie und Infektionsmedizin, Tübingen, Deutschland Candida albicans is the most frequent human fungal pathogen and infections range from superficial mucosal lesions to life-threatening invasive mycoses with a high mortality rate.
T313 85685-85868 Sentence denotes Hyphal growth, biofilm formation and the transcriptional regulation of putative virulence traits for adaptation to the host environment are key issues during C. albicans pathogenesis.
T314 85869-86096 Sentence denotes Hyphal formation is a prerequisite for penetration of host tissues causing systemic candidiasis, and the formation of biofilms allows this pathogen to persist in the host and to increase resistance against antifungal compounds.
T315 86097-86246 Sentence denotes The transcription factor Tec1p, a member of the TEA transcription factor family, plays an important role in morphogenetic development in C. albicans.
T316 86247-86510 Sentence denotes In order to define a structure function relationship for the Tec1p protein, we integrated modified open reading frames (ORFs) of the wild type TEC1 under the control of the C. albicans adapted reverse Tet-dependent transactivator (rtTA) into the tec1/tec1 mutant.
T317 86511-86752 Sentence denotes We screened the series of such C. albicans mutants with modifications of the Tec1p for their ability of in vitro biofilm formation and also examined the hyphal formation and survival of a nematode host using C. elegans as an infection model.
T318 86753-87163 Sentence denotes In biofilm assays, the full length TEC1 ORF was able to restore the ability for biofilm formation ability in the tec1/tec1 mutant background after Tet induced gene expression, and the biofilm mass was similar to the biofilm formed by the wild type SC5314 strain, whereas the TEA domain mutant and mutants lacking the proposed C-terminal motif were impaired in biofilm formation under similar growth conditions.
T319 87164-87323 Sentence denotes Previously, C. elegans has been successfully used as an infection model host to understand the virulence mechanisms of various human pathogenic microorganisms.
T320 87324-87555 Sentence denotes Further in vivo investigations with C. elegans infection model revealed that the C. albicans TEC1 mutant lacking this C-terminal motif shows less virulence against nematodes in terms of hyphal invasion and killing of the nematodes.
T321 87556-87815 Sentence denotes This TEC1 mutant library screening revealed that besides the already known essential regions as the TEA domain and the important N-terminal end of Tec1p, also a C-terminal motif of the transcription factor plays an important role in morphogenetic development.
T322 87816-87985 Sentence denotes It can be demonstrated that this C-terminal motif of Tec1p is essential for yeast-hyphae transition, infection and killing of C. elegans nematodes and biofilm formation.
T323 87986-87990 Sentence denotes V.R.
T324 87991-88005 Sentence denotes Stoldt* 1 , M.
T325 88006-88023 Sentence denotes Abtmeier 1 , K.C.
T326 88024-88038 Sentence denotes Huynh 1 , R.E.
T327 88039-88174 Sentence denotes Scharf 1 1 Universitätsklinikum Düsseldorf, Institut für Hämostaseologie und Transfusionsmedizin, Düsseldorf, Deutschland Introduction:
T328 88175-88263 Sentence denotes Candida albicans is a dimorphic fungus that can switch between the yeast and hypha form.
T329 88264-88348 Sentence denotes The ability to germinate and grow in hypha is an important virulence factor in vivo.
T330 88349-88408 Sentence denotes Platelets can bridge the innate with the adaptive immunity.
T331 88409-88515 Sentence denotes To attack microorganisms, platelets store microbicidal and immune stimulatory proteins in their -granules.
T332 88516-88654 Sentence denotes Platelet integrins and their plasma ligands can govern both binding to the pathogen and activation of platelets to release their granules.
T333 88655-88836 Sentence denotes Here we tested the hypothesis that fibronectin (which is also stored in platelet -granular and bound by platelet integrins) supports the host defense against C. albicans infections.
T334 88837-88858 Sentence denotes Material and methods:
T335 88859-89011 Sentence denotes Platelet-rich plasma from citrated anticoagulated blood samples was washed with phosphate buffered salt solution (PBS) pH 6.5 containing 2 U/ml apyrase.
T336 89012-89151 Sentence denotes Platelet pellet was resuspended in Tyrode buffer pH 7.3 containing MgCl2, CaCl2 and 10 μM of the cell tracker dye CMFDA (Molecular Probes).
T337 89152-89213 Sentence denotes Isolated plasma fibronectin was labeled with alexa fluor 488.
T338 89214-89367 Sentence denotes C. albicans (SC5314) cultured over night in yeast extract pepton glucose medium at 30°C was washed in PBS pH 7.3 and starved for 1 h at room temperature.
T339 89368-89421 Sentence denotes Germination was induced at 37°C by adding 10 % serum.
T340 89422-89467 Sentence denotes Samples were taken over the germination time.
T341 89468-89677 Sentence denotes For flow cytometry C. albicans yeast and hypha forms with an optical density (OD) of 0.4 were labeled for 1h with 40 μg/ml Fn-alexa fluor 488 and 1x10 7 fluorescently stained platelets under static conditions.
T342 89678-89686 Sentence denotes Results:
T343 89687-89728 Sentence denotes Only C. albicans hypha bound fibronectin.
T344 89729-89860 Sentence denotes The mean fluorescence intensity (arbitrary units, AU) increased from 140 AU to 380 AU after 60 min and 120 min time of germination.
T345 89861-89992 Sentence denotes Furthermore, germination of C. albicans increased binding of platelets up to 40 % and 90 %, after 90 min and 180 min, respectively.
T346 89993-90090 Sentence denotes Moreover, pretreatment of C. albicans hypha with fibronectin enhanced their binding to platelets.
T347 90091-90102 Sentence denotes Conclusion:
T348 90103-90181 Sentence denotes Hypha of C. albicans, an important virulence factor, is attacked by platelets.
T349 90182-90318 Sentence denotes Fibronectin, a ligand of the platelet integrins ( 5 1, 5 1, IIb 3) opsonizes the hypha form of C. albicans and fosters platelet binding.
T350 90319-90484 Sentence denotes Hence, platelets and fibronectin, with its intrinsic multivalent binding site can support the platelet mediated host defense and immune response against C. albicans.
T351 90485-90487 Sentence denotes K.
T352 90488-90501 Sentence denotes Dichtl 1 , F.
T353 90502-90513 Sentence denotes Dirr 1 , C.
T354 90514-90532 Sentence denotes Helmschrott 1 , J.
T355 90533-90611 Sentence denotes Wagener* 1 1 Max von Pettenkofer-Institut, Bakteriologie, München, Deutschland
T356 90612-90834 Sentence denotes The organization and biosynthesis of the fungal cell wall, an essential structure that defines the organism's shape and shields it from environmental stress, is regulated by the cell wall integrity (CWI) signaling pathway.
T357 90835-90990 Sentence denotes Its central MAP kinase module was subject of diverse studies and is conserved from Saccharomyces cerevisiae to filamentous fungi, such as Aspergillus spp..
T358 90991-91134 Sentence denotes On the contrary, components upstream of this MAP kinase module are well characterized in S. cerevisiae but largely unknown in Aspergillus spp..
T359 91135-91298 Sentence denotes We have identified, localized and functionally characterized homologues of the S. cerevisiae cell surface sensors ScWsc1 -ScWsc3 and ScMid2/ScMtl1 in A. fumigatus.
T360 91299-91415 Sentence denotes Moreover, we have analyzed the roles of three Rho-GTPases that have been implicated in CWI signaling in other fungi.
T361 91416-91550 Sentence denotes We show that AfRho1 is essential and conditional downregulation of rho1 or deletion of rho2 or rho4 result in a severely impaired CWI.
T362 91551-91691 Sentence denotes In addition, our data indicate fundamental functional differences on the sensor level of the CWI pathways of S. cerevisiae and A. fumigatus.
T363 91692-91775 Sentence denotes Trends in multidrug resistance among Escherichia coli and klebsiellae in Germany M.
T364 91776-91793 Sentence denotes Kresken* 1,2 , D.
T365 91794-92059 Sentence denotes Hafner 2,3 1 Antiinfectives Intelligence GmbH, Rheinbach, Deutschland 2 Paul-Ehrlich-Gesellschaft für Chemotherapie e.V., Rheinbach, Deutschland 3 Institute for Pharmacology and Clinical Pharmacology, Heinrich-Heine-University, Düsseldorf, Deutschland Introduction:
T366 92060-92176 Sentence denotes Escherichia coli (ECO) and klebsiellae are among the most common pathogens causing infections in in-and outpatients.
T367 92177-92334 Sentence denotes Antimicrobial agents such as 3rd generation cephalosporins (3GC), carbapenems (carba) and fluoroquinolones (FQ) are essential drugs to treat serious illness.
T368 92335-92499 Sentence denotes Treatment outcome, however, has increasingly been threatened by the emergence and dissemination of resistant (R) strains (multi-drug R [MDR] strains in particular).
T369 92500-92668 Sentence denotes The objective of this study was to document changes in the R patterns of ECO, Klebsiella pneumoniae (KPN) and Klebsiella oxytoca (KOX) in Germany between 1995 and 2007.
T370 92669-92677 Sentence denotes Methods:
T371 92678-92911 Sentence denotes Susceptibility data of clinical isolates collected in 21, 20, 21, 22 and 21 laboratories that participated in five surveillance studies conducted by the Paul-Ehrlich-Society in 1995 , 1998 , 2001 , 2004 , respectively, were analysed.
T372 92912-93063 Sentence denotes MICs of antimicrobial agents were determined by the broth microdilution procedure according to the standard DIN ISO and interpreted by EUCAST criteria.
T373 93064-93220 Sentence denotes The CLSI MIC method using ceftazidime (CAZ) +/-clavulanic acid (CLA) and cefotaxime (CTX) +/-CLA was employed as screening test for ESBL-producing isolates.
T374 93221-93308 Sentence denotes MDR was defined as antimicrobial R to more than two of the following four drug classes:
T375 93309-93343 Sentence denotes 3-cef, carba, FQ, aminoglycosides.
T376 93344-93352 Sentence denotes Results:
T377 93353-93488 Sentence denotes A total of 3,964 isolates primarily recovered from urine (33%), wounds (20%), respiratory specimens (19%), and blood (11%) were tested.
T378 93489-93556 Sentence denotes There were 890 (24%) ICU isolates and 3,074 (76%) non-ICU isolates.
T379 93557-93742 Sentence denotes Single R to ciprofloxacin (CIP) in ECO, KPN, and KOX increased from 6% (26/449), 4% (9/224) and 2% (2/83), respectively, in 1995 to 28% (115/418), 13% (25/190) and 17% (17/101) in 2007.
T380 93743-93815 Sentence denotes R to CTX in all three species rose from 1-4% in 1995 to 11-15% in 2007 .
T381 93816-94003 Sentence denotes Concurrently, an increase in the percentage of strains showing the ESBL phenotype was observed, while R to carba (ertapenem [ERT] , imipenem [IMP] , meropenem [MEM] ) remained uncommonly.
T382 94004-94077 Sentence denotes MDR rates for each of the species and study years are shown in the table.
T383 94078-94214 Sentence denotes Of the 36 MDR isolates recovered in 2007, none were susceptible (S) to CTX and CIP, 8% to gentamicin and 39% to piperacillin-tazobactam.
T384 94215-94318 Sentence denotes In contrast, carba were active against MDR isolates, except one KOX (MICs of ERT, IMP, MEM 16-64 mg/l).
T385 94319-94331 Sentence denotes Conclusions:
T386 94332-94466 Sentence denotes Single R to CIP and CTX, representing FQ and 3GC, respectively, as well as MDR in ECO and klebsiellae increased between 1995 and 2007.
T387 94467-94539 Sentence denotes In contrast, activity of carba did not change over the last 10-15 years.
T388 94540-94553 Sentence denotes Introduction:
T389 94554-94737 Sentence denotes Following regulatory approval for use in complicated skin and skin structure infections and complicated intraabdominal infections, tigecycline (TGC) was introduced in Germany in 2006.
T390 94738-94960 Sentence denotes Data of the German Tigecycline Evaluation Surveillance Trial (G-TEST) conducted between 2005 and 2009 were analyzed for the susceptibility of most important aerobic Gram-pos. and Gram-neg. pathogens to TGC and comparators.
T391 94961-94969 Sentence denotes Methods:
T392 94970-95206 Sentence denotes The susceptibility of isolates of four Gram-pos. and seven Gramneg. species consecutively collected in three surveillance studies conducted in cooperation with the same 13 laboratories in 2005, 2007, and 2009 (G-TEST I-III) were tested.
T393 95207-95238 Sentence denotes Each laboratory collected appr.
T394 95239-95280 Sentence denotes 250 pathogens from hospitalized patients.
T395 95281-95402 Sentence denotes MICs were determined in a central laboratory using the microdilution method according to the standard ISO 20776- 1:2006 .
T396 95403-95473 Sentence denotes EUCAST breakpoints were applied to all antibiotics for interpretation.
T397 95474-95532 Sentence denotes ESBL-producing organisms were identified by CLSI criteria.
T398 95533-95541 Sentence denotes Results:
T399 95542-95685 Sentence denotes A total of 3,248 isolates recovered from wounds (n=1,573; 48.4%), the peritoneal cavity (n=567; 17.5%), and blood (n=1,108; 34.1%) were tested.
T400 95686-95823 Sentence denotes Based on MIC-50/90 values, TGC demonstrated unchanged (within +/-one dilution) in vitro activity against all species tested (see Table) .
T401 95824-95976 Sentence denotes In contrast, a considerable increase of resistance (R) to beta-lactams and fluoroquinolones (FQ) was noted for members of the Enterobacteriaceae family.
T402 95977-96142 Sentence denotes Between 2004 and 2009, R to FQ in Escherichia coli, Enterobacter cloacae and Klebsiella pneumoniae increased from 20% to 27%, 6% to 11%, and 7% to 23%, respectively.
T403 96143-96301 Sentence denotes The increase of R to cefotaxime in E. coli and K. pneumoniae went along with a rise of ESBLproducing strains, from 6% to 11% and from 5% to 14%, respectively.
T404 96302-96434 Sentence denotes R to imipenem in Acinetobacter baumannii group isolates was not observed in 2005, but was 18% and 8% in 2007 and 2009, respectively.
T405 96435-96538 Sentence denotes The rate of vancomycin-resistant strains among Enterococcus faecium isolates varied between 9% and 19%.
T406 96539-96682 Sentence denotes Conclusion: TGC retained its very good in vitro activity against all Grampos. and Gram-neg. organisms tested after the introduction in Germany.
T407 96683-96820 Sentence denotes Against a background of pathogens that are frequently resistant to various antibiotic classes, TGC remains an important treatment option.
T1 96822-96962 Sentence denotes Pharmacodynamics of three carbapenems considering the current susceptibility patterns of drug-resistant Gram-negative bacteria in Germany M.
T2 96963-96978 Sentence denotes Kresken* 1 , B.
T3 96979-97000 Sentence denotes Körber-Irrgang 1 , S.
T4 97001-97022 Sentence denotes Decker-Burgard 2 , D.
T5 97023-97217 Sentence denotes Hafner 3 1 Antiinfectives Intelligence GmbH, Rheinbach, Neuss, Deutschland 3 Institute of Pharmacology and Clinical Pharmacology, Heinrich-Heine-University, Düsseldorf, Deutschland Introduction:
T6 97218-97471 Sentence denotes The carbapenems (CARBA) are widely used for empiric therapy of serious infections involving drug-resistant Gram-neg. pathogens like P. aeruginosa, A. baumannii or Enterobacteriaceae expressing an ESBL phenotype or a stably de-repressed AmpC ß-lactamase.
T7 97472-97797 Sentence denotes The objective of this study was to predict the probabilities of attaining targeted pharmacodynamic exposure for clinically used dosing regimens of doripenem (DOR), imipenem/cilastatin (IPM), and meropenem (MEM) against a German collection of contemporary drug-resistant Gram-neg. pathogens using Monte Carlo simulation (MCS).
T8 97798-97806 Sentence denotes Methods:
T9 97807-97968 Sentence denotes Isolates from hospitalised patients were collected in 15 medical microbiology laboratories during two resistance surveillance studies conducted in 2007 and 2009.
T10 97969-98113 Sentence denotes Bacterial groups investigated were P. aeruginosa (n=149), A. baumannii group (n=139), and ceftriaxone (CRO)resistant Enterobacteriaceae (n=349).
T11 98114-98449 Sentence denotes MCS modelled 5,000 patients receiving the following dosing regimens: DOR 500 mg every 8 hours (1-h infusions) , DOR 500 mg every 8 hours (4-h infusions) , IPM 1,000 mg every 8 hours (1-h-infusions) , IPM 500 mg every 6 hours (1-h-infusions), MEM 1,000 mg every 8 hours (1-h-infusions) , and MEM 2,000 mg every 8 hours (1-h-infusions) .
T12 98450-98573 Sentence denotes Pharmacodynamic targets to predict efficacy were the free drug concentrations above the MIC for 40% of the dosing interval.
T13 98574-98675 Sentence denotes Simulations were based upon a normal distribution of volume of distribution and total body clearance.
T14 98676-98765 Sentence denotes Mean pharmacokinetic parameters and their distribution were taken from published studies.
T15 98766-98903 Sentence denotes The cumulative fraction of response (CFR) was calculated considering the MIC distribution data for each of the three groups of organisms.
T16 98904-98912 Sentence denotes Results:
T17 98913-99080 Sentence denotes All CARBA regimens achieved CFR rates of >98% against CROresistant Enterobacteriaceae, but varied between 80% and 88% against A. baumannii group isolates (see Table) .
T18 99081-99240 Sentence denotes Against P. aeruginosa, the highest CFR (91.9%) was obtained with DOR 500 mg every 8 hours (4-h infusions) and the lowest (72.7%) with IPM 500 mg every 6 hours.
T19 99241-99252 Sentence denotes Conclusion:
T20 99253-99502 Sentence denotes Considering the current susceptibility patterns of drugresistant Gram-neg. pathogens in Germany, all clinically used dosing regimens of CARBA are sufficient to achieve a probability of 95% or greater success against CRO-resistant Enterobacteriaceae.
T21 99503-99629 Sentence denotes In contrast, only the DOR prolonged-infusion and MEM 2,000 mg dosing regimens achieved at least 90% CFR against P. aeruginosa.
T22 99630-99701 Sentence denotes None of the CARBA regimens achieved 90% CFR against A. baumannii group.
T23 99702-99713 Sentence denotes Background:
T24 99714-99907 Sentence denotes Current regional antibiotic susceptibility data from hospitalized patients and outpatients are needed to support local networks for the defence against the spreading of multiresistant bacteria.
T25 99908-99929 Sentence denotes Material and methods:
T26 99930-100225 Sentence denotes Results of routine suseptibility data for twelve antibiotic agents of a total of 90.043 isolates of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis from four Thuringian microbiology laboratories were analysed retrospectively for the years 2007 to 2010.
T27 100226-100351 Sentence denotes Susceptibility testing was performed by agar diffusion test or breakpoint-testing according to the German standard DIN 58940.
T28 100352-100526 Sentence denotes The results were compared to nationwide German data published online by the Robert Koch-Institute on the Antibiotic Resistance Surveillance website (ARS, https://ars.rki.de).
T29 100527-100535 Sentence denotes Results:
T30 100536-100703 Sentence denotes Resistance rates of S. aureus from outpatients against penicillin, oxacillin, and ciprofloxacine were significantly lower than those of hospitalized patients (p<0.05).
T31 100704-100925 Sentence denotes The rate of methicillin resistant S. aureus (MRSA) in outpatients rose from 6.9% in 2007 to 8.2% in 2010, whereas in both large (>500 beds) and small (<500 beds) hospitals, the MRSA rate remained constant at around 15.0%.
T32 100926-101065 Sentence denotes Resistance rates of E. coli against ampicillin, cefotaxime, gentamicin, and ciprofloxacin were significantly lower in outpatients (p<0.05).
T33 101066-101286 Sentence denotes The resistance rate of E. coli against cefotaxime as a marker for extended spectrum betalactamase (ESBL) production was constant at about 9.0% in hospitals but quadrupled in outpatients from 1.3% in 2007 to 5.9% in 2010.
T34 101287-101517 Sentence denotes Resistance rates of P. aeruginosa against piperacillin, ciprofloxacine, and ceftazidime were significantly lower in outpatients (p<0.05), whereas gentamicin was ineffective against P. aeruginosa in 6.0% of both in-and outpatients.
T35 101518-101581 Sentence denotes E. faecalis isolates remained highly susceptible to ampicillin.
T36 101582-101594 Sentence denotes Conclusions:
T37 101595-101793 Sentence denotes As of December 2010, in Thuringia/Germany, MRSA rates in both outpatients (8.2%) and hospitals (15.0%) remained well below nationwide levels as determined by the ARS (12.7% and 24.6%, respectively).
T38 101794-101897 Sentence denotes However, there was an alarming increase of ESBL rates in outpatients from 1.3% in 2007 to 5.9% in 2010.
T39 101898-102065 Sentence denotes The German EHEC outbreak strain ST678 harbours the CTX-M-15 extended-spectrum -lactamase (ESBL) (EHEC) was observed in Germany with the main focus in northern Germany.
T40 102066-102126 Sentence denotes Probably the outbreak was caused by contaminated vegetables.
T41 102127-102135 Sentence denotes Methods:
T42 102136-102282 Sentence denotes An EHEC outbreak isolate was sent to the National Reference Laboratory for multidrug-resistant gramnegative bacteria for further characterization.
T43 102283-102348 Sentence denotes Susceptibility testing was performed by disk diffusion and Etest.
T44 102349-102405 Sentence denotes Virulence genes of E. coli pathovars were tested by PCR.
T45 102406-102487 Sentence denotes MLST typing and PCR based determination of the phylogenetic group were performed.
T46 102488-102589 Sentence denotes The presence of several -lactamase genes was tested by PCR and the genetic environment characterized.
T47 102590-102664 Sentence denotes Transconjugation and transformation of a resistance plasmid was attempted.
T48 102665-102835 Sentence denotes S1 restriction followed by pulsed-field gel electrophoresis and subsequent Southern Blot hybridization was done to determine a plasmidic localization of resistance genes.
T49 102836-102844 Sentence denotes Results:
T50 102845-102996 Sentence denotes The EHEC outbreak isolate was isolated from a female patient from Northern Germany and tested positive for stx2 and aggR, but negative for stx1 or eae.
T51 102997-103067 Sentence denotes Typing revealed the MLST type ST678 and E. coli phylogenetic group B1.
T52 103068-103165 Sentence denotes The isolated displayed an ESBL phenotype and blaTEM-1 as well as blaCTX-M-15 could be identified.
T53 103166-103262 Sentence denotes The genetic environment of blaCTX-M-15 showed ISEcp1 upstream and ORF477 downstream of the gene.
T54 103263-103324 Sentence denotes No carbapenemase gene like KPC, VIM, OXA-48 or NDM was found.
T55 103325-103411 Sentence denotes Furthermore, resistance to tetracycline and sulfamethoxazoletrimethoprim was observed.
T56 103412-103452 Sentence denotes S1 restriction revealed a plasmid of ca.
T57 103453-103459 Sentence denotes 90 kb.
T58 103460-103547 Sentence denotes Transconjugants and transformants with a plasmid harbouring blaCTX-M-15 where obtained.
T59 103548-103635 Sentence denotes No co-resistance to tetracycline or sulfamethoxazoletrimethoprim could be demonstrated.
T60 103636-103647 Sentence denotes Conclusion:
T61 103648-103838 Sentence denotes Within two days after receipt of the EHEC outbreak isolate the molecular basis of its unusual ESBL phenotype could be clarified and was immediately reported to the German health authorities.
T62 103839-103904 Sentence denotes EHEC strains harbouring CTX-M-15 have never been reported before.
T63 103905-104096 Sentence denotes The presence of resistance genes in diarrheagenic bacterial strains is worrisome since it might contribute to an enhanced spread of resistance determinants, e. g. by horizontal gene transfer.
T64 104097-104156 Sentence denotes Antimicrobial susceptibility of ocular isolates in Germany,
T65 104157-104159 Sentence denotes M.
T66 104160-104175 Sentence denotes Kresken* 1 , B.
T67 104176-104265 Sentence denotes Körber-Irrgang 1 1 Antiinfectives Intelligence GmbH, Rheinbach, Deutschland Introduction:
T68 104266-104370 Sentence denotes Antimicrobial resistance has increased over the past two decades in Germany, as in many other countries.
T69 104371-104473 Sentence denotes Data on the in vitro susceptibility of pathogens isolated from ocular infections, however, are scarce.
T70 104474-104679 Sentence denotes Two multi-centre studies were carried out in order to assess the occurrence of resistance amongst ocular isolates against antibacterial agents used topically in the treatment of superficial eye infections.
T71 104680-104688 Sentence denotes Methods:
T72 104689-104845 Sentence denotes Bacterial isolates were prospectively collected in two surveillance studies conducted in cooperation with the same 23 medical laboratories in 2004 and 2009.
T73 104846-105025 Sentence denotes Minimal inhibitory concentrations were determined in a central laboratory using the broth microdilution method according to the German DIN standard (now DIN-EN-ISO 20776-1:2006) .
T74 105026-105205 Sentence denotes Interpretive criteria applied were EUCAST clinical breakpoints for systemic use of antibacterial agents as well as EUCAST epidemiological cut-off values (ECOFFs), where available.
T75 105206-105450 Sentence denotes Susceptibility data of nine antibacterial agents were analysed: levofloxacin (LVX), moxifloxacin (MXF), ofloxacin (OFX), gentamicin (GEN), kanamycin (KAN), azithromycin (AZI), erythromycin (ERY), chloramphenicol (CHL) and oxytetracycline (OTE).
T76 105451-105535 Sentence denotes Additionally, the susceptibility of staphylococci to oxacillin (OXA) was determined.
T77 105536-105544 Sentence denotes Results:
T78 105545-105663 Sentence denotes A total of 2,006 ocular isolates (2004, n=933; 2009, n=1,073) primarily recovered from conjunctival swabs were tested.
T79 105664-105791 Sentence denotes S. aureus was the predominant pathogen in either study (28-34%), followed by H. influenzae (13-18%) and S. pneumoniae (15-16%).
T80 105792-106051 Sentence denotes Patient related data were comparable in both studies, with two third of the isolates derived from outpatients and more than half of the patients being S. aureus isolates collected in 2004 and 2009, 10.3% and 9.8%, respectively, were OXA-resistant (i.e. MRSA).
T81 106052-106218 Sentence denotes Resistance to CHL, GEN and KAN among S. aureus decreased between 2004 and 2009, while resistance to ERY and the fluoroquinolones (FQ) remained unchanged (see Table) .
T82 106219-106319 Sentence denotes No significant changes in the rates of resistance were observed for S. pneumoniae and H. influenzae.
T83 106320-106499 Sentence denotes In contrast, isolates with reduced susceptibility to LVX and OFX among Enterobacteriaceae increased from 0% and 1% in 2004 (n=103) to 1.9% and 5.8% in 2009 (n=104) , respectively.
T84 106500-106511 Sentence denotes Conclusion:
T85 106512-106692 Sentence denotes Overall, studied aminoglycosides and CHL displayed lower resistance rates in 2009 as compared to 2004, while resistance rates of FQ remained either unchanged or slightly increased.
T86 106693-106706 Sentence denotes Introduction:
T87 106707-106822 Sentence denotes The worldwide increase of multidrug-resistance in gramnegative bacteria has become an important clinical challenge.
T88 106823-106975 Sentence denotes Carbapenem resistance in P. aeruginosa can be caused by a variety of mechanisms, however the worldwide spread of carbapenemases is especially important.
T89 106976-107064 Sentence denotes Most carbapenemases in P. aeruginosa belong to Ambler class B metallo--lactamases (MBL).
T90 107065-107159 Sentence denotes Here we describe a novel IMP-type MBL that was found in two isolates from patients in Germany.
T91 107160-107168 Sentence denotes Methods:
T92 107169-107242 Sentence denotes Susceptibility to antibiotics was determined by disk diffusion and Etest.
T93 107243-107364 Sentence denotes The presence of a MBL was determined by EDTA combineddisc-tests, MBL Etest and by a bioassay based on cell-free extracts.
T94 107365-107401 Sentence denotes A modified Hodge-Test was performed.
T95 107402-107499 Sentence denotes PCRs and subsequent sequencing were performed for VIM, IMP, NDM, GIM, SIM, SPM, AIM, DIM and KHM.
T96 107500-107562 Sentence denotes Clonal relatedness of the isolates was determined by RAPD-PCR.
T97 107563-107619 Sentence denotes Integron structures were analysed by PCR and sequencing.
T98 107620-107628 Sentence denotes Results:
T99 107629-107842 Sentence denotes Two P. aeruginosa isolates from single patients hospitalized in different geographic regions in Germany were sent to the National Reference Laboratory for Multidrug-resistant Bacteria for further characterization.
T100 107843-108118 Sentence denotes The isolates were resistant to piperacillin, piperacillin/tazobactam, ceftazidime, cefepime, aztreonam, imipenem (MICs 32 mg/L and 128 mg/L, respectively), meropenem and doripenem (MICs >32 mg/L) as well as to gentamicin, tobramycin, amikacin, ciprofloxacin and levofloxacin.
T101 108119-108187 Sentence denotes Both isolates were susceptible for colistin with a MIC of 0,38 mg/L.
T102 108188-108263 Sentence denotes The modified Hodge-test was positive for imipenem, meropenem and ertapenem.
T103 108264-108346 Sentence denotes The isolates showed synergy with EDTA in the combined disk test and the MBL Etest.
T104 108347-108461 Sentence denotes An unspecific result due to toxicity of EDTA could be excluded by a positive bioassay based on cell-free extracts.
T105 108462-108539 Sentence denotes With consensus primers for IMP a 587 bp fragment was amplified and sequenced.
T106 108540-108655 Sentence denotes The sequence showed only 88 % similarity to IMP-8 and IMP-24 and revealed 18 amino acid substitutions relative to .
T107 108656-108782 Sentence denotes The isolates also harbour at least one class I integron containing the gene for an aminoglycoside adenylyltransferase (aadA6).
T108 108783-108794 Sentence denotes Conclusion:
T109 108795-108860 Sentence denotes The two clinical isolates harbour a novel IMP-type carbapenemase.
T110 108861-109010 Sentence denotes It clearly shows a MBL phenotype, while the MIC differences for imipenem are likely to be the result of additional mechanism for imipenem resistence.
T111 109011-109167 Sentence denotes With 23 amino acid substitutions compared to IMP-8, the identified IMP-type carbapenemase shows significant differences to any known IMP-type carbapenemase.
T112 109168-109202 Sentence denotes Carbapenemases arrived in Germany:
T113 109203-109295 Sentence denotes Report of the National Reference Laboratory for multidrug-resistant gramnegative bacteria M.
T114 109296-109309 Sentence denotes Kaase* 1 , F.
T115 109310-109325 Sentence denotes Szabados 1 , A.
T116 109326-109339 Sentence denotes Anders 1 , S.
T117 109340-109425 Sentence denotes Gatermann Abteilung für Medizinische Mikrobiologie, Bochum, Deutschland Introduction:
T118 109426-109695 Sentence denotes Multidrug-resistance in Enterobacteriacea, Pseudomonas aeruginosa and Acinetobacter baumannii is of utmost therapeutic importance since no innovative antimicrobial drugs against gramnegative bacteria will be introduced into clinical practice within the next five years.
T119 109696-109801 Sentence denotes Among all resistance mechanisms the worldwide spread of carbapenemases is the most worrisome development.
T120 109802-109906 Sentence denotes However, the correct identification of carbapenemases is challenging for the microbiological laboratory.
T121 109907-109915 Sentence denotes Methods:
T122 109916-110099 Sentence denotes The National Reference Laboratory for Multidrug-Resistant Gramnegative Bacteria offers the free service of carbapenemase detection in bacterial isolates with elevated carbapenem MICs.
T123 110100-110176 Sentence denotes All isolates are tested by a wide array of phenotypic and molecular methods.
T124 110177-110266 Sentence denotes A bioassay based on cell-free extracts allows the detection of still unknown -lactamases.
T125 110267-110275 Sentence denotes Results:
T126 110276-110516 Sentence denotes A total of 808 isolates were sent to the National Reference Laboratory in 2010 mainly for investigation for carbapenemases, but also for clarification of the resistance mechanism to 3 rd generation cephalosporins or molecular strain typing.
T127 110517-110833 Sentence denotes Several different carbapenemases could be detected, including OXA-48 (n = 59), KPC-2 (n = 31), KPC-3 (n = 26), VIM-1 (n = 37), VIM-2 (n = 20), VIM-4 (n = 4), VIM-5 (n = 1), IMP-7 (n = 2), IMP-8 (n = 12), IMP-16 (n = 1), NDM-1 (n = 5), NDM-2 (n = 1), GIM-1 (n = 4), OXA-23 (n = 69), OXA-72 (n = 2) and OXA-58 (n = 6).
T128 110834-110939 Sentence denotes In Enterobacteriacea most Carbapenemases were found in K. pneumoniae, especially OXA-48, KPC-2 and KPC-3.
T129 110940-111026 Sentence denotes VIM-2 was the most frequent carbapenemase in P. aeruginosa and OXA-23 in A. baumannii.
T130 111027-111038 Sentence denotes Discussion:
T131 111039-111105 Sentence denotes Almost all carbapenemases found worldwide have arrived in Germany.
T132 111106-111294 Sentence denotes However, the molecular epidemiology in Germany with a predominance of OXA-48 differs significantly from observations made in other countries like Greece, Israel, USA or the United Kingdom.
T133 111295-111409 Sentence denotes An ongoing surveillance of resistance determinants is necessary, especially for infection control and diagnostics.
T134 111410-111421 Sentence denotes Objectives:
T135 111422-111589 Sentence denotes The successful worldwide dissemination of carbapenemase producing Enterobacteriaceae is of special concern because of limited therapeutic options and higher mortality.
T136 111590-111900 Sentence denotes While the dissemination of carbapenemase KPC is predominantly bound to distinct Klebsiella pneumoniae strains, spread of OXA-48 and NDM-1 enzymes is mainly due to conjugative transfer of different plasmids carrying these carbapenemase genes and facilitating the transmission among many enterobacterial species.
T137 111901-111999 Sentence denotes Here we report on molecular analysis of multidrug-resistant Enterobacteriaceae isolates from Oman.
T138 112000-112008 Sentence denotes Methods:
T139 112009-112196 Sentence denotes Carbapenem resistant isolates of Klebsiella spp. (n=13), E. coli (n=4) and Enterobacter cloacae (n=1) were collected from three hospitals in Oman between September 2010 and February 2011.
T140 112197-112261 Sentence denotes Relevant resistance genes were identified by PCR and sequencing.
T141 112262-112317 Sentence denotes Conjugation experiments and PFGE-typing were performed.
T142 112318-112341 Sentence denotes Results and conclusion:
T143 112342-112399 Sentence denotes The carbapenemase OXA-48 was identified in five isolates.
T144 112400-112471 Sentence denotes Ten Isolates produced NDM-1 and OXA-181 was identified in two isolates.
T145 112472-112567 Sentence denotes Three patients recently travelled to India and Pakistan before their admission to the hospital.
T146 112568-112777 Sentence denotes XbaI-macrorestriction analysis revealed seven closely related NDM-1 isolates and three isolates with OXA-48 indicating several events of clonal transmission of carbapenemase producing strains in two hospitals.
T147 112778-112946 Sentence denotes Furthermore additional ESBL CTX-M-15 (n=14) and CTX-M-24 (n=3) as well as other beta-lactamases (TEM-1, SHV-11, OXA-1, OXA-9) were detected in the majority of isolates.
T148 112947-113032 Sentence denotes The blaOXA-48 genes were successfully transferred into recipients and plasmids of ca.
T149 113033-113078 Sentence denotes 60kb size were isolated from transconjugants.
T150 113079-113153 Sentence denotes The blaNDM-1 genes were located on conjugative plasmids of different size.
T151 113154-113392 Sentence denotes The reduced carbapenem MIC values of transconjugants indicate the occurrence of further carbapenem resistance mechanisms like increased production of efflux pumps or loss of porins (outer membrane proteins, OMPs) in the clinical isolates.
T152 113393-113488 Sentence denotes In Germany the NDM-1 carbapenemase was identiefied in only 13 isolates in the last three years.
T153 113489-113687 Sentence denotes Although carbapenemase producing Enterobacteriaceae are still rare in many European countries there is an urgent need of surveillance to prevent further spread of these multidrugresistant pathogens.
T154 113688-113769 Sentence denotes How should we treat double -and triple -resistant Helicobacter pylori in Germany?
T155 113770-113795 Sentence denotes Background and Objective:
T156 113796-113903 Sentence denotes Enterobacteriaceae have been shown to play a crucial role in the transmission of antimicrobial resistances.
T157 113904-114024 Sentence denotes The most prominent enterobacterial species in humans and animals are Escherichia (E.) coli and Salmonella (S.) enterica.
T158 114025-114306 Sentence denotes Dissemination of antibiotic resistance can be due to the transfer of resistant bacteria between different host organisms or by horizontal transfer of resistance properties between isolates of the same or different species and genera within and beyond the family Enterobacteriaceae.
T159 114307-114629 Sentence denotes Especially, resistance to -lactams mediated by production of Extended-spectrum beta-lactamases (ESBL) and resistance to (fluoro)quinolones influenced by different plasmid-mediated quinolone resistance determinants are of concern since they limit dramatically the therapeutic options in both, veterinary and human medicine.
T160 114630-114871 Sentence denotes Therefore the RESET research consortium aims at assessing the impact of different origins, transmission routes and pathogen attributes on the risk for humans being exposed to ESBL-producing and (fluoro)quinolone-resistant Enterobacteriaceae.
T161 114872-114892 Sentence denotes Studies and Methods:
T162 114893-115071 Sentence denotes The RESET consortium consists of 10 network partners and 5 associated partners from human and veterinary medicine, fundamental and application-oriented research and epidemiology.
T163 115072-115613 Sentence denotes The RESET network will perform different studies to provide indispensable, reliable information on -the pathogen distribution in specific human and animal populations as well as in food, feed and the environment, -the occurrence of ESBL and PMQR (plasmid-mediated quinolone resistance) genes within the pathogen populations, -the transmission pathways between the different populations (animal-, environment-, human-or pathogen-related), and -the impact of the use of antibiotics in animals and humans in respect to the resistance situation.
T164 115614-115625 Sentence denotes Conclusion:
T165 115626-115849 Sentence denotes In 2008, the Federal Ministry of Health (BMG) started the DART (Deutsche Antibiotika-Resistenzstrategie) initiative for prevention and control of further spread of antimicrobial resistances in human and veterinary medicine.
T166 115850-116067 Sentence denotes The RESET network includes different interacting and complementary studies on factors associated with the dissemination of emerging resistance properties in Enterobacteriaceae from humans, animals and the environment.
T167 116068-116152 Sentence denotes As such, the results expected from RESET will be a substantial contribution to DART.
T168 116153-116372 Sentence denotes Nearly all clinical Vancomycin-resistant E. faecium strains are resistant to ampicillin (MICs >8 mg/L) and possess high level resistance to ciprofloxacin (>16 mg/L); typical features of hospital-associated clonal types.
T169 116373-116576 Sentence denotes High level resistance (HLR) to gentamicin (MICs >256 mg/L) occurs in 40 -57% of the strains (with increasing frequencies between 2008 and 2010); HLR to streptomycin (>512 mg/L) in 36 -58% of the strains.
T170 116577-116762 Sentence denotes Resistances to antibiotics of last resort such as linezolid (0 -4%), tigecycline (0 -2%), daptomycin (0%) or quinupristin/dalfopristin (1 -18%) are very seldom or of moderate frequency.
T171 116763-116786 Sentence denotes Discussion/Conclusions:
T172 116787-117048 Sentence denotes The presented frequencies of antibiotic resistances among Vancomycin-resistant E. faecium add important data to existing resistance surveillance systems such as ARS or GERMAP; the latter collect resistance data of all enterococci not especially focusing on VRE.
T173 117049-117231 Sentence denotes Our data show frequent co-resistances to therapeutically relevant antibiotics (ampicillin, aminoglycosides) for enterococci; resistances to antibiotics of last resort are still rare.
T174 117232-117331 Sentence denotes Comparably high rates of vanB VRE suggest a changing epidemiology of van genotypes in recent years.
T175 117332-117343 Sentence denotes Background:
T176 117344-117429 Sentence denotes The prevalence of multidrug-resistant gramnegative pathogens is increasing worldwide.
T177 117430-117584 Sentence denotes Especially the spread of carbapenemase producing Enterobacteriaceae is worrisome since those strains are usually also resistant to most other antibiotics.
T178 117585-117694 Sentence denotes No new antibiotics with activity against gramnegative bacteria will be introduced within the next five years.
T179 117695-117791 Sentence denotes Therefore, testing the activity of older and rarely used antibiotics is of high clinical impact.
T180 117792-117800 Sentence denotes Methods:
T181 117801-118201 Sentence denotes Enterobacteriaceae (n = 107) with elevated carbapenem MICs were sent to the National Reference Laboratory for Multidrug-Resistant Gramnegative Bacteria from several laboratories all over Germany and investigated for the presence of carbapenemases by the modified Hodge test, synergy testing using EDTA and boronic acid, a microbiological assay using cell-free extracts and PCR followed by sequencing.
T182 118202-118346 Sentence denotes Resistance testing of fosfomycin was performed by Etest, disk diffusion and agar dilution on Glucose-6-phosphate containing Mueller-Hinton agar.
T183 118347-118355 Sentence denotes Results:
T184 118356-118614 Sentence denotes In a collection of multidrug-resistant K. pneumoniae (n = 57), E. coli (n = 18), E. cloacae (n = 12), K. oxytoca (n = 7) and further species various -lactamases like OXA-48 (n = 24), KPC-2 (n = 12), KPC-3 (n =11), VIM-1 (n = 15), CTX-M-15 (n = 9) were found.
T185 118615-118711 Sentence denotes Regarding agar dilution as the reference method the MIC50 was 8 mg/L and the MIC90 was 512 mg/L.
T186 118712-118795 Sentence denotes Overall 77 isolates (72%) were susceptible according to EUCAST with an MIC 32 mg/L.
T187 118796-118902 Sentence denotes Susceptibility rates were 66.7% for K. pneumoniae, 88.9% for E. coli and 75% for E. cloacae, respectively.
T188 118903-119011 Sentence denotes Etest gave concordant results in 92.5% and showed seven very major errors (6.5%) and one major error (0.9%).
T189 119012-119194 Sentence denotes Applying a threshold of 16 mm as susceptible in disk diffusion (fosfomycin 200 μg) as recommended by CLSI for E. coli eleven very major errors (10.3%) and no major errors were found.
T190 119195-119207 Sentence denotes Conclusions:
T191 119208-119359 Sentence denotes Using EUCAST criteria and agar dilution a diverse collection of multidrug-resistant Enterobacteriaceae strains showed fosfomycin susceptibility of 72%.
T192 119360-119474 Sentence denotes With less than 10% very major or major errors Etest is an acceptable method for fosfomycin susceptibility testing.
T193 119475-119590 Sentence denotes In this study we report on increasing frequency of resistance and molecular characterization of resistant isolates.
T194 119591-119599 Sentence denotes Methods:
T195 119600-119784 Sentence denotes All isolates sent to the German Reference Centre for Staphylococci were tested for antibiotic susceptibility by broth microdilution and were assigned to clonal complexes by spa-typing.
T196 119785-119916 Sentence denotes For a subset of isolates antibiotic susceptibility was tested by automatic susceptibility testing (VITEK) and E-test in comparison.
T197 119917-120095 Sentence denotes To elucidate the molecular basis of resistance 20 representative isolates from different geographic origin were subjected to Solexa whole genome sequencing of pooled DNA-samples.
T198 120096-120251 Sentence denotes Additionally, the DNA of two clinical isolates obtained before and after unsuccessful Daptomycin treatment was sequenced by Solexa whole genome sequencing.
T199 120252-120325 Sentence denotes All sequence data were mapped to appropriate S. aureus reference strains.
T200 120326-120513 Sentence denotes Based on the whole genome data the mprF locus, which was previously shown to be associated with daptomycin resistance was re-sequenced in 64 daptomycin resistant and susceptible isolates.
T201 120514-120522 Sentence denotes Results:
T202 120523-120659 Sentence denotes Among isolates received at the National Reference Centre frequency of daptomycin resistance increased from 0.7% in 2008 to 1.8% in 2011.
T203 120660-120748 Sentence denotes Most isolates were MRSA, predominantly belonging to clonal complexes CC5/ST225 and CC22.
T204 120749-120885 Sentence denotes Automatic susceptibility testing (VITEK) generally indicated slightly elevated MIC values as compared to broth microdilution and E-test.
T205 120886-120995 Sentence denotes Sequence analysis of whole genome data revealed a small number of SNPs putatively associated with resistance.
T206 120996-121203 Sentence denotes However, after re-sequencing the corresponding loci in a larger number of resistant and susceptible isolates strong association of resistance to mutations in a certain locus could be confirmed only for mprF.
T207 121204-121300 Sentence denotes Resequencing of mprF revealed point mutations in 44 out of 46 resistant isolates (MIC > 2μg/ml).
T208 121301-121312 Sentence denotes Conclusion:
T209 121313-121502 Sentence denotes Results corroborate the role of mprF-mutations in resistance development towards daptomycin; however further studies must elucidate the complete mechanism leading to reduced susceptibility.
T210 121503-121599 Sentence denotes Spread of KPC-3 producing K. pneumoniae belonging to the international clone ST258 in Germany M.
T211 121600-121615 Sentence denotes Kaase* 1 , J.L.
T212 121616-121637 Sentence denotes Schwichtenberg 1 , F.
T213 121638-121650 Sentence denotes Lange 1 , F.
T214 121651-121666 Sentence denotes Szabados 1 , N.
T215 121667-121686 Sentence denotes Pfennigwerth 1 , S.
T216 121687-121770 Sentence denotes Gatermann Abteilung für Medizinische Mikrobiologie, Bochum, Deutschland Background:
T217 121771-121974 Sentence denotes The worldwide spread of carbapenemase producing Enterobacteriaceae is worrisome since therapeutic options for such strains are severely limited and no new antibiotics will be licensed in the near future.
T218 121975-122027 Sentence denotes Data regarding carbapenemases in Germany are scarce.
T219 122028-122130 Sentence denotes This study focuses on the spread of KPC-3 carbapenemases from different geographic regions in Germany.
T220 122131-122139 Sentence denotes Methods:
T221 122140-122300 Sentence denotes Strains with elevated carbapenem MICs were sent to the National Reference Laboratory for multidrug-resistant gramnegative bacteria for further characterization.
T222 122301-122381 Sentence denotes Carbapenemases were detected by phenotypic tests, PCR and subsequent sequencing.
T223 122382-122428 Sentence denotes MICs for carbapenems were determined by Etest.
T224 122429-122507 Sentence denotes The genetic environment of blaKPC was characterized by a set of specific PCRs.
T225 122508-122625 Sentence denotes Plasmid characterization was performed by S1 nuclease restriction and conjungative transfer of the plasmid was tried.
T226 122626-122774 Sentence denotes Results: KPC-3 was detected in K. pneumoniae (n=27) and E. coli (n=1) single patient strains from ten different geographic regions all over Germany.
T227 122775-122911 Sentence denotes The MICs for ertapenem fell in a range from 0.5 to >32 mg/L, for imipenem from 0.38 to >32 mg/L and for meropenem from 0.25 to >32 mg/L.
T228 122912-123009 Sentence denotes The modified Hodge-Test and the combined disk test with boronic acid was positive in all strains.
T229 123010-123125 Sentence denotes PFGE demonstrated relatedness of 26 K. pneumoniae strains and MLST typing of five exemplary strains revealed ST258.
T230 123126-123179 Sentence denotes PCR detected blaOXA-1 in 14 and blaTEM in 25 strains.
T231 123180-123364 Sentence denotes Transconjugation experiments were successful only in one strain, however the presence of blaKPC on plasmids could be demonstrated by restriction with S1 endonuclease and Southern Blot.
T232 123365-123462 Sentence denotes PCRs demonstrated that blaKPC-3 was surrounded by Tn4401a showing a 100-bp-deletion in all cases.
T233 123463-123565 Sentence denotes Conclusions: KPC-3 producing K. pneumoniae belonging to the ST258 clone are spreading also in Germany.
T234 123566-123655 Sentence denotes Similar to reports from the USA and Israel blaKPC-3 is surrounded by the Tn4401a isoform.
T235 123656-123763 Sentence denotes The low MIC for carbapenems in some strains emphasizes that detection of carbapenemases can be challenging.
T236 123764-123793 Sentence denotes Resistance to Zink in MRSA W.
T237 123794-123807 Sentence denotes Witte* 1 , C.
T238 123808-123897 Sentence denotes Cuny* 1 1 Robert Koch Institut, Bereich Wernigerrode, Wernigerode, Deutschland Objective:
T239 123898-124173 Sentence denotes The finding of a gene coding for a Zn 2+ porter (czrC) contained by an SCCmecV element in livestock associated MRSA CC398 and the previous use of zinkoxide together with organic acids in pig feeding raised the more general question of coselection of MRSA by use of zinkoxide.
T240 124174-124394 Sentence denotes In this context it is of interest whether czrC and the already known resistance gene cadA (both code for an ATPase based efflux) are disseminated among different clonal lineages of MRSA prevalent in humans and livestock.
T241 124395-124403 Sentence denotes Methods:
T242 124404-124535 Sentence denotes Defined collection of MRSA from humans (HA-MRSA, CA-MRSA) and from other animals (LA-MRSA) attributed to different clonal lineages.
T243 124536-124635 Sentence denotes Phenotypic susceptibility testing for ZnCL2 by means of microbroth MIC in cation adjusted MH broth.
T244 124636-124694 Sentence denotes PCR demonstration of Zn 2+ resistance genes cadA and czrC.
T245 124695-124910 Sentence denotes Results: czrC was exclusively associated with MRSA ST398 (20/27 from pigs; 21/23 from humans exposed to pigs; 10/20 infections; 10/10 infections in humans) and contained by a defined SCCmec-element with tandem ccrC.
T246 124911-125137 Sentence denotes It was not found in HA-MRSA and CA-MRSA from central Europe clonal lineages. cadA is typical for CA-MRSA ST30 (as to be expected!) and ST80, it was also detected in 2/5 CA-MRSA ST22 but not in CA-MRSA ST1, ST5, ST8, and ST152.
T247 125138-125227 Sentence denotes Among HA-MRSA CC5 cadA was only found in isolate of ST228, not in isolates ST225 and ST5.
T248 125228-125348 Sentence denotes Among HA-MRSA CC8 only the two isolates attributed to ST239 contained cadA, and one among 5 isolates attributed to CC45.
T249 125349-125360 Sentence denotes Discussion:
T250 125361-125536 Sentence denotes It can not be excluded that the previous use of zinkoxide ions in conjunction with organic acids as feed additives has selected for czrC and possibility coselected MRSA ST398.
T251 125537-125637 Sentence denotes In human medicine zinkoxid is widely used in hospitals as constituent of ointments for care of skin.
T252 125638-125780 Sentence denotes As the most widely disseminated MRSA (MRSA ST22, IV, ST225, II) remain susceptible to Zn 2+ there is obviously no selection in favour of MRSA.
T253 125781-125986 Sentence denotes With respect to multiresistant but methicillin susceptible S. aureus ST30 ("80/80 complex") in the past and use of zinkoxide ointments containing also boronic acid this situation might have been different.
T254 125987-126000 Sentence denotes Introduction:
T255 126001-126160 Sentence denotes The emergence of healthcare associated infections caused by multidrug-resistant Gram-negative bacteria has become an increasing problem over the past 20 years.
T256 126161-126296 Sentence denotes Colistin, a 'forgotten' antibiotic, has attracted attention, because of its activity against multidrug-resistant Gramnegative bacteria.
T257 126297-126358 Sentence denotes It is, however, not active against Proteeae and Serratia spp.
T258 126359-126647 Sentence denotes The objective of this study was to evaluate the susceptibilities of clinical isolates of Enterobacter cloacae (ECL), Escherichia coli (ECO), Klebsiella pneumoniae (KPN), Klebsiella oxytoca (KOX), Pseudomonas aeruginosa (PAE) and Acinetobacter baumannii group (ABA) to COL and comparators.
T259 126648-126656 Sentence denotes Methods:
T260 126657-126862 Sentence denotes A total of 1,529 isolates were prospectively collected from 21 microbiology laboratories across Germany, which participated in the surveillance study conducted by the Paul-Ehrlich-Society in November 2007.
T261 126863-127039 Sentence denotes MICs of COL, ciprofloxacin (CIP), ceftazidime (CAZ), gentamicin (GEN) and meropenem(MEM) were determined by the broth microdilution procedure according to the standard DIN ISO.
T262 127040-127095 Sentence denotes MICs were interpreted by EUCAST criteria, if available.
T263 127096-127167 Sentence denotes Breakpoints of COL were <=2 mg/L (susceptible) and >2 mg/L (resistant).
T264 127168-127304 Sentence denotes The CLSI MIC method using CAZ +/-clavulanic acid (CLA) and cefotaxime +/-CLA was employed as screening test for ESBL-producing isolates.
T265 127305-127313 Sentence denotes Results:
T266 127314-127422 Sentence denotes Isolates were primarily recovered from wounds (26%), respiratory specimens (26%) urine (20%) and blood (9%).
T267 127423-127478 Sentence denotes There were 460 ICU isolates and 1,069 non-ICU isolates.
T268 127479-127562 Sentence denotes Of the ECO, KPN and KOX isolates, 11.5%, 10.5% and >15.8% showed an ESBL-phenotype.
T269 127563-127622 Sentence denotes MIC-50 and MIC-90 values (mg/L) are displayed in the table.
T270 127623-127725 Sentence denotes Of the ECO isolates, 100% were susceptible (S) to COL and MEM, while 72% were S to CIP and 87% to GEN.
T271 127726-127800 Sentence denotes Of the ECL isolates, 91% and 9% were S and resistant to COL, respectively.
T272 127801-127893 Sentence denotes Susceptibilities of ECL to CAZ, CIP, GEN and MEM were 52%, 89%, 88%, and >99%, respectively.
T273 127894-128043 Sentence denotes Among KPN and KOX isolates, susceptibilities to COL and MEM were 98-100%, while susceptibilities to CIP and GEN were 82-85% and 94-96%, respectively.
T274 128044-128147 Sentence denotes COL was the most active drug against ABA and PAE, with susceptibility rates of 100% for either species.
T275 128148-128224 Sentence denotes For ABA, susceptibilities of comparators varied from 78% (CIP) to 95% (MEM).
T276 128225-128350 Sentence denotes Susceptibility rates of PAE for comparative agents were as follows: GEN (91%) followed by CAZ (87%), MEM (85%) and CIP (75%).
T277 128351-128363 Sentence denotes Conclusions:
T278 128364-128453 Sentence denotes Overall, susceptibility to COL seems to be high among Gram-negative pathogens in Germany.
T279 128454-128509 Sentence denotes COL may be considered as an important treatment option.
T280 128510-128743 Sentence denotes Bacterial pathogens Neisseria meningitidis and Haemophilus influenzae are infrequent causes for invasive disease manifesting as sepsis or meningitis; their respective incidence rates in Germany were 0.47 and 0.26 per 100,000 in 2010.
T281 128744-128873 Sentence denotes We present antibiotic susceptibility data over a period of nine and three years for Meningococci and H. influenzae, respectively.
T282 128874-129136 Sentence denotes Using gradient test systems (Etest), 3283 non-duplicate meningococcal strains were tested for susceptibility against benzylpenicillin, rifampicin, and ciprofloxacin between 2002 and 2010; 417 meningococcal strains were also assayed for cefotaxime susceptibility.
T283 129137-129242 Sentence denotes Between 2008 and 2010, 298 nonduplicate H. influenzae strains were assayed for ampicillin susceptibility.
T284 129243-129513 Sentence denotes In meningococci, no temporal trends in minimal inhibitory concentrations (MIC) or susceptibility categorization according to EUCAST were observed; 85.5%, 13.8%, and 0.7% of strains were categorized as susceptible, intermediate, and resistant to penicillin, respectively.
T285 129514-129602 Sentence denotes Rate of rifampicin-resistant and -susceptible isolates was 0.4% and 99.6%, respectively.
T286 129603-129716 Sentence denotes Only 0.1% of strains were not susceptible to ciprofloxacin and all tested strains were susceptible to cefotaxime.
T287 129717-129982 Sentence denotes MIC to penicillin depended on serogroup: using serogroup B and rare serogroups (e.g. A, Z, X, 29E) as reference category with a mean MIC of 0.056 mg/L, serogroups C, W-135, and Y showed significant deviations with MICs of 0.074, 0.107, and 0.067 mg/L, respectively.
T288 129983-130075 Sentence denotes Interestingly, 51.9% of serogroup W-135 strains were intermediate susceptible to penicillin.
T289 130076-130191 Sentence denotes Most resistant isolates (82.6%) were of serogroup C, while serogroups W-135 and Y had no resistant isolates at all.
T290 130192-130341 Sentence denotes Although no resistance was observed for cefotaxime, cefotaxime MICs showed a weak yet significant linear correlation with penicillin MICs (r^2 0.23).
T291 130342-130489 Sentence denotes Accordingly, mean cefotaxime MICs were 0.003, 0.005, and 0.011 mg/L for penicillin susceptible, intermediate, and resistant isolates, respectively.
T292 130490-130629 Sentence denotes Due to low rate of nonsusceptibility, no meaningful analyses were possible for antimicrobials rifampicin and ciprofloxacin in meningococci.
T293 130630-130703 Sentence denotes In H. influenzae, 10.4% were resistant to ampicillin according to EUCAST.
T294 130704-130778 Sentence denotes A downard temporal trend was observed between 2008 (16.7%) to 2010 (8.4%).
T295 130779-130928 Sentence denotes Resistance was significantly more common in non-typeable H. influenzae lacking a polysaccharide capsule (13.0%) compared to capsulate strains (3.7%).
T296 130929-130972 Sentence denotes Rate of beta-lactamase production was 9.3%.
T297 130973-131137 Sentence denotes Three ampicillin-resistant strains (1.1% of total) lacked beta-lactamase activity and were thus categorized as beta-lactamase-negative-ampicillin-resistant (BLNAR).
T298 131138-131223 Sentence denotes Antimicrobial resistance in K. pneumoniae -Results from the German ARS System 2009 I.
T299 131224-131236 Sentence denotes Noll* 1 , B.
T300 131237-131255 Sentence denotes Schweickert 1 , M.
T301 131256-131267 Sentence denotes Feig 1 , G.
T302 131268-131281 Sentence denotes Krause 1 , T.
T303 131282-131349 Sentence denotes Eckmanns Infektionsepidemiologie, Berlin, Deutschland Introduction:
T304 131350-131719 Sentence denotes High prevalence of resistance in invasive K. pneumoniae isolates from hospitalised patients to third-generation cephalosporins (3GC), fluoroquinolones (FQ) and aminoglycosides (AG) and a high level of combined resistance to these classes in most European countries was a major result of the European Antimicrobial Resistance Surveillance Network (EARS-Net) Report 2009.
T305 131720-131933 Sentence denotes In Germany, the ARS (Antimicrobial Resistance Surveillance)-System allows to study resistance and resistance patterns in further settings: urine and respiratory samples from inpatients as well as from outpatients.
T306 131934-131942 Sentence denotes Methods:
T307 131943-132084 Sentence denotes Analysis is based on non-duplicate isolates of K. pneumoniae collected in 2009 by nine laboratories covering 160 hospitals and 865 practices.
T308 132085-132225 Sentence denotes Species identification and antimicrobial susceptibility testing is performed by VITEK 2, results are evaluated according to CLSI guidelines.
T309 132226-132331 Sentence denotes Isolates are classified as resistant to an antibiotic class if they show resistance to one of its agents:
T310 132332-132455 Sentence denotes 3GC: ceftazidime or cefotaxime or ceftriaxone; FQ: ciprofloxacin or levofloxacin; AG: gentamicin or tobramicin or amikacin.
T311 132456-132523 Sentence denotes The distinct class resistances are combined to resistance patterns.
T312 132524-132532 Sentence denotes Results:
T313 132533-132625 Sentence denotes The sample is composed of 5,548 non-duplicate K. pneumoniae isolates from inpatients (blood:
T314 132626-132652 Sentence denotes 299; respiratory samples :
T315 132653-132666 Sentence denotes 1,118; urine:
T316 132667-132732 Sentence denotes 4,131 ) and 2,666 isolates from outpatients (respiratory samples:
T317 132733-132744 Sentence denotes 173; urine:
T318 132745-132752 Sentence denotes 2,493).
T319 132753-132786 Sentence denotes Results are displayed in table 1.
T320 132787-133007 Sentence denotes Proportions of resistance are highest against FQ in all subsets ranging from 21.1% in blood cultures from inpatients to 4.6% in respiratory samples from outpatients followed by 3GC (16.4% to 3.5%) and AG (13.4% to 2.3%).
T321 133008-133245 Sentence denotes Regarding resistance against the three antibiotic classes simultaneously reveals that triple resistance is the most frequent pattern in all inpatient subsets reaching 10% in blood cultures, 9.3% in respiratory samples and 6.8% in urines.
T322 133246-133442 Sentence denotes In outpatient samples, single resistance against FQ is the most frequent pattern followed by triple resistance in second place accounting for 3.0% in urine samples and 1.2% in respiratory samples.
T323 133443-133454 Sentence denotes Discussion:
T324 133455-133860 Sentence denotes While the EARS-Net-Surveillance limited to invasive isolates from hospitalised patients captures the sector with highest levels of antimicrobial resistance in K. pneumoniae, the extended approach of ARS reveals an emerging problem in outpatient care that physicians should be aware of, even if resistance proportions might be overestimated as samples are more likely to be taken from pre-treated patients.
T325 133861-133972 Sentence denotes The German multicentre study ResiNet was launched by the National Reference Centre (NRC) for H. pylori in 2001.
T326 133973-134139 Sentence denotes The aims of this ongoing surveillance study are to monitor and to prospectively investigate risk factors for the development of antimicrobial resistance in H. pylori.
T327 134140-134273 Sentence denotes 14 microbiological centres spread over the whole country each collaborating with 2 to 7 gastroenterologists participate in the study.
T328 134274-134338 Sentence denotes At the end of December 2010, 1423 H. pylori strains were tested.
T329 134339-134436 Sentence denotes Most of the patients suffered from gastritis (44%) followed by ulcus ventriculi or duodeni (19%).
T330 134437-134561 Sentence denotes The overall resistance rates are 42% for metronidazole (MZ), 29% for clarithromycin (CLA) and 17% for fluoroquinolones (FQ).
T331 134562-134664 Sentence denotes Double-(MZ & CLA) and triple-resistant strains (MZ & CLA & FQ) were found in 19% and 5%, respectively.
T332 134665-134744 Sentence denotes Antimicrobial pre-treatment is the main risk factor for resistance development.
T333 134745-134857 Sentence denotes After one course of H. pylori eradication therapy resistance rates reach 50% (MZ), 59% (CLA) and 31% (MZ & CLA).
T334 134858-134958 Sentence denotes More than one eradication therapy led to resistance rates of 82% (MZ), 74% (CLA) and 64% (MZ & CLA).
T335 134959-135039 Sentence denotes When we started the study in 2001, 32% of the strains were resistant against MZ.
T336 135040-135105 Sentence denotes At the end of 2010, 42% of the strains were resistant against MZ.
T337 135106-135161 Sentence denotes For CLA the resistance rates increased from 15% to 29%.
T338 135162-135251 Sentence denotes In 2007, we started a follow up study to investigate the outcome of our ResiNet patients.
T339 135252-135291 Sentence denotes 451 patients have been examined so far.
T340 135292-135368 Sentence denotes The majority of the patients (87%) were treated with an eradication therapy.
T341 135369-135427 Sentence denotes The most common scheme used was the French triple therapy.
T342 135428-135533 Sentence denotes After treatment 70% of the patients reported an improvement of their complaints or no complaints anymore.
T343 135534-135632 Sentence denotes Eradication control was performed in 80% of the patients using the urea-breath test predominantly.
T344 135633-135679 Sentence denotes The eradication rate for all patients was 61%.
T345 135680-135847 Sentence denotes 19% of the patients were still H. pylori positive after treatment, which might be explained by the high rate of clarithromycin resistance (29%) within this population.
T346 135848-136040 Sentence denotes In conclusion, these results underline the hypothesis that development of resistance in H. pylori in contrast to other pathogens is dependent on treatment strategies in the individual patient.
T347 136041-136203 Sentence denotes Further, the data clearly demonstrate that culture and susceptibility testing from gastric biopsy specimen is mandatory already after the first treatment failure.
T348 136204-136401 Sentence denotes MRSA-Surveillance in Germany: data from the Antibiotic Resistance Surveillance System (ARS) and the mandatory surveillance of MRSA in blood and cerebrospinal fluid. inhabitants/year, were reported.
T349 136402-136485 Sentence denotes Males >65 years present with the highest incidence (23.5/100,000 inhabitants/year).
T350 136486-136721 Sentence denotes Stratification by federal states shows considerable regional differences (range:1.8-6.4/100,000 inhabitants/year), which were positively correlated to regional MRSA-rates (6.7%-27.3%) generated in ARS (spearman correlation coefficient:
T351 136722-136736 Sentence denotes 0.74; p=0.04).
T352 136737-136748 Sentence denotes Conclusion:
T353 136749-136912 Sentence denotes While mandatory reporting of MRSA-bacteraemia serves as an indicator of the situation in hospital care, ARS additionally provides a view on the outpatient setting.
T354 136913-137004 Sentence denotes Expectedly, in ambulatory care MRSA-rates are considerably lower compared to hospital care.
T355 137005-137117 Sentence denotes Nevertheless, the results indicate that in Germany MRSA plays a relevant role in the outpatient setting as well.
T356 137118-137367 Sentence denotes Particularly, high resistance rates in urine samples suggest that further investigations about the epidemiological context in which infections occur and patient characteristics are necessary to assess possible consequences for antibiotic management.
T357 137369-137507 Sentence denotes Comparable high carriage rates of Extended-spectrum beta-Lactamases producing Escherichia coli in birds of prey from Germany and Mongolia.
T358 137508-137510 Sentence denotes S.
T359 137511-137526 Sentence denotes Günther* 1 , K.
T360 137527-137547 Sentence denotes Aschenbrenner 1 , I.
T361 137548-137560 Sentence denotes Stamm 2 , A.
T362 137561-137573 Sentence denotes Bethe 1 , M.
T363 137574-137589 Sentence denotes Stubbe 3 , L.H.
T364 137590-137603 Sentence denotes Wieler 1 , C.
T365 137604-137669 Sentence denotes Ewers 1 1 Freie Universität Berlin, Institut für Mikrobiologie u.
T366 137670-137770 Sentence denotes Tierseuchen, Berlin, Deutschland 2 Vet Med Labor GmbH, Ludwigsburg, Halle, Deutschland Introduction:
T367 137771-138007 Sentence denotes Several studies give evidence for the carriage of Extendedspectrum beta-Lactamases (ESBL) producing E. coli in the feces of wild birds even in remote areas, where the presence of antibiotic selection pressure is assumed to be very rare.
T368 138008-138245 Sentence denotes In the present study we compared wild avian fecal E. coli isolates from remote areas in Mongolia with those from an intensively agriculturally used area in Germany with respect to the rate of ESBL production and their clonal relatedness.
T369 138246-138266 Sentence denotes Methods and Results:
T370 138267-138385 Sentence denotes 281 cloacal swabs, mostly taken from juvenile birds, were screened for E. coli, resulting in a number of 108 isolates.
T371 138386-138536 Sentence denotes By means of a selective growth medium ESBL-production was confirmed in 13.8% (n=9) German and in 11.6 % (n=5) of the Mongolian avian E. coli isolates.
T372 138537-138707 Sentence denotes In both groups we determined ESBL enzymes of the CTX-M family as the most prominent ones, while blaCTX-M-1 dominated the German samples and blaCTX-M-9 the Mongolian ones.
T373 138708-138909 Sentence denotes Genotyping led to the frequent detection of virulence determinants associated with extraintestinal pathogenicity whereas genes associated with intestinal pathogenic pathovars were only rarely observed.
T374 138910-139171 Sentence denotes Phylogenetic typing via MLST and structure analysis revealed high rates of ancestral group B2 -a group linked with extraintestinal virulence -particularly in the German ESBL strainsmost of the Mongolian ESBL strains were assigned to phylogenetic groups D and A.
T375 139172-139369 Sentence denotes Among the Mongolian samples we identified several STs, such as ST167 (ancestral group A) or ST648 (hybrid group ABD) which have previously been reported from human clinical ESBL producing isolates.
T376 139370-139595 Sentence denotes Macrorestriction patterns revealed a clonal relatedness of wild avian fecal isolates from Asia to animal and human clinical isolates from Europe, indicating the global and interspecies dissemination of multiresistant strains.
T377 139596-139608 Sentence denotes Conclusions:
T378 139609-139732 Sentence denotes Our findings underline the urgent need to unravel the role of wild birds in the spread of antimicrobial resistant bacteria.
T379 139733-139998 Sentence denotes Due to their migration behavior and with respect to the omnipresence of bird´s feces in wild and urban environments, wild birds might represent a non-point source of infection and could by that considerably contribute to the transmission of ESBL producing bacteria.
T380 139999-140001 Sentence denotes M.
T381 140002-140016 Sentence denotes Franke* 1 , R.
T382 140017-140033 Sentence denotes Söller 2 , K.-D.
T383 140034-140218 Sentence denotes Thoben 1 1 BIBA, IKAP, Bremen, Deutschland 2 Qiagen, Hamburg, Deutschland Introduction: qRT-PCR has widely been used for over ten years and has entered into domains with medical needs.
T384 140219-140322 Sentence denotes The detection of microorganisms in industrial environment takes place with the methods of microbiology.
T385 140323-140563 Sentence denotes The issue of reducing the time of detection of microorganisms is not resolved with the usage of the qRT-PCR method , but improved stronglyAll developed PCR analyze methods need an additional pre-processing to reduce the wide range of noise.
T386 140564-140738 Sentence denotes The currently used methods require the use of experts to evaluate the result of noise removal, because the methods need some parameters which correlate to the detected noise.
T387 140739-140805 Sentence denotes The adaption/calibration can only be executed by a manual process.
T388 140806-140898 Sentence denotes This fact prevents the usage of the qRT-PCR-analysis in an automated industrial environment.
T389 140899-141042 Sentence denotes The focus of this article lies in the development of automated pre-processing method which can reduce the significantly influence of the noise.
T390 141043-141049 Sentence denotes Noise:
T391 141050-141134 Sentence denotes Generally, in test results, two different types of noise can occur (Wilhelm, 2003) .
T392 141135-141235 Sentence denotes The first type of noise is the background noise and the second type of noise is called signal trend.
T393 141236-141295 Sentence denotes The possible causes of the occurrence are not yet resolved.
T394 141296-141419 Sentence denotes The effects of background noise and signal trend for the repeated execution of sample analysis may lead to varying results.
T395 141420-141434 Sentence denotes Noise removal:
T396 141435-141559 Sentence denotes The noise removal cannot generally detect and mathematically formulate the background noise and the signal trend of a curve.
T397 141560-141634 Sentence denotes Existing methods do not try to detect the noise but only try to remove it.
T398 141635-141708 Sentence denotes Such methods use parameters to control the strength of the noise removal.
T399 141709-141747 Sentence denotes Pre-processing using the Bézier curve:
T400 141748-141926 Sentence denotes The smoothing with Bézier curve generally changes the curve progression so that the local extramas will be removed and the four phases of an exponential curve will be pronounced.
T401 141927-141938 Sentence denotes Evaluation:
T402 141939-142125 Sentence denotes The project ProDIAP (This project which this report was funded by the Bundesministeriums für Bildung und Forschung under support code 01RI0709A-C) used 640 references for the evaluation.
T403 142126-142277 Sentence denotes The evaluation of the quality of the qualitative analysis with the Bézier smoothing estimated with the criteria: recall, precision and the specificity.
T404 142278-142391 Sentence denotes The results of the recall is 100%, the average of the precision is 96% and the average of the specificity is 97%.
T405 142392-142405 Sentence denotes Introduction:
T406 142406-142544 Sentence denotes Ulifloxacin is a new fluorochinolone approved for the treatment of lower urinary tract infections and exacerbations of chronic bronchitis.
T407 142545-142671 Sentence denotes The prodrug Prulifloxacin is available for oral administration and is metabolized in the liver to its active form Ulifloxacin.
T408 142672-142784 Sentence denotes Ulifloxacin demonstrates a broad spectrum of antimicrobial activity against grampositive and -negative bacteria.
T409 142785-142916 Sentence denotes In this study the susceptibility of Ulifloxacin was compared to other fluorochinolones in 267 isolates of urinary tract infections.
T410 142917-142939 Sentence denotes Materials and Methods:
T411 142940-143081 Sentence denotes During testing period in April 2011 in 267 clinical isolates of urinary tract infections Ulifloxacin was added to routine resistance testing.
T412 143082-143170 Sentence denotes Used method was disc diffusion; interpretation was performed according to CLSI criteria.
T413 143171-143251 Sentence denotes For Ulifloxacin (disc containing 5 μg) interpretative criteria were as followed:
T414 143252-143365 Sentence denotes 15 and 19 mm for Enterobacteriaceae, 16 and 20 mm for Nonfermenters, and 14 and 18 mm for Gram-positive bacteria.
T415 143366-143534 Sentence denotes Tested urinary tract infection strains include mainly E. coli (n= 159), other Enterobacteriaceae (n= 57), Pseudomonas sp (n= 14), Enterococci and Staphylococci (n= 24).
T416 143535-143543 Sentence denotes Results:
T417 143544-143678 Sentence denotes Comparing Ulifloxacin vs. Ciprofloxacin and Norfloxacin concordant results could be obtained in 250 of the 267 (93.6%) tested strains.
T418 143679-143842 Sentence denotes Differences in the interpretation between tested substances were found in some isolates of E. coli (7 of 159), Pseudomonas sp. (2 of 14) and Enterococci (8 of 21).
T419 143843-143957 Sentence denotes In 2 of 17 cases Ciprofloxacin and Norfloxacin were resistant, whereas Ulifloxacin was interpreted as susceptible.
T420 143958-144035 Sentence denotes The other 15 of these 17 cases showed minimal interpretation variations only.
T421 144036-144189 Sentence denotes Regarding tested isolates E. coli (n= 159) revealed a susceptibility of 79.2% for Ciprofloxacin and Norfloxacin, and 80.5% for Ulifloxacin, respectively.
T422 144190-144319 Sentence denotes Pseudomonas sp. (n=14) showed a susceptibility of 71.4% for Ciprofloxacin and Norfloxacin respectively and 85.7% for Ulifloxacin.
T423 144320-144369 Sentence denotes For Enterococci following results could be found:
T424 144370-144464 Sentence denotes Susceptibility to Ciprofloxacin, Norfloxacin and Prulifloxacin was 38%, 42%, 28% respectively.
T425 144465-144476 Sentence denotes Discussion:
T426 144477-144558 Sentence denotes Ulifloxacin is a new fluorochinolone approved for lower urinary tract infections.
T427 144559-144694 Sentence denotes Comparing the results of Ciprofloxacin, Norfloxacin and Ulifloxacin concordant results in 250 of 267 strains (93.6%) could be obtained.
T428 144695-144785 Sentence denotes Ulifloxacin was as active as Ciprofloxacin and Norfloxacin against gram negative isolates.
T429 144786-144947 Sentence denotes Against gram positive strains Ulifloxacin was slightly less active as Ciprofloxacin and Norfloxacin, but this statement is limited by the low number of isolates.
T430 144948-145172 Sentence denotes Because of some minor differences in the interpretation of the susceptibility pattern testing of Ulifloxacin and/or other chinolones resistance testing seems to be noteworthy if therapy with this fluorochinolone in intended.
T1 145173-145339 Sentence denotes Impact of thymidine on the antimicrobial activities of trimethoprim/sulfamethoxazole, 5-iodo-2'-deoxyuridine, and rifampicin against Staphylococcus aureus Objectives:
T2 145340-145503 Sentence denotes A treatment option against S. aureus in general and MRSA in particular is trimethoprim/sulfamethoxazole (SXT) either alone or in combination with rifampicin (RIF).
T3 145504-145666 Sentence denotes SXT inhibits different enzymatic steps of the folic acid pathway, leading to cessation of bacterial synthesis of thymidine monophosphate via thymidylate synthase.
T4 145667-145878 Sentence denotes However, the antimicrobial activity of SXT can be antagonised by bacterial utilisation of thymidine, which can be present in infected and/or inflamed tissues such as the airways of cystic fibrosis (CF) patients.
T5 145879-146132 Sentence denotes The aim of this study was to determine thymidine concentrations in the sputa of CF patients and to analyse the antimicrobial activity of SXT, 5-iodo-2´-deoxyuridine (IdUrd) and RIF alone and in combination against S. aureus in the presence of thymidine.
T6 146133-146154 Sentence denotes Material and Methods:
T7 146155-146250 Sentence denotes Thymidine concentrations in CF sputa were determined by high performance liquid chromatography.
T8 146251-146379 Sentence denotes The antimicrobial activities of SXT, IdUrd and RIF alone and in combination against S. aureus were analysed by time kill assays.
T9 146380-146388 Sentence denotes Results:
T10 146389-146493 Sentence denotes Thymidine concentrations in the sputa of ten different CF patients varied from <100 μg/L to 38 845 μg/L.
T11 146494-146635 Sentence denotes The abolished antimicrobial activity of SXT against 22 S. aureus strains in the presence of thymidine was restored by combination with IdUrd.
T12 146636-146800 Sentence denotes In contrast, SXT combined with RIF in the presence of thymidine did not show a synergistic effect and, furthermore, allowed the emergence of RIF-resistant bacteria.
T13 146801-146905 Sentence denotes Adding RIF to the combination of SXT and IdUrd did not improve antimicrobial activity against S. aureus.
T14 146906-146918 Sentence denotes Conclusions:
T15 146919-147073 Sentence denotes Our data suggest that SXT in combination with rifampicin may fail to cure clinical S. aureus infections in tissues with elevated thymidine concentrations.
T16 147074-147220 Sentence denotes SXT in combination with a thymidine kinase inhibitor, however, may become an interesting antimicrobial combination for this setting in the future.
T17 147221-147281 Sentence denotes Further in vitro and in vivo studies are therefore required.
T18 147282-147400 Sentence denotes Non-nucleoside analogs as bacterial thymidine kinase inhibitors identified by multi-step virtual screening Objectives:
T19 147401-147582 Sentence denotes The antimicrobial activity of folic acid antagonists can be antagonized by thymidine as detected in tissues with necrotic cells such as pus and sputum from cystic fibrosis patients.
T20 147583-147791 Sentence denotes Trimethoprim/sulfamethoxazole (SXT) inhibits different enzymatic steps of the folic acid pathway leading to cessation of the bacterial synthesis of deoxythymidine monophosphate (dTMP) by thymidylate synthase.
T21 147792-148025 Sentence denotes However, several bacterial species including S. aureus possess an alternative pathway for synthesis of intracellular dTMP by uptake of extracellular thymidine and subsequent intracellular phosphorylation to dTMP via thymidine kinase.
T22 148026-148314 Sentence denotes This study was aimed at (i) screening for nonnucleoside analog inhibitors of S. aureus thymidine kinase by multi-step virtual screening, and (ii) determining the in vitro activity of these thymidine kinase inhibitors against S. aureus in combination with SXT in the presence of thymidine.
T23 148315-148336 Sentence denotes Material and Methods:
T24 148337-148471 Sentence denotes An adaptive virtual screening protocol for novel antibiotics using a combination of ligand-and structure-based approaches was applied.
T25 148472-148612 Sentence denotes Two consecutive rounds of virtual screening and in vitro susceptibility testing were performed that resulted in several non-nucleoside hits.
T26 148613-148621 Sentence denotes Results:
T27 148622-148843 Sentence denotes The most potent compound exhibits substantial antimicrobial activity against both methicillin-resistant S. aureus strain ATCC 700699 and nonresistant strain ATCC 29213, when combined with SXT in the presence of thymidine.
T28 148844-148856 Sentence denotes Conclusions:
T29 148857-149023 Sentence denotes Our study demonstrates that multi-step virtual screening can help identify bioactive substances from a large screening compound pool with limited experimental effort.
T30 149024-149182 Sentence denotes Rapid focusing on promising candidate structures was possible, so that inhibitors of bacterial thymidine kinase with non-nucleoside scaffolds were identified.
T31 149183-149384 Sentence denotes These inhibitory compounds exhibit moderate to high antimicrobial activity when combined with folic acid antagonists in the presence of thymidine, and provide rich opportunity for further optimization.
T32 149385-149401 Sentence denotes Drugs from bugs:
T33 149402-149540 Sentence denotes The bacterial protein YopM is an autonomous `self-delivering´ anti-inflammatory agent that reduces inflammation in rheumatoid arthritis C.
T34 149541-149554 Sentence denotes Rüter* 1 , J.
T35 149555-149570 Sentence denotes Bertrand 2 , J.
T36 149571-149587 Sentence denotes Scharnert 1 , C.
T37 149588-149601 Sentence denotes Cromme 2 , T.
T38 149602-149614 Sentence denotes Pap 2 , M.A.
T39 149615-149915 Sentence denotes Schmidt 1 1 ZMBE, Infektiologie, Münster, Deutschland 2 UKM, Institute of Experimental Musculoskeletal Medicine, Münster, Deutschland Cell-permeable proteins such as 'cell-penetrating peptides (CPP)' have the ability to cross cellular membranes, either alone or in association with a bioactive cargo.
T40 149916-150120 Sentence denotes We identified the Yersinia protein YopM as a novel bacterial cell-permeable protein and described the ability of isolated recombinant YopM to enter host cells without a requirement for additional factors.
T41 150121-150243 Sentence denotes Furthermore, treatment of cells with recombinant YopM leads to the down-regulation of inflammatory cytokines such as TNF .
T42 150244-150413 Sentence denotes This might contribute to systemic effects on innate immunity and, furthermore, suggests potential therapeutic applications of YopM e.g. in chronic inflammatory diseases.
T43 150414-150719 Sentence denotes Hence, we were interested to investigate whether YopM might have the potential to act as an immune modulating molecule by exhibiting beneficial effects on inflammation and the production of cytokines that maintain the inflammatory processes linked to autoimmune diseases such as rheumatoid arthritis (RA).
T44 150720-150942 Sentence denotes First, we analyzed the penetration of recombinant YopM into cells that actively drive joint and bone destruction in RA (bone marrow macrophages, osteoclasts, and synovial fibroblasts) by confocal laser scanning microscopy.
T45 150943-151054 Sentence denotes In addition, we studied the effects of YopM on osteoclastogenesis using an in vitro osteoclast formation assay.
T46 151055-151255 Sentence denotes To unravel the signaling pathways involved in the effects of YopM on cellular activation and differentiation, we investigated the activation of MAP-kinases and NF B signaling by Western Blot analysis.
T47 151256-151482 Sentence denotes With respect to a potential in vivo application of YopM, we injected YopM intra-articularly (i.a.) and intra-venously (i.v.) in wild-type and hTNFtg mice and monitored its distribution by fluorescence reflection imaging (FRI).
T48 151483-151612 Sentence denotes Furthermore, the effects of YopM on the disease manifestations were monitored by histopathology and established clinical scoring.
T49 151613-151804 Sentence denotes Our investigations revealed that once inside RA-driver cells, YopM is an effective reducer of the TNF -induced production of pro-inflammatory cytokines and cartilage-degrading enzymes (e.g. .
T50 151805-151970 Sentence denotes Furthermore, YopM is also capable of preventing structural damage by inhibition of RANKL induced osteoclastogenesis of murine bone marrow cells via the NF B-pathway.
T51 151971-152113 Sentence denotes After i.a. injection of the protein into hTNFtg mice, we observed a drastically reduced onset of symptoms and inflammation of skeletal joints.
T52 152114-152358 Sentence denotes These encouraging results combined with the possibility for focused topical applications in skeletal joints underline and strengthen the potential of YopM as a novel pathogen-derived immune modulatory agent for the treatment of inflammatory RA.
T53 152359-152456 Sentence denotes Comparison of genotypic, proteotypic and phenotypic methods for the identification of bacteria M.
T54 152457-152469 Sentence denotes Patel 1 , U.
T55 152470-152693 Sentence denotes Herber* 1 1 Accugenix, Marketing, Newark, Deutschland Accurate classification of unknown bacterial isolates is an essential first step in understanding the impact these organisms have on an environmental monitoring program.
T56 152694-152914 Sentence denotes There are many methods, technologies, and strategies utilized to determine the identity of unknown microorganisms, however, the selection of these methods is often impacted by more than the performance of the technology.
T57 152915-153041 Sentence denotes Cost, time and the amount of expertise required to perform an assay are major points to consider during the selection process.
T58 153042-153227 Sentence denotes Current available methods of identification range from genotypic to phenotypic, with 16S sequencing being universally acknowledged as the standard for routine bacterial identifications.
T59 153228-153330 Sentence denotes Still, there is even variability within this process as not all 16S sequencing methods are comparable.
T60 153331-153471 Sentence denotes When identifications are based on phenotypic characteristics, the methods are more subjective and results can be impacted by many variables.
T61 153472-153649 Sentence denotes The first practical proteotypic identification systems utilize matrix-assisted laser desorption/ionization -time of flight (MALDI-TOF) spectroscopy for microbial identification.
T62 153650-153778 Sentence denotes This technology is based on whole cell protein profiles that are subject to less expression variability than phenotypic systems.
T63 153779-154114 Sentence denotes This study directly compared performance between several of these technologies, including metabolic profiling (bioMerieux VITEK® 2 Compact), MALDI-TOF (Bruker BioTyper™), automated DNA sequence analysis (ABI MicroSEQ® 2.1), and DNA sequencing with a referencequality, customized data analysis process and curated libraries (Accugenix).
T64 154115-154219 Sentence denotes These microbial identification methods were used to analyze 60 unknown environmental bacterial isolates.
T65 154220-154305 Sentence denotes Accuracy, as well as assay cost, time, and ease of use for each method are discussed.
T66 154306-154536 Sentence denotes While 16S rRNA gene sequencing remains the standard for microbial identification of environmental isolates, proteotypic MALDI-TOF technology outperformed more phenotypic methods without compromising assay cost and turnaround time.
T67 154537-154539 Sentence denotes M.
T68 154540-154554 Sentence denotes Poceva* 1 , C.
T69 154555-154568 Sentence denotes Rueter 1 , K.
T70 154569-154581 Sentence denotes Loser 2 , A.
T71 154582-154835 Sentence denotes Schmidt 1 1 WWU Muenster, Institute of Infectiology, ZMBE, Muenster, Department of Dermatology, Muenster, Deutschland Psoriasis is one of the most common immune-mediated chronic inflammatory skin disorders affecting approximately 2-3% of the population.
T72 154836-154945 Sentence denotes It is considered as an incurable, life-long skin condition that affects all races, age groups and both sexes.
T73 154946-155014 Sentence denotes The pathogenesis of psoriasis is complex and not clearly understood.
T74 155015-155129 Sentence denotes Ongoing research indicates that immune T-cells appear to play a dominant role for the development of this disease.
T75 155130-155278 Sentence denotes These cells release TNF-a, IFN-and other pro-inflammatory cytokines that lead to vasodilation, leukocyte migration, and activation of keratinocytes.
T76 155279-155354 Sentence denotes This further activates dendritic cells which in turn leads to inflammation.
T77 155355-155592 Sentence denotes A common disadvantage (besides their side effects) of all drugs currently available for treating this primarily cutaneous disease is that they are only effective when given systemically but are largely ineffective when applied topically.
T78 155593-155776 Sentence denotes In our previous work, we identified the T3SSdependent YopM of Yersinia enterocolitica as a bacterial CPP capable of penetrating eukaryotic cells without a need for additional factors.
T79 155777-155918 Sentence denotes Moreover, we showed that cell-penetrating YopM efficiently downregulates the transcription of several pro-inflammatory cytokines (e.g. TNF ).
T80 155919-156017 Sentence denotes Therefore, YopM might be applicable as a topical therapeutic agent for the treatment of psoriasis.
T81 156018-156183 Sentence denotes To investigate whether YopM might be functional as an immune-modulator for the treatment of psoriasis, we utilized the murine imiquimod (IMQ)induced psoriasis model.
T82 156184-156343 Sentence denotes In this model, daily topical application of imiquimod (IMQ), a TLR7/8 ligand and potent immune activator, on mouse back skin induces and exacerbates psoriasis.
T83 156344-156449 Sentence denotes Therefore, this model could serve for the analysis of pathogenic mechanisms in psoriasis-like dermatitis.
T84 156450-156594 Sentence denotes Here, we applied YopM either topically, intraperitoneally, or subcutaneously (injected every 48 h) to groups of mice over a period of two weeks.
T85 156595-156799 Sentence denotes Our current results confirmed the 'self-delivering' abilities of YopM across the cutaneous barrier for topically applied YopM, and already indicated a remarkable dampening of overt inflammatory reactions.
T86 156800-156993 Sentence denotes Repression of disease symptoms was also found in the group of mice subcutaneously injected with YopM, whereas no significant phenotypic changes were observed in intraperitoneally injected mice.
T87 156994-157044 Sentence denotes Genomic islands of Legionella pneumophila Corby M.
T88 157045-157060 Sentence denotes Lautner* 1 , V.
T89 157061-157076 Sentence denotes Herrmann 1 , K.
T90 157077-157093 Sentence denotes Rydzewski 1 , K.
T91 157094-157251 Sentence denotes Heuner P26, Berlin, Deutschland Exchange of genetic information by horizontal gene transfer is an important mechanism for the evolution of bacterial genomes.
T92 157252-157350 Sentence denotes Horizontal gene transfer was also observed in Legionella pneumophila and other Legionella species.
T93 157351-157521 Sentence denotes In the genome of Legionella pneumophila Corby we identified and characterized a new trb/tra conjugation type IVA secretion system, localized on the genomic islands Trb-1.
T94 157522-157662 Sentence denotes The genomic island is integrated within the tRNA Pro gene (lpc2778) and can exist in an integrated chromosomal or an episomal circular form.
T95 157663-157851 Sentence denotes Additional to all essential trb/tra genes require for conjugation and an oriT region, a site-specific integrase (int-1; lpc2818) and a Legionella vir region (lvr) also had been identified.
T96 157852-157976 Sentence denotes By deleting the int-1 gene, we could demonstrate that the excision and forming of an episomal circle is integrase dependent.
T97 157977-158149 Sentence denotes First experiments with a mutant lacking the vir region, also indicate a growth dependent regulation of the alteration between the integrated and the circular form of Trb-1.
T98 158150-158294 Sentence denotes The whole genomic island can be transferred to other Legionella strains and integrated site-specifically into the genome of the transconjugants.
T99 158295-158432 Sentence denotes Using the int-1 mutant as a donor in conjugation experiments, we obtain reduced conjugation frequencies, compared to the parental strain.
T100 158433-158566 Sentence denotes Further analysis of the L. pneumohila Corby genome led to the identification of two additional genomic islands (Lpc-GI3 and Lpc-GI4).
T101 158567-158646 Sentence denotes GI3 and GI4 are flanked by a tRNA gene and exhibit several putative integrases.
T102 158647-158738 Sentence denotes Both islands exhibit an lvr region, the genes traG, traD, virB4, pilT and a hel gene locus.
T103 158739-158867 Sentence denotes In addition, GI4 contains genes encoding an ATP synthase and various efflux systems for heavy metals and other toxic substances.
T104 158868-158978 Sentence denotes Similar genomic islands are also present in the genomes of L. pneumophila Philadelphia, Paris, Lens and Alcoy.
T105 158979-159114 Sentence denotes We could demonstrate that both genomic islands can exist in an integrated chromosomal and an exercised episomal form, similar to Trb-1.
T106 159115-159202 Sentence denotes Production of L-asparaginase by a protease-deficient Bacillus megaterium strainITBHU01:
T107 159203-159545 Sentence denotes Characterization and production optimization FTP10 Unravelling glycosphingolipid receptors of enterohemorrhagic Escherichia coli by means of solid phase bacterial adhesion assay combined with IR-MALDI mass spectrometry Glycosphingolipids (GSLs) play pivotal roles in the manifestation of infectious diseases as attachment sites for pathogens.
T108 159546-159628 Sentence denotes Many germs are known to exploit GSLs as targets on the surface of mammalian cells.
T109 159629-159753 Sentence denotes This initial protein (pathogen) carbohydrate (host) interaction contributes to colonization and virulence of many pathogens.
T110 159754-159861 Sentence denotes Enterohemorrhagic E. coli (EHEC) causes diarrhea, bloody diarrhea, and hemolytic-uremic syndrome in humans.
T111 159862-160065 Sentence denotes Although a substantial amount of information about the GSLreceptors for Shiga toxins (Stxs), the major virulence factors of EHEC, is available, less is known about the receptors for EHEC bacterial cells.
T112 160066-160355 Sentence denotes The thin-layer chromatography (TLC) overlay assay coupled to infrared matrixassisted laser desorption/ionization orthogonal time-of-flight mass spectrometry (IR-MALDI-o-TOF-MS) represents a powerful tool for the detection and structural characterization of GSL receptors of microorganisms.
T113 160356-160729 Sentence denotes Here we report on the employment of this strategy aimed at the identification and structural characterization of GSL-receptors of the most common EHEC, E. coli O157, which consists of two types: nonsorbitol-fermenting strains of serotype O157:H7, which are spread worldwide, and Sfp-fimbriated sorbitol-fermenting nonmotile strains (O157:NM), which are pathogens in Europe.
T114 160730-160929 Sentence denotes In the present study the potential of GSL recognition by various EHEC O157 strains was investigated employing neutral GSLs and sialylated GSLs (=gangliosides) from different types of mammalian cells.
T115 160930-161087 Sentence denotes We were able to identify certain neutral GSLs carrying oligosaccharides with -configurated terminal galactose residues showing binding activity towards EHEC.
T116 161088-161235 Sentence denotes This type of interaction contrasts the common binding structures of Stxs that specifically recognize neutral GSLs with -linked galactose molecules.
T117 161236-161371 Sentence denotes EHEC-binding GSLs were further characterized by IR-MALDI-o-TOF-MS in parallel to TLC overlay assays using specific anti-GSL antibodies.
T118 161372-161548 Sentence denotes The ganglioside GM3 (=II 3 Neu5Ac-Lc2Cer) and terminally sialylated gangliosides with neolacto-series core were negative indicating that sialylation abrogates EHEC interaction.
T119 161549-161773 Sentence denotes In conclusion, this study may help to better understand host-pathogen interactions and in particular the adhesion process of EHEC toward target cells and, moreover, to develop strategies to prevent and treat EHEC infections.
T120 161774-161969 Sentence denotes This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG)-funded International Graduate School (GRK 1409, collaboration between the projects 3.6. of H.K. and 3.10 of J.M.).
T121 161970-162311 Sentence denotes Glycosphingolipid Shiga toxin receptors in lymphoid and myeloid cell lines: subcellular receptor distribution and expression profiles of glycosyltransferases related to their biosynthesis pathway Glycosphingolipids (GSLs) are amphipathic membrane constituents comprising a hydrophilic oligosaccharide chain and a hydrophobic ceramide moiety.
T122 162312-162474 Sentence denotes The ceramide lipid anchor of GSLs is embedded in the outer leaflet of the plasma membrane and the carbohydrate chain protrudes into the extracellular environment.
T123 162475-162625 Sentence denotes GSLs are preferentially organized in microdomains known as lipid rafts, which play crucial functional roles in GSL-mediated cell-surface interactions.
T124 162626-162799 Sentence denotes GSLs organized in lipid rafts are capable for efficient interaction with pathogens such as viruses and bacteria or pathogen-released virulence factors like bacterial toxins.
T125 162800-163002 Sentence denotes Microdomain association of GSLs is believed to be the linchpin in effective binding, internalization and subcellular routing of GSL-binding AB5-toxins such as Shiga toxin (Stx) to intracellular targets.
T126 163003-163188 Sentence denotes The pentameric B-subunit of Stxs specifically binds with high and low affinity to the neutral GSLs globotriaosylceramide (Gb3Cer/CD77) and globotetraosylceramide (Gb4Cer), respectively.
T127 163189-163347 Sentence denotes Our investigations are focused on the interaction analysis of Stxs with these GSL receptors that are expressed on the surface of leukocyte derived cell lines.
T128 163348-163795 Sentence denotes The structural characterization of the globo-series neutral GSLs by means of electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF-MS) with collision-induced dissociation (CID) in conjunction with Stx as well as anti-Gb3Cer and anti-Gb4Cer antibody binding assays revealed Gb3Cer (d18:1, C24:1/C24:0) and Gb3Cer (d18:1, C16:0) as the prevalent Stx-receptors in cell lines of lymphoid (B cell) and myeloid (monocyte) origin.
T129 163796-163944 Sentence denotes Studies on the subcellular distribution of Stx receptors in both cell types showed their predominant association with and enrichment in lipid rafts.
T130 163945-164061 Sentence denotes Here, we also provide data on the high sensitivity of Stxs to GSL receptor positive B-cell and monocyte descendants.
T131 164062-164219 Sentence denotes As expected from the GSL profiles granulocyte and T-cell derived cell lines lacking Stx-receptors were found to be resistant to the cytotoxic action of Stxs.
T132 164220-164389 Sentence denotes Moreover, studies on the expression of the glycosyltransferases responsible for biosynthesis of GSL receptors showed high compliance with immunostaining of GSL isolates.
T133 164390-164531 Sentence denotes Our findings provide the basis for further exploring the functional role of lipid raft-associated Stx-receptors in cells of leukocyte origin.
T134 164532-164726 Sentence denotes This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG)-funded International Graduate School (GRK 1409, collaboration between the projects 3.10 of J.M. and 3.6 of H.K.).
T135 164727-164789 Sentence denotes Biological activity of Bacillus extracts against Legionella F.
T136 164790-164803 Sentence denotes Frese* 1 , J.
T137 164804-164820 Sentence denotes Tiefenau* 1 , M.
T138 164821-165054 Sentence denotes Rohde 2 1 TU Braunschweig, Institut für Mikrobiologie, Braunschweig, Deutschland 2 Helmholtz Zentrum für Infektionsforschung, Braunschweig, Deutschland Legionella pneumophila is an intracellular parasite of aquatic and soil protozoa.
T139 165055-165265 Sentence denotes Due to man-made water systems like cooling towers or air conditioning systems, L. pneumophila is also able to accidentally infect humans causing a severe, sometimes fatal pneumonia termed Legionnaires´ disease.
T140 165266-165490 Sentence denotes The bacterial-protozoa interaction and sheltering biofilms lead to a remarkable resistance to biocide compounds and chlorination, thus representing ecological niches for legionellae persistence in such man-made environments.
T141 165491-165632 Sentence denotes From air samples we isolated two different Bacillus species that showed strong growth-inhibitory activity against legionellae on agar plates.
T142 165633-165790 Sentence denotes The observed inhibitory activity was Legionella-specific since the growth of a brought spectrum of Gram-positive and Gram-negative bacteria was not affected.
T143 165791-165901 Sentence denotes By 16S rDNA sequencing the two bacilli strains were identified as Bacillus pumilus and Bacillus licheniformis.
T144 165902-166013 Sentence denotes The inhibitory compounds were purified from Bacillus cultures by solvent extraction or binding to XAD-16 resin.
T145 166014-166204 Sentence denotes The compounds revealed resistance to high temperatures (121 °C for 20 min) and remained active in agar growth-inhibition assays after acid (pH 0) and alkali (pH 14) treatments, respectively.
T146 166205-166275 Sentence denotes On human blood agar plates the compounds exhibited hemolytic activity.
T147 166276-166370 Sentence denotes Minimal inhibitory concentrations (MIC) were determined in liquid assay with 10 7 bacteria/ml.
T148 166371-166576 Sentence denotes The crude extracts showed specific bactericidal activity against L. pneumophila (MIC 6.25 μg/ml), whereas concentrations of 800 μg/ml were not able to inhibit other Gram-negative or Gram-positive bacteria.
T149 166577-166694 Sentence denotes The cytotoxicity of the compounds against the human lung epithelial cell line NCI-H292 was analyzed by the MTT assay.
T150 166695-166850 Sentence denotes Compared to the high bactericidal activity, the crude extracts showed only low cytotoxic effects on the eukaryotic cells at concentrations above 200 μg/ml.
T151 166851-167085 Sentence denotes Phenotypic characterization by transmission electron microcopy (TEM) after coincubation of L. pneumophila with B. licheniformis or B. pumilus extracts for 48 h revealed a dose-dependent effect on the L. pneumophila membrane integrity.
T152 167086-167150 Sentence denotes The bacterial cell exhibited increased blebbing and plasmolysis.
T153 167151-167344 Sentence denotes The narrow spectrum of activity and the results of the TEM revealed a specific interaction of the bacilli extracts against the highly hydrophobic and structurally unique membrane of Legionella.
T154 167345-167467 Sentence denotes These promising initial data suggest further characterization of the compounds and an evaluation of putative applications.
T155 167468-167563 Sentence denotes Functional analysis of the translation elongation factor EF-Tu from Chlamydophila pneumoniae S.
T156 167564-167584 Sentence denotes De Benedetti* 1 , A.
T157 167585-167603 Sentence denotes Gaballah 1 , H.-G.
T158 167604-167615 Sentence denotes Sahl 1 , B.
T159 167616-167817 Sentence denotes Henrichfreise 1 1 University of Bonn, Pharmaceutical Microbiology, Bonn, Deutschland The bacterial translation elongation factor EF-Tu is well known to be involveld in prokaryotic protein biosynthesis.
T160 167818-168112 Sentence denotes EF-Tu from Escherichia coli has been shown to polymerize in vitro and a recent study provided evidence that the protein serves besides its function in translation another vital role in Bacillus subtilis: it affects cell morphology, apparently via interaction with the cytoskeleton protein MreB.
T161 168113-168236 Sentence denotes In rod-shaped bacteria the actin-ortholog MreB is thought to direct incorporation of cell wall material into the side wall.
T162 168237-168395 Sentence denotes Surprisingly, chlamydiae harbor, despite their spherical shape and the absence of a cell wall, MreB and we recently proved in vitro activity for this protein.
T163 168396-168455 Sentence denotes Here, we show that chlamydial EF-Tu is functional in vitro.
T164 168456-168606 Sentence denotes The purified, strep-tagged protein polymerized in a concentration, pH and ion strength dependent fashion in light scattering and sedimentation assays.
T165 168607-168732 Sentence denotes Additionally, co-polymerization assays revealed interactions between the elongation factor and the cytoskeletal protein MreB.
T166 168733-168921 Sentence denotes A deeper insight into the functions of EF-Tu and its role in chlamydial cell biology on molecular level will provide valuable information for the design of new anti-chlamydial antibiotics.
T167 168922-169079 Sentence denotes Ecological fitness investigations of antibiotic resistant mutants of Legionella pneumophila strain MTZ OLDA under natural conditions in artificial microcosm.
T168 169080-169082 Sentence denotes T.
T169 169083-169100 Sentence denotes Koshkolda* 1 , C.
T170 169101-169234 Sentence denotes Lück 1 1 TUD, Medical Microbiology and Hygiene, Dresden, Deutschland Legionella pneumophila is a gram-negative, rod-shaped bacterium.
T171 169235-169337 Sentence denotes This ubiquitous bacterium can cause two forms of legionellosis -Legionellapneumonia and Pontiac fiver.
T172 169338-169445 Sentence denotes Newer macrolides and fluoroquinolones are currently the drugs of choice for the treatment of legionellosis.
T173 169446-169649 Sentence denotes Although the failure in the therapy occurs from time to time, there are no reports about the isolation of the macrolide or quinolon-resistant Legionella mutants from the patients and environment to date.
T174 169650-169772 Sentence denotes A number of macrolide resistant mutants of L. pneumophila strain MTZ OLDA were selected in vitro using two-step selection.
T175 169773-170001 Sentence denotes To investigate the interaction of Legionella antibiotic resistant mutants and their protozoan host under the influence of environmental flora, we used a previously established natural microcosm with simulated natural conditions.
T176 170002-170106 Sentence denotes This microcosm was established based on the microbial community of the water supply of a large building.
T177 170107-170191 Sentence denotes The diversity of this community was determinated using the 16S rRNA clone libraries.
T178 170192-170293 Sentence denotes It includes multiple heterotrophic species such as Sphingomonas sp., Chromobacter sp., Flavimonas sp.
T179 170294-170410 Sentence denotes In addition the microcosm contains L. pneumophila strain MTZ OLDA wilde type (SG1) and amoeba strain Hartmanella sp.
T180 170411-170554 Sentence denotes Results showed that the resistant mutants cannot survive within the microcosm and that they disappeared after approx. ten weeks of cultivation.
T181 170555-170614 Sentence denotes Furthermore they are less fit in co-cultivation experiment.
T182 170615-170775 Sentence denotes The reduced ecological fitness of the macrolide resistant mutants might be an explanation for the lack occurrence of resistant strains under natural conditions.
T183 170776-170955 Sentence denotes For more than a hundred years, it has been known that patients with solid tumors who concomitantly suffer from a bacterial infection experience a regression of their tumor burden.
T184 170956-171048 Sentence denotes Still, the phenomenon of bacterial tumor targeting and lysis remains only poorly understood.
T185 171049-171233 Sentence denotes The vision behind the PROTumor research project is not only to explore the mechanisms of bacterial oncolysis, but also, among other things, to find ways to further augment this effect.
T186 171234-171360 Sentence denotes Amid several theoretically interesting approaches, we focused our efforts on immunosuppressive and pro-inflammatory cytokines.
T187 171361-171547 Sentence denotes The immunosuppressive cytokines are supposed to expand the inoculum in the tumor and should enable the bacteria to penetrate the host immune defense shielding the tumor from total lysis.
T188 171548-171712 Sentence denotes On the other hand, the proinflammatory cytokines should attract inflammatory cells, boost NK cell and CD8+ T-cell activity and eventually lead to tumor vaccination.
T189 171713-171845 Sentence denotes Here, we are going to present first results with recombinant Interleukin-10 and Interleukin-12 fusion proteins expressed in E. coli.
T190 171846-172038 Sentence denotes In both cytokines, the corresponding monomeric proteins were linked with a glycine-serinelinker in order to achieve an improved stability and an optimized yield of biologically active protein.
T191 172039-172128 Sentence denotes The genes used were synthesized as codonoptimized DNA sequences by a commercial provider.
T192 172129-172251 Sentence denotes The recombinant proteins are secreted into the periplasmatic fraction with help of the OmpF signaling sequence of E. coli.
T193 172252-172367 Sentence denotes The Interleukin-10 fusion protein was expressed in E. coli BL21 (DE3) and Origami B (DE3) under varying conditions.
T194 172368-172582 Sentence denotes The quantity of expressed Interleukin-10 was determined with a quantitative ELISA technique and its biologic activity was assessed in a cell-based assay measuring phosphorylation of the STAT-3 transcription factor.
T195 172583-172588 Sentence denotes A.-S.
T196 172589-172603 Sentence denotes Gunkel* 1 , S.
T197 172604-172617 Sentence denotes Sachse 1 , W.
T198 172618-172632 Sentence denotes Pfister 1 , E.
T199 172633-172647 Sentence denotes Straube 1 , A.
T200 172648-172988 Sentence denotes Sobke Institute of Medical Microbiology, Jena, Deutschland While the preceeding decade was marked by the efficient spread of enterobacteriaceae carrying the ESBL phenotype in Germany and other European countries, we are now facing an alarming distribution of carbapenemase harbouring isolates (referred to as superbugs in the yellow press).
T201 172989-173110 Sentence denotes These isolates are resistant to all ß-lactams except monolactams andlike ESBL-typically carry further resistance markers.
T202 173111-173270 Sentence denotes On basis of structural differences, carbapenemases are divided into several classes, with superbug enzymes like NDM-1 belonging to the class B metallo-enzymes.
T203 173271-173421 Sentence denotes These require zinc in their catalytic centre and inhibition of the lactamase activity by the cation chelator EDTA is used during screening procedures.
T204 173422-173545 Sentence denotes However, while EDTA has found widespread application as a conservative, its clinical use is hampered by toxic side effects.
T205 173546-173675 Sentence denotes Looking for clinically administrable alternatives, we studied the cation chelator and broadspectrum antibiotic nitroxoline (NIT).
T206 173676-173808 Sentence denotes Mono-disk assays were carried out with carbapenemase expressing strains, applying either 2 μM EDTA or NIT with 10 μg imipenem disks.
T207 173809-174072 Sentence denotes Interaction of NIT with imipenem was further evaluated by checkerboard assays and hydrolytic activity in cell lysates or whole cell suspensions was followed spectrophotometrically at 298 nm in the presence or absence of NIT (150 μM imipenem in PBS, 37°C, 1 hour).
T208 174073-174131 Sentence denotes NIT had comparable effects to EDTA in the mono-disk assay.
T209 174132-174340 Sentence denotes In checkerboard assays synergism was demonstrated for NIT and imipenem in strains harbouring class B enzymes, but additive or indifferent effects were observed with strains expressing class A or D lactamases.
T210 174341-174554 Sentence denotes Hydrolysis of imipenem by VIM and IMP enzymes was inhibited by NIT in a concentration-dependent manner (complete inhibition reached at 100-150 μM NIT), whereas no effect on KPC enzyme activity was noted (class A).
T211 174555-174775 Sentence denotes The inhibitory effect depended on the respective carbapenemase type, but was not influenced by the bacterial species carrying the enzyme or the integrity of the bacteria (no difference between lysate or cell suspension).
T212 174776-174910 Sentence denotes The inhibition was completely resolved by addition of 100 μM Zn 2+ in the incubation mixture, whereas Ni 2+ had only a minimal effect.
T213 174911-174994 Sentence denotes In conclusion, we found NIT to be an effective inhibitor of class B carbapenemases.
T214 174995-175200 Sentence denotes As this antibiotic has been in clinical use over the last 50 years without serious side effects having been reported, it constitutes a promising candidate for the development of new therapeutic strategies.
T215 175201-175284 Sentence denotes Factors influencing the results of microbial surface water testing in South Africa.
T216 175285-175303 Sentence denotes Problem Statement:
T217 175304-175411 Sentence denotes Faecal coliforms, the hydrogen-sulphide test and E. coli are used in monitoring of microbial water quality.
T218 175412-175502 Sentence denotes Rhodococcus coprophilus indicates faecal contamination from animal sources such as cattle.
T219 175503-175610 Sentence denotes Their mutual agreement and factors influencing the results have not been studied in detail in South Africa.
T220 175611-175734 Sentence denotes Thus a study was conducted at 15 sampling sites in the Eastern Cape Province of South Africa to address this knowledge gap.
T221 175735-175757 Sentence denotes Materials and Methods:
T222 175758-175827 Sentence denotes Sampling sites were located in the Makana and Buffalo Municipalities.
T223 175828-176011 Sentence denotes Concentrations of faecal coliforms and E. coli were measured either using the MPN procedure with the A1 medium, the membrane filtration technique on the m-FC agar and the indole test.
T224 176012-176058 Sentence denotes Cultivations were always conducted at 44.5 ºC.
T225 176059-176189 Sentence denotes Concentrations of Rhodococcus coprophilus was measured by spread-plating using the M3 medium and cultivation at 30 ºC for 21 days.
T226 176190-176256 Sentence denotes The hydrogen-sulphide test medium contained 0.5 % of deoxycholate.
T227 176257-176342 Sentence denotes Grab samples of the water column and sediment were transported to laboratory at 4 ºC.
T228 176343-176433 Sentence denotes Sediment samples were extracted by vortexing and the one-quarter strength Ringer solution.
T229 176434-176581 Sentence denotes Model waters from selected sampling sites were prepared and the survival of faecal coliforms and the hydrogen-sulphide test bacteria were measured.
T230 176582-176590 Sentence denotes Results:
T231 176591-176693 Sentence denotes Concentrations of faecal coliforms and E. coli ranged from 2 to above 1.6 x 10 4 cells or CFUs/100 mL.
T232 176694-176781 Sentence denotes Suspension of accumulated sediment led to concentration changes ranging from 0 to 40 %.
T233 176782-176939 Sentence denotes Positive results in the hydrogen-sulphide test were observed after 14 to 60 hours; and these results were susceptible to influence by sediment re-suspension.
T234 176940-177086 Sentence denotes Temperatures varied between 10 and 32 ºC during the sample collection and transport, but did not change the results of the hydrogen-sulphide test.
T235 177087-177163 Sentence denotes Levels of Rhodococcus coprophilus ranged from 0 to 8.18 x 10 6 cells/100 mL.
T236 177164-177302 Sentence denotes If a suspension of the sediment matter on-site took place, this would lead to a change in the bacterial concentrations between 0 and 24 %.
T237 177303-177506 Sentence denotes Survival studies showed that faecal coliforms and the hydrogen-sulphide producing bacteria survived for 22 to 29 days after faecal contamination depending on the chemical composition of the water column.
T238 177507-177519 Sentence denotes Conclusions:
T239 177520-177618 Sentence denotes Re-suspension of sediment particles can compromise the results of microbial water quality testing.
T240 177619-177797 Sentence denotes Temperature fluctuations between 10 and 32 ºC are not expected to influence the applicability of the hydrogen-sulphide test in detection of faecal contamination of surface water.
T241 177798-177931 Sentence denotes Adaption of multiplex PCR analytics for the detection of microorganism contaminants in the industrial production of emulsion paint R.
T242 177932-177949 Sentence denotes Söller* 1 , B.-J.
T243 177950-177965 Sentence denotes Sporkert 2 , M.
T244 177966-178030 Sentence denotes Franke 3 1 QIAGEN Hamburg GmbH, R&D, Hamburg, Deutschland 2 J.W.
T245 178031-178135 Sentence denotes Ostendorf GmbH & Co.KG, Produktion, Coesfeld, Deutschland 3 BIBA GmbH, Bremen, Deutschland Introduction:
T246 178136-178371 Sentence denotes Classic microbiological colony counting on agar plates is the diagnostic gold standard for the detection of microbial contaminants (MC) in raw material, intermediate goods and final products in the industrial emulsion paint production.
T247 178372-178535 Sentence denotes As time-to-result takes up to 3 days, corrective actions like separation and cleaning of contaminated goods or addition of preservatives are usually retrospective.
T248 178536-178766 Sentence denotes A sustainable in-time management of emulsion paint production should rely on fast quantitative MC analysis of products and their effective separation, thereby minimizing preservative input and waste of contaminated emulsion paint.
T249 178767-178992 Sentence denotes Aim of this study was an overall process optimization by introduction of a faster and more sensitive analysis method for tracing microbial contaminations within the production line of a leading German emulsion paint producer.
T250 178993-179128 Sentence denotes A proposed fast and sensitive multiplex PCR assay for the detection of MC was developed and evaluated against the gold standard method.
T251 179129-179230 Sentence denotes Methods: QRTPCR is well established in the medical routine diagnostics of human microbial infections.
T252 179231-179454 Sentence denotes This study focuses on method adaptations for effective DNA preparation from emulsion paint samples and the development of a multiplex qRT PCR assay for the detection of relevant MC in emulsion paint precursors and products.
T253 179455-179641 Sentence denotes In addition a new PCR expert software for automated result analysis of qRT-PCR runs from the RotorGene instrument (RG) was developed in order to standardize the analysis of qRT PCR data.
T254 179642-179650 Sentence denotes Results:
T255 179651-179836 Sentence denotes The functionality of multiplex qRT PCR assays as a central tool of quality control to trace spiked MC in emulsion paint samples was demonstrated and evaluated against the gold standard.
T256 179837-180065 Sentence denotes While the PCR method was clearly faster and equally sensitive to classic microbiology plate counting regarding ready emulsion paint, it failed when applied to some raw materials containing pure acrylate and some other compounds.
T257 180066-180229 Sentence denotes This was clearly due to the insufficient separation of highly PCR inhibitory acrylate compounds which simply passed the DNA preparation columns used in this study.
T258 180230-180357 Sentence denotes All PCR runs were analysed twice, manually using the standard RG software, and automatically using the new PCR expert software.
T259 180358-180391 Sentence denotes No discrepant results were found.
T260 180392-180404 Sentence denotes Conclusions:
T261 180405-180548 Sentence denotes Further development effort would be needed to optimize sample preparation with special focus on removal of inhibitory emulsion paint compounds.
T262 180549-180715 Sentence denotes All methods should run on (or must easily be adaptable to) integrated sample and analysis robotics applying qRT-PCR, automated analysis and interpretation of results.
T263 180716-180922 Sentence denotes This requirement is crucial since a final qRT-PCR based QC strategy should be able to manage the emulsion paint production without interaction of human PCR experts in a model environment at JWO in Coesfeld.
T264 180923-180951 Sentence denotes Introduction and Objectives:
T265 180952-181092 Sentence denotes Pharmaceutical-water systems are known to harbor biofilms, even though these environments are oligotrophic and often contain a disinfectant.
T266 181093-181169 Sentence denotes Control of these biofilms is important for aesthetic and regulatory reasons.
T267 181170-181307 Sentence denotes In pharmaceutical water, bacteria are affected not only by the environment they live in but also by the variety of other species present.
T268 181308-181454 Sentence denotes By performing studies on the interactions present in multispecies biofilms, basic knowledge on several aspects of sociomicrobiology can be gained.
T269 181455-181577 Sentence denotes The aim of this work was to assess the role of interspecies interactions in species biofilm formation and characteristics.
T270 181578-181599 Sentence denotes Material and Methods:
T271 181600-181725 Sentence denotes The microorganisms used throughout this study were isolated from a model laboratory pharmaceutical-water distribution system.
T272 181726-181923 Sentence denotes Preliminary, presumptive bacteria identification was done using selective media Chromocult TBX agar, Pseudomonas isolation agar, and methanol minimum medium, Gram staining, and biochemical methods.
T273 181924-182182 Sentence denotes In order to assess the biofilm formation abilities and specific respiratory activities of the several bacteria isolated from pharmaceutical water, the standard 96-well microtiter plates with Cristal Violet and XTT staining were used to characterize biofilms.
T274 182183-182191 Sentence denotes Results:
T275 182192-182411 Sentence denotes A ranking of biofilm formation was produced according to the method of Stepanovic´ et al., classifying test bacteria as non-biofilm producer, weak biofilm producer, moderate biofilm producer, or strong biofilm producer.
T276 182412-182424 Sentence denotes Conclusions:
T277 182425-182514 Sentence denotes Biofilms are recognized as focal points where bacteria and other microorganisms interact.
T278 182515-182721 Sentence denotes Recent investigations into the microbial ecology of pharmaceutical water systems have found that pathogen resistance to chlorination is affected by the community biodiversity and interspecies relationships.
T279 182722-182875 Sentence denotes A range of interactions has been observed among microorganisms in biofilms, including competitive, mutualistic, antagonistic and commensal relationships.
T280 182876-182981 Sentence denotes In this work, some of the bacterial isolates tested are recognized as problematic opportunistic bacteria.
T281 182982-183065 Sentence denotes The selected bacterial species were also detected in pharmaceutical-water biofilms.
T282 183066-183095 Sentence denotes Biofilm, Pharmaceutical-water
T283 183096-183178 Sentence denotes Are Campylobacter jejuni secretory proteins suitable antigens for serodiagnostics?
T284 183179-183181 Sentence denotes J.
T285 183182-183195 Sentence denotes Corso* 1 , R.
T286 183196-183209 Sentence denotes Lugert 1 , U.
T287 183210-183223 Sentence denotes Groß 1 , A.E.
T288 183224-183399 Sentence denotes Zautner 1 1 Universitätsmedizin Göttingen, Medizinische Mikrobiologie, Göttingen, Deutschland Campylobacter spp. are the worldwide leading cause for bacterial gastroenteritis.
T289 183400-183482 Sentence denotes The main reservoir and therefore the most common cause of an infection is poultry.
T290 183483-183575 Sentence denotes A primary Campylobacteriosis manifests as diarrhea and is mainly diagnosed by stool culture.
T291 183576-183724 Sentence denotes Following an infection, there is a risk of suffering from a sequela, such as reactive arthritis, neuropathia, myositis or a Guillian-Barré-Syndrome.
T292 183725-183802 Sentence denotes These post-infectious complications can only be traced by serological assays.
T293 183803-183966 Sentence denotes The sensitivity and specificity of most of these tests suffer since they are based on crude whole cell lysates which entail cross reactivites with related species.
T294 183967-184043 Sentence denotes Hence, the use of species specific antigens becomes more and more favorable.
T295 184044-184187 Sentence denotes C. jejuni secretes a number of proteins which are amongst others required for cell invasion and therefore confer the pathogenicity of the germ.
T296 184188-184334 Sentence denotes These proteins are presumably highly antigenic due to the exposure to the host's immune system and may be used as antigens in diagnostic purposes.
T297 184335-184556 Sentence denotes In this study could be demonstrated that recombinantely produced Cj0069, one of the secreted proteins, has a similar sensitivity and specificity compared to the already and commercially established antigens OMP18 and P39.
T298 184557-184568 Sentence denotes Background:
T299 184569-184641 Sentence denotes Horizontal gene transfer plays an important role in bacterial evolution.
T300 184642-184740 Sentence denotes DNA acquired by horizontal gene transfer has to be incorporated into existing regulatory networks.
T301 184741-184868 Sentence denotes The histone-like nucleoid structuring protein H-NS acts as a silencer of horizontally acquired genes to avoid potential damage.
T302 184869-185102 Sentence denotes Here we analyzed the influence of H-NS on the transcription of the horizontally acquired pyp regulatory network controlling transcription of the hreP gene encoding a proteinase essential for full virulence of Yersinia enterocolitica.
T303 185103-185111 Sentence denotes Methods:
T304 185112-185287 Sentence denotes Using -galactosidase and biofilm assays the effect of a dominant-negative H-NS variant (H-NS ) on gene expression of hreP and its regulators pypA, pypB and pypC were analyzed.
T305 185288-185436 Sentence denotes Utilizing electrophoretic mobility shift assays (EMSA) the interaction of H-NS with the promoter regions of hreP and the pyp genes was investigated.
T306 185437-185514 Sentence denotes Results: H-NS represses the transcription of pypA, pypB, and pypC in E. coli.
T307 185515-185628 Sentence denotes Moreover, a dominant-negative version of H-NS relieves repression of hreP and the pyp genes in Y. enterocolitica.
T308 185629-185739 Sentence denotes RovA, an antagonist of H-NS in Yersinia, only activates transcription of pypC, but not of pypA, pypB, or hreP.
T309 185740-185839 Sentence denotes With EMSA we show that H-NS interacts directly with the promoter regions of hreP and the pyp genes.
T310 185840-185981 Sentence denotes In addition, the analysis of biofilm formation clearly demonstrates that overproduction of H-NS strongly increases the formation of biofilms.
T311 185982-185993 Sentence denotes Conclusion:
T312 185994-186193 Sentence denotes This study reveals that hreP and the pyp genes are part of a complex regulatory network that is repressed by H-NS, further underlining our hypothesis that these genes have been acquired horizontally.
T313 186194-186420 Sentence denotes These results also underscore that dominant-negative versions of H-NS as the H-NS protein are an important and valuable tool to study H-NS-dependent phenotypes in strains where hns deletions are lethal such as in Yersinia spp.
T314 186421-186496 Sentence denotes Host-specific flagellin recognition in Salmonella entericainfected cells S.
T315 186497-186515 Sentence denotes Maurischat* 1 , P.
T316 186516-186532 Sentence denotes Schwerk 1 , L.H.
T317 186533-186546 Sentence denotes Wieler 1 , K.
T318 186547-186579 Sentence denotes Tedin 1 1 FU Berlin, Institut f.
T319 186580-186892 Sentence denotes Mikrobiologie & Tierseuchen, Fachbereich Veterinärmedizin, Berlin, Deutschland Bacterial flagellin is one of the major pathogen-associated molecular patterns and plays a pivotal role in recognition of Salmonella by Toll-like receptors such as TLR5 and intracellularly by NOD-like receptors Ipaf and Birc1e/NAIP5.
T320 186893-186999 Sentence denotes Both pathways trigger inflammatory processes and though the host immune response to prevent the infection.
T321 187000-187134 Sentence denotes Salmonella is an important zoonotic pathogen, able to infect and cause disease in humans and animals ranging from reptiles to mammals.
T322 187135-187333 Sentence denotes Depending on the serovar, host species, age and immune status the course of disease can range from asymptomatic carriage or localized acute diarrhea to severe systemic infections with fatal outcome.
T323 187334-187673 Sentence denotes To evaluate possible host differences in flagellin recognition in human and animal cells and subsequent induction of immunostimulatory signalling cascades, we have used the activation of transcription factor nuclear factor -B (NF-B) in NF-B reporter cell lines of different species origins that express a luciferase during NF-B activation.
T324 187674-187921 Sentence denotes Mutants of Salmonella enterica with different flagellar gene deletions (fliC, fljB, fliD, flhDC) or expression patterns were used to infect NF-B reporter cell lines of human, porcine and avian origins (THP-1, LoVo, Caco-2, 3D4/31, IPEC-J2, HD-11).
T325 187922-188053 Sentence denotes The loss of flagellar-associated genes affected the invasiveness of Salmonella differently between cells of different host species.
T326 188054-188187 Sentence denotes In addition, loss of flagella affected NF-B activation in human macrophages and epithelial cells but not in porcine or chicken cells.
T327 188188-188370 Sentence denotes The varying behaviour and sensing of Salmonella flagellin in different host species might contribute to the different outcome of infection and host adaptation of Salmonella serovars.
T328 188371-188537 Sentence denotes Shiga toxins (Stxs) are secreted by Stx-producing Escherichia coli (STEC) that cause diarrhea, bloody diarrhea and potentially lethal hemolytic-uremic syndrome (HUS).
T329 188538-188616 Sentence denotes Stxs are AB5 toxins composed of a single A-subunit and 5 identical B-subunits.
T330 188617-188718 Sentence denotes The catalytic A-subunit has rRNA N-glycosidase activity and inhibits eukaryotic protein biosynthesis.
T331 188719-188929 Sentence denotes The pentameric Bsubunit is responsible for binding to the high-and less-effective cell surface glycosphingolipid (GSL) receptors globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer), respectively.
T332 188930-189211 Sentence denotes Although preferential Stxmediated damage of glomerular microvascular endothelial cells (GMVECs) is known as the major mechanism underlying HUS caused by STEC, the exact nature of the Stx receptor GSLs of GMVECs including the expression of different lipoforms remains to be defined.
T333 189212-189457 Sentence denotes Here, we report for the first time the structures of Stx receptors of GMVECs employing a combination of thin-layer chromatography (TLC) immunostaining using anti-Gb3Cer and anti-Gb4Cer antibodies and biophysical mass spectrometry-based analysis.
T334 189458-189631 Sentence denotes Full structural characterization of Stx receptors was achieved by electrospray ionization (ESI) mass spectrometry combined with collisioninduced dissociation (CID) analysis.
T335 189632-189784 Sentence denotes Gb3Cer and Gb4Cer lipoforms with ceramide moieties mainly composed of C24:0/C24:1 or C16:0 fatty acid and sphingosine (d18:1) were determined in GMVECs.
T336 189785-189902 Sentence denotes These lipoforms were comparable to those previously detected in human brain microvascular endothelial cells (HBMECs).
T337 189903-190081 Sentence denotes Thin-layer chromatography (TLC) immunostaining of isolated GSLs demonstrated a 2.0-fold higher content of Gb3Cer and a 1.4-fold higher content of Gb4Cer in GMVECs than in HBMECs.
T338 190082-190190 Sentence denotes Comparative cytotoxicity assays revealed high cytotoxicity of Stx towards GMVECs in a dose-dependent manner.
T339 190191-190322 Sentence denotes Importantly, Stx caused a significantly higher toxic effect on GMVECs than on HBMECs over the whole span of applied concentrations.
T340 190323-190424 Sentence denotes This higher sensitivity of GMVECs correlated with the enhanced expression of Stx-receptors in GMVECs.
T341 190425-190863 Sentence denotes However, as GSLs are believed to localize abundantly in lipid rafts tightly associated with cholesterol and raftassociated membrane proteins and since lipid raft association of Stx receptors is assumed as a functional prerequisite for Stx-mediated cytotoxicity, future investigations on the subcellular distribution of GSLs in GMVECs are required to explore the functional role of the molecular assembly of GSLs in microdomains of GMVECs.
T1 190864-190966 Sentence denotes Influence of the Cag type IV secretion system of Helicobacter pylori on the migration of phagocytes C.
T2 190967-190980 Sentence denotes Weiss* 1 , W.
T3 190981-191195 Sentence denotes Fischer 1 1 Max von Pettenkofer Institut, Helicobacter pylori, München, Deutschland The Cag type IV secretion system (T4SS) is known to be a major virulence factor of the Gram-negative bacteria Helicobacter pylori.
T4 191196-191355 Sentence denotes Together with the CagA effector protein, it is encoded on the cag pathogenicity island (cag-PAI), the presence of which is associated with more severe disease.
T5 191356-191444 Sentence denotes The T4SS is able to translocate CagA directly into host cells, such as epithelial cells.
T6 191445-191585 Sentence denotes As a consequence, the infected host cell is subject to several changes, such as actin cytoskeleton rearrangements and loss of cell polarity.
T7 191586-191696 Sentence denotes Moreover, epithelial cells are stimulated by the T4SS to produce and secrete chemokines such as interleukin-8.
T8 191697-191818 Sentence denotes H. pylori induces a massive inflammatory response in the host, but cannot be effectively eliminated by the immune system.
T9 191819-191915 Sentence denotes Therefore, it is likely that H. pylori is able to modulate the immune response to its advantage.
T10 191916-192000 Sentence denotes In this study, we have examined the interaction between phagocytes and the Cag T4SS.
T11 192001-192149 Sentence denotes It has been shown before that macrophages aggregate under the influence of the Cag T4SS, and that CagA can be translocated into these cells as well.
T12 192150-192338 Sentence denotes We infected phagocytic cell lines (THP-1, HL-60) and freshly isolated human phagocytes (granulocytes, dendritic cells) with H. pylori and examined their migration in chemotactic gradients.
T13 192339-192497 Sentence denotes To do so, we used two methods to examine cell migration, time-lapse microscopy of threedimensional collagen gels and migration in transwell (Boyden) chambers.
T14 192498-192623 Sentence denotes We confirmed that CagA is translocated into the cells examined and subsequently proteolytically processed and phosphorylated.
T15 192624-192774 Sentence denotes The migration assays suggest that H. pylori infection results in a chemotactic defect of phagocytic cells and in an inhibition of phagocyte migration.
T16 192775-192912 Sentence denotes Infection with H. pylori mutants showed that the inhibitory effect on human granulocyte migration is likely to be caused by the Cag T4SS.
T17 192913-193065 Sentence denotes In conclusion, our results suggest that interaction of the Cag T4SS with phagocytic cells may contribute to immune evasion and persistence of H. pylori.
T18 193066-193076 Sentence denotes Objective:
T19 193077-193184 Sentence denotes Clostridium difficile is a pathogen with emerging evidence for nosocomial and community acquired diarrhoea.
T20 193185-193387 Sentence denotes New 027 outbreak strains with global spreading are characterised by toxin A, B and binary toxin expression, mutations of a potential toxin repressor gene (tcdC), and increased toxin production in vitro.
T21 193388-193467 Sentence denotes Recently, high 027 rates have been detected in the region of Saarland, Germany.
T22 193468-193591 Sentence denotes Single locus sequence typing of surface layer protein A (slpAST) is established in our reference laboratory for genotyping.
T23 193592-193600 Sentence denotes Methods:
T24 193601-193691 Sentence denotes The University of Saarland Hospital is a 1300-bed tertiary care facility with150 ICU-beds.
T25 193692-193908 Sentence denotes Between April 2008 and December 2010 C. difficile isolates (n = 792) of 524 consecutive adult and 36 pediatric patients with diarrhoea were propagated by standard anaerobic stool cultures using CCFA media (CLO-Agar).
T26 193909-194080 Sentence denotes Genotypes were detected by slpAST as previously described (Kato et al., JMM 2005) following sequence alignment with BLAST search and with a growing institutional database.
T27 194081-194222 Sentence denotes In the present single centre retrospective study the appearance of different C. difficile genotypes was analyzed in the different age-groups.
T28 194223-194231 Sentence denotes Results:
T29 194232-194339 Sentence denotes Since July 2009 the frequency of 027 significantly decreased from up to 50% to about 6% at the end of 2010.
T30 194340-194432 Sentence denotes The proportion of other highly prevalent genotypes as e.g. 001, 014 and 017 remained stable.
T31 194433-194608 Sentence denotes Interestingly, the rate of various C. difficile genotypes was different between children and adults with high percentage of non-toxigenic strains in the pediatric group (50%).
T32 194609-194844 Sentence denotes Using an institutional database the genetic relationship between the various slpAST was analysed showing clustering in at least three major genetic groups subdivided in different subgroups with close association to known PCR ribotypes.
T33 194845-194871 Sentence denotes Discussion and conclusion:
T34 194872-194928 Sentence denotes 027 is a pandemic strain with fast geographic spreading.
T35 194929-195016 Sentence denotes We could show that 027 rates recently decrease in our previously high-prevalent region.
T36 195017-195201 Sentence denotes We hypothesize that the rates of epidemic strains and presumably also strain specific virulence may change over time e.g. due to host adaptation and possibly also in different regions.
T37 195202-195277 Sentence denotes Emergence of new epidemic C. difficile strains may occur at any time again.
T38 195278-195428 Sentence denotes Therefore, detection of changing C. difficile epidemiology requires regular genotyping of local or more global CDI without restriction to 027 strains.
T39 195429-195569 Sentence denotes SlpAST is a well standardized method to identify current epidemic C. difficile strains but also strains of still unknown epidemic relevance.
T40 195570-195852 Sentence denotes Helicobacter pylori, the causative agent of type B gastritis, peptic ulcers, gastric adenocarcinoma and MALT lymphoma, uses the Cag type IV secretion system to induce a strong proinflammatory response in the gastric mucosa and to inject its effector protein CagA into gastric cells.
T41 195853-196012 Sentence denotes This protein translocation results in altered host cell gene expression profiles and cytoskeletal rearrangements, and it has been linked to cancer development.
T42 196013-196190 Sentence denotes The Cag system is only distantly related to other type IV secretion systems, and it contains several essential components without functional counterparts in these other systems.
T43 196191-196381 Sentence denotes Recently, it has been shown that the Cag type IV secretion apparatus binds to integrin molecules as receptors on host cells, and that several Cag components are involved in this interaction.
T44 196382-196478 Sentence denotes In this study, we have characterized one of the components with integrin binding capacity, CagI.
T45 196479-196722 Sentence denotes Previous studies have identified the cagI gene as an essential gene for translocation of CagA into host cells, but there have been conflicting results as to its requirement for stimulation of interleukin-8 (IL-8) secretion by epithelial cells.
T46 196723-196976 Sentence denotes Since the cagI gene overlaps and is co-transcribed with the cagL gene encoding another integrin-interacting protein essential for Cag functionality, this discrepancy might be due to transcriptional effects of the resistance markers used for mutagenesis.
T47 196977-197131 Sentence denotes Using a marker-free gene deletion approach and genetic complementation, we show here that CagI is required for both CagA translocation and IL-8 induction.
T48 197132-197324 Sentence denotes The absence of several components of the secretion apparatus has an impact on CagI and CagL protein levels, indicating that (partial) assembly of the apparatus is required for their stability.
T49 197325-197424 Sentence denotes Our data provide evidence for a localization of both CagI and CagL at the bacterial outer membrane.
T50 197425-197523 Sentence denotes Furthermore, we demonstrate by co-immunoprecipitation that CagI and CagL interact with each other.
T51 197524-197651 Sentence denotes Taken together, our results are consistent with the assumption that CagI and CagL form a complex at the bacterial cell surface.
T52 197652-197744 Sentence denotes Still increasing incidence of Clostridium difficile-infections in Germany-need for action B.
T53 197745-197757 Sentence denotes Weiß* 1 , A.
T54 197758-197774 Sentence denotes Friedrich 2 , D.
T55 197775-197794 Sentence denotes Weitzel-Kage 3 , T.
T56 197795-198065 Sentence denotes Eckmanns Infektionsepidemiologie, Berlin, Deutschland 2 Landesuntersuchungsanstalt für das Gesundheits-und Veterinärwesen (LUA) Sachsen, Chemnitz, Deutschland 3 Charite Universitätsmedizin Berlin, Institut für Hygiene und Umweltmedizin, Berlin, Deutschland Introduction:
T57 198066-198250 Sentence denotes Since 2003, the United States, Canada and various countries in Europe have reported an increasing incidence of individual cases and outbreaks of Clostridium difficile infections (CDI).
T58 198251-198416 Sentence denotes This was thought to be associated with the emergence of a more virulent strain, Clostridium difficile PCR ribotype 027, which was first reported in Germany in 20007.
T59 198417-198532 Sentence denotes Since then severe cases of CDI (definition below) are notifiable according to the Protection against Infection Act.
T60 198533-198654 Sentence denotes In order to gain a more comprehensive picture of all CDI over time an analysis of various data sources is presented here.
T61 198655-198663 Sentence denotes Methods:
T62 198664-199007 Sentence denotes We described the incidence of CDI in Germany over time using 4 different data sources: a) notifications of laboratory-confirmed CDI in the federal state of Saxony, identified through additional mandatory surveillance from 2002-2010, b) national surveillance data of severe CDI from 2008-2010 (defined by at least one of the following criteria:
T63 199008-199548 Sentence denotes 1. readmission to a hospital due to a relapse, 2.admission to an intensive care unit due to CDI or its complication 3. surgery for toxic megacolon, perforation or refractory colitis, 4. death within 30 days after diagnosis and CDI as primary or contributive cause) c) nosocomial infection surveillance system for CDI (CDAD-KISS) from 2007-2010 and d) hospital discharges with diagnosis code A04.7: "enterocolitis due to Clostridium difficile" according to the International Classification of Diseases, 10th revision (ICD-10) from 2000-2009.
T64 199549-199623 Sentence denotes Time trends were analyzed with negative binomial regression using STATA10.
T65 199624-199902 Sentence denotes Results: CDI incidences have reached a peak in 2010 with 113 notified cases/100.000 inhabitants in Saxony, 0.6 severe CDI cases/100.000 inhabitants according to the national surveillance system and an incidence density of 0.72 CDI cases/1000 patient days generated by CDAD-KISS.
T66 199903-199974 Sentence denotes Hospital discharge diagnosis data for 2010 are not yet available (2009:
T67 199975-200033 Sentence denotes 120.8 CDI diagnoses/100.000 discharged hospital patients).
T68 200034-200327 Sentence denotes The negative binomial regression models revealed a significant yearly increase in CDI incidence rates for hospital discharges (Incidence rate ratios (IRR) 1.43, p= 0.000), for CDI notifications in Saxony (IRR 1.39, p=0.000), and for the national surveillance of severe CDI (IRR 1.16, p=0.003).
T69 200328-200339 Sentence denotes Discussion:
T70 200340-200430 Sentence denotes The continuing increase of CDI cases that peaked in 2010 shows the urgent need for action.
T71 200431-200481 Sentence denotes The nature of this increase is not yet understood.
T72 200482-200540 Sentence denotes An improved characterization of cases is therefore needed.
T73 200541-200741 Sentence denotes This includes the monitoring of disease origin like proportion of healthcare-and community-associated CDI, the distribution and characterization of strains and a continuing evaluation of risk factors.
T74 200742-200786 Sentence denotes This will help to focus preventive measures.
T75 200787-200800 Sentence denotes Introduction:
T76 200801-200920 Sentence denotes Clostridium difficile associated disease has, in the last few years, increased in incidence and in severity world wide.
T77 200921-201113 Sentence denotes The "epidemic strain" 027/BI/NAP1 causing severe disease in North America and parts of Europe was not observed in large numbers in Germany although the incidence of CDAD has greatly increased.
T78 201114-201192 Sentence denotes Ribotype 001 has been implicated in severe disease in some studies in Germany.
T79 201193-201215 Sentence denotes Methods and Materials:
T80 201216-201531 Sentence denotes Over the period from 2007 to 2011 a large number of isolates has been collected from the greater Düsseldorf area and subjected to ribotyping, PCR analysis of the presence of the toxin genes; tcdA, tcdB and cdtA and mutation causing truncation of the tcdC gene as well as resistance to erythromycin and moxifloxacin.
T81 201532-201540 Sentence denotes Results:
T82 201541-201609 Sentence denotes A total of 254 isolates were analysed for the above characteristics.
T83 201610-201722 Sentence denotes Resistance to erythromycin was observed in 132 [53.7 %] isolates and resistance to moxifloxacin in 194 [78.9 %].
T84 201723-201787 Sentence denotes Resistance to both antibiotics together was detected in 122 [49.
T85 201788-201862 Sentence denotes 6 %] isolates and no resistance to either substance in 42 [17 %] isolates.
T86 201863-201901 Sentence denotes Ribotyping of the isolates is ongoing.
T87 201902-202016 Sentence denotes Preliminary analysis of the data showed that the isolates can be grouped into approximately 50 distinct ribotypes.
T88 202017-202122 Sentence denotes One ribotype contained approximately 80 isolates and the next largest ribotype approximately 20 isolates.
T89 202123-202165 Sentence denotes Confirmation of the ribotyping is awaited.
T90 202166-202177 Sentence denotes Discussion:
T91 202178-202284 Sentence denotes Analysis of a large number of c. difficile from an expansive urban area has not been described in Germany.
T92 202285-202620 Sentence denotes It is interesting to scrutinise such data as one may be able to determine if the increasing incidence of CDAD is perhaps, unlike the case in North Anmerica, not associated with a specific clonal spread and therby also determine if the change is perhaps due to changes in medical practice rather than a shift in the bacterial virulence.
T93 202621-202952 Sentence denotes Our results show that the vast majority of the isolates possesed both toxin A and B genes and that only few [12.6 the binary toxin gene, a disproportionate number of these [60.6 %] however were located in isolates positive for tcdA and negative for tcdB genes, which only accounted for 7.8 % of the total number of isolates tested.
T94 202953-203038 Sentence denotes As previously reported most isolates were resistant to erythromycin and moxifloxacin.
T95 203039-203077 Sentence denotes Giardia duodenalis Arginine deiminase:
T96 203078-203131 Sentence denotes Exploring its function for host-parasite interplay S.
T97 203132-203145 Sentence denotes Banik* 1 , A.
T98 203146-203270 Sentence denotes Aebischer FG 16 Parasitologie, Berlin, Deutschland The protozoa Giardia duodenalis (Gd) is an intestinal parasite of humans.
T99 203271-203339 Sentence denotes It causes diarrhoeal and gastrointestinal disease, named giardiasis.
T100 203340-203402 Sentence denotes Annually, around 300 million people become infected worldwide.
T101 203403-203498 Sentence denotes Gd is ubiquitously distributed and also infects other vertebrates, including companion animals.
T102 203499-203573 Sentence denotes Thus, giardiasis is a major public and veterinary health concern globally.
T103 203574-203673 Sentence denotes It remains unclear to what degree it is a zoonosis and what determines its pathogenicity to humans.
T104 203674-203865 Sentence denotes To assess these issues a Gd biobank shall be generated to link epidemiological data with functional characteristics of clinical isolates and as a means to identify virulence associated genes.
T105 203866-204029 Sentence denotes We hypothesize that potential virulence markers are enzymes released by Giardia upon colonization of the small instestine and interaction with the host epithelium.
T106 204030-204285 Sentence denotes Especially the Gd arginine deiminase (ADI) that converts arginine to citrulline and ammonia shall be investigated as possible virulence and pathogenicity factor of Gd, because arginine plays a critical role in antimicrobial mechanisms and immune response.
T107 204286-204508 Sentence denotes The sequencevariation of ADI will be determined by cloning adi from Gd isolates being collected in the biobank and sequence variants will be functionally characterized by expressing the respective recombinant ADI proteins.
T108 204509-204652 Sentence denotes Finally, correlation between enzyme function and clinical pathology shall be investigated on basis of Gd isolates related epidemiological data.
T109 204653-204704 Sentence denotes We will present the establishment of this workflow.
T110 204705-204833 Sentence denotes Gd clone WB 6 ADI as well as a mutated ADI (C424A) were cloned, recombinantly expressed and purified by affinity chromatography.
T111 204834-204970 Sentence denotes An enzyme activity test was adapted for colorimetric measurement of arginine to citrulline conversion to verify active and inactive ADI.
T112 204971-205144 Sentence denotes Subsequently, in vitro generated human dendritic cells (DCs) were treated with recombinant ADI or mutADI (C424A) in absence and presence of Toll-like receptor (TLR) ligands.
T113 205145-205288 Sentence denotes Investigations of surface molecule expression and cytokine release shall give first evidences for ADI and its role for host-parasite interplay.
T114 205289-205526 Sentence denotes What makes Campylobacter jejuni thrive: the contribution of amino acid metabolism to the efficient proliferation of a widespread food-borne pathogen Campylobacter jejuni is a major cause of food-borne disease in industrialized countries.
T115 205527-205733 Sentence denotes We found recently that the highly pathogenic strain C. jejuni 81-176 has an extended capacity to metabolize amino acids in comparison to several C. jejuni isolates including the reference strain NCTC 11168.
T116 205734-205954 Sentence denotes The amino acids aspartate, glutamate, proline and serine are growth-promoting substrates for both isolates, but only C. jejuni 81-176 possesses the additional capability to utilize asparagine, glutamine, and glutathione.
T117 205955-206288 Sentence denotes Strikingly, the additional glutamine or glutathione metabolism of C. jejuni 81-176 contributes to its enhanced capability to colonize the intestine in a mouse model for colonization, whereas the asparagine utilization results in a more efficient persistence of C. jejuni 81-176 in the liver of infected mice (Hofreuter et al., 2008).
T118 206289-206388 Sentence denotes We have continued our studies and examined the effect of amino acids on the viability of C. jejuni.
T119 206389-206578 Sentence denotes By transposon and targeted mutagenesis we recognized the involvement of previously uncharacterized genes in the amino acid catabolism as well as amino acid biosynthesis of C. jejuni 81-176.
T120 206579-206760 Sentence denotes We tested the impact of amino acid metabolism on the survival of wild-type C. jejuni and several mutants defective in the utilization of amino acids under various stress conditions.
T121 206761-206910 Sentence denotes Further, the contribution of amino acid catabolism by C. jejuni 81-176 on its interaction with cultured intestinal epithelial cells was investigated.
T122 206911-207081 Sentence denotes Using a murine infection model we discovered distinct correlations between the metabolism of the tested amino acids and the persistence of C. jejuni in different tissues.
T123 207082-207218 Sentence denotes The results broaden our knowledge about the amino-acid-driven growth of C. jejuni and its role during the colonization of various hosts.
T124 207219-207347 Sentence denotes Gaussia princeps luciferase (Gluc) can be used to monitor type I protein secretion in Salmonella enterica serovar Typhimurium T.
T125 207348-207362 Sentence denotes Wille 1 , R.G.
T126 207363-207455 Sentence denotes Gerlach* 1 1 Robert Koch-Institut, Nachwuchsgruppe 3, Wernigerode, Deutschland Introduction:
T127 207456-207605 Sentence denotes Gaussia princeps luciferase (Gluc) is a highly active reporter exhibiting approximately 1000-fold higher luminescence compared to Renilla luciferase.
T128 207606-207671 Sentence denotes Data about the applicability of Gluc in bacteria is very limited.
T129 207672-207817 Sentence denotes Given the small size, high activity and co-factor independence of Gluc, it might be especially suited to monitor secretion of bacterial proteins.
T130 207818-207954 Sentence denotes Salmonella Pathogenicity Island 4 (SPI-4) encodes for SiiCDF which forms a type I secretion system (T1SS) and the secreted adhesin SiiE.
T131 207955-208081 Sentence denotes SiiE mediates intimate contact of Salmonella to polarized epithelial cells, thus facilitating the subsequent invasion process.
T132 208082-208090 Sentence denotes Methods:
T133 208091-208171 Sentence denotes A codon-optimized Gluc gene was efficiently expressed in Salmonella enterica sv.
T134 208172-208187 Sentence denotes Typhimurium (S.
T135 208188-208201 Sentence denotes Typhimurium).
T136 208202-208309 Sentence denotes Fusion proteins of Gluc and different C-terminal portions of the SPI-4-encoded adhesin SiiE were generated.
T137 208310-208330 Sentence denotes After transfer in S.
T138 208331-208436 Sentence denotes Typhimurium WT and siiF, luminescence was detected in bacterial lysates, supernatants and whole cultures.
T139 208437-208445 Sentence denotes Results:
T140 208446-208580 Sentence denotes We detected luciferase activity of Gluc-SiiE fusion proteins in supernatants as well as whole cultures depending on a functional T1SS.
T141 208581-208741 Sentence denotes We could show that a C-terminal moiety of SiiE including immunoglobulin (Ig) domain 53 is essential and sufficient mediating type I-dependent secretion of Gluc.
T142 208742-208846 Sentence denotes Secretion efficiency of fusion proteins increased with increasing length of the C-terminal SiiE portion.
T143 208847-208858 Sentence denotes Discussion:
T144 208859-208929 Sentence denotes In conclusion, Gluc can be used to monitor protein secretion via T1SS.
T145 208930-209047 Sentence denotes This is the first demonstration of enzymatic detection of secreted proteins without stable periplasmic intermediates.
T146 209048-209242 Sentence denotes Gluc might open a venue for the systematic identification and analysis of type I-secreted proteins and represents a valuable addition to the toolbox of modern molecular biology and microbiology.
T147 209243-209368 Sentence denotes Helicobacter pylori BCS 100 genome variability and potential virulence determinants during an experimental human infection L.
T148 209369-209386 Sentence denotes Kennemann* 1 , T.
T149 209387-209403 Sentence denotes Aebischer 2 , S.
T150 209404-209417 Sentence denotes Kuhn 1 , T.F.
T151 209418-209430 Sentence denotes Meyer 2 , C.
T152 209431-209456 Sentence denotes Josenhans 1 Introduction:
T153 209457-209620 Sentence denotes Helicobacter pylori is one of the bacterial species with highest allelic diversity, due to a high mutation rate and frequent recombination during mixed infections.
T154 209621-209818 Sentence denotes In a recent whole genome comparison study we demonstrated extensive genomic changes in four sets of sequential isolates obtained from chronically infected Colombians (Kennemann et al., PNAS 2011) .
T155 209819-209932 Sentence denotes Much less is known about genomic adaptations that may occur in the first months of infection of a new human host.
T156 209933-210210 Sentence denotes Here, we present the complete genome sequence of H. pylori strain BCS 100 (H1), a cagPAI deficient isolate that has been used for several human challenge studies in the US and Germany, and draft sequences of reisolates obtained from four volunteers after 3 months of infection.
T157 210211-210219 Sentence denotes Results:
T158 210220-210361 Sentence denotes The genome sequence of H. pylori challenge strain H1 was obtained by 454 sequencing and subsequent gap closure by targeted Sanger sequencing.
T159 210362-210572 Sentence denotes Draft genome sequences were obtained by 454 sequencing of one additional input isolate (H3) and four isolates from four volunteers belonging either to the control group or to the Ty21a(UreA/B) vaccinated group.
T160 210573-210666 Sentence denotes Additional reisolates from 29 volunteers were analyzed at selected loci by Sanger sequencing.
T161 210667-210679 Sentence denotes Conclusions:
T162 210680-210828 Sentence denotes In contrast to the sequential isolates from Colombia, no evidence of recombination (import) was detected in the reisolates from infected volunteers.
T163 210829-210983 Sentence denotes 1-2 novel SNPs were identified in each of the reisolated genomes, permitting the calculation of a robust estimate for the H. pylori in vivo mutation rate.
T164 210984-211200 Sentence denotes Unexpectedly, a previously unknown variant of the input strain was detected in several volunteers which differed from the 454 sequenced input clone H1 by 86 SNPs and three clusters of nucleotide polymorphisms (CNPs).
T165 211201-211368 Sentence denotes This variant was commonly detected among the additional reisolates from the study, suggesting strong in vivo selection favoring this subpopulation in some human hosts.
T166 211369-211489 Sentence denotes Helicobacter pylori Type IV secretion system exploits distinct domains of the human 1-Integrin for CagA translocation C.
T167 211490-211502 Sentence denotes Ertl* 1 , L.
T168 211503-211522 Sentence denotes Jimenez-Soto 1 , R.
T169 211523-211802 Sentence denotes Haas 1 1 Max von Pettenkofer Institut, Bakteriologie, München, Deutschland Translocation of the cytotoxin-associated antigen A (CagA) via the Type IV Secretion System (T4SS) into the host cell is a major risk factor for gastric diseases, like chronic gastritis or gastric cancer.
T170 211803-211915 Sentence denotes The immunodominant protein CagA is encoded, together with the cag-T4SS, on the cagpathogenicity island (cagPAI).
T171 211916-212105 Sentence denotes After translocation into the host cell, CagA subsequently is tyrosine-phosphorylated and modifies different eukaryotic signalling pathways in phosphorylation-dependent and independent ways.
T172 212106-212283 Sentence denotes We and others showed that CagA translocation is dependent on interaction of the pilus-like needle structure of the cag-T4SS with the 1-Integrin (ITGB1) on the host cell surface.
T173 212284-212439 Sentence denotes Integrins are a family of heterodimeric cell surface adhesion molecules, which mediate cell-cell, cell-extracellular matrix and cell-pathogen interactions.
T174 212440-212615 Sentence denotes Three components of the cag-T4SS, CagY and CagI, as well as the translocated effector protein CagA, are binding to ITGB1, as originally identified in a Yeast Two Hybrid Assay.
T175 212616-212808 Sentence denotes We attempt to localize the distinct binding regions of these proteins within the receptor and we want to determine, whether binding of CagA to ITGB1 is an essential step in CagA translocation.
T176 212809-212998 Sentence denotes The N-terminal integrin binding region of CagA was further narrowed down to a stretch of about 100 amino acids and the corresponding domains on the 1-Integrin for CagA binding are verified.
T177 212999-213196 Sentence denotes These data can help to propose a new model of interaction between the pathogenic H. pylori and the eukaryotic cell, mediated by specific interactions of the cag-T4SS and the ITGB1 of the host cell.
T178 213197-213205 Sentence denotes Results:
T179 213206-213290 Sentence denotes A total of 59 ACI outbreaks, 109 PAE outbreaks, and 57 ESBL outbreaks were included.
T180 213291-213419 Sentence denotes As shown in figure 1a , the median duration of the outbreak was 183 days (ACI), 92 days (PAE), and 210 days (ESBL) respectively.
T181 213420-213520 Sentence denotes The average number of affected patients in the three groups was 25 (ACI outbreaks; thereof infected:
T182 213521-213547 Sentence denotes 19 patients; thereof died:
T183 213548-213614 Sentence denotes 4 patients), 23 (PAE; 16; 3), and 57 (ESBL; 46; 2) (figure 1b -d).
T184 213615-213747 Sentence denotes ACI outbreak specific findings included an association to university hospitals, to intensive care units, and to surgery departments.
T185 213748-213902 Sentence denotes PAE outbreaks were associated with significantly less use of isolation precautions, less frequently closure of wards, and less use of protective clothing.
T186 213903-214033 Sentence denotes ESBL outbreaks took place especially in neonatology departments and caused significantly high numbers of urinary tract infections.
T187 214034-214222 Sentence denotes Additional information on the source of pathogen, its route of transmission, type of infection, age groups of patients, and implemented infection control measures is provided in the table.
T188 214223-214234 Sentence denotes Conclusion:
T189 214235-214428 Sentence denotes The Outbreak Database served as a valuable tool for the characterization of nosocomial outbreaks as several significant differences between the three groups of pathogens examined were detected.
T190 214429-214571 Sentence denotes Thus, from the findings of the present study we would recommend adjusting the infection control measures specifically on the type of pathogen.
T191 214572-214694 Sentence denotes However, from our point of view currently there is no MRGN bacterium that should be totally disregarded with this respect.
T192 214695-214793 Sentence denotes Essen, Deutschland 3 University Hospital Essen, Hospital Hygiene, Essen, Deutschland Introduction:
T193 214794-214891 Sentence denotes Postoperative wound infections generally cause considerable extra morbidity, mortality and costs.
T194 214892-214980 Sentence denotes The prevalence of total wound infections after cardiac surgery ranges from 0.47 to 3.5%.
T195 214981-214990 Sentence denotes Question:
T196 214991-215150 Sentence denotes To describe the frequency of surgical site infections (SSIs) following open heart surgery and to identify the most frequent risks factors for these infections.
T197 215151-215159 Sentence denotes Methods:
T198 215160-215329 Sentence denotes A prospective surveillance was performed during the period from January 2011 to April 2011 at the Intensiv CardioSurgery Unit of the Universitätsklinikum Essen, Germany.
T199 215330-215460 Sentence denotes Patients were evaluated daily during hospitalization and when re-admitted to Surgical Unit for any SSIs occurring after discharge.
T200 215461-215569 Sentence denotes The wound infections were defined according to the Centers for Disease Control and Prevention (CDC) and U.S.
T201 215570-215637 Sentence denotes National Nosocomial Infections Surveillance (NNIS) system criteria.
T202 215638-215686 Sentence denotes The descriptive epidemiological method was used.
T203 215687-215695 Sentence denotes Results:
T204 215696-215789 Sentence denotes In total 305 patients were included, mean age was 67.2 years, 25.6 % of patients were female.
T205 215790-215882 Sentence denotes 82.6% were elective procedures, 61.3% represented by coronary artery bypass grafting (CABG).
T206 215883-216092 Sentence denotes Methicillin-resistant Staphylococcus aureus (MRSA) screening was performed by 92.1% of patients, and perioperative antibiotic prophylaxis with a first-generation cephalosporin was administered in 93% of cases.
T207 216093-216133 Sentence denotes SSIs were observed in 5 patients (1.6%):
T208 216134-216198 Sentence denotes 2 (0.6%) were superficial infections and 3 (1%) deep infections.
T209 216199-216295 Sentence denotes All infections occurred in CABG procedures and were diagnosed more than 14 days after operation.
T210 216296-216334 Sentence denotes All patients had BMI >30, ASA score 3.
T211 216335-216419 Sentence denotes Three patients had blood-glucose levels >180 mg/dl during the postoperative 24hours.
T212 216420-216431 Sentence denotes Conclusion:
T213 216432-216510 Sentence denotes Preventing SSIs in the operating room is a primary goal for the surgical team.
T214 216511-216642 Sentence denotes Attention should be paid to antibiotic prophylaxis and Methicillin-resistant Staphylococcus aureus (MRSA) nasal carriage treatment.
T215 216643-216733 Sentence denotes In this study BMI >30 and high blood-glucose levels were associated with wound infections.
T216 216734-216803 Sentence denotes Overall the rate of SSIs was low, but further surveillance is needed.
T217 216804-216839 Sentence denotes Changes in hand hygiene compliance:
T218 216840-216892 Sentence denotes Are professional status and discipline risk factors?
T219 216893-216895 Sentence denotes K.
T220 216896-216908 Sentence denotes Graf* 1 , E.
T221 216909-216919 Sentence denotes Ott 1 , N.
T222 216920-216932 Sentence denotes Tramp 1 , M.
T223 216933-216945 Sentence denotes Wolny 1 , G.
T224 216946-216960 Sentence denotes Porep 1 , I.F.
T225 216961-216978 Sentence denotes Chaberny 1 1 Med.
T226 216979-217014 Sentence denotes Hochschule Hannover, Inst. für Med.
T227 217015-217084 Sentence denotes Mikrobiologie u.Krankenhaushygiene, Hannover, Deutschland Background:
T228 217085-217176 Sentence denotes Hand hygiene is the most effective measure for preventing infections related to healthcare.
T229 217177-217510 Sentence denotes This study aims to evaluate the hand hygiene compliance in a German university hospital overall and in addiction to the professional status after implementation of the national hand hygiene campaign "Aktion Saubere Hände in 2008.This is the largest nationwide survey on intensive care units (ICU) in a university hospital in Germany.
T230 217511-217519 Sentence denotes Methods:
T231 217520-217638 Sentence denotes During our investigation period (January 2008-December 2010), education and training of hand hygiene were implemented.
T232 217639-217709 Sentence denotes Consumption rates of hand rub and gloves were collected and evaluated.
T233 217710-217951 Sentence denotes Changes in healthcare workers behaviour were observed and compared between different professional groups, compliance rates in hand hygiene of Healthcare personnel of twelve ICUs were evaluated before and after implementation of the campaign.
T234 217952-218077 Sentence denotes Implementation included extensive education and training of all health care workers (at least three teaching units per ward).
T235 218078-218211 Sentence denotes Five indications of hand hygiene were classified according to WHO definitions, compliance rates were evaluated for these indications.
T236 218212-218312 Sentence denotes The health care workers were divided in three groups: doctors, nurses and other health care workers.
T237 218313-218321 Sentence denotes Results:
T238 218322-218411 Sentence denotes Consumption rates of hand rub increased from 27 to 32 hand disinfections per patient day.
T239 218412-218511 Sentence denotes No significant correlation between hand rub consumption and hand hygiene compliance was calculated.
T240 218512-218564 Sentence denotes No significant change in use of gloves was detected.
T241 218565-218628 Sentence denotes Overall 4040 opportunities of hand disinfection were monitored.
T242 218629-218801 Sentence denotes A significant improvement of hand hygiene compliance was assessed (from 54 to 61%, p>0.05), the risk of no disinfection was reduced significantly (p>0.05, 95%CI 0.74-0.85).
T243 218802-218975 Sentence denotes The risk of no disinfection decreased significantly in the nurses group, but not in the doctors group (other health care workers were not evaluated due to the small number).
T244 218976-219203 Sentence denotes There was no significant difference in hand hygiene compliance before the intervention between doctors and nurses, after the intervention, there was a significant difference in hand hygiene compliance (p>0.05, 95%CI 1.42-1.85).
T245 219204-219346 Sentence denotes Hand hygiene compliance is higher in the nurses group, the risk of no hand disinfection in doctors is decreased, but there is no significance.
T246 219347-219359 Sentence denotes Conclusions:
T247 219360-219445 Sentence denotes A significant change of hand hygiene behaviour was recognized for doctors and nurses.
T248 219446-219578 Sentence denotes The hand hygiene compliance of nurses increased significantly higher, hence other training strategies for doctors will be essential.
T249 219579-219721 Sentence denotes Effective and rapid analysis of nosocomial outbreaks using the DiversiLab semi-automated repetitive sequence-based PCR test system Objectives:
T250 219722-219824 Sentence denotes Rapid and sensitive methods are essential for typing and monitoring of hospital associated infections.
T251 219825-219962 Sentence denotes The new DiversiLab system (BioMerieux) exhibits a semiautomated repetitive-sequence-based polymerase chain reaction (rep-PCR) for typing.
T252 219963-220252 Sentence denotes Aim of the study was to evaluate the usefulness and accuracy of the DiversiLab system for bacterial strain typing and determination of genetic relatedness of strains associated with nosocomial outbreaks and integration of the system in the workflow of a routine microbiological laboratory.
T253 220253-220261 Sentence denotes Methods:
T254 220262-220599 Sentence denotes For evaluation 20 Methicillin-resistant Staphylococcus aureus (MRSA), 26 multi-drug resistant (MDR)-Acinetobacter baumannii strains and 43 extended-spectrum-ß-lactamase producing Klebsiella spp. strains (30 K. pneumoniae and 13 K. oxytoca strains) from well-defined nosocomial outbreaks were typed using rep-PCR on the DiversiLab system.
T255 220600-220703 Sentence denotes As reference method for bacterial strains-typing pulsed-field gel electrophoresis (PFGE) was performed.
T256 220704-220712 Sentence denotes Results:
T257 220713-220829 Sentence denotes Concerning the 20 MRSA isolates, there was identical cluster formation in both, the DiversiLab and the PFGE systems.
T258 220830-221017 Sentence denotes The same result could be observed in the 26 multi-resistant A. baumannii strains, where strains showed identical cluster formation (into 3 separate clusters) and very similar dendrograms.
T259 221018-221264 Sentence denotes The 30 ESBL-positive K. pneumoniae strains originated from two chronologically separated nosocomial outbreaks: PFGE placed 28 of the ESBL-positive K. pneumoniae strains into the same cluster, whereas PFGE defined all strains as indistinguishable.
T260 221265-221405 Sentence denotes The DiversiLab-system also formed a cluster of these 28 strains, but the strains in this cluster were not defined as identical, but similar.
T261 221406-221478 Sentence denotes Both systems identified 2 K. pneumoniae strains as non-outbreak strains.
T262 221479-221707 Sentence denotes Results from the K. oxytoca outbreak provide similar minor differences between PFGE and results obtained with the DiversiLab-system, whereas rep-PCR on the DiversiLab system exhibits slightly more discriminatory power than PFGE.
T263 221708-221871 Sentence denotes Conclusion: -The DiversiLab system is a rapid, semi-automated repetitive sequencebased PCR test system for typing and analysing bacterial strains, including fungi.
T264 221872-222130 Sentence denotes -Compared to PFGE, this study confirms the good discriminatory power of the DiversiLab system. -Results on the DiversiLab system could be obtained in a short period (8-24h), which is an essential advantage in rapid analysis of hospital associated infections.
T265 222131-222142 Sentence denotes Background:
T266 222143-222248 Sentence denotes There are several products that claim to sanitize surfaces quickly and easily by the use of UV radiation.
T267 222249-222379 Sentence denotes Those products are for use in households or during travel (e.g., on toilet facilities, hotel mattresses, and personal care items).
T268 222380-222496 Sentence denotes However, often their actual ability of killing germs and their overall practicability in daily life remains unknown.
T269 222497-222504 Sentence denotes Method:
T270 222505-222615 Sentence denotes We tested the disinfection ability of the Verilux® CleanWave® Sanitizing Wand (Verilux Inc., Waitsfield, USA).
T271 222616-222721 Sentence denotes This is a portable device that emits UV-C radiation from a UV light bulb (3 Watt) when held horizontally.
T272 222722-222861 Sentence denotes According to the manufacturer's instructions the wand should be used as close as possible and no more than one-half inch above the surface.
T273 222862-222978 Sentence denotes As stated in the operating manual, a 90% elimination of S. aureus would be expected within 12 sec by this procedure.
T274 222979-223104 Sentence denotes S. aureus (ATCC 29213), E. faecium (ATCC 19434) , and E. coli (ATCC 25922) were each suspended in sterile 0.9% NaCl solution.
T275 223105-223207 Sentence denotes A 100 μL sample containing 5*10 6 CFU were spateled on standard sheep blood agar (surface = 56.7 cm²).
T276 223208-223344 Sentence denotes Continuous UV-C radiation was then applied at a distance of 0.7 cm (FIGURE 1A) for various time intervals (10, 20, 30, 45 , and 60 sec).
T277 223345-223434 Sentence denotes Reduction of bacterial growth was determined visually after 18 hours incubation at 37 °C.
T278 223435-223599 Sentence denotes For this purpose, the number of CFU was counted on a defined area of 4 cm² per plate (FIGURE 1B) and compared to the number of expected CFU in an area of that size.
T279 223600-223637 Sentence denotes All experiments were performed twice.
T280 223638-223646 Sentence denotes Results:
T281 223647-223702 Sentence denotes The corresponding killing curves are shown in FIGURE 2.
T282 223703-223859 Sentence denotes A sufficient reduction ( 5 log steps) was achieved for S. aureus and E. coli after 10 sec already, but 20 sec were needed for the same effect on E. faecium.
T283 223860-223871 Sentence denotes Discussion:
T284 223872-223968 Sentence denotes Our results confirm that UV-C light from the device is capable of killing bacteria in principle.
T285 223969-224196 Sentence denotes However, when using such a device for disinfection purpose in daily life, one has to keep in mind that 20 sec UV-C light exposition time was necessary in order to properly disinfect a rather small surface area from enterococci.
T286 224197-224289 Sentence denotes Thus, much longer time intervals will be necessary for larger surfaces such as toilet seats.
T287 224290-224453 Sentence denotes So there might be few opportunities when such a device could be a useful tool, but it seems less helpful on larger exterior due to time necessary for disinfection.
T288 224454-224548 Sentence denotes Noteworthy, new batteries allowed an overall device working time of less than 15 minutes only.
T289 224550-224642 Sentence denotes Killing kinetic curves of S. aureus, E. faecium, and E. coli after UV-C exposition over time
T290 224643-224723 Sentence denotes Influence of water flow rate on the presence of enteric viruses in Ruhr river L.
T291 224724-224738 Sentence denotes Jurzik* 1 , I.
T292 224739-224751 Sentence denotes Hamza 1 , F.
T293 224752-224766 Sentence denotes Remmler 2 , M.
T294 224767-224814 Sentence denotes Wilhelm Abt. für Hygiene, Bochum, Deutschland 2
T295 224816-225027 Sentence denotes In the last years we have demonstrated the presence of enteric viruses in the Ruhr river, which is a drinking water source for about 5 Mio. people (Hamza et al., 2009 , Hamza et al., 2011 , Jurzik et al., 2010 .
T296 225028-225168 Sentence denotes Different factors can control the survival of enteric viruses in aquatic environments such as temperature, sunlight, pH, and organic matter.
T297 225169-225277 Sentence denotes However, little is known about the effect of the flow rate on the concentration of viruses in surface water.
T298 225278-225398 Sentence denotes Therefore, the present study aims to investigate the impact of flow rate on the presence of enteriv virus in Ruhr river.
T299 225399-225510 Sentence denotes Water samples were collected nearly weekly at five different sampling sites from January 2008 to December 2009.
T300 225511-225607 Sentence denotes Water samples were concentrated using VIRADEL method with a high-salt alkaline phosphate buffer.
T301 225608-225674 Sentence denotes For the quantification of viruses, qReal-Time PCR was carried out.
T302 225675-225746 Sentence denotes Somatic coliphages were quantified using the double layer plaque assay.
T303 225747-225888 Sentence denotes The quantification of E. coli and intestinal Enterococci were done according to the standard reference method (DIN EN ISO 9308-3 and 7899-1).
T304 225889-225988 Sentence denotes The flow rate was daily measured at the Ruhr river in Villigst (Schwerte) for the same time period.
T305 225989-226196 Sentence denotes Results showed that Ruhr river was highly contaminated with human adenovirus (2.6x10 3 gen. equ./l, n=175), human polyomavirus (1.0x10 3 gen. equ./l, n=123), and human rotavirus (8.2x10 3 gen. equ./l, n=93).
T306 226197-226390 Sentence denotes The median concentrations for bacterial parameters were as follows: E. coli (1.0x10 3 cfu/l, n=155), total coliforms (1.4x10 3 cfu/l, n=177), and intestinal enterococci (6.2x10 1 cfu/l, n=172).
T307 226391-226475 Sentence denotes Moreover, no correlation between microbiological parameters and flow rate was found.
T308 226476-226590 Sentence denotes Further investigations are needed to characterize hydrological factors influencing enteric viruses in river water.
T309 226591-226606 Sentence denotes Hamza, I.A., L.
T310 226607-226617 Sentence denotes Jurzik, A.
T311 226618-226627 Sentence denotes Stang, K.
T312 226628-226636 Sentence denotes Sure, K.
T313 226637-226651 Sentence denotes Uberla, and M.
T314 226652-226660 Sentence denotes Wilhelm.
T315 226661-226667 Sentence denotes 2009 .
T316 226668-226810 Sentence denotes Detection of human viruses in rivers of a densly-populated area in Germany using a virus adsorption elution method optimized for PCR analyses.
T317 226812-226952 Sentence denotes Homogeneous distribution of phenotypic characteristics in clonally unrelated Pseudomonas aeruginosa isolates from technical water systems H.
T318 226953-226973 Sentence denotes Petry-Hansen* 1 , J.
T319 226974-226986 Sentence denotes Hanke 1 , J.
T320 226987-227002 Sentence denotes Heise 1 , H.-C.
T321 227003-227018 Sentence denotes Flemming 1 , J.
T322 227019-227217 Sentence denotes Wingender Biofilm Centre, Abteilung Aquatische Mikrobiologie, Essen, Deutschland P. aeruginosa is an opportunistic pathogen ubiquitously found in water environments where it can persist in biofilms.
T323 227218-227372 Sentence denotes Cells released from biofilms contaminate the water phase and pose a potential threat to human health, especially when occurring in man-made water systems.
T324 227373-227469 Sentence denotes The genotypic and phenotypic variability of P. aeruginosa in technical water systems is unknown.
T325 227470-227694 Sentence denotes During a period of seven years (2003 -2010) , 177 P. aeruginosa strains were isolated from technical water systems (drinking water distribution systems, plumbing systems, public swimming pools, and industrial water systems).
T326 227695-227870 Sentence denotes Using pulsed field gel electrophoresis of SpeI restricted genomic DNA of these isolates and seven P. aeruginosa reference strains 42 different clonal variants were identified.
T327 227871-228255 Sentence denotes For 77 representative strains and all seven reference strains the following characteristics were analyzed in order to reveal possible correlations between water source and genotypic as well as phenotypic characteristics: colony morphology, pigment production, hemolysis, cell surface hydrophobicity, biofilm formation, resistance against antibiotics and occurrence of virulence genes.
T328 228256-228531 Sentence denotes 64% of these 77 strains and the seven reference strains showed the typical colony morphology for P. aeruginosa, 92.8 % produced at least one of the pigments pyocyanin, pyoverdin, pyorubin and pyomelanin, 98.8 % displayed b-hemolysis and 90.5 % had a hydrophilic cell surface.
T329 228532-228674 Sentence denotes All strains were sensitive to ciprofloxacin, imipenem, meropenem, piperacillintazobactam, gentamycin, and except of one strain to ceftazidime.
T330 228675-228813 Sentence denotes An intermediate sensitivity to aztreonam was determined for all strains and except of one strain all strains were resistant to ampicillin.
T331 228814-229003 Sentence denotes Compared to reference strain PAO1, 53 strains (63 %) showed a higher biofilm formation on abiotic (polystyrene) surface, with 13 strains (15.5%) forming a cluster of strong biofilm formers.
T332 229004-229171 Sentence denotes In all 84 strains, the virulence gene lasB was detected as well as either the virulence genes exoS (63 %) or exoU (38 %); only one strain contained both exoS and exoU.
T333 229172-229335 Sentence denotes For the first time an extensive pool of P. aeruginosa isolates from different types of technical water systems were characterized phenotypically and genotypically.
T334 229336-229470 Sentence denotes No correlation between phenotypic and genotypic traits, and between these characteristics and the origin of the strains were detected.
T335 229471-229670 Sentence denotes The phenotypic characteristics and the occurrence of virulence genes seemed to be distributed in a relatively homogeneous way among the clonally unrelated strains from diverse man-made water systems.
T336 229671-229791 Sentence denotes There seem to be no selection of a specific genotypic or phenotypic variant of P. aeruginosa in technical water systems.
T337 229792-229794 Sentence denotes E.
T338 229795-229806 Sentence denotes Ott* 1 , S.
T339 229807-229822 Sentence denotes Saathoff 1 , A.
T340 229823-229837 Sentence denotes Legarth 1 , K.
T341 229838-229849 Sentence denotes Graf 1 , F.
T342 229850-229863 Sentence denotes Schwab 2 , I.
T343 229864-229866 Sentence denotes F.
T344 229867-229918 Sentence denotes Chaberny 1 1 Medizinische Hochschule Hannover, Med.
T345 229919-230025 Sentence denotes Mikrobiologie und Krankenhaushygiene, Hannover, Hygiene und Umweltmedizin, Berlin, Deutschland Background:
T346 230026-230115 Sentence denotes The incidence of nosocomial infections (NI) increased considerably during the last years.
T347 230116-230195 Sentence denotes Knowledge about the real frequency of nosocomial infections in Germany is rare.
T348 230196-230341 Sentence denotes The purpose of this study was to investigate the current frequency of nosocomial infections in a university hospital by a point-prevalence study.
T349 230342-230350 Sentence denotes Methods:
T350 230351-230432 Sentence denotes In March 2010, a systematic investigation was conducted in all adult in-patients:
T351 230433-230552 Sentence denotes Patients suffering from nosocomial infections were detected via an intensive chart review and interviews of physicians.
T352 230553-230649 Sentence denotes A standardized protocol was used for definition and categorization of all nosocomial infections.
T353 230650-230714 Sentence denotes Nosocomial infections were categorized by using CDC-definitions.
T354 230715-230853 Sentence denotes Information about using of antimicrobial agents, invasive devices, causative organism, comorbidities and invasive procedures was recorded.
T355 230854-230862 Sentence denotes Results:
T356 230863-230921 Sentence denotes Overall, 1,047 in-patients could be included in the study.
T357 230922-231002 Sentence denotes 125 NIs were detected in 118 patients for a prevalence of 11.3 per 100 patients.
T358 231003-231126 Sentence denotes The majority of NIs (37%) were identified in surgery, followed by intensive care medicine (30%) and general medicine (14%).
T359 231127-231441 Sentence denotes Surgical site infection was the most common NI (36 (29%) of all NI), followed by gastrointestinal tract infection (32 (27%) of all NI), respiratory tract infection (24 (19%) of all NI), urinary tract infection (20 (16%) of all NI), bloodstream infection (5 (3%) of all NI), and other infections (8 (6%) of all NI).
T360 231442-231632 Sentence denotes Escherichia coli was isolated most frequently (16%) followed by coagulase-negative Staphylococci (14%), Enterococcus faecium (9%), Enterococcus faecalis (8%) and Pseudomonas aeruginosa (8%).
T361 231633-232005 Sentence denotes Furthermore, the multivariate analysis identified gastrointestinal disease (OR=2.5; CI95% 1.6-3.8), disturbances of the electrolyte and fluid balance (OR=3.3; CI95% 1.1-10.5), history of hospital admission within 12 months (OR=1.6; CI95% 1.0-2.5), history of surgery within 12 months (OR=1.8; CI95% 1.1-3.1), history of antibiotic therapy within 6 months (OR=2.7; CI95% 1.
T362 232006-232127 Sentence denotes 4-5.1) , and for each device (OR=1.4; CI95% 1.2-1.7) as independent risk factors for occurrence of nosocomial infections.
T363 232128-232139 Sentence denotes Conclusion:
T364 232140-232229 Sentence denotes This study provides basic information about the current situation of NIs in our hospital.
T365 232230-232379 Sentence denotes This data is useful to provide an estimate of the health burden of NI and will also be helpful to initiate infection prevention and control measures.
T366 232380-232458 Sentence denotes Thorax-, Transplantationsund Gefäßchirurgie, Hannover, Deutschland Background:
T367 232459-232550 Sentence denotes Hand hygiene is the most effective measure for preventing infections related to healthcare.
T368 232551-232700 Sentence denotes This study aims to evaluate the hand hygiene behaviour of health care workers in a German university hospital in treatment of special patient groups.
T369 232701-232827 Sentence denotes In addiction compliance rates of different professional groups (doctors, nurses, other health care workers) were investigated.
T370 232828-232885 Sentence denotes Transplanted patients were chosen for this investigation.
T371 232886-233043 Sentence denotes They are high risk patients for developing healthcare associated infections due to the high number of devices, the long hospital stays and immunosuppression.
T372 233044-233052 Sentence denotes Methods:
T373 233053-233302 Sentence denotes During our investigation period (January 2008-December 2010), extensive education and training of all health care workers was implemented (at least three teaching units per ward).Consumption rates of hand rub and gloves were collected and evaluated.
T374 233303-233437 Sentence denotes Changes in healthcare workers hand hygiene compliance were calculated and compared between transplanted and not transplanted patients.
T375 233438-233507 Sentence denotes Therefore, the five WHOindications of hand hygiene were investigated.
T376 233508-233516 Sentence denotes Results:
T377 233517-233581 Sentence denotes Overall, 1607 opportunities of hand disinfection were evaluated.
T378 233582-233683 Sentence denotes A significant difference in hand disinfection behaviour was detected between the occupational groups.
T379 233684-233740 Sentence denotes Nurses hand hygiene compliance was significantly higher.
T380 233741-233881 Sentence denotes There was no significant difference in hand hygiene compliance between transplanted or not transplanted patients (OR 1.16, 95%CI 0.95-1.42).
T381 233882-233972 Sentence denotes The same effect was calculated for all healthcare workers and for the professional groups.
T382 233973-234087 Sentence denotes The disinfection was performed significantly more often after intervention than before (p>0.05, 95% CI 1.24-1.84).
T383 234088-234099 Sentence denotes Conclusion:
T384 234100-234223 Sentence denotes There is no difference in hand hygiene behaviour of health care workers between transplanted and not transplanted patients.
T385 234224-234280 Sentence denotes Nurses hand hygiene compliance was significantly higher.
T386 234281-234455 Sentence denotes The risk of no hand disinfection was significantly higher before the intervention, therefore more education and training will be needed to protect this special patient group.
T387 234456-234502 Sentence denotes Knowing where your infectious patients reside:
T388 234503-234563 Sentence denotes An early warning infection information system for hospitals.
T389 234564-234646 Sentence denotes Infectious disease outbreaks in hospitals are a severe threat to patients' safety.
T390 234647-234792 Sentence denotes Once the number of infected persons including hospital staff climbs to a threshold hospital departments may be closed or even the whole hospital.
T391 234793-235004 Sentence denotes Medical and administrative staff members especially those responsible for hospital hygiene management and infection control have a need to know where infectious patients are situated or should be transferred to.
T392 235005-235177 Sentence denotes An Infection Information System (IIS) was developed for the hospital operator MTG Malteserträgergesellschaft to support hospital professionals to monitor infected patients.
T393 235178-235320 Sentence denotes In case of an outbreak admission, discharge and transfer (ADT) operations can be monitored, controlled and analysed retrospectively over time.
T394 235321-235426 Sentence denotes The core component is a monitoring dashboard which visualizes all beds in hospital on a single PC screen.
T395 235427-235531 Sentence denotes Infections of patients are presented in one hospital with a colour code, in two others with a text code.
T396 235532-235660 Sentence denotes The text code allows for displaying multiple infections and is likely to become the preferred standard of the hospital operator.
T397 235661-235762 Sentence denotes So far Norovirus, ESBL, MRSA, Clostriods and other infections can be differentiated in the dashboard.
T398 235763-235887 Sentence denotes The data basis of IIS is collected data in its own data warehouse loaded from the central hospital information system (HIS).
T399 235888-236057 Sentence denotes Every 30 minutes a complete copy of the ADT situation is loaded into the database of the IIS including the notes on infections entered permanently by the hospital staff.
T400 236058-236181 Sentence denotes The IIS is operational since january 2011 at MTG and first feedback from users was encouraging to continue the development.
T401 236182-236333 Sentence denotes Especially the linkage between types of infection, localization of the patient and the overview in one screen is seen as a major benefit in daily work.
T402 236334-236404 Sentence denotes Monitoring dashboard using colour code to visualize infected patients.
T403 236405-236473 Sentence denotes Monitoring dashboard using text code to visualize infected patients.
T404 236474-236587 Sentence denotes Review of European guidelines for the prevention of C. difficile infections results form PROHIBIT WP2 Objectives:
T405 236588-236757 Sentence denotes The European Commission (FP-7) funded project "Prevention of Hospital Infections by Intervention and Training" (PROHIBIT; www.prohibit.unige.ch) was established in 2010.
T406 236758-237009 Sentence denotes PROHIBIT aims to analyse existing guidelines and practices to prevent healthcare associated infections (HAI) in Europe, to identify factors that influence compliance with best practices and to test the effectiveness of interventions of known efficacy.
T407 237010-237227 Sentence denotes WP 2 provides a review of current national and subnational (regional) European guidelines for the prevention of C. difficile infections (CDI) as well as an overview on surveillance systems and public reporting of CDI.
T408 237228-237511 Sentence denotes Methods: HAI surveillance National Contact Points (NCP) of the ECDC and other HAI experts in 34 countries (27 EU member states -whereby UK counts as 4 countries, Croatia, Iceland, Norway and Switzerland) were asked to provide their national/subnational guidelines for CDI prevention.
T409 237512-237605 Sentence denotes A review of the guidelines as well as a comparison with existing ECDC guidance was conducted.
T410 237606-237740 Sentence denotes In addition a questionnaire was developed to gather information about national surveillance systems and practices of public reporting.
T411 237741-237749 Sentence denotes Results:
T412 237750-237841 Sentence denotes Eighteen of 34 European countries have a national/subnational guideline for CDI prevention.
T413 237842-237904 Sentence denotes England, Northern Ireland and Wales follow the same guideline.
T414 237905-237996 Sentence denotes The oldest guideline dates back to 1995, all others were published within the past 4 years.
T415 237997-238104 Sentence denotes Seven of the 16 guidelines rate the scientific level of supporting evidence and strength of recommendation.
T416 238105-238212 Sentence denotes All but one guideline explicitly recommend placing symptomatic patients in a single room whenever possible.
T417 238213-238302 Sentence denotes Cleaning or disinfection of frequently touched surfaces is recommended in all guidelines.
T418 238303-238463 Sentence denotes However, only 13 include specific recommendation about disinfectants; out of these 11 prefer chlorine based solutions as specified in the ECDC guidance of 2008.
T419 238464-238515 Sentence denotes There is variation in hand hygiene recommendations.
T420 238516-238572 Sentence denotes Most guidelines favour hand washing with soap and water.
T421 238573-238663 Sentence denotes Alcoholic hand rub is mostly mentioned as an additional task to eradicate other pathogens.
T422 238664-238765 Sentence denotes Thirteen countries providing a CDI guideline also run a national surveillance system for the disease.
T423 238766-238875 Sentence denotes In Belgium, England, Northern Ireland, Scotland, Wales and Ireland participation is compulsory for hospitals.
T424 238876-239007 Sentence denotes In the UK and Ireland public reporting of CDI rates of individual hospitals has been established since 2007 and 2008, respectively.
T425 239008-239020 Sentence denotes Conclusions:
T426 239021-239175 Sentence denotes Although CDI is recognized as a major challenge in infection control only half of the European countries presently provide a guideline for CDI prevention.
T427 239176-239264 Sentence denotes Furthermore, there is important variation in scope and detailing of the recommendations.
T428 239265-239355 Sentence denotes The majority of guidelines do not rate the scientific level or strength of recommendation.
T429 239356-239489 Sentence denotes Countries with mandatory participation in a national surveillance system have introduced public reporting of CDI rates for hospitals.
T1 239490-239584 Sentence denotes Blended learning Kurs für Hygienebeauftragte Ärzteinnovativ, aktiv, zertifiziert Introduction:
T2 239585-239686 Sentence denotes There is a world-wide trend using wipes systems for the surface disinfection in health care settings.
T3 239687-239797 Sentence denotes However, some technical challenges have to be taken into consideration to assure a safe use of these products.
T4 239798-240010 Sentence denotes Recently, it has been shown, that active ingredients of surface disinfectants e.g. quaternary ammonium compounds can adsorp to the fabrics of wipes and impair the efficacy of the disinfectants significantly (1) .
T5 240011-240109 Sentence denotes Currently only methods are published testing the efficacy of liquid surface disinfectants (2, 3) .
T6 240110-240206 Sentence denotes Therefore, it was our aim to develop an easy and reproducible test method for this product type.
T7 240207-240228 Sentence denotes Material and Methods:
T8 240229-240314 Sentence denotes Basis of the new test method are the DGHMstandard method 14 (2) and the EN 13697 (3).
T9 240315-240466 Sentence denotes OR-tiles according to DGHM (2) were used as test surfaces and a standardized technique for direct application of the soaked wipes has been established.
T10 240467-240560 Sentence denotes In addition, the amount of disinfectant applied to the surface has been investigated Results:
T11 240561-240679 Sentence denotes Our results show that the suggested assay is suitable to test the practice like efficacy of disinfectant wipe systems.
T12 240680-240847 Sentence denotes In addition, the results show that not only the type of fabric (1) but also the amount of impregnation solution is determining the sufficient efficacy of the products.
T13 240848-241027 Sentence denotes There is a strong correlation between the amount of impregnation solution and the amount that can be transferred on the surface which is mandatory for the efficacy of the product.
T14 241028-241251 Sentence denotes We were able to show that the amount of disinfectant that can be transferred from a wipe onto a surface is significant lower than the amounts that are applied on the test carriers in the current published standards (2, 3) .
T15 241252-241263 Sentence denotes Discussion:
T16 241264-241389 Sentence denotes Now, a reproducible and easy standard method is available that can be used to show the microbicidal efficacy of wipe systems.
T17 241390-241572 Sentence denotes The authors strongly recommend testing the microbicidal efficacy of the wipe systems before practical use and before introducing these systems in positive lists such as VAH and SFHH.
T18 241573-241771 Sentence denotes Only wipes with the correct choice of fabrics (1) that are thoroughly impregnated with disinfectant solution are able to assure a sufficient efficacy and a safe use of the disinfecting wipe systems.
T19 241772-241785 Sentence denotes Introduction:
T20 241786-241931 Sentence denotes Shiga toxin-producing E. coli (STEC) are pathogens causing foodborne diarrhea, Hemorrhagic Colitis (HC), and the Hemolytic Uremic Syndrome (HUS).
T21 241932-242178 Sentence denotes In May 2011, a large outbreak of an O104:H4 STEC strain (HUSEC 41) expressing an ESBL-positive phenotype occurred with a hot spot in Schleswig-Holstein leading to great challenges in the laboratory diagnosis and infection control in the hospital.
T22 242179-242200 Sentence denotes Laboratory diagnosis:
T23 242201-242327 Sentence denotes From May 17 th patients with STEC diarrhea and HUS were admitted to the University Hospital Schleswig-Holstein, Campus Lübeck.
T24 242328-242541 Sentence denotes Having confirmed an ESBL-producing and Sorbitole-fermenting STEC clone as the causing agent within 24 hours, a chromogenic selective media for ESBL screening was immediately introduced in the diagnostic algorithm.
T25 242542-242638 Sentence denotes Shiga toxin-expression was analyzed from all E. coli isolates grown on media selective for ESBL.
T26 242639-242783 Sentence denotes For all stool specimens negative for ESBL-producing E. coli, Shiga toxin-expression was analyzed from an enrichment broth incubated in parallel.
T27 242784-242893 Sentence denotes From 80 of 129 (62 %) of patients with probable STEC infection stool specimens were tested in our laboratory.
T28 242894-242971 Sentence denotes In 58 stool specimen (72.5 %) STEC ESBL was confirmed as the causative agent.
T29 242972-243085 Sentence denotes None of the ESBL E. coli culture negative stool specimens was positive for Shiga toxin from the enrichment broth.
T30 243086-243178 Sentence denotes For all patients displaying symptoms of HUS, STEC ESBL infection could be confirmed (18/18).
T31 243179-243268 Sentence denotes Norovirus (3), Rotavirus (1) and Campylobacter jejuni (1) were detected in five patients.
T32 243269-243327 Sentence denotes No causative pathogen could be identified in 16 specimens.
T33 243328-243355 Sentence denotes Infection control measures:
T34 243356-243454 Sentence denotes A regular ward was immediately defined as quarantine ward for the management of affected patients.
T35 243455-243535 Sentence denotes Only few severely ill patients were treated on an ICU under barrier precautions.
T36 243536-243658 Sentence denotes During the peak of infections patients were directly examined at the quarantine ward to unburden the emergency department.
T37 243659-243790 Sentence denotes For patient management, barrier precautions with single usage coats and gloves for direct medical care procedures were established.
T38 243791-243879 Sentence denotes Medical face masks were suggested only for procedures with potential aerosol production.
T39 243880-243982 Sentence denotes No nosocomial transmission of STEC ESBL was observed indicating successful infection control measures.
T40 243983-243995 Sentence denotes Conclusions:
T41 243996-244193 Sentence denotes Introduction of selective chromogenic media for ESBLproducing Enterobacteriaceae proved to be a rapid and highly sensitive diagnostic procedure for the detection of the outbreak causing STEC clone.
T42 244194-244273 Sentence denotes Timely infection control measures circumvented nosocomial transmission of STEC.
T43 244274-244594 Sentence denotes Rapid screening method for multiple gastro-enteric pathogens also detects novel EHEC O104:H4 Patients suspected to be infected with EHEC O104:H4 and suffering from bloody diarrhoea bear the risk of developing the serious and life threatening haemolytic-uremic syndrome (HUS) and thus need a rapid differential diagnosis.
T44 244595-244789 Sentence denotes To avoid misdiagnosis potentially guiding false initial therapy or infection control measures rapid screening diagnostic methods are required and have been developed by specialized laboratories.
T45 244790-244993 Sentence denotes However, taking into account the inflation-like increasing number of newly suspected cases, the majority of laboratories has become overloaded, and the risk of delayed diagnosis increases proportionally.
T46 244994-245002 Sentence denotes Methods:
T47 245003-245074 Sentence denotes We thus tested the novel xTAG GPP assay from Luminex (Toronto, Canada).
T48 245075-245218 Sentence denotes This assay was developed to simultaneously detect the most important and serious viral, bacterial and parasitic pathogens in a single reaction.
T49 245219-245408 Sentence denotes Pathogens to be detected include Shiga-like Toxin producing E.coli, thus we assumed that also the novel variant which is Shigatoxin 1 negative but Shigatoxin 2 positive could be identified.
T50 245409-245417 Sentence denotes Results:
T51 245418-245558 Sentence denotes Until now a total number of 20 patients suffering from bloody diarrhoea or suspected to be infected with the novel EHEC variant were tested.
T52 245559-245720 Sentence denotes The multiplex testing revealed 4 patients positive for the novel EHC O104:H4 variant of which two were already independently confirmed by an external laboratory.
T53 245721-245914 Sentence denotes Two further patients suffered from severe Campylobacter infections and were tested negative for EHEC, one patient was tested positive for another EHEC variant that produced Shigatoxins 1 and 2.
T54 245915-245926 Sentence denotes Conclusion:
T55 245927-246025 Sentence denotes We conclude that the assay is useful to pre-screen patients suffering from the novel EHEC variant.
T56 246026-246141 Sentence denotes Patients can be monitored more closely by the clinicians and test results associated to the clinical course of HUS.
T57 246142-246458 Sentence denotes Additional beneficiary effects are that only a pre-selected cohort of clinical samples has to be analysed by the specialized laboratories and that those patients that are negative for EHC but positive for pathogens like Campylobacter spec. or Clostridium difficile can be administered the correct antibiotic therapy.
T58 246459-246767 Sentence denotes The Luminex technology is widely available and the assays procedure is rather simple with a total hands-on time of less than 5 hours and a sample capacity of up to 96 samples per run, the assays is suitable for high throughput analyses and thus will cover the peaks in the epidemiological outbreak situation.
T59 246768-246781 Sentence denotes Introduction:
T60 246782-246904 Sentence denotes Staphylococcal Scalded Skin Syndrome (SSSS) is a toxinmediated disease caused by exfoliative toxins A and B (ETA and ETB).
T61 246905-247021 Sentence denotes In December 2010/January 2011 five neonates developed bullous skin lesions in our neonatal department within a week.
T62 247022-247172 Sentence denotes Index patients were twins one of whom suffered from generalized exfoliation and thus had to be transferred to our neonatal intensive care unit (NICU).
T63 247173-247295 Sentence denotes The other one showed a localized desquamation at the rump and the occipital region and remained on the neonatal ward (NW).
T64 247296-247452 Sentence denotes Two additional cases occurred on the NICU thereafter with localized bullous lesions periorally as well as periorally, in the groin, and hands, respectively.
T65 247453-247561 Sentence denotes On the NW, one further case was identified presenting with a desquamation around a peripheral venous access.
T66 247562-247693 Sentence denotes In 4 of the cases microbiological samples showed Staphylococcus aureus isolates, which were sensitive to all antimicrobials tested.
T67 247694-247702 Sentence denotes Methods:
T68 247703-247764 Sentence denotes Contact precautions were initiated for the children affected.
T69 247765-247919 Sentence denotes Swabs of the nose and perineum were taken from all patients on both wards at three different time points within ten days in order to screen for S. aureus.
T70 247920-248027 Sentence denotes Environmental samples were performed on different sites and items on both wards (e. g. skin care products).
T71 248028-248146 Sentence denotes S. aureus isolates were sent to the German Reference Centre for Staphylococci for further characterization and typing.
T72 248147-248211 Sentence denotes They were subjected to susceptibility testing by microbroth MIC.
T73 248212-248300 Sentence denotes The presence of the genes for the exfoliative toxins A and B (eta and etb) was analyzed.
T74 248301-248341 Sentence denotes Furthermore the spa-type was determined.
T75 248342-248350 Sentence denotes Results:
T76 248351-248398 Sentence denotes All skin lesions resolved without any scarring.
T77 248399-248497 Sentence denotes No more newborns with desquamations of the skin were observed after precautions had been expanded.
T78 248498-248549 Sentence denotes No S. aureus was detected in environmental samples.
T79 248550-248622 Sentence denotes 36 children were hospitalized on both wards within the screening period.
T80 248623-248743 Sentence denotes Among those 12 S. aureus carriers were identified, two isolates of whom showed the same phenotype compared to the cases.
T81 248744-248895 Sentence denotes All four available isolates from the children affected and one isolate detected by screening a child without any symptoms were tested positive for eta.
T82 248896-248951 Sentence denotes All eta-positive strains belonged to the spa-type t084.
T83 248952-249067 Sentence denotes We observed an outbreak of SSSS caused by an exfoliative toxin A producing S. aureus strain within our institution.
T84 249068-249188 Sentence denotes The phenotype with all antimicrobials tested sensitive may indicate a community rather than a nosocomial primary source.
T85 249189-249334 Sentence denotes We assume that after introduction of the strain -maybe by the family of the twins first affected -further nosocomial transmissions have occurred.
T86 249335-249372 Sentence denotes However, the parents were not tested.
T87 249373-249543 Sentence denotes After initiation of expanded infection control measures and screening and eventually discharge of the last child that carried the strain no more cases have been observed.
T88 249544-249555 Sentence denotes Background:
T89 249556-249654 Sentence denotes Salmonella species are commonly isolated as pathogens of gastroenteritis in hospitalised patients.
T90 249655-249706 Sentence denotes Mostly, salmonella infection is community-acquired.
T91 249707-249717 Sentence denotes Objective:
T92 249718-249866 Sentence denotes To perform a prospective in-hospital surveillance of salmonella infection in order to detect possible nosocomial cases in an acute hospital setting.
T93 249867-249875 Sentence denotes Methods:
T94 249876-250063 Sentence denotes In three tertiary care hospitals all patients with detection of salmonella species were investigated in respect to whether the detection occurred later than 48 h after hospital admission.
T95 250064-250178 Sentence denotes Patients´charts were investigated for clinical symptoms and patients with nosocomial acquisition were interviewed.
T96 250179-250260 Sentence denotes Health care authorities (HCA) were informed about relevant interview information.
T97 250261-250269 Sentence denotes Results:
T98 250270-250355 Sentence denotes 2010 in 20, 4 and 31 patients Salmonella species were detected in hospital A, B or C.
T99 250356-250440 Sentence denotes In March 2010 two nosocomial cases with S. enteritidis occurred in hospital A and C.
T100 250441-250498 Sentence denotes Both hospitals were served by different canteen kitchens.
T101 250499-250580 Sentence denotes Patients reported having eaten a cheesecake in two different hospital-near cafés.
T102 250581-250671 Sentence denotes Informed HCA detected the use of raw eggs in the bakery serving both cafés and stopped it.
T103 250672-250734 Sentence denotes In September 2010 two nosocomial cases occurred in hospital A.
T104 250735-250784 Sentence denotes Again, patients reported having eaten cheesecake.
T105 250785-250906 Sentence denotes Involved HCA identified further two cases reporting consumption of cheesecake during a visit of a relative in hospital A.
T106 250907-250953 Sentence denotes The same bakery as in March had used raw eggs.
T107 250954-251048 Sentence denotes The baker and one person in the café were tested positive for S. enteritidis in stool samples.
T108 251049-251125 Sentence denotes For a defined period both persons were forbidden to work in food production.
T109 251126-251163 Sentence denotes No further cases occurred since then.
T110 251164-251175 Sentence denotes Conclusion:
T111 251176-251259 Sentence denotes A systematic surveillance of all salmonella-cases identified rare nosocomial cases.
T112 251260-251374 Sentence denotes Food available at food providers near the hospital should be considered if nosocomial salmonella infection occurs.
T113 251375-251536 Sentence denotes The interaction with HCA is essential for the identification and control of pathogens transmitted to hospitalised patients if acquired from outside the hospital.
T114 251537-251596 Sentence denotes Cluster of Adenovirus in a pediatric haemato-oncology ward:
T115 251597-251617 Sentence denotes Is this an outbreak?
T116 251618-251628 Sentence denotes Objective:
T117 251629-251732 Sentence denotes Human Adenovirus (HAdV) can cause severe infections especially in haemato-oncologic pediatric patients.
T118 251733-251836 Sentence denotes In our hospital contactand droplet-precautions are recommended when infections with HAdV are suspected.
T119 251837-252026 Sentence denotes Despite this recommendation HAdV was identified in stool and nasopharyngeal secretion samples of five children in a pediatric haematooncology ward within a two week period in February 2010.
T120 252027-252092 Sentence denotes Nosocomial transmissions were suspected and precautions expanded.
T121 252093-252238 Sentence denotes As a high compliance with precautions in ward staff was observed, the theory of an outbreak with patient to patient transmissions was questioned.
T122 252239-252246 Sentence denotes Method:
T123 252247-252307 Sentence denotes A retrospective epidemiological investigation was conducted.
T124 252308-252486 Sentence denotes All children seen in the children's hospital and tested positive for HAdV in a four week period (one week before and after the cluster on the haematooncology ward) were included.
T125 252487-252523 Sentence denotes HAdV was diagnosed via realtime-PCR.
T126 252524-252585 Sentence denotes Molecular typing was conducted with type-specific sequencing.
T127 252586-252593 Sentence denotes Result:
T128 252594-252716 Sentence denotes A total of eight children between seven months and 14 years of age were tested positive for HAdV in this four week period.
T129 252717-252796 Sentence denotes All had symptoms of gastroenteritis or upper respiratory tract infection (URI).
T130 252797-252910 Sentence denotes Five of eight children were hospitalized on the haemato-oncology ward, all at least two weeks prior to diagnosis.
T131 252911-253076 Sentence denotes Two children were seen only in the out-patient services; one child was hospitalized with URI on another ward and developed HAdV-diarrhoea three days after admission.
T132 253077-253155 Sentence denotes The molecular typing of seven isolates of HAdV showed four different genotyps:
T133 253156-253184 Sentence denotes 2xC1, 1xC6, 2xA31 und 1xF41.
T134 253185-253253 Sentence denotes Two children on the haemato-oncology ward had the same genotype A31.
T135 253254-253265 Sentence denotes Conclusion:
T136 253266-253411 Sentence denotes Immediate symptom-related implementation of specific precautions in children is successful in the prevention of patient-to-patient transmissions.
T137 253412-253551 Sentence denotes Possible reasons for the observed cluster may be transmission from parents or siblings or the reactivation of HAdV under immunosuppression.
T138 253552-253678 Sentence denotes Improved education and information of parents and other family members might be a useful additional effort in pediatric wards.
T139 253679-253690 Sentence denotes Objectives:
T140 253691-253818 Sentence denotes The role of infected and/or colonized health care workers (HCWs) as a potential source of a nosocomial outbreak is yet unclear.
T141 253819-253971 Sentence denotes The present study summarizes data from such outbreaks in order to elucidate areas of special interest and to clarify potential reasons for transmission.
T142 253972-253980 Sentence denotes Methods:
T143 253981-254154 Sentence denotes We conducted a systematic review of the medical literature based on PubMed, the Outbreak Database (www.outbreak-database.com), and on a hand search of so retrieved articles.
T144 254155-254285 Sentence denotes We extracted data on the setting, type of infection, number of patients, duration of the epidemic, and infection control measures.
T145 254286-254294 Sentence denotes Results:
T146 254295-254375 Sentence denotes A total of 152 outbreaks and 1,449 patients got included thereof 51 fatal cases.
T147 254376-254461 Sentence denotes The main route of transmission was direct or indirect contact (53% of all outbreaks).
T148 254462-254521 Sentence denotes Duration of the outbreaks ranged from 1 to 287 weeks (mean:
T149 254522-254539 Sentence denotes 28 weeks; median:
T150 254540-254552 Sentence denotes 10.5 weeks).
T151 254553-254625 Sentence denotes The number of affected patients per outbreaks ranged from 1 to 75 (mean:
T152 254626-254638 Sentence denotes 9.5; median:
T153 254639-254642 Sentence denotes 7).
T154 254643-254759 Sentence denotes Surgical (40%) and neonatological departments (24%) were most often involved in outbreaks primarily caused by staff.
T155 254760-254852 Sentence denotes Transmission took frequently place in surgical theaters (40%) and on peripheral wards (38%).
T156 254853-254942 Sentence denotes The primary types of infection were surgical site infections (27%) and hepatitis B (22%).
T157 254943-255075 Sentence denotes The corresponding causative main agents were Staphylococcus aureus (32%), hepatitis B virus (14%), and Streptococcus pyogenes (12%).
T158 255076-255214 Sentence denotes As shown in the figure, physicians (59 thereof 30 surgeons) and nurses (56) were the predominant staff spreading the pathogen to patients.
T159 255215-255272 Sentence denotes Nine per cent of them were aware of their carrier status.
T160 255273-255455 Sentence denotes When checking for the compliance to hand hygiene (HH) of the person causing the outbreak, we found that HH was considered adequate in 14% of the outbreaks but poor in 10.5% (table) .
T161 255456-255787 Sentence denotes The infection control measures most often implemented in order to terminate the outbreak included screening of personnel (90%) and patients (77%), education of staff and disinfection / sterilization procedures (53% each), application of antimicrobial substances (47%), improvement in HH (46%), and use of protective clothing (39%).
T162 255788-255799 Sentence denotes Conclusion:
T163 255800-255913 Sentence denotes Nosocomial outbreaks caused by staff are an important problem in the field of epidemiology and infection control.
T164 255914-256038 Sentence denotes In the next step we will check for characteristics of HCWs that may predispose for an increased pathogen spread to patients.
T165 256039-256226 Sentence denotes Staphylococcus (S.) aureus is both a prominent cause of nosocomial infections with significant morbidity and mortality and a commensal with nasal carriage in around 30% of the population.
T166 256227-256416 Sentence denotes The rapid spread of multiresistant strains necessitates novel therapeutic strategies; a challenging task, because the species S. aureus and the host response against it are highly variable.
T167 256417-256550 Sentence denotes Although S. aureus nasal carriers show an increased risk of endogenous S. aureus infection, they have a significantly better outcome.
T168 256551-256657 Sentence denotes We propose that humoral memory, acquired during colonization, shapes the response to S. aureus bacteremia.
T169 256658-256774 Sentence denotes In a prospective study among 2023 surgical and non-surgical patients twelve patients developed S. aureus bacteremia.
T170 256775-256892 Sentence denotes The infecting strains were isolated and patients' sera were obtained at diagnosis and during the course of infection.
T171 256893-256952 Sentence denotes They were analysed in detail using a personalized approach.
T172 256953-257117 Sentence denotes For each patient, the extra-cellular proteins of the infecting S. aureus strain were identified and the developing antibody response was assessed on 2D immunoblots.
T173 257118-257194 Sentence denotes S. aureus carriers showed clear evidence of strain specific preimmunisation.
T174 257195-257312 Sentence denotes In all immune-competent bacteremia patients, antibody binding increased strongly, in most cases already at diagnosis.
T175 257313-257369 Sentence denotes This observation may be exploited for earlier diagnosis.
T176 257370-257469 Sentence denotes In endogenous infections, the pattern of antibody binding was similar to the pre-infection pattern.
T177 257470-257598 Sentence denotes In exogenous infections, in contrast, the pre-infection pattern was radically altered with the acquisition of new specificities.
T178 257599-257649 Sentence denotes These were characteristic for individual patients.
T179 257650-257791 Sentence denotes Nevertheless, a common signature of ten conserved S. aureus proteins, recognized in at least half of the bacteremic patients, was identified.
T180 257792-257871 Sentence denotes All patients mounted a dynamic antibody response to a subset of these proteins.
T181 257872-258036 Sentence denotes 2D-analysis of the humoral S. aureus immunome proved to be a powerful tool for the coordinated analysis of the variable encounters of S. aureus with its human host.
T182 258037-258115 Sentence denotes Staphylococcus (S.) aureus is a frequent colonizer and also a common pathogen.
T183 258116-258185 Sentence denotes Furunculosis, a severe skin infection, is mainly caused by S. aureus.
T184 258186-258332 Sentence denotes Interestingly, the clinical isolates inducing furunculosis frequently harbour the pvl-genes, encoding the toxin Panton-Valentine-Leukocidin (PVL).
T185 258333-258426 Sentence denotes Even at low concentrations PVL is known to induce the lysis of human neutrophil granulocytes.
T186 258427-258484 Sentence denotes Neutrophils are essential for the clearance of S. aureus.
T187 258485-258659 Sentence denotes Besides degranulation and phagocytosis, neutrophils are able to kill microbes by releasing their DNA and antimicrobial peptides to form neutrophil extracellular traps (NETs).
T188 258660-258832 Sentence denotes To test whether invasive S. aureus strains that cause chronic furunculosis differ from colonizing strains we investigated their ability to induce cell death in neutrophils.
T189 258833-258943 Sentence denotes Neutrophils were stimulated with living bacteria or bacterial supernatants obtained in different growth media.
T190 258944-259035 Sentence denotes DNA release indicating cell death was quantified by exclusively staining extracellular DNA.
T191 259036-259103 Sentence denotes The morphology of cell death was determined by immune fluorescence.
T192 259104-259226 Sentence denotes Supernatants from S. aureus isolates from furunculosis patients but not from healthy nasal carriers lysed the neutrophils.
T193 259227-259315 Sentence denotes This was dependent on the presence of the pvl-genes and on the bacterial culture medium:
T194 259316-259437 Sentence denotes Only supernatants obtained in nutrient-rich media contained measurable amounts of PVL and caused necrosis of neutrophils.
T195 259438-259538 Sentence denotes At sublytic concentrations, PVL did not elicit NETosis (neutrophil cell death during NET formation).
T196 259539-259626 Sentence denotes Sublytic concentrations of bacterial supernatants did not induce neutrophil cell death.
T197 259627-259747 Sentence denotes In contrast, the life span of neutrophils was extended, while their ability of proinflammatory activation was conserved.
T198 259748-259829 Sentence denotes We can exclude indirect effects by modulating cytokines of contaminating T cells.
T199 259830-259887 Sentence denotes When exposed to living S. aureus cells, neutrophils died.
T200 259888-260037 Sentence denotes Depending on the multiplicity of infection we observed rapid DNA release, predominantly necrosis, or delayed cell death, mainly apoptosis or NETosis.
T201 260038-260147 Sentence denotes However, there were no differences between S. aureus cells from colonizing or furunculosis inducing isolates.
T202 260148-260366 Sentence denotes Opsonization of the S. aureus cells with human serum followed by coculture with neutrophils delayed the DNA release of neutrophils, caused higher IL-8 production and substantially reduced the numbers of vital bacteria.
T203 260367-260604 Sentence denotes We conclude that S. aureus isolates from nasal carriage and furunculosis infection differed in their ability to cause PVL-mediated neutrophil lysis, but their capacity to induce NET formation or apoptosis in neutrophils was very similar.
T204 260605-260708 Sentence denotes The Yersinia effector protein YopM uses the endocytic pathway to reach its intracellular destination J.
T205 260709-260726 Sentence denotes Scharnert* 1 , L.
T206 260727-260740 Sentence denotes Greune 1 , D.
T207 260741-260757 Sentence denotes Zeuschner 2 , C.
T208 260758-260772 Sentence denotes Rüter 1 , M.A.
T209 260773-260828 Sentence denotes Schmidt 1 1 ZMBE, Infektiologie, Münster, Deutschland 2
T210 260830-260954 Sentence denotes Cell-penetrating proteins (CPP) are able to cross cellular membranes, either alone or in association with a bioactive cargo.
T211 260955-261064 Sentence denotes We identified the Yersinia outer protein M (YopM) as a novel bacterial CPP with anti-inflammatory properties.
T212 261065-261162 Sentence denotes This implicates a potential use of YopM as an immunmodulatory agent for therapeutic applications.
T213 261163-261322 Sentence denotes To provide a significant advancement in understanding of YopM´s intracellular and molecular function the routes of internalization were analyzed in this study.
T214 261323-261416 Sentence denotes HeLa cells were incubated with YopM and analyzed by cryo-immunogold electron microscopy (EM).
T215 261417-261583 Sentence denotes For quantification fifty cell profiles were randomly selected and antibody signals were evaluated by number of gold particles linked to certain cellular compartments.
T216 261584-261710 Sentence denotes Furthermore, colocalization studies with specific marker proteins were performed using confocal immunofluorescence microscopy.
T217 261711-261803 Sentence denotes Additionally, localization of YopM was confirmed by cell fractionation and Western Blotting.
T218 261804-261909 Sentence denotes Cryo-immunogold EM revealed YopM throughout the endosomal compartments, but never in secretory membranes.
T219 261910-261982 Sentence denotes Free YopM could be detected throughout the cytoplasm and in the nucleus.
T220 261983-262058 Sentence denotes Additionally, the nuclear localization was confirmed by cell fractionation.
T221 262059-262180 Sentence denotes To further characterize the nature of the YopM-positive endosomal compartments immunofluorescence studies were performed.
T222 262181-262309 Sentence denotes We found that YopM colocalizes with specific endosomal markers characteristic for early and late endosomes as well as lysosomes.
T223 262310-262431 Sentence denotes Additionally, the absence of YopM at secretory membranes was confirmed, excluding the secretory pathway as a YopM target.
T224 262432-262542 Sentence denotes Our results show that recombinant YopM enters eukaryotic cells predominantly in an endosome-associated manner.
T225 262543-262702 Sentence denotes After escaping from the endosomes it localizes to the cytosol and partially in the nucleus, indicating an intracellular endosomal-independent movement of YopM.
T226 262703-262705 Sentence denotes O.
T227 262706-262722 Sentence denotes Witzke* 1 , F.M.
T228 262723-262741 Sentence denotes Heinemann 2 , P.A.
T229 262742-262753 Sentence denotes Horn 2 , M.
T230 262754-262767 Sentence denotes Lindemann 2 1
T231 262769-262782 Sentence denotes Introduction:
T232 262783-262963 Sentence denotes Vaccination against Streptococcus pneumoniae is recommended by the German advisory board for vaccination (STIKO) in immunocompromized patients such as kidney transplant recipients.
T233 262964-263065 Sentence denotes S. pneumoniae mainly infects the upper respiratory tract; it can cause lobar pneumonia or meningitis.
T234 263066-263139 Sentence denotes Morbidity and mortality are both increased in immunocompromized patients.
T235 263140-263276 Sentence denotes It was the aim of the present study to define the long-term efficiency of vaccination in clinically stable kidney transplant recipients.
T236 263277-263285 Sentence denotes Methods:
T237 263286-263387 Sentence denotes 49 patients (21 female, 28 male, median age 55, range 29-74 years) were immunized using Pneumovax 23.
T238 263388-263572 Sentence denotes Antibodies against 14 pneumococcal capsular polysaccharide antigens (serotypes) were determined prior to, one month and 15 months after vaccination by multiplexed bead assay (Luminex).
T239 263573-263581 Sentence denotes Results:
T240 263582-263821 Sentence denotes One month after vaccination, patients displayed a significant increase (p<0.0001) in the total antibody concentration against 14 pneumococcal serotypes from a median of 18.2 mg/l (range 2.9-55.5) prior to vaccination to 53.6 mg/l (range 4.
T241 263822-263832 Sentence denotes 5-132.4) .
T242 263833-264003 Sentence denotes Fifteen month after vaccination, the total antibody concentration was still significantly higher (p<0.0001) than prior to vaccination (median 41.3, range 4.9-105.0 mg/l).
T243 264004-264234 Sentence denotes In addition, the kidney transplant recipients showed a significant increase in the number of serotypes recognized from a median of 8 (range 0-13) to 13 (range 0-14, p<0.0001) at month 1 and to 11 (range 0-14, p=0.006) at month 15.
T244 264235-264488 Sentence denotes Antibody responses after vaccination were slightly, but significantly (p<0.0001, both at month 1 and 15) lower than in a published cohort of vaccinated, healthy controls [14 (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) , Borgers et al., 2009) ].
T245 264489-264500 Sentence denotes Conclusion:
T246 264501-264651 Sentence denotes Our results demonstrate that kidney transplant recipients can produce almost normal concentrations of antibodies against pneumococcal polysaccharides.
T247 264652-264754 Sentence denotes Fifteen month post vaccination, 77% of the initial antibody response (at month 1) remained detectable.
T248 264755-264877 Sentence denotes Cystic fibrosis is caused by a defective expression of the Cystic Fibrosis Transmembraneconductance Regulator (Cftr) gene.
T249 264878-264975 Sentence denotes Pulmonary symptoms of the disease are in particularchronic pulmonary inflammation and infections.
T250 264976-265108 Sentence denotes The pathophysiological mechanisms bywhich these changes are induced in the lungs of patients with cystic fibrosis requiredefinition.
T251 265109-265205 Sentence denotes This study found that Cftr deficiency in mice results in the upregulation andactivation of CD95.
T252 265206-265379 Sentence denotes CD95 activation is caused by increased ceramide concentrations in cysticfibrosis lungs, as revealed by genetic modifications that normalize pulmonary ceramideconcentrations.
T253 265380-265531 Sentence denotes However, the activation of CD95 further increases pulmonary ceramide levelsand results in a vicious cycle of CD95 activation and ceramide accumulation.
T254 265532-265744 Sentence denotes Genetic studies reveal that CD95 is crucially involved in the induction of aseptic inflammation, an increase inthe bronchial cell death rate, and an increased susceptibility to infection among Cftr-deficientmice.
T255 265745-265851 Sentence denotes All of these pathologies are at least partially corrected by heterozygosity of CD95 inCftr-deficient mice.
T256 265852-266016 Sentence denotes These findings identify CD95 as an important regulator of lung functionsin cystic fibrosis and suggest that CD95 may be a novel target for treating cystic fibrosis.
T257 266017-266028 Sentence denotes Objectives:
T258 266029-266093 Sentence denotes Staphylococcus aureus plays a role as commensal and as pathogen.
T259 266094-266192 Sentence denotes About 20 % of the human population is persistently colonized with S. aureus in the anterior nares.
T260 266193-266304 Sentence denotes We have shown that carriage tunes the adaptive immune system and results in a strain-specific humoral response.
T261 266305-266394 Sentence denotes The adaptive immune system can mobilize different effector mechanisms to a given antigen.
T262 266395-266585 Sentence denotes E.g., Ig class switch to IgG1 is driven by pro-inflammatory Th1 cells, whereas IgG4 production requires help from Th2 or Tregs that play a role in allergy and immune tolerance, respectively.
T263 266586-266702 Sentence denotes We have determined the Ig subclass composition of human anti-S. aureus antibodies, focussing on the IgG1/IgG4 ratio.
T264 266703-266711 Sentence denotes Methods:
T265 266712-266857 Sentence denotes Colonizing and invasive S. aureus strains and sera of carriers and S. aureus bacteremia patients were investigated using a personalized approach.
T266 266858-267025 Sentence denotes The extra-cellular proteins of each strain were separated by 2D SDS-Page, blotted, decorated with serum and binding of S. aureus-specific IgG1 and IgG4 was quantified.
T267 267026-267034 Sentence denotes Results:
T268 267035-267141 Sentence denotes In carriers, the IgG1 and IgG4 binding patterns to the colonizing S. aureus strain were highly individual.
T269 267142-267272 Sentence denotes This applied to the number of protein spots recognized by IgG1 and IgG4 and to the relative intensity of IgG1 versus IgG4 binding.
T270 267273-267385 Sentence denotes Some S. aureus antigens were bound by both IgGsubclasses, whereas others were selectively bound by IgG1 or IgG4.
T271 267386-267464 Sentence denotes The investigation of the S. aureus bacteremia patients showed similar results.
T272 267465-267602 Sentence denotes Overall, antibody binding increased during infection in immune competent patients, and new anti-S. aureus antibody specificities emerged.
T273 267603-267690 Sentence denotes The relative increase of binding differed between the IgG subclasses and protein spots.
T274 267691-267703 Sentence denotes Conclusions:
T275 267704-267788 Sentence denotes In summary, humans differ strongly in their antibody subclass response to S. aureus.
T276 267789-267899 Sentence denotes On the other hand, individual S. aureus antigens elicit heterogenous subclass profiles in the same individual.
T277 267900-268019 Sentence denotes We are currently determining the antigen signatures of IgG1-versus IgG4dominance in the antibody response to S. aureus.
T278 268020-268123 Sentence denotes Yersinia-induced apoptosis involves cleavage of MyD88 and concomitant deregulation of TLR signalling K.
T279 268124-268143 Sentence denotes Ruckdeschel* 1 , L.
T280 268144-268159 Sentence denotes Novikova 1 , N.
T281 268160-268175 Sentence denotes Czymmeck 1 , A.
T282 268176-268243 Sentence denotes Deuretzbacher 1 1 Universitätsklinikum Eppendorf, Institut für Med.
T283 268244-268453 Sentence denotes Mikrobiologie, Hamburg, Deutschland A wide variety of pathogenic microorganisms has evolved tactics to modulate host cell death or survival pathways for colonizing the host organism and establishing infection.
T284 268454-268594 Sentence denotes Yersinia enterocolitica triggers apoptosis in macrophages by injecting the YopP effector protein through type III secretion into host cells.
T285 268595-268781 Sentence denotes YopP deactivates several TLR-induced signaling pathways which impedes the development of a coordinated immune response and finally affects cell survival, leading to macrophage apoptosis.
T286 268782-268918 Sentence denotes Here, we show that Yersinia-induced apoptosis of human macrophages involves caspase-dependent cleavage of the TLR adapter protein MyD88.
T287 268919-269016 Sentence denotes In the same manner, overexpressed MyD88 is cleaved in HEK293 cells following apoptosis induction.
T288 269017-269112 Sentence denotes The processing of MyD88 occurs in the advanced stages of apoptosis when caspase-3 is activated.
T289 269113-269178 Sentence denotes Accordingly, caspase-3 cleaves MyD88 in in-vitro protease assays.
T290 269179-269304 Sentence denotes The processing of MyD88 is not restricted to Yersinia infection, but is also observed during staurosporine-induced apoptosis.
T291 269305-269396 Sentence denotes It appears that the cleavage of MyD88 uncouples the activation of TLR4 from NF-B signaling.
T292 269397-269528 Sentence denotes Thus, the disruption of MyD88 signaling in the execution of apoptosis may render the dying cells less sensitive to TLR stimulation.
T293 269529-269704 Sentence denotes This suggests that the cleavage of MyD88 is implicated in apoptotic death signaling by conferring immunogenic tolerance to the dying cell in response to microbial stimulation.
T294 269705-269707 Sentence denotes M.
T295 269708-269723 Sentence denotes Hausner* 1 , R.
T296 269724-269736 Sentence denotes Dumke 1 , E.
T297 269737-269745 Sentence denotes Jacobs 1
T298 269746-269873 Sentence denotes The cell wall-less bacterium Mycoplasma pneumoniae is a leading cause of both upper and lower respiratory infections in humans.
T299 269874-270002 Sentence denotes A crucial step in host colonization is the adhesin-mediated binding of this pathogen to the cells of the respiratory epithelium.
T300 270003-270199 Sentence denotes For cytadherence M. pneumoniae exhibits a complex membrane extension called attachment organelle which includes amongst other proteins the major adhesin P1 and the adherenceassociated protein P30.
T301 270200-270360 Sentence denotes In the present study we investigated the immune response of guinea pigs against M. pneumoniae over a time period of 21 days after a single intranasal infection.
T302 270361-270616 Sentence denotes ELISA, Western blot and FACS analysis have shown an early and strong serum immune response against the major adhesin P1 and a time-dependent reduction of adhesion of mycoplasmas to HeLa cells after pretreatment of the bacteria with the different antisera.
T303 270617-270752 Sentence denotes In a previously published work we created a chimeric protein (HP14/30) derived from functional adhesion regions of proteins P1 and P30.
T304 270753-270886 Sentence denotes A polyclonal antiserum against HP14/30 was able to decrease M. pneumoniae adherence to three different human cell lines to around 5%.
T305 270887-271075 Sentence denotes We used this chimeric protein as antigen for an immunization strategy to elicit an antibody response in the respiratory tract of guinea pigs which can reduce the adhesion of M. pneumoniae.
T306 271076-271233 Sentence denotes For this purpose, we combined the adherence inhibitory properties of HP14/30 and the mucoadhesive characteristics of the polysaccharide chitosan as adjuvant.
T307 271234-271331 Sentence denotes Guinea pigs were immunized intranasally (i.n.) with an optimized mixture of HP14/30 and chitosan.
T308 271332-271457 Sentence denotes Laboratory experiments confirmed the stability of the protein-chitosan mixture and the time-dependent release of the antigen.
T309 271458-271584 Sentence denotes The immune responses in the i.n. immunized animal group were compared with those obtained by subcutaneous (s.c.) immunization.
T310 271585-271736 Sentence denotes Preliminary tests have shown that i.n. immunization with chitosan and HP14/30 was able to induce higher mucosal IgA titre than s.c. immunization alone.
T311 271737-271998 Sentence denotes The results are promising to achieve increased levels of M. pneumoniae-specific secretory IgA in comparison to the conventional systemic immunization by using the advantages of chitosan for the nasal vaccination of host organisms with the characterized antigen.
T312 271999-272134 Sentence denotes The fibrinogen binding protein Fbl of Staphylococcus lugdunensis is not involved in the invasion into epithelial and endothelial cells.
T313 272135-272257 Sentence denotes Bacterial invasion of non-phagocytic host cells is an important pathogenicity factor for escaping the host defense system.
T314 272258-272488 Sentence denotes Several Staphylococci, such as Staphylococcus aureus, Staphylococcus saprophyticus and Staphylococcus epidermidis, are internalized in eukaryotic cells and this mechanism is discussed as an important part of the infection process.
T315 272489-272611 Sentence denotes Fibronectin binding proteins have been discussed as an important prerequisite for internalization described for S. aureus.
T316 272612-272710 Sentence denotes For S. epidermidis a fibronectin independent mechanism via HSP70 has been also recently described.
T317 272711-272793 Sentence denotes Two fibrinogen binding adhesins have been previously described for S. lugdunensis.
T318 272794-272962 Sentence denotes Fibrinogen binding proteins support the adhesion of bacteria to eukaryotic cells, but were not considered as prerequisite for the internalization process for S. aureus.
T319 272963-273043 Sentence denotes To date a fibronectin binding protein for S. lugdunensis has not been described.
T320 273044-273236 Sentence denotes We sought to investigate whether S. lugdunensis is internalized into the epithelial and endothelial cell line cells (5637 and EA.hy 926) using a previously described FACS-based invasion assay.
T321 273237-273402 Sentence denotes We could show that clinical isolates of S. lugdunensis were internalized into human urinary bladder carcinoma cell line 5637 and the endothelial cell line EA.hy 926.
T322 273403-273515 Sentence denotes Interestingly, all strains that invade into human urinary bladder carcinoma cell line 5637 bound to fibronectin.
T323 273516-273606 Sentence denotes In contrast to this, strains not binding to solid-phase fibronectin were not internalized.
T324 273607-273799 Sentence denotes The fibrinogen binding protein Fbl was not involved in this internalization process, since an isogenic fbl knockout mutant also invade into eukaryotic cells and unaltered bound to fibronectin.
T325 273800-274020 Sentence denotes The discovery of the internalization attribute of S. lugdunensis and its linkage to fibronectin binding suggests that the internalization of this important pathogen also occurs via a fibronectin mediated surface contact.
T326 274021-274023 Sentence denotes J.
T327 274024-274037 Sentence denotes Elias* 1 , A.
T328 274038-274052 Sentence denotes Tremmel 1 , U.
T329 274053-274293 Sentence denotes Vogel 1 1 Universität Würzburg, Institut für Hygiene und Mikrobiologie, Würzburg, Deutschland Laboratory staff working with Neisseria meningitidis should be offered immunization with available vaccines against serogroups A, C, W-135, and Y.
T330 274294-274690 Sentence denotes As part of our biological safety management we have performed a crosssectional study with 20 individuals between 18 and 50 years, who were vaccinated once with quadrivalent polysaccharide vaccine; the aims of the study were: estimation of the duration of protection by polysaccharide vaccine; and assessment whether revaccination with newly available quadrivalent conjugate vaccine was necessary.
T331 274691-274788 Sentence denotes Timepoints of immunization were retrospectively obtained from immunization cards of participants.
T332 274789-275131 Sentence denotes Serum-bactericidal activity testing is the most widely accepted surrogate parameter for assessment of vaccine effectiveness against meningococci; we tested sera using the tilt method (Maslanka et al., 1997) with reference strains M99 243594 ("CDC strain F8238"), C11, M01 240070, and M00 242975 for serogroups A, C, W135, and Y, respectively.
T333 275132-275389 Sentence denotes The minimal duration of protection was estimated by the intersection of the lower 95% prediction band of a linear model describing the temporal decline of the titre's logarithm (to the base 2) with a horizontal line representing the protective titre of 1:8.
T334 275390-275505 Sentence denotes Minimal duration of protection was 33.7, 25.2, 0.0, and 55.2 months for serogroups A, C, W135, and Y, respectively.
T335 275506-275580 Sentence denotes Mean protective duration, however, exceeded 100 months for all serogroups.
T336 275581-275736 Sentence denotes Individuals with titres below 1:8 were vaccinated with a quadrivalent conjugate vaccine and protective titres were restored after 4 weeks (data not shown).
T337 275737-275864 Sentence denotes In conclusion, our analyses might point to less effective immunization against serogroup W135 by a polysaccharide-only vaccine.
T338 275865-276005 Sentence denotes To corroborate this finding, additional measurements of the concentration of anticapsular IgG present in the sera of vaccinees are underway.
T339 276006-276232 Sentence denotes Haemophilus influenzae uses the PE surface protein to acquire human plasminogen and to evade innate immunity Haemophilus influenzae is an important respiratory pathogen and is responsible for a variety of infections in humans.
T340 276233-276419 Sentence denotes The nontypeable H. influenzae (NTHi) strain 3655 binds human plasminogen and we identify the adhesion protein Protein E (PE) of NTHi 3655 as an new and major plasminogen binding protein.
T341 276420-276536 Sentence denotes Plasminogen is a 92 kDa human plasma protein that is expressed by liver cells and is the key enzyme in fibrinolysis.
T342 276537-276650 Sentence denotes Plasminogen is composed of five homologous triple loop "kringle" domains and a C-terminal serine protease domain.
T343 276651-276745 Sentence denotes Plasminogen bound dose dependently to intact bacteria and also to recombinant PE of NTHi 3655.
T344 276746-276825 Sentence denotes Plasminogen also bound to ten different tested clinical H. influenzae isolates.
T345 276826-276922 Sentence denotes The plasminogen-PE interaction is mediated by lysine residues and is affected by ionic strength.
T346 276923-277046 Sentence denotes Plasminogen binding to the PE knock out strain (NTHi 3655 pe) is reduced by 65 %, as compared with the wild-type NTHi 3655.
T347 277047-277142 Sentence denotes In addition, plasminogen bound to PE which was ectopically expressed on the surface of E. coli.
T348 277143-277238 Sentence denotes The zymogen plasminogen can be converted by specific activators to the serine protease plasmin.
T349 277239-277371 Sentence denotes Activated plasmin controls complement attack by degrading C3b and also degrades extracellular matrix components, such as fibrinogen.
T350 277372-277565 Sentence denotes Plasminogen, when bound to intact NTHi 3655 or to PE is converted by human uPa to active plasmin which cleaved the synthetic substrate S-2251 and also the natural substrates C3b and fibrinogen.
T351 277566-277803 Sentence denotes The plasminogen-binding region were localized within the PE protein by using synthetic linear peptides and the major plasminogen-binding region was localized to a linear peptide that represents the 28 aa of the N-terminus, i.e. aa 41-68.
T352 277804-277905 Sentence denotes PE binds plasminogen as well as vitronectin and the two human plasma proteins compete for PE binding.
T353 277906-278094 Sentence denotes Thus, PE of H. influenzae is a major plasminogen-binding protein and PE bound human plasminogen and when converted to plasmin aids in immune evasion and contributes to bacterial virulence.
T354 278095-278189 Sentence denotes Hepatocyte-specific knockout of A20 augments defense against malaria liver stage infections X.
T355 278190-278202 Sentence denotes Wang* 1 , J.
T356 278203-278216 Sentence denotes Introduction:
T357 278217-278398 Sentence denotes During a malaria infection, Plasmodium sporozoites migrate through several host cells before finally infecting hepatocytes, where they develop into erythrocyte-infecting merozoites.
T358 278399-278607 Sentence denotes Cytosolic factors released from the traversed hepatocytes activate the NF-B pathway in adjacent hepatocytes, which, in turn, leads to the reduction in parasite load by inducing the expression of TNF and iNOS.
T359 278608-278794 Sentence denotes A20, an ubiquitin-editing enzyme, can block the proximal NF-B signalling pathway under various stimulations and A20 knockout mice succumb prematurely from uncontrolled hyperinflammation.
T360 278795-278962 Sentence denotes In order to explore the function of A20 in Plasmodium liver stage development, hepatocyte-specific A20-null mice were generated and investigated for malaria infection.
T361 278963-278984 Sentence denotes Material and methods:
T362 278985-279109 Sentence denotes Hepatocyte-specific A20 knockout mice were generated by utilizing the Cre/loxP system under control of the albumin promoter.
T363 279110-279269 Sentence denotes A20 was efficiently and specifically deleted in hepatocytes of the Alb-creA20fl/fl (KO) mice, while its expression was not affected in A20fl/fl (control) mice.
T364 279270-279436 Sentence denotes We applied blood stage parasitemia and quantitative RT-PCR of liver parasite loads as two complementary primary endpoints of a malaria sporozoite challenge infection.
T365 279437-279532 Sentence denotes Infiltrating leukocytes were isolated from livers and subpopulations were investigated by FACS.
T366 279533-279619 Sentence denotes Expression of cytokines and chemokines in infected livers was also assessed by qRTPCR.
T367 279620-279628 Sentence denotes Results:
T368 279629-279728 Sentence denotes Forty-two hours post infection, KO mice had lower liver parasite load as compared to A20fl/fl mice.
T369 279729-279917 Sentence denotes In the first few days after infection, parasitemia of KO mice was also significantly lower as compared to control mice, but this difference gradually disappeared as the disease progressed.
T370 279918-280029 Sentence denotes FACS analysis showed more infiltrating CD8 T cells, B cells, macrophages and inflammatory monocytes in KO mice.
T371 280030-280173 Sentence denotes Besides, higher levels of TNF, IL6, IP10, MCP1 and MCP5 were also detected in the liver of KO mice indicating a stronger inflammatory response.
T372 280174-280314 Sentence denotes Interestingly, mRNA levels of iNOS, which is responsible for producing anti-malarial NO, were also significantly higher in infected KO mice.
T373 280315-280508 Sentence denotes Our study demonstrates that A20 deletion in hepatocytes is beneficial for parasite clearance during malaria liver stage infection, presumably by inducing a stronger local inflammatory response.
T374 280509-280616 Sentence denotes Augmenting anti-liver stage defense mechanisms may further improve experimental malaria vaccine strategies.
T375 280617-280630 Sentence denotes Introduction:
T376 280631-280690 Sentence denotes Tuberculosis (TB) still is a major threat to global health.
T377 280691-280801 Sentence denotes According to the WHO nearly 1.7m people died in 2009 whereas nearly 10m people are infected with TB worldwide.
T378 280802-280942 Sentence denotes Since drug-resistant strains of Mycobacterium tuberculosis (Mtb) continue to rise, new ways of treating this disease have to be established.
T379 280943-281019 Sentence denotes Therefore a basic understanding of the infection mechanisms of TB is needed.
T380 281020-281144 Sentence denotes It has been shown that Mycobacteria infect their host by various mechanisms, eg. phagosome maturation inhibition (Via et al.
T381 281145-281223 Sentence denotes 1997 ) and inhibition of the inflammation after infection (Guenin-Mace, 2009).
T382 281224-281453 Sentence denotes Our group has found a lipid (Phenolic glycolipid, PGL) in the outer cell wall of Mycobacterium marinum (Mmar, a model organism for the research of Mtb) which might interfere with the innate immune system (Robinson et al., 2008) .
T383 281454-281561 Sentence denotes We are currently focused on establishing the link between PGL and the pathways of the innate immune system.
T384 281562-281648 Sentence denotes Material/Method: PGL was isolated from 7H9 cultures of Mmar and subsequently purified.
T385 281649-281764 Sentence denotes Human monocyte derived macrophages (HMDM) were isolated from Buffy Coats and cultured over 6 days in RPMI, 10% FCS.
T386 281765-281892 Sentence denotes PGL was coated onto RPC18 paramagnetic beads or heat-killed Bacillus Calmette-Guerin (HK-BCG) and chased into HMDM for 5 hours.
T387 281893-281956 Sentence denotes In case of the magnetic beads HMDM were co-stimulated with LPS.
T388 281957-281990 Sentence denotes After that the RNA was extracted.
T389 281991-282059 Sentence denotes The cytokines IL-6, IL-12p40 and TNF-alpha were analysed by qRT-PCR.
T390 282060-282196 Sentence denotes HMDM were transfected with a luciferase based reporter system for promotors of different signaling pathways of the innate immune system.
T391 282197-282273 Sentence denotes After infection with PGL-coated HK-BCG the luciferase activity was analysed.
T392 282274-282282 Sentence denotes Results:
T393 282283-282402 Sentence denotes Upon infection of HMDM with PGL-coated HK-BCG the amount of IL-12p40 RNA is greatly reduced after prolonged incubation.
T394 282403-282459 Sentence denotes Other cytokines like IL-6 or TNF-a show the same effect.
T395 282460-282575 Sentence denotes Even after the stimulation of HMDM with LPS and PGL at the same time cytokine RNA levels are significantly reduced.
T396 282576-282754 Sentence denotes The promotor activity for NF-kB and AP-1 after stimulation of HMDM with PGL-coated HK-BCG is greatly reduced in comparison to the activity after stimulation with uncoated HK-BCG.
T397 282755-282766 Sentence denotes Discussion:
T398 282767-282881 Sentence denotes Our findings suggest that PGL is indeed able to interfere with the signaling pathways of the innate immune system.
T399 282882-283007 Sentence denotes We could show that PGL supresses the production of IL-6, IL-12p40 and TNF-a RNA in HMDM after stimulation with HK-BCG or LPS.
T400 283008-283128 Sentence denotes The reduced promotor activity of NF-kB and AP-1 indicates that PGL interferes with pathways of the innate immune system.
T401 283129-283203 Sentence denotes It still remains elusive where exactly PGL interferes with these pathways.
T402 283204-283319 Sentence denotes These results show the importance of PGL for Mycobateria to be able to undermine the response of the immune system.
T403 283320-283418 Sentence denotes Our work could therefore help to complete the picture of the infection mechanisms of Mycobacteria.
T404 283419-283559 Sentence denotes The intracellular protozoan parasite Toxoplasma gondii can invade the brain and cause a persistent infection, toxoplasmic encephalitis (TE).
T405 283560-283697 Sentence denotes IFNproducing CD8 + T cells, activated microglia, macrophages, and astrocytes appear to be the major effector mechanisms in host immunity.
T406 283698-283830 Sentence denotes Infiltration of the brain depends on the expression of different adhesion molecules and on the integrity of the blood brain barrier.
T407 283831-283948 Sentence denotes The adhesion and degranulationpromoting adapter protein (ADAP) is expressed in T cells, myeloid cells, and platelets.
T408 283949-284098 Sentence denotes This adapter protein is involved in T cell receptor (TCR)dependent inside-out signaling and thereby regulates the activation and adhesion of T cells.
T409 284099-284153 Sentence denotes Little is known about the role of ADAP in brain cells.
T410 284154-284369 Sentence denotes Since we could demonstrate that ADAP-deficient (ADAP -/-) mice showed ameliorated disease progression in the experimental autoimmune encephalomyelitis (EAE) model, a function of ADAP in brain cells was hypothesized.
T411 284370-284488 Sentence denotes In this study, we performed low dose infection of ADAP -/mice and wildtype mice with T. gondii and monitored survival.
T412 284489-284691 Sentence denotes Furthermore, we examined the cyst load and inflammatory infiltrates using immunohistochemistry as well as the distribution of microglia cells, macrophages, and lymphocytes in the brain by FACS analysis.
T413 284692-284840 Sentence denotes Strikingly, ADAP -/mice showed significantly reduced survival in line with increased cyst load at day 44 after infection compared to wild-type mice.
T414 284841-285019 Sentence denotes Although the number of IFN-producing lymphocytes was increased in the brains of ADAP -/mice, all ADAP -/mice died until 54 days post infection, while all wild-type mice survived.
T415 285020-285128 Sentence denotes In contrast, the number of activated microglia cells and astrocytes was reduced in the brain of ADAP -/mice.
T416 285129-285350 Sentence denotes These data indicate an important role of ADAP for the activation of microglia cells during toxoplasmosis, whereas the integrin-mediated migration of T cells through the blood-brain-barrier seems to be independent of ADAP.
T417 285351-285468 Sentence denotes ADAP contributes to survival from TE and therefore plays a major role in maintaining immune homeostasis in the brain.
T418 285469-285567 Sentence denotes The staphylococcal protein Efb binds plasminogen and enhances plasmin-mediated C3 degradation T.K.
T419 285568-285584 Sentence denotes Enghardt* 1 , D.
T420 285585-285599 Sentence denotes Barthel 1 , M.
T421 285600-285615 Sentence denotes Reuter 1 , P.F.
T422 285616-285631 Sentence denotes Zipfel 1,2 , C.
T423 285632-285640 Sentence denotes Skerka 1
T1 285642-285779 Sentence denotes Staphylococcus aureus is a human pathogen that lives as a human commensal but can also cause superficial and life-threatening infections.
T2 285780-285968 Sentence denotes Upon infection, S. aureus induces host innate immune reactions, such as the activation of the complement system, recruiting of phagocytic cells and the secretion of antimicrobial peptides.
T3 285969-286083 Sentence denotes However S. aureus, as master of immune evasion, interferes particularly with the actions of the complement system.
T4 286084-286210 Sentence denotes Complement activation normally leads to opsonization and lysis of pathogenic microorganisms as well as inflammatory reactions.
T5 286211-286327 Sentence denotes S. aureus has developed multiple strategies to target and inhibit C3, the central molecule of the complement system.
T6 286328-286489 Sentence denotes S. aureus secretes Efb (extracellular fibrinogen binding protein), which directly binds the precursor C3 and the opsonin C3b to block the C3 convertase activity.
T7 286490-286629 Sentence denotes Upon binding to C3, Efb induces a conformational change in C3 leading to an enhanced susceptibility to degrading proteases such as trypsin.
T8 286630-286857 Sentence denotes Since S. aureus also recruits the human serum protein plasminogen, and subsequently activates plasminogen by Staphylokinase (SAK) to the C3/C3b-degrading plasmin, we asked whether Efb modulates the plasmin-mediated C3 cleavage.
T9 286858-286901 Sentence denotes Here we show that Efb recruits plasminogen.
T10 286902-287055 Sentence denotes Efb bound plasminogen is accessible for the human plasminogen activator uPa as well as for the bacterial SAK, both generating proteolytic active plasmin.
T11 287056-287146 Sentence denotes The interaction of Efb with C3 and plasmin leads to an enhances C3 degradation by plasmin.
T12 287147-287311 Sentence denotes Thus Efb targets C3 in two ways, directly by binding and inducing a conformational change in C3, and indirectly by facilitating the proteolytic activity of plasmin.
T13 287312-287504 Sentence denotes Type I and type III interferons (IFN) bind to different cell surface receptors but induce identical signal transduction pathways leading to the expression of antiviral host effector molecules.
T14 287505-287710 Sentence denotes Although IFN-l has been shown to predominantly act on mucosal organs such as the lung and gastrointestinal tract, studies using IFN-l receptor-deficient mice have failed to define a non-redundant function.
T15 287711-287893 Sentence denotes Here we show a protective role of IFN-signalling during rotavirus infection using suckling wild-type, type I IFN receptordeficient, IFN-l receptor-deficient and double-knockout mice.
T16 287894-288086 Sentence denotes Enhanced virus replication and tissue damage was found in the absence of IFN-l signalling, whereas no significant contribution of IFN-a/b signalling to the antiviral host defence was detected.
T17 288087-288344 Sentence denotes Quantitative RT-PCR and simultaneous immunostaining of virus-infected cells and the antiviral Mx1 protein highlighted the distinct responsiveness of intestinal epithelial cells to IFN-l and its critical role to restrict epithelial viral replication in vivo.
T18 288345-288479 Sentence denotes Thus, the antiviral host protection of the intestinal epithelium in response to rotavirus infection relies on intact IFN-l signalling.
T19 288480-288683 Sentence denotes Recognition of pathogenic microbes by intestinal epithelial cells results in transcriptional activation and secretion of soluble mediators that attract professional immune cells to the site of infection.
T20 288684-288837 Sentence denotes This defence mechanism works very efficiently despite the often low number of pathogens and the limited amount of mediators secreted per epithelial cell.
T21 288838-288974 Sentence denotes We have recently identified horizontal intercellular communication as critical component of an efficient epithelial innate host defence.
T22 288975-289166 Sentence denotes Chemokine production after L. monocytogenes infection was primarily observed in neighboring, noninfected cells despite the invasion-dependent nature of Listeria-induced epithelial activation.
T23 289167-289365 Sentence denotes Cell-to-cell communication was independent of gap junction formation, cytokine secretion, ion fluxes or nitric oxide synthesis, but required NADPH oxidase (Nox) 4-dependent oxygen radical formation.
T24 289366-289604 Sentence denotes In conclusion, our results provide a novel concept of a coordinated epithelial host response upon microbial challenge to maintain mucosal homeostasis and provide an efficient protection from infection with enteropathogenic microorganisms.
T25 289605-289817 Sentence denotes In human cells stimulation with IFN-results in a strong activation of the tryptophan degrading enzyme Indoleamine 2,3-dioxygenase (IDO) which is an important antimicrobial-and immunoregulatory effector mechanism.
T26 289818-289933 Sentence denotes IDO converts the essential amino acid tryptophan to formyl-kynurenine, which is channelled into kynurenine pathway.
T27 289934-290010 Sentence denotes IDO mediates its antimicrobial effects mainly by a starvation of tryptophan.
T28 290011-290125 Sentence denotes However, tryptophan metabolites produced in the kynurenine pathway are able to inhibit the growth of immune cells.
T29 290126-290369 Sentence denotes In this study we analysed the effect of kynurenine (Kyn), 3-hydroxy-kynurenine (3-HK), 3-hydroxy-anthranilic acid (3-HAA) and of quinolinic acid (QA) on the proliferation of T-cells, Staphylococcus aureus, Toxoplasma gondii and of tumor cells.
T30 290370-290528 Sentence denotes We found that Kyn, 3-HK and 3-HAA do not influence the growth of S. aureus, while these metabolites are able to inhibit the proliferative response of T-cells.
T31 290529-290642 Sentence denotes Additional experiments showed an influence of these metabolites on the growth of T. gondii in glioblastoma cells.
T32 290643-290774 Sentence denotes Detailed analyses indicate that this antiparasitic effect is due to a toxic effect of the tryptophan metabolites on the host cells.
T33 290775-290866 Sentence denotes This astonishing result was furthermore investigated, using different types of tumor cells.
T34 290867-291020 Sentence denotes We found that the strongest toxic effect was mediated by 3-HAA followed by 3-HK and Kyn, while QA did not influence the growth of all tumor cells tested.
T35 291021-291107 Sentence denotes In addition we detected a synergistic effect of Kyn and 3-HAA on tumor cell viability.
T36 291108-291269 Sentence denotes Quantitative analysis of the toxicity indicates that some tumor cells e.g. A549 cells (lung cancer cells) are more resistant against 3-HK and 3-HAA than T-cells.
T37 291270-291368 Sentence denotes In vivo many human tumor cells express IDO activity and are therefore able to produce kynurenines.
T38 291369-291585 Sentence denotes These metabolites might inhibit the activity of tumor specific T-cells, while the enhanced resistance of some tumor cells against the toxic effect of tryptophan metabolites might represent an immune escape mechanism.
T39 291586-291597 Sentence denotes Background:
T40 291598-291769 Sentence denotes Fibroblasts regulate epithelial cell differentiation, are responsible for wound healing, control inflammation process and seem to play an important role in cancerogenesis.
T41 291770-291913 Sentence denotes Bacteria and their products play as well are involved in prevention/promotion of chronic inflammatory diseases, in wound healing and in cancer.
T42 291914-291966 Sentence denotes The linkage might be the hyaluronan (HA) metabolism.
T43 291967-292109 Sentence denotes Bacteria produce hyaluronidases and bacterial products interact with Toll-like-receptors (TLRs) which also seem to be linked to HA metabolism.
T44 292110-292250 Sentence denotes We investigated how far microorganisms/microbial products influence the metabolism of HA in human dermal fibroblasts through TLR activation.
T45 292251-292526 Sentence denotes We stimulated human dermal fibroblasts with the staphylococcal toxins TSST-1 and enterotoxin B (SEB), with lipopolysaccharide (LPS), peptidoglycan (PGN) as well as with soluble products from clinical Staphylococcus aureus, S. epidermidis and Lactobacillus crispatus isolates.
T46 292527-292633 Sentence denotes The release of the chemokine IL-8 was analyzed by ELISA as well as by quantitative real-time PCR (RT-PCR).
T47 292634-292773 Sentence denotes Activation of TLRs (TLR [2] [3] [4] [5] [8] [9] , hyaluronan acid synthases (HAS 1-3) and hyaluronidases (HYAL 1-3) was analyzed by RT-PCR.
T48 292774-292782 Sentence denotes Results:
T49 292783-292966 Sentence denotes Fibroblasts responded to LPS, TSST-1, to soluble products of S. epidermidis and of L. crispatus with the release of high amounts of IL-8 and an up to 200-fold increase in mRNA levels.
T50 292967-293165 Sentence denotes The most pronounced changes in TLR expression were seen for TLR 4, TLR 5 and TLR 9 by S. epidermidis, for TLR4 by the staphylococcal superantigens TSST-1, SEB and by PGN, for TLR-2 and TLR-4 by LPS.
T51 293166-293291 Sentence denotes PGN activated IL-8 release predominantly through NOD1 activation, while the additional stimuli used MYD88 signalling as well.
T52 293292-293474 Sentence denotes In unstimulated fibroblasts the highest mRNA expression was seen for HAS 2 and HYAL 2, moderate expression was seen for HAS 3 and HYAL 3, no signal was obtained for HAS 1 and HYAL 1.
T53 293475-293573 Sentence denotes However, all microorganisms and the microbial products under study did not modulate HA metabolism.
T54 293574-293693 Sentence denotes Breakdown of hyaluronan by staphylococcal hyaluronidases did not change mRNA expression of HAS 1-3 or HYAL 1-3 as well.
T55 293694-293705 Sentence denotes Conclusion:
T56 293706-293834 Sentence denotes Our data clearly show that fibroblasts interact with microorganisms/microbial products to a different degree, e.g. through TLRs.
T57 293835-293933 Sentence denotes However, a possible role of HA metabolism in host-microorganism interacting needs further studies.
T58 293934-293936 Sentence denotes J.
T59 293937-293953 Sentence denotes Kehrmann* 1 , R.
T60 293954-293960 Sentence denotes Tatura
T61 293962-294096 Sentence denotes effector T cells is accused to influence the origin and course of several chronic infections, autoimmune diseases and certain tumours.
T62 294097-294309 Sentence denotes There is also evidence, that in septic patients, the percentage of Treg cells referred to all T cells increases during sepsis and that these patients show a sepsis-induced immunoparalysis and have a poor outcome.
T63 294310-294464 Sentence denotes Besides, an exact quantification of Treg cells is of great interest for elucidating the role of these cells for development and progress of many diseases.
T64 294465-294596 Sentence denotes There are even concepts of T cell therapy for appointed diseases to re-establish a balance between Treg cells and effector T cells.
T65 294597-294737 Sentence denotes Up to now, most studies quantified Treg cells by the analysis of surface markers and of the transcription factor Foxp3 using flow cytometry.
T66 294738-294898 Sentence denotes As to date, a constitutive expressed specific unique marker for human Treg cells is not available, an exact quantification on protein or RNA level is imprecise.
T67 294899-295091 Sentence denotes Recently it has been found that epigenetic differences in DNA methylation between Treg cells and effector T cells exist and that these differences can be used for quantification of Treg cells.
T68 295092-295223 Sentence denotes Treg specific demethylated region (TSDR) of FOXP3 has been demonstrated to be an applicable genomic region for Treg quantification.
T69 295224-295459 Sentence denotes Here, we present data of the quantification of Treg cells by a newly developed methylation sensitive real-time PCR assay using FOXP3-TSDR and compare these data with the analysis of Treg cells by flow cytometry (CD4 + CD25 ++ CD127 -).
T70 295460-295600 Sentence denotes Data of the quantification of Treg cells from blood of septic patients are compared with quantified Treg cells from blood of healthy people.
T71 295601-295603 Sentence denotes W.
T72 295604-295833 Sentence denotes Abdel Halim* 1 1 Osnabrück, Microbiology, Osnabrück, Deutschland Salmonella enterica serovar Typhimurium is a facultative intracellular pathogen which can enter host cells and replicates within a specialized membrane compartment.
T73 295834-295963 Sentence denotes Salmonella is also an interesting vehicle for the display of recombinant antigens to professional antigen presenting cells (APC).
T74 295964-296064 Sentence denotes Heterologous antigens can be expressed in Salmonella as fusions with recombinant or native proteins.
T75 296065-296311 Sentence denotes This approach has been used mainly to direct the expression of the desired antigen to a particular location of the bacterial cell and to increase the immunogenicity of foreign antigens by fusing them to proteins that could exert a carrier effect.
T76 296312-296522 Sentence denotes The type III secretion system (T3SS)-mediated translocation by Salmonella can be used for efficient delivery of heterologous antigens into the cytosol of APC, leading to stimulation of both CD4 and CD8 T-cells.
T77 296523-296648 Sentence denotes In this work, we investigated the use of a subset of effector proteins of the SPI2-encoded T3SS as SseJ, SifA, SseL and SteC.
T78 296649-296734 Sentence denotes These effectors proteins are associated with endosomal membranes after translocation.
T79 296735-296848 Sentence denotes Our In-vitro and in-vivo experiments for vaccination show that effector SseJ is the most suitable fusion partner.
T80 296849-296948 Sentence denotes In previous work it was shown that sifB promoter was the most efficient in-vivo inducible promoter.
T81 296949-297135 Sentence denotes Here we show that SseJ antigen fusion protein under control of the sifB promoter is most efficient in comparison with effector fusions under control of other in-vivo inducible promoters.
T82 297136-297414 Sentence denotes By comparison of various attenuated carrier strains, we observed that the htrA/purD double mutant strain can be used efficiently as attenuated carrier for vaccination and observed the efficient stimulation invitro T-cell proliferation by the delta sifA mutant strain as carrier.
T83 297415-297528 Sentence denotes The production of cytokines in response to microbial danger signals is regulated at the level of MAPK activation.
T84 297529-297641 Sentence denotes Dual specificity phosphatases (Dusp) control MAPK activity by dephosphorylating threonine and tyrosine residues.
T85 297642-297810 Sentence denotes Differential expression and inducibility of Dusp gene expression suggest that these phosphatases specify the outcome of MAPK activation in terms of cytokine production.
T86 297811-297900 Sentence denotes Here, we have investigated the role of Dusp16 (also known as MKP-7) in the immune system.
T87 297901-297992 Sentence denotes Dusp16 expression in macrophages and dendritic cells was inducible by TLR stimuli in vitro.
T88 297993-298140 Sentence denotes In vivo, Dusp16 expression was constitutive in some organs and in B lymphocytes; in contrast, LPS challenge up-regulated Dusp16 mRNA in the spleen.
T89 298141-298308 Sentence denotes A gene trap ES cell clone, bearing an insertion in the Dusp16 locus that abrogates the expression of full length Dusp16 mRNA, was used to generate mice lacking Dusp16.
T90 298309-298410 Sentence denotes Matings of heterozygous Dusp16trap/+ mice failed to yield homozygous Dusp16trap/trap mice at weaning.
T91 298411-298546 Sentence denotes However, embryonic development of homozygous mice was grossly normal until E18.5, indicating that Dusp16trap/trap mice die perinatally.
T92 298547-298674 Sentence denotes Fetal liver cells were used to generate macrophages in vitro and to reconstitute lethally irradiated mice for in vivo analysis.
T93 298675-298827 Sentence denotes Dusp16trap/trap macrophages responded normally to TLR stimulation for most cytokines analysed, but showed a significantly higher production of IL-12p40.
T94 298828-298925 Sentence denotes In vivo, Dusp16trap/trap fetal liver cells reconstituted T and B cell compartments similar to WT.
T95 298926-299097 Sentence denotes Following injection of LPS, Dusp16trap/trapreconstituted mice significantly over-produced IL-12p40, consistent with the phenotype of Dusp16-deficient macrophages in vitro.
T96 299098-299242 Sentence denotes Taken together, the production of the cytokine IL-12p40 in response to TLR stimulation in vitro and in vivo is selectively controlled by Dusp16.
T97 299243-299358 Sentence denotes Ongoing work will address the functional consequences of Dusp16-deficiency in models of inflammation and infection.
T98 299359-299489 Sentence denotes The cellular response of human monocyte derived macrophages to infection with genetically distinct M. tuberculosis complex strains
T99 299491-299508 Sentence denotes Aim of the study:
T100 299509-299754 Sentence denotes The Mycobacterium tuberculosis complex (MTBC) was initially regarded as a highly homogeneous population; however, recent data suggest the causative agents of tuberculosis are genetically and functionally more diverse than appreciated previously.
T101 299755-299877 Sentence denotes The impact of this natural variation on the virulence of the pathogen and clinical manifestations remains largely unknown.
T102 299878-299886 Sentence denotes Methods:
T103 299887-300031 Sentence denotes In this study we analyzed the host response of human primary monocyte derived macrophages upon infection with genetically distinct MTBC strains.
T104 300032-300191 Sentence denotes The study included strains that have caused larger outbreaks in Hamburg, Germany as well as attenuated strains, all of which belonging to the Haarlem genotype.
T105 300192-300299 Sentence denotes Supernatants were collected after 16h and monitored for the release of pro-and anti-inflammatory mediators.
T106 300300-300324 Sentence denotes Results and Conclusions:
T107 300325-300448 Sentence denotes All strains induced a comparable release of Tumor necrosis factor (TNF)-alpha, Interleukin (IL)-12, IL-6, IL-10 and RANTES.
T108 300449-300541 Sentence denotes A significant difference between clustered isolates and attenuated strains was not observed.
T109 300542-300756 Sentence denotes These data demonstrate that differences in strains success monitored in a longitudinal epidemiological study are not reflected at the level of mediator release after infection of human monocyte-derived macrophages.
T110 300757-301151 Sentence denotes However, additional data also demonstrate that a particular MTBC genotype translates into a distinct biological response profile when getting into contact with human immune cells. (Supp. in part by BMBF grant Project 01KI0784, WP D: "Pathogenic variability of clinical M. tb isolates: implications for the modulation of the immune system, infectivity and disease progression" to S.N. and N.R.).
T111 301152-301154 Sentence denotes J.
T112 301155-301172 Sentence denotes Strebovsky 1 , P.
T113 301173-301186 Sentence denotes Walker 1 , A.
T114 301187-301386 Sentence denotes Dalpke* 1 1 Universität Heidelberg, Dept. für Infektiologie, Heidelberg, Deutschland Activation of innate immune cells during infections has to be tightly controlled to avoid overshooting reactivity.
T115 301387-301573 Sentence denotes Suppressor of cytokine signaling (SOCS) proteins are well known feedback inhibitors of janus kinases (Jak) and signal transducer and activator of transcription (STAT) signaling pathways.
T116 301574-301692 Sentence denotes Unexpectedly, we observed that SOCS1, but no other members of the SOCS family, localized predominantly in the nucleus.
T117 301693-301786 Sentence denotes Nuclear localization was due to a so far unknown bipartite nuclear localization signal (NLS).
T118 301787-301892 Sentence denotes Using photoactivable SOCS1 constructs we confirmed rapid translocation from the cytoplasm to the nucleus.
T119 301893-301972 Sentence denotes Within the nucleus SOCS1 was highly mobile as demonstrated by FRAP experiments.
T120 301973-302123 Sentence denotes To further characterize the novel function of SOCS1 within the cell nucleus we generated NLS-mutants whose expression was restricted to the cytoplasm.
T121 302124-302231 Sentence denotes Thereby we identified genes that were differently regulated depending on the nuclear availability of SOCS1.
T122 302232-302319 Sentence denotes We were able to identify the NFkB component p65 as novel interaction partner for SOCS1.
T123 302320-302419 Sentence denotes SOCS1 bound to p65 within the nuclear compartment and increased p65 ubiquitination and degradation.
T124 302420-302528 Sentence denotes The SH2 domain of SOCS1 contributed to p65 binding whereas ubiquitination was mediated through the SOCS box.
T125 302529-302627 Sentence denotes Limitation of p65 availability resulted in reduced expression of a subset of NFkB dependent genes.
T126 302628-302686 Sentence denotes Mass spectrometry identified further interaction partners.
T127 302687-302870 Sentence denotes The results show that SOCS1 regulates the duration of NFkB signaling by means of its ubiquitin ligase activity thereby exerting a so far unrecognized function within the cell nucleus.
T128 302871-302996 Sentence denotes The nuclear function of SOCS1 is important to limit proinflammatory stimulation of innate immune cells by microbial products.
T129 302997-303177 Sentence denotes The mycobacterial cord factor trehalose-6,6-mycolate (TDM) is a major virulence factor of M. tuberculosis that is recognized by innate immune cells in a Syk-Card9 dependent manner.
T130 303178-303328 Sentence denotes Recently, we identified the C-type lectin receptor (CLR) Mincle as the receptor for the TDM and its synthetic analogue trehalose-6,6-dibehenate (TDB).
T131 303329-303494 Sentence denotes These two glycolipids are potent adjuvants that elicit mixed Th1/Th17 responses, whereas Toll-like receptor ligands such as CpG DNA induce exclusively Th1 responses.
T132 303495-303596 Sentence denotes CLR and TLR ligands trigger distinct profiles of cytokines and effectors in antigen presenting cells.
T133 303597-303734 Sentence denotes How these unique transcriptional responses are generated, and which role they play for Th1 and Th17 induction is incompletely understood.
T134 303735-303834 Sentence denotes TLR and CLR signaling activates the MAP-Kinase pathway and the transcription factor (TF) NF-kappaB.
T135 303835-303919 Sentence denotes In contrast, the latent TF NF-AT is activated upon CLR but not upon TLR stimulation.
T136 303920-304023 Sentence denotes However, little is known about the TF profile induced downstream of these constitutively expressed TFs.
T137 304024-304232 Sentence denotes Here we assessed the expression kinetics of inducible TFs in macrophages after TDB and CpG stimulation to understand similarities and differences between expression profiles elicited by the two types of PRRs.
T138 304233-304345 Sentence denotes TDB rapidly and directly induces early growth response genes Egr1, Egr2 and Egr3 expression dependent on Mincle.
T139 304346-304436 Sentence denotes Cebpbeta and Hif1alpha are upregulated at protein level after stimulation with TDB or CpG.
T140 304437-304512 Sentence denotes In ChIP experiments we observed Hif1alpha recruitment to the iNOS promoter.
T141 304513-304639 Sentence denotes Furthermore Hif1alpha deficiency in macrophages results in a strong reduction of NO release after TDB but not CpG stimulation.
T142 304640-304811 Sentence denotes Taken together, our data shed new light on the regulatory network controlling transcriptional activation induced by the CLR Mincle in response to microbial Mincle ligands.
T143 304812-304933 Sentence denotes The type I interferon induced protein IFIT-2 contributes to type I interferon mediated proinflammatory immune response A.
T144 304934-304951 Sentence denotes Siegfried* 1 , S.
T145 304952-304968 Sentence denotes Berchtold 1 , B.
T146 304969-304983 Sentence denotes Manncke 1 , T.
T147 304984-304994 Sentence denotes Ott 2 , I.
T148 304995-304997 Sentence denotes B.
T149 304998-305015 Sentence denotes Autenrieth 1 , E.
T150 305016-305305 Sentence denotes Bohn 1 1 Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Tübingen, Deutschland 2 IZKF-Serviceeinrichtung Transgene Tiere, Tübingen, Deutschland IFIT-2, also called ISG54, is a type I interferon induced protein and belongs to the family of tetratricopeptide repeat proteins.
T151 305306-305480 Sentence denotes IFIT2 is upregulated in various virus infections, as well as it is known that upregulation of IFIT2 leads to better survival of tumor patients after resection of tumors [1] .
T152 305481-305609 Sentence denotes It was reported that IFIT2 binds to the translation initiation factor eIF3c and therefore inhibits translation of proteins [2] .
T153 305610-305722 Sentence denotes Specifically IFIT2 may recognize and may prevent translation of capped viral mRNA lacking 2'-O methylation [3] .
T154 305723-306010 Sentence denotes Stimulation of interferon / Receptor (IFNAR) -/bone marrow derived macrophages (BMDM) demonstrates that TLR3 and TLR4 ligands as well IFN-induced IFIT-2 protein expression is type I interferon receptor dependent, indicating that IFIT-2 can be directly induced by type I interferons only.
T155 306011-306130 Sentence denotes In this study we want to elucidate the role of IFIT-2 as an effector of type I interferon induced biological processes.
T156 306131-306189 Sentence denotes For this purpose IFIT-2 knockout mice have been generated.
T157 306190-306322 Sentence denotes In order to investigate a potential role of IFIT-2 in inflammatory responses, IFIT-2 -/and IFIT-2 +/+ BMDM were stimulated with LPS.
T158 306323-306527 Sentence denotes IFIT-2 -/-BMDM show significantly reduced TNF-, IL-6, MCP-1 and MIP-2 secretion, indicating that IFIT-2 contributes to type I interferon mediated amplification of the LPS induced proinflammatory response.
T159 306528-306695 Sentence denotes In line with these findings, it could be demonstrated that IFIT-2 -/mice are less susceptible to LPS mediated endotoxin shock compared to littermate wildtype controls.
T160 306696-306844 Sentence denotes Taken together, this data demonstrate, that IFIT-2 contributes as an effector to type interferon I mediated amplification of inflammatory responses.
T161 306845-306930 Sentence denotes The vertebrate coagulation system plays a major role in hemostasis and wound healing.
T162 306931-307100 Sentence denotes Coagulation factor XIII (fXIII), a plasma transglutaminase, finally crosslinks a formed fibrin network to generate and stabilize a plasma clot and seal the injured site.
T163 307101-307231 Sentence denotes Evidence is accumulating which implies that coagulation can also contribute to the early innate immune response against pathogens.
T164 307232-307369 Sentence denotes Activation of the intrinsic pathway of coagulation, also known as the contact system, seems to play an important role in these processes.
T165 307370-307558 Sentence denotes Previous work has shown that it can be activated on the surface of many pathogenic bacterial species, which in turn leads to the generation of antimicrobial peptides and bacterial killing.
T166 307559-307727 Sentence denotes Here, we present data which demonstrate that contact system activation and especially human fXIII have beneficial effects during infections with Streptococcus pyogenes.
T167 307728-307887 Sentence denotes We find that the system is activated on the surface of these bacteria after exposure to human plasma, which is followed by an activation of thrombin and fXIII.
T168 307888-307977 Sentence denotes As a consequence of this interaction bacteria are immobilized and killed within the clot.
T169 307978-308183 Sentence denotes In vivo experiments using a streptococcal skin infection model revealed that fXIII -/mice develop more severe signs of pathologic inflammation at the focus of infection when compared with wildtype animals.
T170 308184-308329 Sentence denotes When analyzing skin biopsies from infected wildtype mice, bacteria were found clustered within the fibrin network at the local site of infection.
T171 308330-308434 Sentence denotes However, in infected fXIII -/animals, bacteria were widespread and scattered over a large affected area.
T172 308435-308620 Sentence denotes Treatment of wildtype mice with a human fXIII-concentrate three hours after bacterial inoculation dampened the systemic dissemination of S. pyogenes during the early phase of infection.
T173 308621-308796 Sentence denotes To proof that our in vitro and in vivo results also apply to the clinical situation we analyzed tissue biopsies of patients suffering from streptococcal necrotizing fasciitis.
T174 308797-309038 Sentence denotes As seen in the murine biopsies, we detected fXIII-mediated bacterial crosslinking to the fibrin network in these biopsies supporting the concept that coagulation and fXIII contribute to the early innate immune response against this pathogen.
T175 309039-309129 Sentence denotes A role for the lysosomal phospholipase A2 in immune responses to intracellular bacteria B.
T176 309130-309149 Sentence denotes Schneider* 1,2 , K.
T177 309150-309163 Sentence denotes Hagens 1 , J.
T178 309164-309178 Sentence denotes Shayman 3 , U.
T179 309179-309194 Sentence denotes Schaible 1, 2 1
T180 309196-309332 Sentence denotes Phospholipases form a ubiquitous class of enzymes that catalyse the cleavage of membrane phospholipids into smaller bioactive molecules.
T181 309333-309516 Sentence denotes Recently, a lysosomal phospholipase A2 (LPLA2) has been identified and shown to be highly enriched in alveolar macrophages, where it plays an important role in surfactant degradation.
T182 309517-309692 Sentence denotes LPLA2 knockout (KO) mice develop a phospholipidosis phenotype as demonstrated by a marked accumulation of phospholipids in alveolar and peritoneal macrophages, and the spleen.
T183 309693-309901 Sentence denotes Given the high expression of LPLA2 in macrophages and its importance in membrane homeostasis and phospholipid turnover, we hypothesized that LPLA2 contributes to host responses against intracellular bacteria.
T184 309902-310096 Sentence denotes In order to study the role of macrophage LPLA2 in response to intracellular bacteria, we performed functional in vitro analyses with peritoneal macrophages from wild type (WT) and LPLA2 KO mice.
T185 310097-310203 Sentence denotes In addition, we study the role of LPLA2 in antibacterial host responses against lung tuberculosis in vivo.
T186 310204-310449 Sentence denotes While LPLA2 deficient macrophages do not show defects in the killing of intracellular bacteria such as non-pathogenic E. coli or pathogenic Mycobacterium tuberculosis, we found that they are impaired in digesting intracellular bacteria in vitro.
T187 310450-310568 Sentence denotes Even non-pathogenic E. coli are not readily cleared and persist long after those in WT macrophages have been degraded.
T188 310569-310684 Sentence denotes Furthermore, absence of functional LPLA2 strongly affects the ability of macrophages to stimulate T cells in vitro.
T189 310685-310820 Sentence denotes In line with this, adaptive immunity is impaired in LPLA2 deficient mice upon infection with the lung pathogen M. tuberculosis in vivo.
T190 310821-311005 Sentence denotes In the absence of LPLA2, both the number and activation of M. tuberculosis specific CD4 T cells as well as Th1 cytokine levels are significantly reduced upon M. tuberculosis infection.
T191 311006-311185 Sentence denotes LPLA2 deficient mice present with a mild pathology in both lungs and spleens due to reduced cellular infiltration in both organs while bacterial loads are significantly increased.
T192 311186-311374 Sentence denotes Taken together, our data indicate that LPLA2 is involved in the clearance of bacteria by macrophages and the processing of bacterial antigens for subsequent generation of T cell responses.
T193 311375-311681 Sentence denotes Though (IL)-17A/F-producing T helper (TH)17 cells are potent inducers of tissue inflammation and have been associated with several chronic and autoimmune inflammatory diseases, the main function of TH17 cells appears to be the clearance of pathogens, which are not appropriate affected by TH1 or TH2 cells.
T194 311682-311946 Sentence denotes To evaluate the role of IL-17A or IL-17F and its protective role during Mycobacterium tuberculosis (Mtb) infection, we analyzed the outcome of experimental tuberculosis (TB) in IL-17Adeficient ( -/-) and IL-17RA -/mice, which is the receptor for IL-17A and IL-17F.
T195 311947-312135 Sentence denotes Interestingly, Mtb-infected IL-17A -/mice efficiently generated interferon-gamma (IFNgamma)-producing TH1 cells and IFNgammadependent effector responses after low dose (100 CFU) infection.
T196 312136-312361 Sentence denotes However, IL-17A -/mice were not able to control mycobacterial replication during the chronic phase of experimental TB after low dose infection and died earlier (200 days post infection (dpi)) than corresponding wildtype mice.
T197 312362-312485 Sentence denotes After high dose infection (1000 CFU), IL-17A -/mice rapidly died already during the early stage of Mtb-infection at 30 dpi.
T198 312486-312689 Sentence denotes The breakdown of controlling mycobacterial replication after low and high dose infection was associated with a modulation of gene expression of pro-inflammatory cytokines, chemokines and their receptors.
T199 312690-312898 Sentence denotes Especially the highly increased gene expression of IL-17F in IL-17A -/mice lead to the hypothesis that hypercompensation of IL-17A deficiency by IL-17F induce neutrophilia and susceptibility to Mtb infection.
T200 312899-313047 Sentence denotes In fact and in contrast to IL-17A -/mice, mice deficient for IL-17RA (the receptor for IL-17A and F) were resistant to high dose infection with Mtb.
T201 313048-313162 Sentence denotes So far, our findings lead to the conclusion that IL-17F accounts for susceptibility of Mtb-infected IL-17A -/mice.
T202 313163-313507 Sentence denotes Hence, IL-17A appears to play no direct role for protection against Mtb infection. (Supported by the Inflammation Research Excellence Cluster) Staphylococcus (S.) aureus is a frequent cause of severe skin and soft tissue infections in humans, the treatment of which is complicated by the emergence of multi-resistant and hyper-virulent strains.
T203 313508-313644 Sentence denotes The ability to control the infection is largely dependent on the rapid recruitment of polymorphonuclear neutrophilic granulocytes (PMN).
T204 313645-313828 Sentence denotes To gain more insight into PMN migration and host-pathogen interactions in vivo, we investigated a mouse model of S. aureus flank skin infection using intravital two-photon microscopy.
T205 313829-313938 Sentence denotes For this purpose we generated S. aureus reporter strains, which expressed codon-adapted fluorescent proteins.
T206 313939-314038 Sentence denotes PMN migration in response to infection in vivo was then visualized using LysM-EGFP transgenic mice.
T207 314039-314212 Sentence denotes After injection of the bacteria, we observed the rapid appearance of PMN in the extravascular space of the dermis and their directed movement towards the focus of infection.
T208 314213-314332 Sentence denotes This resulted in the accumulation of large numbers of cells, which led to the delineation of an abscess within one day.
T209 314333-314443 Sentence denotes Surprisingly, the direct encounter of bacterial cells by PMN did not always lead to an uptake of the pathogen.
T210 314444-314562 Sentence denotes Depletion of PMN or blocking G-protein coupled receptors (GPCR) lead to an uncontrolled proliferation of the bacteria.
T211 314563-314800 Sentence denotes Moreover, tracking of transferred labeled bone-marrow derived neutrophils showed that PMN recruitment to the site of infection is dependent on GPCR on the cells themselves, whereas IL-1-receptor was required on host cells other than PMN.
T212 314801-314959 Sentence denotes Finally, by using a reporter of the staphylococcal Agr quorum-sensing system, we were able to follow bacterial gene regulation in vivo on a single cell level.
T213 314960-315067 Sentence denotes Here, we observed that Agr activity could be localized to small areas at the margin of the infection focus.
T214 315068-315292 Sentence denotes Our results establish that two-photon microscopy is a powerful tool to investigate the dynamics of the immune response, bacterial cell location, and gene expression in vivo on a single cell level during S. aureus infections.
T215 315293-315374 Sentence denotes Absence of CYLD is essential for survival during murine listeriosis Introduction:
T216 315375-315553 Sentence denotes In infectious diseases, activating as well as inhibitory transcription factors are required to enable pathogen control and to prevent over-shooting pathological immune reactions.
T217 315554-315646 Sentence denotes CYLD is a deubiquitylating enzyme which plays a pivotal inhibitory role in immune responses.
T218 315647-315789 Sentence denotes CYLD downregulates NF-kB activity by the proteolysis of K63-linked ubiquitin from signal transducing molecules like TNFR, TLR2, TLR4 and CD40.
T219 315790-315919 Sentence denotes In addition to NF-B, CYLD is also known to inhibit MAPK and the tissue protective factor plasminogen activator inhibitor (PAI-1).
T220 315920-316073 Sentence denotes To gain insight into the function of CYLD in listeriosis, we infected C57BL/6 CYLD -/and wildtype (WT) mice with a lethal dose of Listeria monocytogenes.
T221 316074-316246 Sentence denotes The bacterial burden in both the mice groups were determined on days 1, 3 and 5 p.i., On day 5 p.i., tissues from liver and spleen were obtained for RT-PCR and WB analysis.
T222 316247-316402 Sentence denotes To further characterize the role of CYLD, functional assays like inhibition of fibrin production by Warfarin and in vivo IL-6 neutalisation were performed.
T223 316403-316491 Sentence denotes Finally CYLD was knocked down in WT mice by siRNA to show its importance in listeriosis.
T224 316492-316515 Sentence denotes Results and Discussion:
T225 316516-316672 Sentence denotes Whereas all WT mice succumbed to the infection up to day 7 p.i., CYLD -/mice survived the infection indicating that CYLD inhibits protective host responses.
T226 316673-316856 Sentence denotes A macroscopic examination revealed that WT mice suffered from multi-organ bleeding and succumbed to liver failure as shown by increased AST and ALT levels compared to the CYLD -/mice.
T227 316857-316928 Sentence denotes In contrast, the bacterial burden was only partially increased WT mice.
T228 316929-317077 Sentence denotes Additional WB analysis showed an increased activation of NF-B and MAPK as well as an increased production of protective IL-6 and IFN in CYLD -/mice.
T229 317078-317398 Sentence denotes Importantly, CYLD -/mice had increased fibrin deposition in the liver and upon inhibition of fibrin production by warfarin CYLD -/mice succumbed to listeriosis indicating that (i) CLYD inhibits fibrin production via inhibition of PAI-1 and (i) illustrating the importance of fibrin in limiting the spread of listeriosis.
T230 317399-317636 Sentence denotes In good agreement, IL-6 stimulation of CYLD -/hepatocytes resulted in a stronger activation of the transcription factors STAT1 and STAT3 and fibrinogen production as compared to WT hepatocytes in CLYD -/hepatocytes upon IL-6 stimulation.
T231 317637-317763 Sentence denotes Neutralization of IL-6 abolished the protective effect of CYLD-deficiency and resulted in multi-organ bleeding in CYLD -/mice.
T232 317764-317775 Sentence denotes Conclusion:
T233 317776-317844 Sentence denotes Absence of CYLD is essential for survival during murine listeriosis.
T234 317845-318027 Sentence denotes Our observation that neutralisation of CYLD in WT mice by siRNA partially protected WT mice from lethal listerioses identifies CYLD as a potential therapeutic target in listerioisis.
T235 318028-318152 Sentence denotes The complement system directs immune effector functions and modulates the intensity of innate and specific immune responses.
T236 318153-318214 Sentence denotes It is activated by extracellular pathogens and damaged cells.
T237 318215-318471 Sentence denotes Almost nothing is known about the role of the complement system in infections with intracellular bacteria.This project demonstrates its important role in C. psittaci lung infection using several complement factor and receptor knock-out (or deficient) mice.
T238 318472-318606 Sentence denotes Mice were intranasally infected with the non-avian C. psittaci strain DC 15 and sacrificed on days 4, 9, and 21 for detailed analysis.
T239 318607-318655 Sentence denotes Weight and clinical score were determined daily.
T240 318656-318732 Sentence denotes Complement activation occurred before symptoms of pneumonia became apparent.
T241 318733-318853 Sentence denotes Surprisingly, following a phase of partial protection, all complement factor C3 -/mice died until day 17 post infection.
T242 318854-318999 Sentence denotes Compared to wildtype controls, the mice lacking most complement effector functions were 100 times more susceptible to the intracellular bacteria.
T243 319000-319212 Sentence denotes During the phase of partial protection of the C3 -/mice until day 9, more granulocytes migrated into their inflamed lungs, proinflammatory cytokines were more elevated, Th1polarization seemed to be even stronger.
T244 319213-319353 Sentence denotes The bacterial load in the lung of the knock-out mice was significantly decreased, they lost less weight and their clinical score was better.
T245 319354-319479 Sentence denotes Experiments on additional knock-mice are performed to identify the responsible complement effector function downstream of C3.
T246 319480-319491 Sentence denotes In summary:
T247 319492-319653 Sentence denotes Our data indicate that C3-effector functions are harmful early in C. psittaci infection; increased Chlamydia uptake mediated by C3b might be the reason for that.
T248 319654-319845 Sentence denotes However, later in infection, when specific immunity becomes essential for defence, complement activation seems to be necessary for the fine-tuning of an otherwise deleterious immune response.
T249 319846-319984 Sentence denotes Our data show for the first time the strong influence of complement activation on the outcome of an infection with intracellular bacteria.
T250 319985-320315 Sentence denotes Lipoamide dehydrogenase (Lpd) of Pseudomonas aeruginosa binds vitronectin, the human terminal complement pathway regulator for complement evasion The Gram-negative bacterium Pseudomonas aeruginosa is an opportunistic human pathogen and a major cause of hospital-acquired infections, particularly in immuno-compromised individuals.
T251 320316-320461 Sentence denotes In order to cross the first line of host innate immune defence, P. aeruginosa has developed multiple strategies to escape host complement attack.
T252 320462-320633 Sentence denotes Complement resistance is crucial for bacterial virulence and several pathogens acquire host complement inhibitors such as vitronectin and Factor H to modulate this attack.
T253 320634-320956 Sentence denotes In the present study we show that P. aeruginosa binds human vitronectin, a plasma protein that regulates the terminal complement pathway and when bound to the surface of P. aeruginosa, vitronectin blocks the insertion and assembly of the terminal complement complex and thereby increases bacterial survival in human serum.
T254 320957-321125 Sentence denotes Upon incubation in vitronectin-depleted, complement active human serum, survival of P. aeruginosa was decreased as compared to bacterial survival in normal human serum.
T255 321126-321203 Sentence denotes Thus, confirming the protective role of vitronectin at the bacterial surface.
T256 321204-321306 Sentence denotes We identified Lipoamide dehydrogenase (Lpd) as the first vitronectin binding protein of P. aeruginosa.
T257 321307-321478 Sentence denotes Although Lpd was initially identified as a cytoplasmic protein, flow cytometry and electron microscopy experiments revealed that Lpd is localized on the bacterial surface.
T258 321479-321644 Sentence denotes In addition, Lpd was found in the outer membrane fraction of twenty different P. aeruginosa strains when analyzed by Western blotting using a specific Lpd antiserum.
T259 321645-321821 Sentence denotes Lpd bound vitronectin in a dose-dependent manner and heparin inhibited the Lpd/vitronectin binding, suggesting that Lpd binds one of the heparin binding domains of vitronectin.
T260 321822-322034 Sentence denotes By using recombinant fragments, the major Lpd interaction site within human vitronectin was located to amino acid residues 353-363, which are positioned within the Cterminal heparin binding domain of vitronectin.
T261 322035-322224 Sentence denotes Taken together, the surface exposed Lpd of P. aeruginosa binds human vitronectin, the terminal complement pathway regulator and thereby protect the bacteria from complement-mediated attack.
T262 322225-322286 Sentence denotes Yersinia enterocolitica causes an enteric food-borne disease.
T263 322287-322484 Sentence denotes In the mouse model of infection, Yersiniae cause systemic disease with replication in the small intestine, invading Payer's Patches (PPs) of the distal ileum, and disseminating to liver and spleen.
T264 322485-322727 Sentence denotes Y. enterocolitica carries a 70kb-virulence plasmid encoding for Type III secretion system (T3SS) and Yersinia outer Proteins (Yops), which paralyse phagocytes of the innate immunity enabling Yersiniae to survive and replicate extracellularly.
T265 322728-323041 Sentence denotes Further Yersinia is expected to release typical pathogen-associated molecular patterns (PAMPs) such as lipopolysaccharide (LPS), peptidoglycan fragments (PG), lipoprotein (LP) and the Yersinia V antigen (LcrV), which are recognized by the respective host receptors (PRRs) such as diverse TLRs or NOD1/2 receptors.
T266 323042-323239 Sentence denotes PRR signaling leads to release of pro-inflammatory cytokines (TNF, IL-6) as the first step in host defense activation followed by an antiinflammatory response such as secretion of IL-10 and IL-1Ra.
T267 323240-323510 Sentence denotes From this, we suggest that the infection process can be separated in two different modes of pathogen -host cell communication: short-distance effect = silencing of contacted cells and long-distance effect = release of PAMPs with recruitment and activation of phagocytes.
T268 323511-323798 Sentence denotes With the aim to unravel which PAMP-receptor is involved in control of Yersinia-infection, we used different ko-mice with defined defects in innate immunity (TLRs-und NODs -signaling pathway and defects in IL-10 and IL1-Ra production) for susceptibility testing towards Y. enterocolitica.
T269 323799-323893 Sentence denotes The results enable us to define 3 different groups of susceptibility: high-susceptible (Ye-HS:
T270 323894-324098 Sentence denotes MyD88-/-and TLR2-/-/4-/-), susceptible (Ye-S: C57BL/6, TLR2-/-, TLR4-/-, CD14-/-, TLR2-/-/CD14-/-, NOD1-/-, NOD2-/-, NOD1-/-/NOD2-/-, RIP2-/-) and high-resistant mice strains (Ye-HR: IL10-/-and IL1Ra-/-).
T271 324099-324359 Sentence denotes Histological analysis of spleen cryosections of C57BL/6 and IL10-/-mice revealed that Yersinia microcolonies grow within marginal zone of the white pulp in the T-cell areas and are tightly surrounded by infiltrated neutrophils, what leads to abscess formation.
T272 324360-324526 Sentence denotes Histological analysis of spleen cryosections of MyD88-/-mice revealed a homogenous bacterial spread through the whole red pulp and damage of the splenic architecture.
T273 324527-324724 Sentence denotes Further more, we obtained evidence that Yersinia susceptibility of mice is closely associated with dynamics of neutrophil recruitment to the site of infection due to host cell chemokine production.
T274 324725-324940 Sentence denotes IL10-/-mice recruit high amount of neutrophils into the peritoneal space in the first 4h of infection due to the high chemokine production (G-CSF, KC, MIP-2) what leads to initial inhibition of Yersinia replication.
T275 324941-325071 Sentence denotes TLR2-/-/4-/-and MyD88-/-mice are characterized by delayed neutrophil infiltration what leads to uncontrolled Yersinia replication.
T276 325072-325157 Sentence denotes These findings elicit the decisive role of PRRsignalling in Yersinia-infection model.
T277 325158-325312 Sentence denotes Peptides derived from virulence factor of E.coli inhibit TLR signaling and are new promising candidates for treatment of autoimmune diseases and sepsis F.
T278 325313-325329 Sentence denotes Roemmler* 1 , C.
T279 325330-325341 Sentence denotes Cirl 1 , T.
T280 325342-325566 Sentence denotes Miethke 1 1 Technical University Munich, Institute of Medical Microbiology, Immunology and Hygiene, München, Deutschland Toll like receptor signaling is known to be part of the first line defense of the innate immune system.
T281 325567-325741 Sentence denotes Over the last decade, the importance of TLR7 and 9 in autoimmune diseases, such as systemic lupus erythematosus (SLE), psoriasis and rheumathoid arthritis, has been revealed.
T282 325742-325956 Sentence denotes In SLE, inappropriate or excessive activation of TLR7 and 9 by self RNA or DNA sequences is involved in the observed activation of the pDC IFN-alpha pathway and autoreactive B-cell mediated autoantibody production.
T283 325957-326085 Sentence denotes Furthermore, the stimulation of TLR4 plays a key role in hyper inflammatory situations such as systemic inflammation and sepsis.
T284 326086-326311 Sentence denotes Up to date, there are rather unspecific immune modulators available for treatment, such as corticosteroids and cytostatic reagents (e.g. cyclophosphamide), which are limited in their efficiency and cause immense side effects.
T285 326312-326418 Sentence denotes Therefore, there is a need for new therapeutic strategies with a more specific inhibition of certain TLRs.
T286 326419-326572 Sentence denotes With TcpC (TIR domain containing protein C), we identified a virulence factor of uropathogenic E.coli (CFT 073) inhibiting MyD88-dependent TLR responses.
T287 326573-326749 Sentence denotes 1 To make it suitable for therapeutic approaches, we developed peptides derived from exposed loop regions of the TIR-domain of TcpC, mainly the BB-loop and the DD-loop peptide.
T288 326750-326896 Sentence denotes The BB-loop of other Tcps is known to inhibit TLR-signaling and the DD-loop is a highly conserved domain among bacterial pathogens harboring Tcps.
T289 326897-327124 Sentence denotes Upon stimulation of TLR4, TLR7 and TLR9, both peptides inhibit cytokine production, including TNF-alpha, KC, and IL-12, in bone marrow derivedmacrophages (BMDM) and -dendritic cells (BMDC) in vitro without showing any toxicity.
T290 327125-327251 Sentence denotes Moreover, peptides show promising effects in a murine LPS shock-model leading to significantly reduced TNF-alpha serum levels.
T291 327252-327444 Sentence denotes In conclusion, the TcpC-derived peptides are able to inhibit TLR signaling without showing any side effects and are promising candidates for new therapeutics in autoimmune diseases and sepsis.
T292 327445-327456 Sentence denotes Objectives:
T293 327457-327542 Sentence denotes Invariant human Natural Killer T Cells (iNKT-Cells) are innate immune effector cells.
T294 327543-327695 Sentence denotes They are characterized by their expression of both, T-and NK cell markers and therefore they are a connecting link between innate and adaptive immunity.
T295 327696-327844 Sentence denotes These cells are further defined by their T-Cell receptor, consisting of a specific, invariant -chain and a -chain derived from a limited repertoire.
T296 327845-328097 Sentence denotes They strongly react upon stimulation with lipid antigens presented in the context of the MHC-like molecule CD1d on dendritic cells (DCs) and thus they can be expanded in vitro by stimulation with their prototypic agonist -galactosylceramide ( -GalCer).
T297 328098-328245 Sentence denotes A protective role of iNKT cells in autoimmune diseases and cancer immunity, as well as their action against different pathogens is well documented.
T298 328246-328352 Sentence denotes However, little is known about their direct interaction with fungi, especially with Aspergillus fumigatus.
T299 328353-328509 Sentence denotes A. fumigatus, an omnipresent mold, is the most prevalent cause of a highly devastating opportunistic infection, affecting mainly immunocompromised patients.
T300 328510-328518 Sentence denotes Methods:
T301 328519-328626 Sentence denotes In order to expand human iNKT cells (iNKT CD3 ), PBMCs were treated with -GalCer and rhIL-2 for 15-20 days.
T302 328627-328686 Sentence denotes After expansion, purity of the iNKT+CD3+ fraction was >90%.
T303 328687-328775 Sentence denotes To further purify this fraction, a positive selection with CD3 microbeads was performed.
T304 328776-328921 Sentence denotes After that the purity increased above 95%. iNKT cells were cocultured with different morphologies of A. fumigatus, resting conidia and germlings.
T305 328922-329066 Sentence denotes Gene induction was evaluated by microarray analysis (Affymetrix U219) and the protein release by using multiplex ELISA assays (BioRad Bio-Plex).
T306 329067-329157 Sentence denotes Additionally, the induction of IFN-, a major Th1 cytokine, was analyzed by flow cytometry.
T307 329158-329247 Sentence denotes Finally, XTT assays were used to examine the toxic effects of iNKT cells on A. fumigatus.
T308 329248-329256 Sentence denotes Results:
T309 329257-329442 Sentence denotes Stimulating iNKT cells with different A. fumigatus morphologies, we observed a time-and morphotype-dependent induction of INF-, with germlings to be shown more immunogenic than conidia.
T310 329443-329587 Sentence denotes Furthermore, challenging iNKT cells with A. fumigatus germlings at an MOI=1 for 6h, we found that iNKT cells caused a significant fungal damage.
T311 329588-329715 Sentence denotes We could also observe defined patterns of gene induction, especially regarding cytokine and chemokine gene expression profiles.
T312 329716-329786 Sentence denotes By multiplex ELISA assays gene expression patterns could be confirmed.
T313 329787-329798 Sentence denotes Conclusion:
T314 329799-329949 Sentence denotes We found that there is a specific interaction of iNKT cells with A. fumigatus revealed, leading to defined cytokine induction and a fungicidal effect.
T315 329950-330074 Sentence denotes INF-, a cytokine, which has a known protective role against IA, is produced by iNKT cells when confronted with A. fumigatus.
T316 330075-330271 Sentence denotes The course of Plasmodium yoelii-infection is affected by CD4 + Foxp3 + regulatory T-cells Regulatory T-cells (Tregs) play a critical role in the control of a variety of different immune responses.
T317 330272-330385 Sentence denotes However, studies on the impact of Tregs during the course of Plasmodium-infection have shown conflicting results.
T1 330386-330739 Sentence denotes Here we demonstrate that Plasmodium yoelii (P. yoelii)-infection of BALB/c mice leads to an expansion of highly suppressive CD4 + Foxp3 + Tregs, while its diphtheria toxin-mediated depletion in P. yoelii-infected DEREG mice resulted in a significant increased activation of CD4 + and CD8 + T-cells, accompanied by a significant decrease in parasitaemia.
T2 330740-331000 Sentence denotes Gene expression analysis of CD4 + T-cells from P. yoelii-infected mice revealed that molecules described in the context of Tregs are not only up-regulated in CD4 + CD25 + Foxp3 + Tregs but also in CD4 + CD25 + Foxp3 -T-cells, e.g. CTLA-4, PD-1, Lag-3 or IL-10.
T3 331001-331287 Sentence denotes Due to the fact, that P. yoelii-infection induce a strong up-regulation of anti-inflammatory IL-10 in both CD4 + CD25 + Foxp3 + Tregs and CD4 + CD25 -Foxp3 -T cells, we analyzed the impact of CD4 + T-cell derived IL-10 in more detail by using the IL-10 fl/fl x CD4cre (CD4IL10-KO) mice.
T4 331288-331547 Sentence denotes Interestingly, P. yoelii-infection of CD4IL10-KO mice leads to a reduced suppressive capability of CD4 + CD25 + Tregs and a significant enhanced inflammatory CD8 + T-cell response compared to WT mice, whereas the parasitic burden is only marginallyl affected.
T5 331548-331733 Sentence denotes The protozoan parasite Trypanosoma cruzi, the causative agent of human Chagas' disease, is an obligatory intracellular parasite that replicates in the cytoplasm of the parasitized cell.
T6 331734-331906 Sentence denotes Although T. cruzi is able to infect almost any nucleated cell in the body, macrophages represent an important site for T. cruzi replication in the acute phase of infection.
T7 331907-332134 Sentence denotes However, when a T helper (Th)1-based immune response develops, interferon (IFN)-gammaactivated macrophages are the primary immune cells that control T. cruzi replication by the production of reactive nitric intermediates (RNI).
T8 332135-332271 Sentence denotes In this study, we analyzed the role of interleukin (IL)-23 in protection against T. cruzi infection in a mouse model of Chagas' disease.
T9 332272-332373 Sentence denotes Compared to wildtype mice, IL-23p19 -/mice developed a higher parasitemia and an increased mortality.
T10 332374-332450 Sentence denotes However, this susceptibility was not due to an impaired Th1 immune response.
T11 332451-332621 Sentence denotes Because IL-17A production was reduced in IL-23p19 -/mice, we infected IL-17A -/mice with T. cruzi to study the relevance of this cytokine for protective immune responses.
T12 332622-332743 Sentence denotes Like IL-23p19 -/mice, IL-17A -/mice exhibited a higher parasitemia, an elevated mortality and an altered liver pathology.
T13 332744-332829 Sentence denotes Moreover, Th1 immune responses were not affected by the absence of endogenous IL-17A.
T14 332830-332980 Sentence denotes In vitro studies revealed that IL-17A directly stimulated trypanocidal activities in macrophages leading to the eradication of internalized parasites.
T15 332981-333291 Sentence denotes In contrast to unstimulated macrophages, in which trypanosomes quickly escaped from the parasitophorous vacuole into the cytoplasm, IL-17A-treatment of macrophages led to a prolonged residence of parasites in endosomal/lysosomal compartments, where they were vulnerable to the lytic activity of the macrophage.
T16 333292-333470 Sentence denotes Taken together, our results suggest that the IL-23-IL-17A-axis is necessary for host protection against T. cruzi infection by stimulating antimicrobial mechanisms in macrophages.
T17 333471-333482 Sentence denotes Background:
T18 333483-333738 Sentence denotes The health beneficial effects of Resveratrol and Curcumin, compounds found in the red wine grape and curry spices, respectively, as well as of the cholesterol-lowering drug Simvastatin have been demonstrated in various experimental models of inflammation.
T19 333739-333953 Sentence denotes We investigated the potential anti-inflammatory and immunomodulatory mechanisms of the above mentioned compounds in a murine model of hyper-acute Th1-type ileitis following peroral infection with Toxoplasma gondii.
T20 333954-333985 Sentence denotes Methodology/Principal Findings:
T21 333986-334206 Sentence denotes Here we show that after peroral administration of Resveratrol, Curcumin or Simvastatin, mice were protected from ileitis development and survived the acute phase of inflammation whereas all Placebo treated controls died.
T22 334207-334277 Sentence denotes In particular, Resveratrol treatment resulted in longer-term survival.
T23 334278-334520 Sentence denotes Resveratrol, Curcumin or Simvastatin treated animals displayed significantly increased numbers of regulatory T cells and augmented intestinal epithelial cell proliferation/ regeneration in the ileum mucosa compared to placebo control animals.
T24 334521-334621 Sentence denotes In contrast, mucosal T lymphocyte and neutrophilic granulocyte numbers in treated mice were reduced.
T25 334622-334924 Sentence denotes In addition, levels of the anti-inflammatory cytokine IL-10 in ileum, mesenteric lymph nodes and spleen were increased whereas pro-inflammatory cytokine expression (IL-23p19, IFN-g, TNF-a, IL-6, MCP-1) was found to be significantly lower in the ileum of treated animals as compared to Placebo controls.
T26 334925-335099 Sentence denotes Furthermore, treated animals displayed not only fewer pro-inflammatory enterobacteria and enterococci but also higher anti-inflammatory lactobacilli and bifidobacteria loads.
T27 335100-335283 Sentence denotes Most importantly, treatment with all three compounds preserved intestinal barrier functions as indicated by reduced bacterial translocation rates into spleen, liver, kidney and blood.
T28 335284-335308 Sentence denotes Conclusion/Significance:
T29 335309-335531 Sentence denotes Oral treatment with Resveratrol, Curcumin or Simvastatin ameliorates acute small intestinal inflammation by downregulating Th1-type immune responses and prevents bacterial translocation by maintaining gut barrier function.
T30 335532-335654 Sentence denotes These findings provide novel and potential prophylaxis and treatment options of patients with inflammatory bowel diseases.
T31 335655-335841 Sentence denotes Helminths modulate the immune system of their hosts and induce a regulatory, anti-inflammatory milieu that enables not only parasite survival within the host, but also benefits the host.
T32 335842-336019 Sentence denotes Thus, helminth infections prevent or ameliorate autoimmune diseases like Type I diabetes, rheumatoid arthritis, multiple sclerosis, and inflammatory bowel disease (IBD) (1, 2) .
T33 336020-336144 Sentence denotes With regard to IBD, this concept led to the FDA approval to treat patients with eggs of the porcine whipworm Trichuris suis.
T34 336145-336558 Sentence denotes As sepsis is one of the major causes of mortality that is partly due to an exaggerated pro-inflammatory immune response (3), we tested whether natural infection with the rodent filarial nematode Litomosoides sigmodontis ameliorates a systemic inflammatory immune response that is induced via i.p. injection of a sublethal dose of E. coli LPS in C57BL/6 mice 35 dpi or vital E. coli bacteria 90 dpi in BALB/c mice.
T35 336559-336677 Sentence denotes L. sigmodontis infected C57BL/6 mice had a reduced hypothermia 6 hours post LPS injection compared to uninfected mice.
T36 336678-336978 Sentence denotes This correlated with significantly increased amounts of peritoneal bioactive TGF and IL-10 in infected mice compared to uninfected controls, whereas levels of the proinflammatory cytokines IL-6, IL-1 , IFN , and MIP2 were not decreased, rather trended to be increased in L. sigmodontis infected mice.
T37 336979-337180 Sentence denotes Similarly, increased expression of CD80 and CD86 on peritoneal macrophages and neutrophils suggest a stronger activation of these cell types in L. sigmodontis infected mice 6 hours after LPS challenge.
T38 337181-337268 Sentence denotes Using a lethal dose of LPS, survival rate in L. sigmodontis infected mice was improved.
T39 337269-337463 Sentence denotes As L. sigmodontis infected animals had a stronger initial pro-inflammatory immune response, we evaluated whether challenge with vital E. coli may be more efficiently controlled in infected mice.
T40 337464-337719 Sentence denotes Indeed, L. sigmodontis infected BALB/c mice regained their initial body temperature 3h post E. coli challenge, whereas uninfected controls had a progressive drop of their body temperature and a significantly increased bacteremia 6h post E. coli challenge.
T41 337720-338043 Sentence denotes Our results suggest that L. sigmodontis induced modulation of the immune response may increase the survival rate after bacterial challenge by inducing a strong pro-inflammatory immune response that controls bacteremia, followed by a potent anti-inflammatory mechanism that reduces pathology due to exaggerated inflammation.
T42 338044-338057 Sentence denotes Introduction:
T43 338058-338149 Sentence denotes Effective therapy is vital in reducing mortality of Pneumocystis jirovecii pneumonia (PCP).
T44 338150-338295 Sentence denotes Because of the inability to grow the fungus in vitro there are currently no reliable methods of assessing P. jirovecii antimicrobial sensitivity.
T45 338296-338451 Sentence denotes To identify treatment failure, physicians have to rely solely on the clinical presentation, radiology results and unspecific parameters such as LDH levels.
T46 338452-338541 Sentence denotes (1 3)--D-Glucan (BG) is a cell wall component of P. jirovecii and of various other fungi.
T47 338542-338669 Sentence denotes In a previous study we have shown that BG is an excellent biomarker for PCP with a sensitivity of 98% and a specificity of 94%.
T48 338670-338813 Sentence denotes However, at the moment nothing is known about its usefulness for monitoring the patient's clinical course or for identifying treatment failure.
T49 338814-338822 Sentence denotes Methods:
T50 338823-338976 Sentence denotes We conducted a retrospective case control study to investigate whether consecutive serum BG-measurements can be used to assess treatment response in PCP.
T51 338977-339144 Sentence denotes All patients presenting at the University Medical Centre of Freiburg between January 2003 and July 2010 with a confirmed PCP and 5 sera during follow-up were enrolled.
T52 339145-339205 Sentence denotes Serum BG levels were measured by use of the Fungitell assay.
T53 339206-339326 Sentence denotes The study patients were grouped according to the pattern of their BG kinetic and independently by their clinical course.
T54 339327-339394 Sentence denotes BG kinetic and clinical course were then compared for each patient.
T55 339395-339403 Sentence denotes Results:
T56 339404-339518 Sentence denotes 18 patients (7 HIV-patients, 11 non-HIV patients) fulfilled the inclusion criteria and were enrolled in the study.
T57 339519-339585 Sentence denotes A median of seven followup sera (IQR 6-9) per patient were tested.
T58 339586-339695 Sentence denotes The median serum BG level was 2353 pg/ml (IQR 790-3710) and therefore was highly elevated (cut-off 85 pg/ml).
T59 339696-339918 Sentence denotes During follow-up 8/18 patients had constantly decreasing BG levels, 6/18 patients had increasing BG levels and 4/18 patients had complex BG kinetics with one or more episodes of decreasing and increasing BG concentrations.
T60 339919-340020 Sentence denotes Comparison of the BG kinetic with the clinical course gave an overall correlation in 61% of patients.
T61 340021-340166 Sentence denotes Separate analysis of the different subgroups showed that decreasing BG-levels are associated with a favourable outcome in 88% of cases (Fig. 1 ).
T62 340167-340305 Sentence denotes In contrast, increasing BG-levels reflected death or treatment failure in only 44%, while the remaining 56% responded well to PCP-therapy.
T63 340306-340485 Sentence denotes One patient with a complex BG-kinetic had a clinically, radiologically and microbiologically proven treatment failure and his BG level increased 7 fold at the same time (Fig. 2) .
T64 340486-340497 Sentence denotes Conclusion:
T65 340498-340586 Sentence denotes Decreasing BG-levels show a good correlation (88%) with a favourable treatment response.
T66 340587-340695 Sentence denotes This may assist in deciding whether to continue first-line therapy in cases of missing clinical improvement.
T67 340696-340801 Sentence denotes Under special circumstances, consecutive BG-measurements might be suitable to identify treatment failure.
T68 340802-340952 Sentence denotes However, the overall correlation of increasing BG-levels with an adverse outcome is poor (44%) and its usefulness for follow-up in severe PCP limited.
T69 340953-340966 Sentence denotes Introduction:
T70 340967-341159 Sentence denotes Infections caused by methicillin-resistant Staphylococcus aureus (MRSA), ranging from skin and soft tissue infections to sepsis and toxic shock, have become a worldwide challenge in hospitals.
T71 341160-341311 Sentence denotes 2010, a prospective multicentre prevalence screening of patients admitted to 24 hospitals was performed in the region of Saarland, Germany (MRSAarNet).
T72 341312-341320 Sentence denotes Methods:
T73 341321-341509 Sentence denotes In the present evaluation 47 MRSA-isolates collected in the study period at the University Hospital of Saarland were selected for a more detailed phenotypic and genotypic characterization.
T74 341510-341785 Sentence denotes As a control group, 47 matched methicillin-sensitive isolates (MSSA) of colonized patients of the same risk-factors treated at the University Hospital of Saarland were selected (according to patient characteristics such as time period since hospitalization, gender, and age).
T75 341786-341857 Sentence denotes Isolates were collected by nasopharyngeal swabs and of infection sites.
T76 341858-341919 Sentence denotes Spa types were assigned using the Ridom StaphType spa server.
T77 341920-342112 Sentence denotes Matrix-assisted laser desorption ionization/time-offlight mass spectrometry (MALDI-TOF MS) based spectra were used to detect discriminative protein profiles of different MRSA and MSSA strains.
T78 342113-342298 Sentence denotes Antimicrobial resistance genes, pathogenicity associated genes and virulence factors were examined using a microarray based system which covers 333 S. aureus gene sequences and alleles.
T79 342299-342491 Sentence denotes The spa typing of the MRSA collection analyzed here identified spa type t003 as predominant spa type which was found in 28 isolates (60%), followed by t504 which was found in 11% (5 isolates).
T80 342492-342616 Sentence denotes 38 isolates (81%) were positive for the ß-lactamase structural gene (blaZ) and its transcriptional regulators blaI and blaR.
T81 342617-342757 Sentence denotes The resistance gene ermA, conferring resistance to macrolide-, lincosamide-, and streptogramine antibiotics, was found in 42 isolates (89%).
T82 342758-342877 Sentence denotes Nearly all (96%) of the strains possessed a capsule type 5 genotype, and only one strain exhibited a capsule of type 8.
T83 342878-343014 Sentence denotes The predominant accessory gene regulator (agr) type identified in this collection was type 2, which was found in 42 MRSA isolates (89%).
T84 343015-343066 Sentence denotes 5 MRSA isolates were positive for agr type 1 (11%).
T85 343067-343277 Sentence denotes All isolates were negative for Panton-Valentine leukocidin (PVL) ant tst (encoding the toxic shock syndrome toxin), but harbored the genes for the staphylococcal superantigen-like proteins C, 6/SSL1 and 7/SSL7.
T86 343278-343447 Sentence denotes The enterotoxin gene cluster egc (comprising the genes seg, sei, sem, sen, seo, seu) as well as genes coding for enterotoxins M, N and U were found in 44 isolates (94%).
T87 343448-343688 Sentence denotes The results of the genotypic and phenotypic characterization of the MRSA collection analyzed here will be compared with those of matched controls of patients of the same risk-factors colonized with methicillin-susceptible S. aureus strains.
T88 343689-343699 Sentence denotes Backround:
T89 343700-343866 Sentence denotes Rapid detection of causative microorganisms in patients with bloodstream infections is crucial for the early initiation or adaption of adequate antimicrobial therapy.
T90 343867-343877 Sentence denotes Objective:
T91 343878-344118 Sentence denotes The goal of this prospective study was to evaluate a commercial PCR-based kit (SeptiFast, SF) for the detection of 25 clinically important pathogens in septic patients after liver transplantation (LTX) and major abdominal surgery (non-LTX).
T92 344119-344127 Sentence denotes Methods:
T93 344128-344336 Sentence denotes Overall 225 blood samples from 170 patients (46.5% after LTX and 53.5% non-LTX) with suspected sepsis were obtained in parallel for blood culture and SF over a period of three years at a tertiary care centre.
T94 344337-344345 Sentence denotes Results:
T95 344346-344453 Sentence denotes In total, SF and BC yielded concurrent results, negative in 110 (48.8%) samples and positive in 51 (25.3%).
T96 344454-344508 Sentence denotes 35 (15.5%) blood samples were SF positive/BC negative.
T97 344509-344640 Sentence denotes 23 (10.2%) samples were BC positive/SF negative of which in twelve cases the detected pathogens were considered to be contaminants.
T98 344641-344822 Sentence denotes Using BC as gold standard the sensitivity, specificity, positive and negative predictive value of SF was for LTX 81%, 70%, 68%, 82% and for non-LTX 58%, 80%, 53%, 83%, respectively.
T99 344823-344834 Sentence denotes Conclusion:
T100 344835-344905 Sentence denotes In LTX and non-LTX patients SF yielded a higher positive rate than BC.
T101 344906-345083 Sentence denotes The SF performance had a higher sensitivity and positive predictive value for the LTX patients, but lower specificity and negative predictive value compared to non-LTX patients.
T102 345084-345094 Sentence denotes Objective:
T103 345095-345313 Sentence denotes There is growing evidence that microgaps between dental implants and their abutments lead to bacterial contamination of the interior of the implants that might provide a reservoir for pathogens causing periimplantitis.
T104 345314-345484 Sentence denotes The aim of this study was to investigate the intraimplant bacterial spectrum of two stage implants, and to compare the results of two different sample collection methods.
T105 345485-345493 Sentence denotes Methods:
T106 345494-345680 Sentence denotes Ten healthy patients without clinical symptoms of periimplantitis and with two-stage implants on each side wearing ball-attached removable prosthodontics took part in this investigation.
T107 345681-345896 Sentence denotes Abutments were removed 56-105 months (median 63.1 months) after connection, and samples were obtained by i). collecting the intra-implant fluid (planctonic bacteria) and ii). by brushing the cavity (biofilm growth).
T108 345897-345994 Sentence denotes Bacterial colonization was evaluated by aerobic and anaerobic incubation on agar and broth media.
T109 345995-346146 Sentence denotes Isolated bacterial colonies were identified by means of MALDI-TOF identification, classical biochemical differentiation tests, and 16S rDNA sequencing.
T110 346147-346155 Sentence denotes Results:
T111 346156-346239 Sentence denotes Specimen from a total of 20 implants were investigated, resulting in 3 -25 (median:
T112 346240-346328 Sentence denotes 16) different bacterial species per implant, representing a total of 105 different taxa.
T113 346329-346599 Sentence denotes Streptococcus anginosus, Actinomyces naeslundii, and Veillonella parvula were found most frequently while periimplantitis marker bacteria such as Prevotella intermedia or Fusobacteria were only sporadically detected or not at all (Aggregatibacter actinomycetemcomitans).
T114 346600-346705 Sentence denotes Comparison of specimen collection by rinsing and by brushing showed a low consistency of results (median:
T115 346706-346712 Sentence denotes 21 %).
T116 346713-346783 Sentence denotes Most taxa per implant found were detected by brush collection (median:
T117 346784-346901 Sentence denotes 62 % of species), however, only combination of both methods allowed complete analysis of the intra-implant microbiom.
T118 346902-346914 Sentence denotes Conclusions:
T119 346915-347053 Sentence denotes Our results suggest ubiquitous heavy contamination of internal spaces of dental implants, revealing a broad spectrum of bacterial species.
T120 347054-347263 Sentence denotes For analysis of the intra-implant microflora the combined examination of both intra-implant fluid and cavity surface samples is recommended in order to cover biofilm-grown as well as planktonic microorganisms.
T121 347264-347335 Sentence denotes Genomic Staphylococcus aureus adaptation to the cystic fibrosis lung N.
T122 347336-347356 Sentence denotes Hirschhausen* 1 , C.
T123 347357-347371 Sentence denotes Neumann 1 , A.
T124 347372-347387 Sentence denotes Goesmann 2 , G.
T125 347388-347401 Sentence denotes Peters 1 , B.
T126 347402-347700 Sentence denotes Kahl 1 1 Universitätsklinikum Münster, Medizinische Mikrobiologie, Münster, Deutschland 2 Universität Bielefeld, CeBiTec, Bielefeld, Deutschland The airways of cystic fibrosis (CF) patients are frequently colonized and infected by Staphylococcus aureus, often accompanying severe course of disease.
T127 347701-347779 Sentence denotes Many patients are persistently colonized with one predominant S. aureus clone.
T128 347780-347992 Sentence denotes Although we do not have detailed knowledge about genomic alterations, S. aureus long-time persistence in the CF lung assumedly selects for phenotypes that are tightly adapted to this specific environmental niche.
T129 347993-348106 Sentence denotes A unique CF-derived S. aureus collection allows us to track genomic adaptation due to persistence in the CF lung.
T130 348107-348294 Sentence denotes Clonal early and late isolates from three CF patients, isolated 12-14 years apart, were sequenced on a genome analyzer (Illumina, Solexa) and screened for SNPs, insertions, and deletions.
T131 348295-348400 Sentence denotes We identified several mutations in the late S. aureus isolates compared to the respective early isolates.
T132 348401-348723 Sentence denotes By Sanger sequencing we could confirm mutations in a number of genes that are associated with virulence or antibiotic resistance, such as protein A (spa), the autoinducer sensor kinase (agrC), the allelic variants of bone sialoprotein-binding protein (bbp/sdrE), and DNA topoisomerases type II and IV (gyrA and parC/grlA).
T133 348724-348833 Sentence denotes All of these mutations are associated with amino acid changes and we currently analyze the potential outcome.
T134 348834-349021 Sentence denotes The altered gyrA corresponds to an acquired resistance towards fluoroquinolones, whereas altered agrC is assumedly associated with different characteristics affecting S. aureus virulence.
T135 349022-349144 Sentence denotes We furthermore include the sequencing of intermediate S. aureus isolates to determine the time when the mutation occurred.
T136 349145-349286 Sentence denotes Our preliminary data suggest that adaptation of persistent CF isolates include changes on DNA level and the regulation of protein expression.
T137 349287-349425 Sentence denotes However, detected mutations did not occur in all patients indicating that the host environment promotes individual adaptational processes.
T138 349426-349558 Sentence denotes Resulting altered virulence may favor precise camouflage in response to the hostile environment under a specific clinical situation.
T139 349559-349693 Sentence denotes Our results may help to understand the complex pathogenesis of chronic infection against the background of host-pathogen interactions.
T140 349694-349833 Sentence denotes Staphylococcus pettenkoferi is a coagulase-negative staphylococcus (CNS) that has been recently described as a rarely isolated new species.
T141 349834-349932 Sentence denotes Whether this new species is a harmful pathogen or an harmless bystander could not be answered yet.
T142 349933-350012 Sentence denotes Nevertheless, a few case reports suggested an invasive role of S. pettenkoferi.
T143 350013-350143 Sentence denotes Staphylococcus lugdunensis and S. pettenkoferi have been described as potentially related species as shown by biochemical methods.
T144 350144-350333 Sentence denotes To determine whether this new species could be a putative pathogen similar to S. lugdunensis, the strains were characterized for the presence of fibrinogen and fibronectin binding adhesins.
T145 350334-350484 Sentence denotes In addition, the internalization of the bacteria into eukaryotic cells were invesitgated using a previously published FACS-based internalizatio assay.
T146 350485-350618 Sentence denotes The occurrence of putative S. lugdunensis adhesin and hemolysin genes were evalutated using primer pairs designed for S. lugdunensis.
T147 350619-350770 Sentence denotes An estimated 30 % of the S. lugdunensis strains bound to fibrinogen and none of the S. pettenkoferi isolates (n=23) bound to fibrinogen or fibronectin.
T148 350771-350894 Sentence denotes In addition, an estimated 30-40 % of S. lugdunensis isolates were internalized into bladder carcinoma 5637 cell line cells.
T149 350895-350981 Sentence denotes In contrast to this, only one out of 23 isolates of S. pettenkoferi were internalized.
T150 350982-351211 Sentence denotes The lack of fibrinogen and fibronectin binding adhesins, and the lack of an internalization ability indicate that S. pettenkoferi is more likely a harmless colonizer of the human skin flora more than a harmful infective pathogen.
T151 351212-351225 Sentence denotes Introduction:
T152 351226-351347 Sentence denotes Chlamydia pneumoniae (Cpn) is an obligate intracellular bacterium and causes mild-to-moderate pharyngitis and bronchitis.
T153 351348-351461 Sentence denotes In addition, Cpn infection are supposed to play a role in chronic diseases like asthma, COPD and atherosclerosis.
T154 351462-351611 Sentence denotes Systemic dissemination of Cpn from the lung to peripheral organs and arteric vessels has been shown to be mediated by blood mononuclear cells (PBMC).
T155 351612-351741 Sentence denotes Using whole genome sequencing we recently found genetic differences between isolates that were cultivated from different tissues.
T156 351742-351907 Sentence denotes It has been shown previously that intracellular morphology and growth characteristics of chlamydiae signficantly differ between human monocytes and epithelial cells.
T157 351908-352025 Sentence denotes However, phenotypic differences between isolates from different anatomical origins have not been investigated so far.
T158 352026-352136 Sentence denotes We therefore compared growth characteristics and progeny of Cpn isolates from three different infection sites.
T159 352137-352158 Sentence denotes Material and methods:
T160 352159-352304 Sentence denotes Cpn strains, CW029, CV6 and PB2 which were isolated from the respiratory tract, coronary arteries and PBMC, respectively were used in this study.
T161 352305-352444 Sentence denotes Infection doses for HEp-2 cells of the different isolates were adjusted by determination of the amount of genome encoded 16S ribosomal RNA.
T162 352445-352671 Sentence denotes Rate of infection, number of intracellular inclusion, production of infectious progeny, and developmental pace of each isolate were investigated by fluorescence microscopy, transmission electron microscopy and recovery assays.
T163 352672-352680 Sentence denotes Results:
T164 352681-352979 Sentence denotes The coronary artery strain CV6 showed the highest infection and multiple inclusion rates among the three different isolates.The isolate PB2 that was derived from PBMC showed dramatically less production of infectious progeny and slower developmental speed compared to the respiratory strain CWL029.
T165 352980-353002 Sentence denotes Conclusion/Discussion:
T166 353003-353288 Sentence denotes Clinical isolates of Cpn from the bood circulation (PBMC, coronary arteries) can be differentiated from lung isolates by nonsynonymous SNPs in 9 ORFs related with chlamydial metabolisms, bacterial transition, inclusion membrane formation, LPS synthesis and regulation of sigma factors.
T167 353289-353482 Sentence denotes Further studies will have to show whether these genetic differences are responsible for the observed phenotypic differences between the isolates with regard to intracellular growth and progeny.
T168 353483-353559 Sentence denotes Detection of carbapenemases has only very rarely been reported from Germany.
T169 353560-353742 Sentence denotes In order to assess the presence of carbapenemases in Hamburg a number of carbapenem resistant gram-negative bacterial isolates collected during a period of eight month were analysed.
T170 353743-353944 Sentence denotes VIM-1, VIM-2 and KPC-2 were detected in diverse enterobacterial species and Pseudomonas aeruginosa isolates suggesting a more widespread prevalence of carbapenemases in Germany than currently believed.
T171 353945-354045 Sentence denotes Proteomic analysis in early and late Staphylococcus aureus isolates from cystic fibrosis patients D.
T172 354046-354059 Sentence denotes Block* 1 , N.
T173 354060-354079 Sentence denotes Hirschhausen 1 , S.
T174 354080-354096 Sentence denotes Engelmann 2 , G.
T175 354097-354110 Sentence denotes Peters 1 , B.
T176 354111-354517 Sentence denotes Kahl 1 1 Universitätsklinikum Münster, Medizinische Mikrobiologie, Münster, Deutschland 2 Universität Greifswald, Institut für Mikrobiologie, Greifswald, Deutschland Due to mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator, lungs of cystic fibrosis (CF) patients exhibit dehydration of the airway mucus leading to decreased mucociliary clearance and chronic infections.
T177 354518-354621 Sentence denotes One of the earliest pathogens which can be isolated from lungs of CF patients is Staphylococcus aureus.
T178 354622-354714 Sentence denotes Often the lungs of CF patients are infected by a predominant S. aureus clone for many years.
T179 354715-354805 Sentence denotes During long-term colonization the pathogen needs to adapt to the environment of the lungs.
T180 354806-355007 Sentence denotes This project is aiming to further reveal the mechanisms underlying the adaptation of S. aureus during persistent colonization and infection to the hostile niche of CF patients using proteomic analysis.
T181 355008-355096 Sentence denotes In addition the virulence potential was assessed in in vitro, as well as in vivo assays.
T182 355097-355261 Sentence denotes From a unique longitudinal collection of S. aureus strains isolated from the airways of CF patients early and late S. aureus isolates of three patients were chosen.
T183 355262-355402 Sentence denotes The comparison of the proteome of early and late isolates was performed using 2D-gelelectrophoresis followed by MALDI-TOF mass spectrometry.
T184 355403-355503 Sentence denotes S. aureus strains were cultured in artificial sputum medium in order to mimic the in vivo situation.
T185 355504-355629 Sentence denotes Virulence potential of the strains was investigated in vitro by means of invasion assays using the epithelial cell line A549.
T186 355630-355729 Sentence denotes Furthermore, the virulence potential was studied in vivo in a Caenorhabditis elegans killing assay.
T187 355730-355857 Sentence denotes Preliminary results of invasion assays revealed different alterations in invasive properties comparing early and late isolates.
T188 355858-356049 Sentence denotes For late isolates of two patients an increase of invasive properties compared to the early isolates could be observed whereas the properties of the late isolate of one patient were decreased.
T189 356050-356205 Sentence denotes First results using isolates of one patient in the C. elegans killing assay revealed no significant difference in virulence between early and late isolate.
T190 356206-356300 Sentence denotes The investigation of the proteomic changes during long-term colonization is still in progress.
T191 356301-356521 Sentence denotes In this project we studied the adaptive mechanisms of S. aureus during persistent colonization within the airways CF patients on a functional level to gain insights into the interaction between the pathogen and the host.
T192 356522-356834 Sentence denotes Since adaptation of bacteria can occur on genomic as well as on regulatory levels, it is of importance to investigate the alterations in late isolates compared to early isolates in a proteomic approach which allows the analysis of differences in protein expression as well as of post-translational modifications.
T193 356835-356919 Sentence denotes Here we describe the therapy and outcome of ODRI with a difficult-to-treat organism:
T194 356920-356942 Sentence denotes Enterococcus faecalis.
T195 356943-356963 Sentence denotes Patient and methods:
T196 356964-357164 Sentence denotes Five patients with a chronic hip implant infection caused by E. faecalis were treated by a two-stage replacement and with use of antimicrobial cement and long term treatment with antimicrobial agents.
T197 357165-357282 Sentence denotes All five isolates were sensitive to ampicillin and four out of five isolates were high-level resistant to gentamicin.
T198 357283-357405 Sentence denotes All patients recieved a systemic therapy with ampicillin or amoxycillin for six weeks plus a vancomycin containing cement.
T199 357406-357480 Sentence denotes The second device implantation took place after a mean time of 3,5 months.
T200 357481-357489 Sentence denotes Outcome:
T201 357490-357568 Sentence denotes After 24 months there were no further signs of infection in all five patients.
T202 357569-357678 Sentence denotes Two patients needed revision surgery because of surgical site infections prior to second device implantation.
T203 357679-357690 Sentence denotes Conclusion:
T204 357691-357750 Sentence denotes Chronic infections with E. faecalis are difficult to treat.
T205 357751-357898 Sentence denotes As most patients recieve gentamicin containing cement for their first implantion many E. faecalis isolates show a high-level gentamicin resistance.
T206 357899-358023 Sentence denotes The combination of vancomycin containig cement with systemic ampicillin therapy might be a useful option for these patients.
T207 358024-358037 Sentence denotes Introduction:
T208 358038-358132 Sentence denotes In the infected host, Mycobacterium tuberculosis (Mtb) faces oxidative and nitrosative stress.
T209 358133-358218 Sentence denotes Thus, the pathogen needs an effective antioxidative system to maintain the infection.
T210 358219-358420 Sentence denotes Due to the lack of the glutathione system in Mtb the thioredoxin reductase (TrxR) system is an essential mechanism to reduce reactive oxidative and nitrosativ species and assure the bacterial survival.
T211 358421-358567 Sentence denotes Similarity between the mycobacterial and the eukaryotic TrxR system is low, therefore it may be used as a novel drug target to fight tuberculosis.
T212 358568-358576 Sentence denotes Methods:
T213 358577-358722 Sentence denotes To identify new substances which inhibit the interaction between thioredoxin (Trx) and TrxR an in silico high throughput screening was conducted.
T214 358723-358836 Sentence denotes Four altered low mass scaffolds were chosen to be tested in vitro to examine their inhibitory effect on Mtb TrxR.
T215 358837-358977 Sentence denotes After optimization of these compounds, whole cell testing was performed in Mtb liquid cultures using the MGIT 960 system (Becton Dickinson).
T216 358978-358986 Sentence denotes Results:
T217 358987-359078 Sentence denotes The interference of 170 low mass compounds with the mycobacterial TrxR was tested in vitro.
T218 359079-359164 Sentence denotes 10 percent of the chosen low mass compounds showed activity in low μM concentrations.
T219 359165-359289 Sentence denotes After optimization of these compounds some of them showed bacteriostatic effects on Mtb cultures even within a low μM range.
T220 359290-359301 Sentence denotes Conclusion:
T221 359302-359459 Sentence denotes Effecting the interaction between Trx and TrxR with small molecule inhibitors at micromolar levels leads to bacteriostatic effects in Mtb cultures. (ExPEC) .
T222 359460-359703 Sentence denotes Although several important UPEC virulence factors and their role during pathogenesis have been described, many UPEC isolates cannot be unambiguously distinguished from commensal E. coli based on the presence of UPEC virulence-associated genes.
T223 359704-359866 Sentence denotes In contrast, intestinal pathogenic E. coli (IPEC) which cause diarrhoea are characterized by typical sets of virulence factors which are usually absent from UPEC.
T224 359867-360078 Sentence denotes The majority of virulence-associated genes of ExPEC and IPEC are located on mobile and accessory genetic elements, e.g. plasmids, bacteriophages or pathogenicity islands and can thus be horizontally transferred.
T225 360079-360164 Sentence denotes We analyzed 283 E. coli isolates from hospital patients with urinary tract infection.
T226 360165-360265 Sentence denotes These isolates were screened by multiplex PCR for important virulence determinants of UPEC and IPEC.
T227 360266-360419 Sentence denotes Interestingly, about 10 % of these isolates carried one or more known IPEC virulence genes, sometimes in combination with classical UPEC virulence genes.
T228 360420-360557 Sentence denotes In addition, molecular epidemiological and phenotypic analyses were performed to assess the relatedness and similarity of these isolates.
T229 360558-360680 Sentence denotes Notably, among the atypical urine E. coli isolates were strains carrying marker genes of enteroaggregative E. coli (EAEC).
T230 360681-360801 Sentence denotes In addition, isolates were typed as atypical enteropathogenic E. coli (aEPEC), and Shiga toxin-producing E. coli (STEC).
T231 360802-360996 Sentence denotes Our results indicate that nosocomial UTI due to E. coli may be caused by a diverse group of E. coli variants, some of which represent IPEC or carry virulence properties of diarrheagenic E. coli.
T232 360997-361241 Sentence denotes The combination of typical ExPEC and IPEC virulence determinants also further supports the finding that E. coli exhibits a marked genome plasticity and that gene transfer may promote an evolutionary transition from IPEC to ExPEC -or vice versa.
T233 361242-361462 Sentence denotes The relatively high prevalence of EAEC marker genes among urine isolates also demonstrates the heterogeneity of this pathotype and raises the possibility that some EAEC strains may have the potential to be uropathogenic.
T234 361463-361465 Sentence denotes I.
T235 361466-361479 Sentence denotes Aibinu 1 , T.
T236 361480-361495 Sentence denotes Odugbemi 1 , W.
T237 361496-361509 Sentence denotes Koenig 2 , B.
T238 361510-361532 Sentence denotes Ghebremedhin* 2 1 Uni.
T239 361533-361544 Sentence denotes Lagos, Med.
T240 361545-361648 Sentence denotes Microbiol. & Parasitol., Lagos, Nigeria 2 Uniklinik Magdeburg, IMMB, Magdeburg, Deutschland Background:
T241 361649-361862 Sentence denotes We investigated the association between the prevalent extended-spectrum beta-lactamase and the genetic lineage of Escherichia coli from two tertiary hospitals in Lagos, Nigeria within a period of 8 months in 2009.
T242 361863-361871 Sentence denotes Methods:
T243 361872-361968 Sentence denotes Identification and susceptibility testing of E.coli isolates (n=109) were by Vitek 2 and E-test.
T244 361969-362153 Sentence denotes Extended-spectrum beta-lactamase and quinolone resistance genes (the aac-(6')-lb-cr and plasmid-mediated quinolone resistance [PMQR] genes) were characterized using PCR and sequencing.
T245 362154-362229 Sentence denotes Clonality was determined by ERIC PCR and multilocus sequence typing (MLST).
T246 362230-362238 Sentence denotes Results:
T247 362239-362299 Sentence denotes Of 109 isolates, 14 (12.8%) were found to be ESBL-producers.
T248 362300-362388 Sentence denotes All the ESBL isolates were resistant to ceftazidime and cefotaxime with MIC of >16μg/ml.
T249 362389-362492 Sentence denotes Resistance to ciprofloxacin (MIC 32μg/ml) and an aminoglycoside co-existed also in these ESBL isolates.
T250 362493-362545 Sentence denotes They were however, all sensitive to the carbapenems.
T251 362546-362605 Sentence denotes The only ESBL gene detected in these isolates was CTX-M-15.
T252 362606-362812 Sentence denotes All the CTX-M-15 producers carried the insertion sequence ISEcpI which has recently been demonstrated to mobilize 3'-adjacent genes to transfer between DNA replicons and is involved in spread of such ESBLs.
T253 362813-362869 Sentence denotes ERIC-PCR identified 11 types among these ESBL-producers.
T254 362870-362914 Sentence denotes A diversity of MLST types was also observed.
T255 362915-363018 Sentence denotes Clonal group ST131 (35.7%) was the predominant ST, closely followed by the clonal complex ST10 (28.6%).
T256 363019-363153 Sentence denotes The other sequence types established in these strains included ST23 complex (n=1), ST 448 complex (n=1), ST295 (n=1) and ST 501 (n=1).
T257 363154-363272 Sentence denotes Plasmid-mediated quinolone resistance qnrA1 gene (n=3) and qnrB1 (n=1) were detected among the ESBL isolates of ST131.
T258 363273-363340 Sentence denotes All the ESBL isolates with ST131 harboured the aac-(6')-lb-cr gene.
T259 363341-363352 Sentence denotes Conclusion:
T260 363353-363514 Sentence denotes This is the first report of E. coli ST131 and clonal complex 10 in West-Africa (Nigeria) found predominant among hospital isolates of CTX-M-15-producing E. coli.
T261 363515-363592 Sentence denotes The high diversity of STs is remarkable among the CTX-M-15 positive isolates.
T262 363593-363708 Sentence denotes This study highlights the need to adopt appropriate control measures to overcome the ESBL dissemination in Nigeria.
T263 363709-363767 Sentence denotes Bacteriocin production by staphylococcal nasal isolates D.
T264 363768-363781 Sentence denotes Janek* 1 , A.
T265 363782-363796 Sentence denotes Peschel 1 , B.
T266 363797-363966 Sentence denotes Krismer 1 1 Uniklinik Tübingen, Medizinische Mikrobiologie, Tübingen, Deutschland Staphylococcus aureus is a major pathogen in hospital-and communityacquired infections.
T267 363967-364081 Sentence denotes Colonisation of the anterior nares in about 30% of the population is a major risk factor for S. aureus infections.
T268 364082-364154 Sentence denotes Recently the composition of the nasal microbiota has been inverstigated.
T269 364155-364264 Sentence denotes Interestingly, the bacterial diversity in the human nose reaches from aerobic to strictly anaerobic bacteria.
T270 364265-364404 Sentence denotes The most frequently occurring species are Corynebacterium accolens, C. macginleyi, Staphylococcus. epidermidis and Propionibacterium acnes.
T271 364405-364570 Sentence denotes In order to investigate if bacteriocin production might play a role during nasal colonisation we analysed the bacteriocin production of nasal Staphylococcus strains.
T272 364571-364700 Sentence denotes The test strains (Micrococcus luteus or S. aureus) were casted in an agar plate and the nasal isolates were stamped on the plate.
T273 364701-364802 Sentence denotes Various isolates showed growth inhibition zones of the test strains indicating baceriocin production.
T274 364803-364951 Sentence denotes Transposon plasmids could be transformed into various strains and mutagenesis was performed to identify corresponding bacteriocinbiosynthetic genes.
T275 364952-365078 Sentence denotes Analysis of 93 stapylococcal nasal isolates revealed that various strains produce bacteriocins against M. luteus or S. aureus.
T276 365079-365185 Sentence denotes The bacteriocin production of some nasal isolates turned out to be inducible by certain stress conditions.
T277 365186-365345 Sentence denotes One of these bacteriocins, produced by an S. epidermidis strain, could be characterized as a nukacin-like lantibiotic with activity against Micrococcos luteus.
T278 365346-365519 Sentence denotes Extending our knowledge about the various interactions between staphylococcal and other nasal species could be important for finding new effective anti-S. aureus strategies.
T279 365520-365672 Sentence denotes Humanisation of the murine anti-IsaA antibody -a potential target for an antibody-based therapy against multi-resistant Staphylococcus aureus strains B.
T280 365673-365789 Sentence denotes Oesterreich* 1 1 Institut für Molekulare Infektionsbiologie, Universität Würzburg, Würzburg, Deutschland Background:
T281 365790-365884 Sentence denotes The Gram-positive bacterium Staphylococcus aureus is the major cause of nosocomial infections.
T282 365885-366179 Sentence denotes In particular, diseases caused by methicillin-resistant S. aureus (MRSA) are associated with higher morbidity, mortality and medical costs due to showing resistance to several classes of established antibiotics and their ability to develop resistance mechanisms against new antibiotics rapidly.
T283 366180-366243 Sentence denotes Therefore, there is an urgent need for novel treatment options.
T284 366244-366360 Sentence denotes Immunotherapy has the potential to close the gap in the treatment of infections caused by multi-resistant S. aureus.
T285 366361-366551 Sentence denotes The immunodominant staphylococcal antigen A (IsaA) has been identified as a putative target for immunotherapy due to its expression by all clinical strains in vivo, and the surface exposure.
T286 366552-366713 Sentence denotes Preclinical experiments revealed protective properties of a monoclonal mouse anti-IsaA antibody (UK-66) in different experimental models of S. aureus infections.
T287 366714-366840 Sentence denotes In addition, in vitro phagocytosis assays showed enhancement of opsonophagocytic killing of S. aureus in the presence of UK66.
T288 366841-366960 Sentence denotes Therefore, the mouse monoclonal antibody was selected for humanization as a prerequisite of clinical studies in humans.
T289 366961-367068 Sentence denotes The hybridoma clone UK-66 was the basis for the identification of the antibody binding domain against IsaA.
T290 367069-367224 Sentence denotes The coding sequence was used to construct recombinant scFv and scFvFc fragments towards IsaA ( Fig. 1 ) and to humanised the murine antigen binding domain.
T291 367225-367373 Sentence denotes The murine and humansed scFv fragment was expressed in E.coli and the murine and humanised scFvFc fragment was expressed in eucaryotic HEK293 cells.
T292 367374-367548 Sentence denotes The scFv-and scFvFc fragments were characterized in their function and specificity by Western Blot analysis, ELISA-studies, immuno-fluorescence analysis and FACS experiments.
T293 367549-367756 Sentence denotes The results revealed that all constructed fragments showed a high specificity towards IsaA and the property of the antigen binding fragments to detect IsaA on the cell surface of different S. aureus strains.
T294 367757-367768 Sentence denotes Conclusion:
T295 367769-367849 Sentence denotes IsaA is an antigenic structure expressed in vivo in different S. aureus strains.
T296 367850-367911 Sentence denotes The murine anti-IsaA antibody is protective in animal models.
T297 367912-368002 Sentence denotes The murine and humanised scFv-and scFvFc fragments showed a high specificity towards IsaA.
T298 368003-368136 Sentence denotes For an improved treatment of infectious diseases the identification of optimized combinations of antibiotics is a promising approach.
T299 368137-368301 Sentence denotes A major requirement for such investigations is the rapid testing of a moderate number of different compound combinations with a limited number of infectious agents.
T300 368302-368444 Sentence denotes We developed a device for the combinatorial generation of compound mixtures by diffusion with concentrations over several orders of magnitude.
T301 368445-368541 Sentence denotes Diffusion takes place in cell compatible polymer matrices such as agarose, gelatine or AB3 agar.
T302 368542-368643 Sentence denotes Mixtures of test compounds are generated by a superposition of concentration gradients (2D-gradient).
T303 368644-368826 Sentence denotes Only two pipetting steps are required to generate a series of assays that only depends on the spatial resolution by which the diffusion gradient is converted into a biological assay.
T304 368827-368963 Sentence denotes The diffusion device is compatible with 12-well microtiter plates enabling the efficient testing of combinations of different compounds.
T305 368964-369081 Sentence denotes The device is applicable for all diffusible compounds such as small molecules, peptides, lipopeptides and antibodies.
T306 369082-369256 Sentence denotes Here we present the investigation of antibiotic combinations against different staphylococcal strains to define antibiotic combinations with additive and synergistic effects.
T307 369257-369480 Sentence denotes The miniaturization, parallelization and especially the reduction of the number of steps required for the preparation of samples enabled the rapid and efficient screening of drug combinations on different microbial strains.
T308 369481-369657 Sentence denotes Invasive fungal infections are a main cause for morbidity and mortality in transplant recipients and immunocompromised patients in the clinic and thus a growing health problem.
T309 369658-369789 Sentence denotes Regarding the constantly growing number of resistant strains the discovery of new classes of antimycotic drugs is highly necessary.
T310 369790-369969 Sentence denotes In a high throughput activity-selectivity screening a series of (S)-2aminoalkyl benzimidazole derivatives with strong antimycotic activity against Candida albicans was identified.
T311 369970-370087 Sentence denotes A collection of analogues for a SAR-study was prepared in order to evaluate and improve the antimicrobial properties.
T312 370088-370326 Sentence denotes We examined the activity of five of the (S)-2-aminoalkyl benzimidazoles against a set of twenty-one fungal strains including different type strains and significant clinical iso-lates of Candida spp. and confirmed the antimycotic activity.
T313 370327-370420 Sentence denotes The most active com-pound 120B12 in this study showed potencies equal to that of fluconazole.
T314 370421-370572 Sentence denotes Consequently, the novel non-toxic (S)-2-aminoalkyl benzimidazoles represent promising candidates for future developments in antimycotic drug discovery.
T315 370573-370636 Sentence denotes Currently, studies with Caenorhabditis elegans are carried out.
T316 370637-370836 Sentence denotes The Bavarian Health and Food Safety authority (Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, LGL) provides several microbiological services for local health authorities in Bavaria.
T317 370837-371097 Sentence denotes According to the German Protection Act (Infektionsschutzgesetz, IFSG) § § 29-31 the public health system is responsible to ensure an adequate care of TB cases and in the following the performance of contact investigations to actively identify infected persons.
T318 371098-371205 Sentence denotes In this context, the LGL maintains a TBlaboratory facility for the diagnosis of M. tuberculosis infections.
T319 371206-371507 Sentence denotes As the interferon-gamma-release-assays (IGRA) have been shown to improve the diagnosis (better specificity and sensibility than TST) especially of latent tuberculosis infections (LTBI) this test has been increasingly requested with more than 8500 samples in 2010 by health care workers and clinicians.
T320 371508-371642 Sentence denotes IGRAs (e.g. QuantiFeron®-TB Gold In-Tube test and T-SPOT®-TB) measure a person's immune reactivity to specific mycobacterial antigens.
T321 371643-371726 Sentence denotes Specimens are mixed either with peptides that simulate antigens of M. tuberculosis.
T322 371727-371921 Sentence denotes These M. tuberculosis specific antigens (ESAT-6, CFP-10 und TB7.7) are lacking in all BCG strains and almost all non-tuberculous mycobacteria (NTM) except M. kansasii, M. szulgai and M. marinum.
T323 371922-372039 Sentence denotes In a person infected with M. tuberculosis, the white blood cells recognize the peptides and release interferon-gamma.
T324 372040-372102 Sentence denotes Test result is based on the amount of interferon-gamma releas.
T325 372103-372257 Sentence denotes At present the LGL supply the QuantiFeron®-TB Gold In-Tube test (QFT) mainly due to operational advantages regarding the high-throughput of blood samples.
T326 372258-372338 Sentence denotes The poster will give an overview about laboratory performance of QFT at the LGL.
T327 372339-372427 Sentence denotes Salmonella enterica is one of the most common causes of human gastroenteritis worldwide.
T328 372428-372513 Sentence denotes It is also responsible for nosocomial outbreaks particularly in developing countries.
T329 372514-372665 Sentence denotes In Tunisia, many nosocomial outbreaks, occurred in pediatric services, have been involved various different serotypes, especially serotype Livingstone.
T330 372666-372765 Sentence denotes In Belgium, the reported sporadic human cases of S. enterica serotype Livingstone infection is low.
T331 372766-372831 Sentence denotes In this study, 43 S. enterica serotype Livingstone were analyzed.
T332 372832-372978 Sentence denotes Most of them were isolated from human (31 strains from Tunisia and 10 from Belgium) and tow strains came from food products (pork and beef meats).
T333 372979-373110 Sentence denotes Tunisian strains were isolated in the same hospital but during different periods (2002, n=1; 2004, n=1; 2005, n=27 and 2009, n=2) .
T334 373111-373210 Sentence denotes All strains were characterized by antibiogramm, Multilocus sequence Typing (MLST) and virulotyping.
T335 373211-373362 Sentence denotes This last technique was carried out by simple PCR of chromosomic genes (agfA, hin/H2, iroB, phoP/Q and sly A) and plasmids genes (spvA, spvB and spvC).
T336 373363-373528 Sentence denotes Tunisian strains were resistant to amoxicillin, amoxicillin-clavulanic acid, ticarcillin, gentamicin, kanamycin and showed a particular -lactam resistance phenotype.
T337 373529-373598 Sentence denotes Belgium strains were susceptible for the majority of the antibiotics.
T338 373599-373684 Sentence denotes Further to MLST analyses, tunisian strains belonged to the same sequence type, ST543.
T339 373685-373810 Sentence denotes For belgium isolates, 3 strains had a ST638 profile, 8 strains had ST543 profile and one strain had an unknown sequence type.
T340 373811-374005 Sentence denotes Analyses of the virulence genes content showed that strains isolated in different years, in different origins (human and food products) and in different countries had the same virulence profile.
T341 374006-374102 Sentence denotes They carried the same chromosomic virulence genes and the plasmidic virulence genes were lacked.
T342 374103-374358 Sentence denotes Combination of different typing methods showed that the majority of belgium strains (8/12) and all tunisian strains isolated in different time periods are closely related; they belonged to the same sequence type (ST543) and had the same virulence profile.
T343 374359-374361 Sentence denotes M.
T344 374362-374636 Sentence denotes Höck* 1 1 DRK Kliniken Berlin / Westend, Institut für Mikrobiologie und Krankenhaushygiene, Berlin, Deutschland Objectives: ESBL producing E.coli strains have become a serious problem, as these strains often leave few therapeutic options, especially in intensive care units.
T345 374637-374674 Sentence denotes The therapeutic options seem limited.
T346 374675-374898 Sentence denotes Furthermore the former EUCAST expert rules were changed so that the definitions of sensible to beta lactams were changed to intermediate and intermediate to resistant whenever an ESBL was recorded for the respective strain.
T347 374899-374925 Sentence denotes The change for 2010 reads:
T348 374926-375047 Sentence denotes Cephalosporin breakpoints for Enterobacteriaceae will detect clinically important resistance mechanisms (including ESBL).
T349 375048-375324 Sentence denotes Some strains that produce beta-lactamases are susceptible or intermediate to 3rd or 4th generation cephalosporins with these breakpoints and should be reported as found, i.e. the presence or absence of an ESBL does not in itself influence the categorization of susceptibility.
T350 375325-375432 Sentence denotes We analysed isolates in order to see if the change in the EUCAST expert rule opens new therapeutic options.
T351 375433-375441 Sentence denotes Methods:
T352 375442-375538 Sentence denotes The laboratory participating in the network use the automated BD PHOENIX-systems measuring MICs.
T353 375539-375666 Sentence denotes The BD EPICENTER Data-Management-System is used for evaluation of lab data and for the transfer of the data for joint analysis.
T354 375667-375758 Sentence denotes For this study we analysed the MICs of ESBL producing E. coli isolates during 2006 to 2010.
T355 375759-375897 Sentence denotes ESBL production was checked with the associated BD-expert system, as the Phoenix shows excellent performance in ESBL detection (Maurine A.
T356 375898-375925 Sentence denotes Leverstein-van Hall, et al:
T357 375926-375943 Sentence denotes J Clin Microbiol.
T358 375944-375966 Sentence denotes 2002 October; 40(10) :
T359 375967-376000 Sentence denotes 3703) Copy strains were excluded.
T360 376001-376049 Sentence denotes Quality control assays were routinely performed.
T361 376050-376058 Sentence denotes Results:
T362 376059-376263 Sentence denotes Using the MICs of strains producing ESBL producing strains to ceftazidime (CAZ), cefotaxime (CTX), cefepime (FMC), and aztreonam (ATM) we calculated the sensitivity using the new EUCAST breakpoints. [2] .
T363 376264-376475 Sentence denotes For the dendrogram creation with a Microflex mass spectrometer (Bruker Daltonik, Bremen, Germany) and the MALDI Biotyper 2.0 software, VRE isolates underwent an extraction procedure, as described elsewhere [3] .
T364 376476-376661 Sentence denotes Spectra were recorded in the positive linear mode (laser frequency 20 Hz; ion source 1 voltage at 20 kV ion source 2 voltage at 18.5 kV; lens voltage 8.5 kV; mass range 2000 -20137 Da).
T365 376662-376762 Sentence denotes For each spectrum 6x480 shots from different positions of 6 target spots were collected per isolate.
T366 376763-376872 Sentence denotes Spectra were cleaned and calibrated with the peak of the 50S ribosomal protein L36 of E. faecium at 4427 kDa.
T367 376873-376928 Sentence denotes Subsequently minimum spanning trees (MSP) were created.
T368 376929-376997 Sentence denotes The creation of the dendrogram was based on cross-wise MSP matching.
T369 376998-377160 Sentence denotes The topology of the dendrogram (Figure 1 ) generated from the spectra of the VRE isolates was in agreement with that inferred from the PFGE analysis ( Figure 2 ).
T370 377161-377213 Sentence denotes Three different strains could be clearly identified.
T371 377214-377307 Sentence denotes C1 was the index case and was held responsible for the transmission to all others beforehand.
T372 377308-377439 Sentence denotes Epidemiological analysis revealed actually two transmissions; the first from patient C1 to C2 and the second from patient A1 to A2.
T373 377440-377489 Sentence denotes Patient B was probably an already prevalent case.
T374 377490-377585 Sentence denotes In the dendrogram, strain B is separated from the others but closer related to strain C than A.
T375 377586-377705 Sentence denotes Discussio: MALDI-TOF MS provided results concordant to the PFGE gold standard and that in a fraction of time and costs.
T376 377706-377844 Sentence denotes These results show great promise for MALDI-TOF mass spectrometry as an epidemiological tool for the real-time management of VRE outbreaks.
T377 377845-377967 Sentence denotes In Cologne, the most prevalent spa type was t003 (32/38); t001, t018, t030, t032, t038, and t068 each in a single patient.
T378 377968-378059 Sentence denotes Conclusion: SAB is a common infection in German hospitals associated with a high mortality.
T379 378060-378179 Sentence denotes In Cologne, the most prevalent spa type is t003 which currently is also one of the most prevalent spa-types in Germany.
T380 378180-378182 Sentence denotes D.
T381 378183-378387 Sentence denotes The control of the infectious agent is usually not the determining factor for a favourable outcome of an infectious disease in its final stadium, but rather the control of the over-reactive immune system.
T382 378388-378456 Sentence denotes Complement is an important part of the humoral innate immune system.
T383 378457-378627 Sentence denotes Inherited mutations in this system, and particularly in its regulatory proteins, lead to such an overreactive state and eventually to a haemolytic uraemic syndrome (HUS).
T384 378628-378981 Sentence denotes This type of HUS is termed "atypical", as it is unrelated to an infection with enterohaemorrhagic E. coli (EHEC Eculizumab was recently very successfully used to block complement in the treatment of paroxysmal nocturnal haemoglobinuria (PNH), in which red blood cells are lysed by complement due to an inherited lack of inhibitors on their cell surface.
T385 378982-379253 Sentence denotes As Eculizumab works for PNH, and atypical HUS also shows an uncontrolled complement activation, this humanised antibody was also successfully used by us in a boy who suffered from an inherited mutation leading to this clinical condition (Zimmerhackl et al., N Engl J Med.
T386 379254-379273 Sentence denotes 2010; 362:1746-8) .
T1 379274-379699 Sentence denotes Last, but not least we provided the rationale for a treatment of the typical HUS with a complement inhibitory antibody: although it is well established that the released EHEC shiga toxins have a direct deleterious effect on kidney cells, they also activate complement and bind factor H, its main regulator, leading to both a stronger activation and a weaker control of complement (Orth et al., J Immunol 2009; 182:6394-400) .
T2 379700-379874 Sentence denotes Several other labs have now corroborated the role of complement in typical HUS (Thurman et al., Clin J Am Soc Nephrol 2009; 4:1920-4; Stahl et al., Blood 2011; 117:5503-13) .
T3 379875-380070 Sentence denotes This opened the field for using Eculizumab in typical HUS, as recently and successfully performed by Lapeyraque and coworkers in three patients (Lapeyraque et al., N Engl J Med, 2011, in press ).
T4 380071-380239 Sentence denotes The evaluation of the outcome of the many dozens patients treated during the O104 outbreak will eventually demonstrate the clinical impact of complement in typical HUS.
T5 380240-380248 Sentence denotes Results:
T6 380249-380369 Sentence denotes 39.9% (n=59) of the blood samples (n=148) were positive using multiplex-PCR and 20.3% (n=30) using conventional culture.
T7 380370-380445 Sentence denotes In 11.4% of all samples multiplex-PCR detected more than one microorganism.
T8 380446-380661 Sentence denotes Among the 77 microorganisms identified by multiplex-PCR, only 25 (32.5%) could be confirmed by blood culture, additional 17 could be confirmed by microbiological test results from other significant patient specimen.
T9 380662-380951 Sentence denotes Positive blood samples independent of the detection method were characterized by significant elevated levels of procalcitonin ( Infection with S. aureus can result in a clinical condition ranging from Systemic Inflammatory Response Syndrome to Septic Shock with multiple organ dysfunction.
T10 380952-381104 Sentence denotes One commonality of this spectrum is an evolving and eventually overwhelming host immune response that is often manifested clinically in pulmonary edema.
T11 381105-381242 Sentence denotes Ceramide is a biologically active sphingolipid created through hydrolysis of sphingomyelin by the inducible enzyme acid sphingomyelinase.
T12 381243-381368 Sentence denotes As a membrane constituent and signaling molecule, ceramide functions as an integral component in multiple signaling pathways.
T13 381369-381550 Sentence denotes In the present study we sought to demonstrate that inducible ceramide is an important mediator of the overwhelming immune response leading to septic shock from S. aureus bacteremia.
T14 381551-381693 Sentence denotes C57Bl6 (wt) mice and acid sphingomyelinase knockout (Asm(-/-)) littermates were inoculated intravenously with a clinical isolate of S. aureus.
T15 381694-381805 Sentence denotes Dosing of the organism was such as to induce a state of septic shock and death within 24-48 hours for all mice.
T16 381806-382004 Sentence denotes Using immunohistochemical techniques, quantification of GR1 positive staining cells in the lung revealed >50% decrease in neutrophil trafficking at 24 hours in Asm(-/-) mice compared to wt controls.
T17 382005-382132 Sentence denotes Measurement of Evan's Blue dye extravasation in the lung displayed a two-fold reduction of pulmonary edema in Asm(-/-) animals.
T18 382133-382384 Sentence denotes While sequestering of neutrophils in the lung and pulmonary edema was diminished in Asm(-/-) mice, these animals failed to clear the infection, as indicated by a progressive increase in S. aureus CFUs isolated from saponin treated splenic homogenates.
T19 382385-382473 Sentence denotes Wild type mice, in comparison, began to clear the infection at 6 hours post-inoculation.
T20 382474-382630 Sentence denotes These data indicate that membrane ceramide is an important mediator of immune system activation and pulmonary dysfunction in S. aureus induced septic shock.
T21 382631-382794 Sentence denotes While the pulmonary manifestations of septic shock are reduced in Asm(-/-) mice, they demonstrate impaired bacterial clearance and ultimately succumb to infection.
T22 382795-382927 Sentence denotes This outcome implicates acid sphingomyelinase dependent ceramide production as an important component of the innate immune response.
T23 382928-383020 Sentence denotes Wolbachia endbacteria of filarial nematodes are sensitive to Acyldepsipeptides Introduction:
T24 383021-383097 Sentence denotes Resistance to carbapenems in Gram-negative Bacteria is increasing worldwide.
T25 383098-383194 Sentence denotes In Pseudomonas aeruginosa resistance can be mediated by production of metallo--lactamases (MBL).
T26 383195-383292 Sentence denotes MBLs are able to hydrolyse penicillins, cephalosporines, and even carbapenems, but not aztreonam.
T27 383293-383399 Sentence denotes In Germany members of the Verona integron-encoded (VIM) family of MBLs have been reported most frequently.
T28 383400-383569 Sentence denotes Here we report the detection of an IMP-13-type MBL (IMP for "active on imipenem") in two clinical isolates of P. aeruginosa at a university hospital in northern Germany.
T29 383570-383689 Sentence denotes To the best of our knowledge, this is the first report of IMP-13producing organisms from clinical specimens in Germany.
T30 383690-383702 Sentence denotes Case report:
T31 383703-383806 Sentence denotes Two P. aeruginosa strains were isolated from clinical samples in February and March 2010, respectively.
T32 383807-383929 Sentence denotes One Isolate was detected in the urine from a 70-year old male patient with a neurogenic disturbance of bladder and rectum.
T33 383930-384022 Sentence denotes The patient was admitted to our hospital for surgical treatment of a sacral decubital ulcer.
T34 384023-384195 Sentence denotes A second P. aeruginosa isolate was found in an inguinal wound swab from a 61-year old male patient who was admitted to hospital because of a type B dissection of the aorta.
T35 384196-384292 Sentence denotes Both patients received multiple antibiotics before detection of the IMP-producing P. aeruginosa.
T36 384293-384400 Sentence denotes More over, both patients developed Clostridium difficile-associated diarrhea during their stay at hospital.
T37 384401-384490 Sentence denotes Bacterial isolates were cultivated and identified by standard microbiological procedures.
T38 384491-384554 Sentence denotes Susceptibility testing was done by the Vitek2 automated system.
T39 384555-384628 Sentence denotes Production of MBL was confirmed by Etest and PCR from whole cell lysates.
T40 384629-384706 Sentence denotes Amplified PCR-products were sequenced and analyzed using the BLAST algorithm.
T41 384707-384870 Sentence denotes The MICs for both P. aeruginosa isolates were (isolate 1/isolate2, [ g/ml]): piperacillin/tazobactam 128/ 128, ceftazidime 64/ 64, imipenem 8/ 16, meropenem 4/ 16.
T42 384871-384934 Sentence denotes The production of an MBL was confirmed by Etest phenotypically.
T43 384935-385073 Sentence denotes Sequencing of a 661 bp fragment (isolate 1) and a 622 bp fragment (isolate 2), respectively, revealed an IMP-13-type MBL in both isolates.
T44 385074-385085 Sentence denotes Discussion:
T45 385086-385306 Sentence denotes Although IMP-13 has been detected in isolates from other countries like Italy, Argentina or Austria, to the best of our knowledge, this is the first report of detection of IMP-13 in clinical isolates from Germany so far.
T46 385307-385426 Sentence denotes This report underscores the sustaining spread of multidrug-resistant bacteria producing broad-spectrum beta-lactamases.
T47 385427-385534 Sentence denotes It stresses the need for more effective infection control measures and development of new antibiotic drugs.
T48 385535-385693 Sentence denotes The urinary antibiotic nitroxoline blocks septic stone formation by multiple mechanisms: a better and non-toxic alternative to acetohydroxamic acid treatment?
T49 385694-385986 Sentence denotes Urinary tract infections with P. mirabilis, P. vulgaris or P. rettgeri are often complicated and refactory to therapy, as the urease activity of these organisms is unique in mediating urine pH rises to 8 and higher, thereby leading to the formation of precipitates containing viable bacteria.
T50 385987-386100 Sentence denotes Biofilm formation on catheter or epithelial surfaces will further enhance the retention of these septic crystals.
T51 386101-386238 Sentence denotes For the treatment of such infections the nickel chelator and urease inhibitor acetohydroxamic acid (AHA) is in clinical use in the U.S.A.
T52 386239-386311 Sentence denotes However, due to severe side effects AHA has not been approved in Europe.
T53 386312-386447 Sentence denotes The purpose of this study was to test the urinary antiseptic and cation chelator nitroxoline (NIT) as a possible non-toxic alternative.
T54 386448-386643 Sentence denotes UTI isolates of P. mirabilis, P. vulgaris, P. rettgeri were collected during routine diagnostic procedures and MIC determined according to CLSI recommendations or in artificial urine with 2% TSB.
T55 386644-386735 Sentence denotes MIC values for NIT were found to lie between 7.8 and 31.3 μM being slightly lower in urine.
T56 386736-386820 Sentence denotes In contrast, no inhibitory effect was noted for AHA at concentrations of up to 1 mM.
T57 386821-387027 Sentence denotes Effects on enzymatic urease activity were examined by suspending bacteria in artificial urine with or without NIT or AHA and taking samples at regular intervals during a one hour incubation period at 37 °C.
T58 387028-387229 Sentence denotes At 1 mM, the urine concentration found during oral treatment, NIT nearly completely blocked the pH rise caused by ammonia formation (measured by glas electrode and the indophenol method, respectively).
T59 387230-387340 Sentence denotes The effect was concentration dependent and partially resolved by addition of 100 μM Ni 2+ , but not by Fe 2+ .
T60 387341-387487 Sentence denotes Similarly, precipitate formation was completely blocked by 1 mM NIT (determined by ARS staining and subsequent spectrophotometric quantification).
T61 387488-387541 Sentence denotes In comparison, AHA was slightly weaker in all assays.
T62 387542-387705 Sentence denotes When precipitate formation in preformed P. mirabilis biofilms was examined, complete blockage was obtained already with 1 μM NIT, the lowest concentration studied.
T63 387706-387817 Sentence denotes In contrast, results observed with AHA correlated well with the results obtained during urease activity assays.
T64 387818-388005 Sentence denotes However, when biofilm was preformed in the presence of urea, the subsequent NIT effect on precipitate formation was lower, thus indicating an additional effect of NIT on urease induction.
T65 388006-388162 Sentence denotes Finally, we observed a reduction of biofilm in the presence of NIT (determined by crystal violet staining and subsequent spectrophotometric quantification).
T66 388163-388268 Sentence denotes Taken together, NIT was thus found to be as good or better than AHA in the inhibition of urease activity.
T67 388269-388547 Sentence denotes Moreover, it was found to exhibit potent antimicrobial effects against all clinical isolates examined, and reduced precipitate formation in P. mirabilis biofilms, presumably by a combination of biofilm degradation, inhibition of urease induction and blockage of urease activity.
T68 388548-388559 Sentence denotes Background:
T69 388560-388702 Sentence denotes This study aimed at assessing the risk of Cronobacter sakazakii contamination of spray drying processes within infant milk formula production.
T70 388703-388736 Sentence denotes Three main aspects were examined:
T71 388737-388919 Sentence denotes The bacteria specific D-value for thermic sterilisation properties, biofilm formation on certain filter materials, and efficiency of cleaning agents for the cleaning in place system.
T72 388920-389106 Sentence denotes The D-value was analyzed in CASO bouillon and reconstituted infant formula, both were inoculated with C. sakazakii (DSM 4485) in a concentration about 10 7 cfu/ml and inoculated at 58°C.
T73 389107-389144 Sentence denotes Samples were taken every two minutes.
T74 389145-389360 Sentence denotes The bacteria count was determined by the cast plate method on CASO agar.For optical detection of biofilm formation, cellulose and polypropylene filter were inoculated with C. sakazakii and exposed to dry conditions.
T75 389361-389468 Sentence denotes The filter material was fixed in 4% Glutaraldehyde and then dried in ethanol and in a critical point dryer.
T76 389469-389545 Sentence denotes The pictures were made by a scanning electron micrograph LEO Stereoscan 420.
T77 389546-389667 Sentence denotes An agar diffusion test and a qualitative suspension test were made to test the effect of acids and bases to C. sakazakii.
T78 389668-389773 Sentence denotes Sodium hydroxide and nitric acid were used in different concentrations (2 %, 2.5 %, 3 %, 4 %, 5 %, 10 %).
T79 389774-389862 Sentence denotes The bacteria count was determined by cast plate method on CASO agar with an inactivator.
T80 389863-389958 Sentence denotes The D58°C of C. sakazakii was 2.2 to 6.6 in CASO bouillon and 1.8 to 4.3 in the infant formula.
T81 389959-390056 Sentence denotes Influencing factors were heat conductivity of media, culture conditions and state of cell growth.
T82 390057-390153 Sentence denotes C. sakazakii is able to form biofilm on cellulose and polypropylene fibres under dry conditions.
T83 390154-390249 Sentence denotes On cellulose fibres the bacteria were rod-shaped while on polypropylene coccoid forms occurred.
T84 390250-390303 Sentence denotes C. sakazakii is approvingly sensitive to nitric acid.
T85 390304-390438 Sentence denotes Sodium hydroxide (2.5 -3%) treatment leads to aggregate formations of the bacteria and to a total depletion after 60s incubation time.
T86 390439-390450 Sentence denotes Conclusion:
T87 390451-390594 Sentence denotes As C. sakazakii adheres to organic and synthetic filter materials, spray drying processes of milk based products are at risk for contamination.
T88 390595-390771 Sentence denotes Therefore, pasteurisation process validation for all kind of heat exchanger and cleaning regimes with nitric acid and sodium hydroxide reduces this specific contamination risk.
T89 390772-391051 Sentence denotes Cronobacter bacteria, formerly classified as Enterobacter sakazakii, have been implicated in several incidents as the cause of meningitis and enterocolitis with high mortality rates in premature infants resulting from feeding with contaminated powdered infant formula (PIF) [1] .
T90 391052-391205 Sentence denotes PIF therefore is strictly recommended to be "sakazakii-free" which is defined as the absence of any colony forming unit in 30 samples of 10g of PIF [2] .
T91 391206-391478 Sentence denotes In cooperation with the Food Process Engineering section of our institute and the Milchwerke "Mittelelbe" GmbH (an industrial producer of PIF), we are developing a modified production process assuring the inactivation of Cronobacter by pasteurization of all raw materials.
T92 391479-391631 Sentence denotes Detection of viable cells is accomplished by quantitative RealTime PCR as well as by selective growth on chromogenic media following enrichment culture.
T93 391632-391751 Sentence denotes We are going to present our first results of heat inactivation kinetics of Cronobacter species in raw materials of PIF.
T94 391752-391828 Sentence denotes Based upon these data, a modified pasteurization process is to be developed.
T95 391829-391900 Sentence denotes Clean rooms are essential in aseptic pharmaceutical or food production.
T96 391901-392016 Sentence denotes Monitoring microbial distribution and identifying the predominant isolates is part of Good Manufacturing Practices.
T97 392017-392187 Sentence denotes Aims of this research were to screen for the predominant bacteria strains distributed in clean rooms and to analyze their microbiological and molecular characterizations.
T98 392188-392209 Sentence denotes Material and Methods:
T99 392210-392384 Sentence denotes Bacteria strains were isolated in the routine aseptic monitoring courses from the clean rooms of a pharmaceutical factory, using air sampler and with air settle plate method.
T100 392385-392458 Sentence denotes Representative bacteria strains were selected based on colony morphology.
T101 392459-392549 Sentence denotes On divergent colonies, Gram staining, as well as oxidize and catalase tests was performed.
T102 392550-392596 Sentence denotes The method of these tests followed John et al.
T103 392597-392603 Sentence denotes 1994 .
T104 392604-392732 Sentence denotes With this process, and based on the distribution frequencies of these isolates, 3 predominant strains named S1~S3 were selected.
T105 392733-392991 Sentence denotes Their morphology and physiological/ biochemical characteristics were tested according to John et al. (1994) Strains of the Gram-positive human pathogenic species Listeria monocytogenes have caused several food-borne outbreaks of disease including fatalities.
T106 392992-393109 Sentence denotes The species is characterized by its ubiquitous occurrence as well as a wide range in growth temperature and pH-value.
T107 393110-393182 Sentence denotes Risk foods are especially raw milk and raw milk products, meat and fish.
T108 393183-393280 Sentence denotes Especially the ready to eat-foods smoked and graved salmon have been identified as risk products.
T109 393281-393377 Sentence denotes Listeria may be spread through the fishprocessing equipment and machinery and may persist there.
T110 393378-393517 Sentence denotes Today, insufficient data exist whether the H1-lubricants applied in these machines are putative reservoirs and sources of L. monocytogenes.
T111 393518-393681 Sentence denotes In this project, we investigated the survival of three L. monocytogenes strains in H1-lubricants with different chemical compositions applied in the fish industry.
T112 393682-393773 Sentence denotes In a first step, the presence of L. monocytogenes in the native H1-lubricants was assessed.
T113 393774-393981 Sentence denotes Upon a negative result, in the second step of the studies the lubricants were contaminated with the three L. monocytogenes strains and their survival in the lubricants was monitored over a period of 14 days.
T114 393982-394104 Sentence denotes Furthermore, the influence of the water content of the lubricants (0, 1 and 5%) on the survival of Listeria was evaluated.
T115 394105-394154 Sentence denotes None of the native lubricants contained Listeria.
T116 394155-394314 Sentence denotes Upon contamination of the lubricants with the three L. monocytogenes strains their viable counts decreased significantly by at least 99.995% within seven days.
T117 394315-394439 Sentence denotes The reduction rates were found to depend on the chemical composition of the lubricant as well as on the investigated strain.
T118 394440-394559 Sentence denotes Interestingly, the isolate from smoked salmon tolerated this environment better than the reference and the type strain.
T119 394560-394628 Sentence denotes The water content of the lubricant had no influence on the survival.
T120 394629-394738 Sentence denotes Based on the results of this study we conclude that the investigated strains cannot survive in H1-lubricants.
T121 394739-394924 Sentence denotes Cronobacter sakazakii is an emerging food-borne pathogen which has been involved in life-threatening infections such as meningitis, enterocolitis and septicemia in neonates and infants.
T122 394925-395066 Sentence denotes Outbreaks are rare, but in several of these cases contaminated powdered infant formula was identified as a source of C. sakazakii infections.
T123 395067-395212 Sentence denotes The detection of C. sakazakii with the current ISO/TS standard method 22964:2006 requires five to six days to confirm the species identification.
T124 395213-395356 Sentence denotes Therefore, a rapid, specific and sensitive method for the detection and identification of C. sakazakii in powdered infant formula is necessary.
T125 395357-395513 Sentence denotes This study describes the development of a real-time PCR (RT-PCR) based assay for the detection of C. sakazakii in powdered milk products and infant formula.
T126 395514-395599 Sentence denotes The proposed method combines an unspecific enrichment, sample preparation and RT-PCR.
T127 395600-395686 Sentence denotes All bacterial strains were grown in brain heart infusion broth at 37°C with agitation.
T128 395687-395810 Sentence denotes Genomic DNA was extracted from overnight cultures using an illustra TM bacteria genomic Prep Mini Spin Kit (GE-Healthcare).
T129 395811-395937 Sentence denotes Two primers and a TaqMan probe were designed with the Beacon Designer software to target outer membrane protein A gene (ompA).
T130 395938-396044 Sentence denotes The assay was validated with 40 C. sakazakii strains and 17 strains of closely related species and genera.
T131 396045-396172 Sentence denotes A TaqMan RT-PCR assay was successfully developed with a detection limit of 0,001 μg DNA/ml and a reaction efficiency of 96.5 %.
T132 396173-396324 Sentence denotes All tested C. sakazakii strains were identified in 100 min with reproducible amplification curves and all non C. sakazakii strains were not detectable.
T133 396325-396414 Sentence denotes The developed TaqMan RT-PCR assay reliably detects and identifies C. sakazakii very fast.
T134 396415-396522 Sentence denotes The analysis time of the complete assay including enrichment and sample preparation was approximately 22 h.
T135 396523-396639 Sentence denotes The described method is a solid basis for the fast and reliable detection of C. sakazakii in powdered milk products.
T136 396640-396697 Sentence denotes Differential proteomic expression of enterohemorrhagic E.
T137 396698-396795 Sentence denotes Enterohemorrhagic Escherichia coli (EHEC) are enteric pathogens causing severe illness in humans.
T138 396796-396911 Sentence denotes EHEC food-borne infections may lead to nonbloody diarrhea, bloody diarrhea and the hemolytic uremic syndrome (HUS).
T139 396912-397132 Sentence denotes The survival of EHEC in the food chain and in the human intestine is dependent of environmental conditions such as available nutrients and/or the exposure to stressful agents such as bile salts and/or pancreatic enzymes.
T140 397133-397208 Sentence denotes Exposure to different environments may cause a shift in metabolic pathways.
T141 397209-397292 Sentence denotes In EHEC, this may also lead to a shift in the expression of virulence determinants.
T142 397293-397561 Sentence denotes In order to test the behaviour of EHEC O157:H7 strain EDL933 grown in the gut of the human host, we used simulated ileal environment medium (SIEM) and simulated colon environment medium (SCEM) and compared it to tryptic soy broth (TSB), used as a standard rich medium.
T143 397562-397716 Sentence denotes With two-dimensional gel electrophoresis, we differentiated cytosolic proteins with at least 2-fold higher expression in SIEM and SCEM as compared to TSB.
T144 397717-397862 Sentence denotes After growth under aerobic and anaerobic conditions, the proteins were detected and evaluated by a software and identified by MALDI-TOF-analysis.
T145 397863-397978 Sentence denotes In TSB, we found a high amount of the flagellin protein FliC, suggesting that it is downregulated in SIEM and SCEM.
T146 397979-398077 Sentence denotes In contrast, several proteins involved in biofilm synthesis were up-regulated in the latter media.
T147 398078-398228 Sentence denotes We also found modulated "locus of enterocyte effacement"-associated proteins such as EspP and EspB as well as overexpressed phage-associated proteins.
T148 398229-398397 Sentence denotes In cooperation with the group of Wolfgang Eisenreich, TU Munich, we tested the incorporation rates of 13 C6labelled glucose with respect to the synthesised amino acids.
T149 398398-398457 Sentence denotes Therefore, EDL933 was cultured in SIEM, SCEM and TSB media.
T150 398458-398609 Sentence denotes The results of these analyses gave us a hint how environmental conditions affect the metabolism and virulence properties of EHEC O157:H7 strain EDL933.
T151 398610-398718 Sentence denotes Antifungal activity of lactic acid bacteria Lactobacillus plantarum IBL-2 AND TSD-10 AGAINST Penicillium sp.
T152 398719-398884 Sentence denotes The staple food such as rice, wheat, corn, peanuts and greenbeans often stored in warehouse as stockpiled supplies, but it is often damage in large numbers by fungi.
T153 398885-398937 Sentence denotes One of fungi causes of the damage is Penicillium sp.
T154 398938-399019 Sentence denotes Several species of Penicillium can grow in food that was given potassium sorbate.
T155 399020-399092 Sentence denotes The using of synthetic preservatives antifungal often induce resistance.
T156 399093-399220 Sentence denotes Food industry has been using lactic acid bacteria widely as biocontrol agent to improve food safety at minimum food processing.
T157 399221-399300 Sentence denotes It's important to exploring lactic acid bacteria as biopreservative antifungal.
T158 399301-399445 Sentence denotes The aim of this research is to know antifungal activity of lactic acid bacteria Lactobacillus plantarum 1BL-2 and TSD-10 against Penicillium sp.
T159 399446-399611 Sentence denotes Colonies of Lactobacillus plantarum isolates 1BL-2 and TSD-10 cultured in GYP liquid medium and then incubated at 37 °C for two days, then cultures were centrifuged.
T160 399612-399690 Sentence denotes Some supernatant was done ammonium sulfate precipitation and then centrifuged.
T161 399691-399740 Sentence denotes The precipitate obtained resuspended with buffer.
T162 399741-399837 Sentence denotes 50 L of sample were added to paper disk and put into MRS agar containing 0.5% of Penicillium sp.
T163 399838-399943 Sentence denotes Antifungal activity of Lactobacillus plantarum stains against Penicillium sp. was range from 15 to 18 mm.
T164 399944-400086 Sentence denotes Samples that have been neutralized still produce clear zone. this suggests that the antifungal activity not only because of acidic conditions.
T165 400087-400203 Sentence denotes Lactobacillus plantarum 1BL-2 and TSD-10 produces plantaricin that have antifungal activity against Penilcillium sp.
T166 400204-400256 Sentence denotes Lactobacillus plantarum, antifungal, Penicillium sp.
T167 400257-400506 Sentence denotes Antibacterial effects of probiotics isolated from yoghurts containing probiotics against some common bacterial pathogens Most technologies and databases designed for rapid identification of microorganisms are designed with clinical isolates in mind.
T168 400507-400737 Sentence denotes When adapted for use in the microorganism monitoring programs of pharmaceutical, nutraceutical, and sterile manufacturing environments, these technologies underperform due to the diversity of organisms found in these environments.
T169 400738-400928 Sentence denotes Whole cell proteotypic analysis with matrix-assisted laser desorption/ionization -time of flight (MALDI-TOF) spectroscopy is an emerging rapid and inexpensive method of identifying bacteria.
T170 400929-401209 Sentence denotes This study tests the accuracy, robustness, and reproducibility of this technology when applied to an extensive panel of known culture collection strains and frequently seen species found in pharmaceutical, nutraceutical and sterile manufacturing environmental monitoring programs.
T171 401210-401389 Sentence denotes The MALDI-TOF manufacturer's recommended methods for sample processing and data analysis are directly compared to methods optimized in our laboratory for environmental monitoring.
T172 401390-401482 Sentence denotes Advantages and limitations of using MALDI-TOF technology for this application are discussed.
T173 401483-401645 Sentence denotes This study showed that fewer than 70% of frequently seen environmental isolates could be accurately identified with the MALDI Biotyper™ system (Bruker Daltonics).
T174 401646-401761 Sentence denotes The >30% that failed to be identified were largely due to limitations in the clinically focused Biotyper™ database.
T175 401762-401900 Sentence denotes At Accugenix, the methods were optimized and additions made to the library such that the number of samples that were identified increased.
T176 401901-402140 Sentence denotes While a custom database and method modifications are required to optimize this technology for environmental monitoring applications, the low cost and rapid turnaround time of MALDI-TOF analysis remain strong advantages for this technology.
T177 402141-402340 Sentence denotes The gram-negative bacterium Pseudomonas aeruginosa (PA) is a frequent opportunistic human pathogen and the leading cause of morbidity and mortality among patients suffering from cystic fibrosis (CF).
T178 402341-402582 Sentence denotes For long-term persistence in the dehydrated and highly viscous sputum of CF airways PA has to adapt its phenotype remarkably in response to prevailing requirements of the CF lung environment, including a loss of acute virulence determinants.
T179 402583-402637 Sentence denotes In contrast, PA gains obviously metabolic adaptations.
T180 402638-402803 Sentence denotes Previous studies of our group have shown that in end stage CF isolates the icd and idh genes are significantly up-regulated in comparison to early-stage CF isolates.
T181 402804-403014 Sentence denotes Interestingly, PA possesses two isoenzymes of isocitrate dehydrogenase, namely dimeric Icd and the monomeric Idh that both catalyze the oxidative decarboxylation of isocitrate to ketoglutarate, CO2 and NAD(P)H.
T182 403015-403158 Sentence denotes Beside PA, only very few bacteria have two isoenzymes of isocitrate dehydrogenase suggesting a role during adaptation to environmental changes.
T183 403159-403315 Sentence denotes Additionally, isocitrate dehydrogenase is a metabolic key enzyme that controls the metabolic flux between tricarboxic acid (TCA) cycle and glyoxylate shunt.
T184 403316-403460 Sentence denotes By use of icd and idh single transposon mutants and the construction of double mutant we tried to figure out the metabolic role of both enzymes.
T185 403461-403535 Sentence denotes By Blue-Native-PAGE we were able to display the activities of each enzyme.
T186 403536-403706 Sentence denotes Furthermore, the double mutant showed a significant growth defect and a modified growth behavior in minimal media supplemented with different TCA intermediates/C-sources.
T187 403707-403802 Sentence denotes Further, we constructed a malate dehydrogenase mutant, which is also involved in the TCA-cycle.
T188 403803-403974 Sentence denotes Moreover, there are many lines of evidence that NAD(P) + -dependent isocitrate dehydrogenase and malate dehydrogenase are involved in the defense against oxidative stress.
T189 403975-404070 Sentence denotes Therefore, we will investigate these metabolic enzymes with a view to oxidative stress as well.
T190 404071-404243 Sentence denotes Campylobacter jejuni, an important food-borne bacterial pathogen in industrialized countries and in the developing world, is one of the major causes of bacterial diarrhoea.
T191 404244-404427 Sentence denotes To identify new genes which are important for the invasion of host cells by the pathogen, we screened a transposon generated mutant library based on the clinical C. jejuni isolate B2.
T192 404428-404515 Sentence denotes Thereby, we identified clones with a transposon insertion in genes cj0952c and cj0005c.
T193 404516-404622 Sentence denotes The gene encoding cj0952c together with the adjacent gene cj0951c were supposed to act as a chemoreceptor.
T194 404623-404875 Sentence denotes We could show that these genes are not translated as one protein, diminish the motility of the pathogen, alter the chemotactical behaviour of C. jejuni towards formic acid, but are not related to the utilization of formic acid by formate dehydrogenase.
T195 404876-404966 Sentence denotes The second gene cj0005c represents one subunit of a sulphite: cytochrome c oxidoreductase.
T196 404967-405109 Sentence denotes Further characterization showed the cj0005c-deficient genotype to possess a clearly reduced motility and a diminished adherence to host cells.
T197 405110-405291 Sentence denotes Furthermore, the transcription of genes responsible for the synthesis of legionaminic acid (LegAm) were downregulated and the mutant possessed a reduced capacity to autoagglutinate.
T198 405292-405425 Sentence denotes In contrast, neither the proliferation of the mutant, nor its intracellular ATP quantity was altered compared to the parental strain.
T199 405426-405518 Sentence denotes Transposons are mobile genetic elements capable of random insertion into their host genomes.
T200 405519-405660 Sentence denotes This fundamental feature is employed since decades for the inactivation of genes, leaving a detectable signature at the locus of integration.
T201 405661-405910 Sentence denotes Different modifications of the traditional transposon mutagenesis facilitated the development of new techniques, such as signature target mutagenesis, transposon mediated differential hybridization, genetic footprinting and gene expression analysis.
T202 405911-406006 Sentence denotes The crucial step of all of these methods is the localization of the transposon insertion sites.
T203 406007-406108 Sentence denotes This process is usually a time-consuming experimental procedure that requires intensive optimization.
T204 406109-406339 Sentence denotes InFiRe-Insertion Finder via Restriction digest -is a novel software tool which allows for the computational identification of transposon insertion sites in known bacterial genome sequences after transposon mutagenesis experiments.
T205 406340-406394 Sentence denotes The strategy of this procedure is outlined in Fig. 1 .
T206 406395-406496 Sentence denotes It is based on restriction digestions of genomic DNA in combination with Southern blot hybridization.
T207 406497-406584 Sentence denotes In the first step, the genomic DNA is cleaved with different restriction endonucleases.
T208 406585-406671 Sentence denotes Each restriction enzyme produces a unique pattern of DNA fragments with defined sizes.
T209 406672-406825 Sentence denotes In the second step the sizes of the fragments with an inserted transposon are determined by Southern blot hybridization with a transposon-specific probe.
T210 406826-406943 Sentence denotes The size of the fragment in the original genome equals the obtained fragment size minus the length of the transposon.
T211 406944-407087 Sentence denotes Finally, using the derived fragment size pattern the most probable genomic position of the transposon can be calculated by the InFiRe software.
T212 407088-407186 Sentence denotes Afterwards, confirmation of the accurate determination of insertion sites can be performed by PCR.
T213 407187-407288 Sentence denotes The method is potentially applicable to any virus or sequenced organisms with known genomic sequence.
T214 407289-407392 Sentence denotes The outlined method provides the solid basis for the establishment of a new high-throughput technology.
T215 407393-407449 Sentence denotes The software was implemented as a stand alone R-package.
T216 407450-407538 Sentence denotes In addition, a web interface to the program is available at http://www.infire.tu-bs.de/.
T217 407540-408066 Sentence denotes The alternative Sigma factor B modulates virulence gene expression in a murine Staphylococcus aureus infection model but does not influence kidney gene expression pattern of the host RT-qPCR of mixed eukaryotic and prokaryotic RNA from infected tissue resulted in the detection of SigB-dependent higher expression levels of asp23 and clfA in the wild type strain S. aureus RN1HG, whereas, in agreement with in vitro data, expression of hla and aur was higher in kidney tissue from animals infected with the sigB mutant strain.
T218 408067-408240 Sentence denotes Although the virulence of sigB deficient strains is often reported to be similar to that of wild type strains, the pathomechanism of different infection settings might vary.
T219 408241-408384 Sentence denotes Therefore, the rationale of this study was to investigate whether the deletion of sigB would lead to a different reaction of the infected host.
T220 408385-408492 Sentence denotes Gene expression profiling indicated a highly reproducible host kidney response to infection with S. aureus.
T221 408493-408610 Sentence denotes The comparison of infected with non-infected samples at 120 h post infection revealed a strong inflammatory reaction.
T222 408611-408630 Sentence denotes This included e. g.
T223 408631-408754 Sentence denotes TLR signaling, complement system, antigen presentation, IFN and IL-6 signaling, but also counterregulatory IL-10 signaling.
T224 408755-409054 Sentence denotes However, the results of this study did not provide any hints for differences in the pathomechanism of the S. aureus strains RN1HG Outer membrane vesicles are released from the outer membrane of pathogenic Neisseria and previous works suggested a role of OMVs in invasive meningococcal disease (IMD).
T225 409055-409163 Sentence denotes In more recent studies, OMVs are viewed as long distance delivery tools for bacterial pathogenicity factors.
T226 409164-409344 Sentence denotes Because meningococcal OMV can be purified from plasma of IMD patients, we asked whether they impact polymorphonuclear neutrophils (PMN), which are a first line defence against IMD.
T227 409345-409579 Sentence denotes By flow cytometry and fluorescence microscopy we detected fusion of Neisseria meningitidis (Nm) OMV to PMN, both in whole blood assays and in isolated cell fractions, and to differentiated HL-60 preparations, but not to T lymphycytes.
T228 409580-409657 Sentence denotes As result of fusion of OMV, phagocytosis and ROS release by PMN was impaired.
T229 409658-409724 Sentence denotes Live imaging revealed that PMN motility was also affected by OMVs.
T230 409725-409821 Sentence denotes These results suggested reduced PMN mediated killing of Nm in the presence of meningococcal OMV.
T231 409822-410005 Sentence denotes As a possible causal effector protein we identified a ribosyl transferase that proved to be present in OMVs and whose knock-out reduced the above mentioned OMV effects on neutrophils.
T232 410006-410162 Sentence denotes Taken together, our data provide evidence that Nm is able to modulate cellular immune response mechanisms by release of OMV which fuse to host immune cells.
T233 410163-410364 Sentence denotes This might aid the rapid spread of the bacteria once the blood vessels are invaded and thus support the rapid development of septic shock in many patients suffering from invasive meningococcal disease.
T234 410365-410709 Sentence denotes Cell invasion by N. meningitidis requires as common interaction between focal adhesion kinase, c-Src and cortactin Entry of Neisseria meningitidis (the meningococcus) into human brain microvascular endothelial cells (HBMEC) is mediated by fibronectin or vitronectin bound to the surface protein Opc forming a bridge to the respective integrins.
T235 410710-410790 Sentence denotes This interaction leads to cytoskeletal rearrangement and uptake of meningococci.
T236 410791-410981 Sentence denotes Here we report that binding of pathogenic N. meningitidis triggered the recruitment of the focal contact associated proteins vinculin, paxillin and zyxin to the site of bacterial attachment.
T237 410982-411153 Sentence denotes We determined that the focal adhesion kinase (FAK), which directly associates with integrins, is involved in integrin-mediated internalization of N. meningitidis in HBMEC.
T238 411154-411277 Sentence denotes Overexpression of the dominant-negative version of FAK (FRNK) blocked integrin-mediated internalization of N. meningitidis.
T239 411278-411466 Sentence denotes Importantly, FAKdeficient fibroblasts were resistant to N. meningitidis invasion and showed reduced phosphorylation of the cytoskeleton proteins and decreased development of stress fibers.
T240 411467-411606 Sentence denotes Furthermore N. meningitidis induced tyrosine phosphorylation of several host proteins including the c-Src /FAK complex substrate cortactin.
T241 411607-411862 Sentence denotes Inhibition of cortactin expression by siRNA silencing and mutation of critical amino acid residues within cortactin significantly reduced meningococcal invasion into brain endothelial cells indicating a critical role for cortactin in meningococcal uptake.
T242 411863-412004 Sentence denotes Moreover we demonstrate that the phosphorylation status of cortactin is enhanced during infection involving both FAK and Src kinase activity.
T243 412005-412242 Sentence denotes Together, these results indicate that N. meningitidis exploits the integrin signal pathway for its entry and that FAK is an important signalling molecule that mediates the transfer of signals from activated integrins to the cytoskeleton.
T244 412243-412366 Sentence denotes A cooperative interplay between FAK, c-Src and cortactin then enables endocytosis of N. meningitidis into eukaryotic cells.
T245 412367-412519 Sentence denotes Despite antibiotic therapy, infections with Neisseria meningitidis still demonstrate a high rate of morbidity and mortality even in developed countries.
T246 412520-412734 Sentence denotes The fulminant septicemic course, named Waterhouse-Friderichsen syndrome, with massive hemorrhage into the adrenal glands and widespread petechial bleeding suggest pathophysiological inhibition of platelet function.
T247 412735-412941 Sentence denotes Our data show that N. meningitidis produces the important physiological platelet inhibitor and cardiovascular signalling molecule nitric oxide (NO), also known as endothelium-derived relaxing factor (EDRF).
T248 412942-413222 Sentence denotes N. meningitidis-derived NO inhibited ADP induced platelet aggregation through the activation of soluble guanylyl cyclase (sGC) followed by an increase in platelet cyclic nucleotide levels and subsequent activation of platelet cGMP-and cAMP-dependent protein kinases (PKG and PKA).
T249 413223-413430 Sentence denotes Furthermore, direct measurement of horseradish peroxidase (HRP) passage through a vascular endothelial cell monolayer revealed that N. meningitidis significantly increased endothelial monolayer permeability.
T250 413431-413588 Sentence denotes Immunfluorescence analysis demonstrated NO dependent disturbances in the structure of endothelial adherens junctions after coincubation with N. meningitidis.
T251 413589-413689 Sentence denotes In contrast to platelet inhibition, the NO effects on HBMEC were not mediated by cyclic nucleotides.
T252 413690-413811 Sentence denotes Our study provides evidence that NO plays an essential role in the pathophysiology of septicemic meningococcal infection.
T253 413812-413919 Sentence denotes Legionella pneumophila is a Gram-negative intracellular pathogen that can cause a severe form of pneumonia.
T254 413920-413998 Sentence denotes This bacterium naturally inhabits freshwaters and parasitizes protozoan hosts.
T255 413999-414112 Sentence denotes After aerosol formation in man-made water systems, L. pneumophila can enter, colonize and destroy the human lung.
T256 414113-414223 Sentence denotes During infection the pathogen employs sophisticated machineries to deliver proteins to host cells and tissues.
T257 414224-414332 Sentence denotes Besides the secretion of individual proteins, L. pneumophila sheds vesicles derived from its outer membrane.
T258 414333-414591 Sentence denotes Outer membrane vesicles (OMVs) are spherical bilayer structures and consist of characteristic outer membrane constituents including outer membrane proteins, phospholipids and lipopolysaccharides (LPS; Shevchuk et al., 2011) as well as periplasmic components.
T259 414592-414704 Sentence denotes The group has described a comprehensive proteome reference map for OMVs of L. pneumophila (Galka et al., 2008) .
T260 414705-414797 Sentence denotes A functional classification of the proteome showed that OMVs contain many virulence factors.
T261 414798-414974 Sentence denotes Zymography and enzyme assays demonstrate that OMVs display proteolytic and lipolytic activities which may contribute to the destruction of the alveolar lining during infection.
T262 414975-415095 Sentence denotes Confocal laser scanning microscopy revealed a spatial association between L. pneumophila OMVs and the host cell surface.
T263 415096-415200 Sentence denotes It remains unclear if this indicates adhesion, binding or fusion events between OMVs and host membranes.
T264 415201-415281 Sentence denotes The role of L. pneumophila OMVs in interbacterial communication is also unknown.
T265 415282-415451 Sentence denotes To address the question if OMV material is incorporated into target cell membranes, human macrophages and different bacteria were co-incubated with OMVs in PBS at 25 °C.
T266 415452-415507 Sentence denotes Samples were taken and analysed at various time points.
T267 415508-415648 Sentence denotes The presence of the L. pneumophila major outer membrane protein (MOMP) could not be detected in any of the target cells by Western blotting.
T268 415649-415765 Sentence denotes This finding hints towards a weak interaction with cell surfaces or rapid ingestion and degradation of OMV material.
T269 415766-415909 Sentence denotes Ongoing studies address the effect of L. pneumophila OMVs on human macrophages in regard to metabolic activity and cytoskeleton rearrangements.
T270 415910-416023 Sentence denotes Interbacterial effects of L. pneumophila OMVs are dissected by immunofluorescence microscopy and FACS analysis. .
T271 416024-416132 Sentence denotes While B of S. aureus has been studied extensively, only little is known about the latter two sigma subunits.
T272 416133-416269 Sentence denotes S belongs to the group of ECF-type sigma factors, and was recently shown to modulate the stress response and virulence of this pathogen.
T273 416270-416450 Sentence denotes H , on the other hand, shares homology to H of Bacillus subtilis and ComX of Streptococcus pneumoniae, which are both involved in regulating genetic competence and stress response.
T274 416451-416639 Sentence denotes In S. aureus, this alternative sigma factor seems to control a very small regulon, which includes the operons SA1374-SA1371, SA1418-SA1416, and the prophageencoded integrase gene int only.
T275 416640-416738 Sentence denotes However, nothing is known to date, whether and under which condition H is active in this pathogen.
T276 416739-416847 Sentence denotes To address this question we analyzed the expression of sigH and the Hdepended genes by different approaches.
T277 416848-417035 Sentence denotes We first constructed a gfp/lux based reporter system under the control of the SA1418 promoter (pTF9), and a vector harboring sigH under the control of a cadmium-inducible promoter (pTF8).
T278 417036-417156 Sentence denotes Co-transforming these plasmids into S. aureus strain RN4220 allowed confirming the H -dependence of the SA1418 promoter.
T279 417157-417495 Sentence denotes Monitoring the luciferase activity of a S. aureus Newman strain harboring pTF9 during growth in TSB, however, failed to identify significant levels of luciferase activity or Gfp formation at nearly any growth stage analyzed, confirming previous findings indicating that H is essentially not active during growth under in vitro conditions.
T280 417496-417677 Sentence denotes Only at the early lag phase, immediately after the inoculation of the fresh medium with cells of an overnight culture, we detected a short but clear increase in luciferase activity.
T281 417678-417906 Sentence denotes Real-time PCR experiments allowed confirming this finding by showing clear increases in sigH, SA1374 and SA1418 transcription during the early lag phase in strain Newman, while no such effect was visible in a Newman sigH mutant.
T282 417907-418257 Sentence denotes Interestingly, neither the overexpression of sigH in strain Newman did allow to enhance the transcription of the int gene, nor did the inoculation of Newman and its sigH-derivative into fresh medium reveal clear differences in int transcription between the wild-type and the mutant, strongly suggesting that the int gene is not a direct target of H .
T283 418258-418402 Sentence denotes Highly pathogenic Staphylococcus aureus and the opportunistic pathogen Staphylococcus epidermidis secrete phenol-soluble modulin (PSM) peptides.
T284 418403-418575 Sentence denotes Virulence of S. epidermidis depends mostly on the PSM peptides, which induce chemotaxis in neutrophils and cytokine induction in peripheral blood mononuclear cells (PBMCs).
T285 418576-418656 Sentence denotes The regulation of PSM secretion and production occurs through the agr regulator.
T286 418657-418799 Sentence denotes While chemotaxis and cytokine induction are crucial for infections, the molecular basis of the recognition by leucocytes has remained unknown.
T287 418800-418923 Sentence denotes Here we demonstrate that the human formyl peptide receptor 2 (FPR2) senses S. epidermidis PSMs at nanomolar concentrations.
T288 418924-419198 Sentence denotes Specific blocking of FPR2 or the down regulation of the PSM genes in the agr mutant led to severely diminished capacities of neutrophils to detect S. epidermidis PSMs. Moreover, Staphylococci developed the quorum sensing system agr to control their detection via human FPR2.
T289 419199-419284 Sentence denotes Thus, the innate immune system uses a global mechanism to detect bacterial pathogens.
T290 419285-419368 Sentence denotes Targeting FPR2 may help to manage severe infections induced by different pathogens.
T291 419369-419465 Sentence denotes Endothelial cytokine-expression in response to Streptococcus gallolyticus subsp. gallolyticus M.
T292 419466-419483 Sentence denotes Weinstock* 1 , J.
T293 419484-419497 Sentence denotes Dreier 1 , C.
T294 419498-419511 Sentence denotes Knabbe 1 , T.
T295 419512-419561 Sentence denotes Vollmer Bad Oeynhausen, Deutschland Introduction:
T296 419562-419671 Sentence denotes Infective endocarditis (IE) is an infection of endocardium with a broad spectrum of causative microorganisms.
T297 419672-419786 Sentence denotes Streptococcus gallolyticus subsp. gallolyticus (SGG) was identified in up to 20% of streptococcal-caused IE cases.
T298 419787-420035 Sentence denotes In this study, the influences of different SGG strains on endothelial cell damage was analyzed on the basis of the expression profile of interleukin-1 (IL-1 ), IL-6, IL-8, macrophage inflammatory protein-1 (MIP-1 ) and tumor necrosis factor-(TNF-).
T299 420036-420057 Sentence denotes Material and Methods:
T300 420058-420151 Sentence denotes The endothelial cell line EA.hy926 was inoculated with 6 isolates of SGG (10 5 -10 6 CFU/ml).
T301 420152-420247 Sentence denotes Cells were lysed after 2 h and 6 h of incubation, followed by RNA-isolation and cDNA-synthesis.
T302 420248-420353 Sentence denotes Gene expression of inflammatory cytokines (IL-1 , IL-6, IL-8, MIP-1 ) was quantified using real-time PCR.
T303 420354-420451 Sentence denotes TNF-gene expression of EA.hy926 cells was incapable of measurement by quantitative real-time PCR.
T304 420452-420625 Sentence denotes Furthermore, 27 different SGG-isolates (10 5 -10 6 CFU/ml) were compared concerning the potential for stimulation the TNF--synthesis in whole blood during 4 h of incubation.
T305 420626-420699 Sentence denotes TNF-measurement was conducted using the IMMULITE 1000 Immunoassay System.
T306 420700-420708 Sentence denotes Results:
T307 420709-420887 Sentence denotes Overall, different SGG isolates showed significant differences in the stimulation-intensity for IL-6, IL-8 and MIP-1 (e.g. IL-6 relative gene expression after 6 h: isolate 10672:
T308 420888-420911 Sentence denotes 1.43 vs. isolate 05950:
T309 420912-420918 Sentence denotes 2.05).
T310 420919-421062 Sentence denotes The relative gene expression of IL-8 was significant increased even after 2 h inoculation SGG with endothelial cells (relative gene expression:
T311 421063-421080 Sentence denotes 2.31 up to 3.14).
T312 421081-421175 Sentence denotes After 6 h incubation, the gene expression level was still increased (relative gene expression:
T313 421176-421193 Sentence denotes 1.27 up to 2.16).
T314 421194-421310 Sentence denotes In contrast, the gene expression of IL-6 was significant increased after 6 h, but not markedly after 2 h incubation.
T315 421311-421414 Sentence denotes MIP-1 gene expression was stimulated only marginal after 2 h inoculation of endothelial cells with SGG.
T316 421415-421497 Sentence denotes After 6 h inoculation, two isolates caused an increased gene expression of MIP-1 .
T317 421498-421591 Sentence denotes SGG caused no increase of IL-1 gene expression in endothelial cells during incubation period.
T318 421592-421807 Sentence denotes The comparison of the potential of different strains to stimulate TNF--synthesis in whole blood revealed significant differences (e.g. isolate ATCC9809 c (TNF-) = 2800 pg/ml vs. isolate AC1181 c (TNF-) = 640 pg/ml).
T319 421808-421970 Sentence denotes Conclusions: SGG isolates revealed differences in the potential to stimulate IL-6, IL-8, MIP-1 and TNF-gene expression in both, endothelial cells and whole blood.
T320 421971-422062 Sentence denotes These divergences could affect the dimension and progress of endothelial cell damage in IE.
T321 422063-422230 Sentence denotes Further characterization of the inflammatory process will focus on orchestration of inflammatory parameters during interaction of SGG, endothelial cells and monocytes.
T322 422231-422244 Sentence denotes Introduction:
T323 422245-422402 Sentence denotes Streptococcus pneumoniae is colonizing the nasopharyngeal cavity of humans and is the causative agent of otitis media, meningitis, pneumonia and septicaemia.
T324 422403-422532 Sentence denotes Pneumococci bind human plasminogen on the bacterial cell surface via surface exposed glycolytic enzymes (Bergmann et al., 2001) .
T325 422533-422716 Sentence denotes The phosphoglycerate kinase (PGK) is present in the bacterial cytoplasm and converting 1,3-bisphoshoglycerate into 3-phosphoglycerate within glycolytic pathway thereby generating ATP.
T326 422717-422865 Sentence denotes Plasminogen is a fibrinolytic proenzyme of the serine protease plasmin, which is a key enzyme of the fibrinolytic pathway (Miyashita et al., 1988) .
T327 422866-422983 Sentence denotes The first four kringle domains of plasminogen act as inhibitory molecule for angiogenesis and are termed angiostatin.
T328 422984-423134 Sentence denotes The pgk gene was cloned using pQE30expression system (Qiagen) and the expressed recombinant His-tagged protein was purified by affinitychromatography.
T329 423135-423350 Sentence denotes Plasminogen binding was analysed using Western blot overlay, dot spot analyses and spot membranes representing the amino acid sequence of PGK by overlapping peptides of 16 amino acids each immobilized on a membrane.
T330 423351-423443 Sentence denotes Localization of PGK on the pneumococcal surface was performed by immune electron microscopy.
T331 423444-423546 Sentence denotes Binding kinetic parameters of plasminogen-PGK interaction was determined by surface plasmon resonance.
T332 423547-423555 Sentence denotes Results:
T333 423556-423667 Sentence denotes In plasminogen binding analyses, PGK of Streptococcus pneumoniae was identified as plasminogen-binding protein.
T334 423668-423831 Sentence denotes Electron microscopic studies using PGK-specific antibodies visualized its localization on the pneumococcal surface of both encapsulated and non capsulated strains.
T335 423832-423900 Sentence denotes Protein overlay analyses demonstrated that PGK binds to angiostatin.
T336 423901-424001 Sentence denotes The dissociation constants for PGK-plasminogen interaction revealed a kD within the nanomolar range.
T337 424002-424215 Sentence denotes Peptide spot array indicated the presence of two plasminogen/angiostatin binding sites located within the Nterminal PGK fragment and detected lysines 14/15 and arginines 53 as crucial for plasminogen interactions.
T338 424216-424411 Sentence denotes Protein structure of the pneumococcal PGK was determined by crystal structure analysis performed at the CSIC in Madrid and confirmed localization of these amino acids on the PGK molecule surface.
T339 424412-424423 Sentence denotes Discussion:
T340 424424-424562 Sentence denotes Pneumococcal glycolytic enzymes provide a group of multifunctional proteins displayed on the bacterial surface and binding to plasminogen.
T341 424563-424791 Sentence denotes Interestingly, the amino acid sequences, which are crucial for plasminogen binding seem to vary with respect to localization within the glycolytic proteins and with respect to charge properties required for specific interaction.
T342 424792-424965 Sentence denotes Since PGK binds preferentially to the N-terminal kringle domains of plasminogen, the interaction between pneumococcal PGK and angiostatin displays a new mode of interaction.
T343 424966-425005 Sentence denotes References Bergmann, S., et al., 2001 .
T344 425006-425010 Sentence denotes Mol.
T345 425011-425021 Sentence denotes Microbiol.
T346 425022-425037 Sentence denotes 40:1273 -1287 .
T347 425038-425055 Sentence denotes Miyashita, C., E.
T348 425056-425070 Sentence denotes Wenzel, and M.
T349 425071-425085 Sentence denotes Heiden (1988 .
T350 425086-425106 Sentence denotes Haemostasis 18:7-13.
T351 425107-425347 Sentence denotes Streptococcus pneumoniae (pneumococci) colonize the human mucosal respiratory epithelium and are able to cause severe local infections such as otitis media and life-threatening diseases including community-acquired pneumonia and meningitis.
T352 425348-425502 Sentence denotes Pneumococci have evolved sophisticated strategies to encounter its host and produce a wide variety of virulence factors contributing to the pathogenicity.
T353 425503-425725 Sentence denotes The pneumococcal surface protein C (PspC) is a major virulence factor and adhesin interacting with the complement regulator Factor H and the polymeric immunoglobulin receptor (pIgR) of respiratory mucosal epithelial cells.
T354 425726-425829 Sentence denotes The human specific PspC-pIgR interaction mediates pneumococcal adherence to and invasion of host cells.
T355 425830-425925 Sentence denotes However, the mechanism of pneumococcal endocytosis via the PspC-hpIgR interaction is not known.
T356 425926-426151 Sentence denotes Here, we demonstrated that PspC-mediated endocytosis requires clathrin coated vesicle formation, which is regulated by dynamin, and visualized the subsequent endosomal trafficking of pneumococci upon ingestions by host cells.
T357 426152-426306 Sentence denotes Pretreatment of epithelial cells with dynasore, which is a potent inhibitor of dynamin, reduced pneumococcal uptake up to 60% compared to untreated cells.
T358 426307-426458 Sentence denotes Similar, the knock-down of dynamin by shRNA also impaired pneumococcal internalization suggesting that pneumococcal uptake requires functional dynamin.
T359 426459-426715 Sentence denotes Pneumococcal internalization was also reduced significantly by inhibiting clathrin with chlorpromazine and clathrin shRNA, indicative of the important role of the clathrin machinery for pneumococcal internalization by pIgR expressing host epithelial cells.
T360 426716-426828 Sentence denotes Furthermore, confocal laser scanning microscopy was performed to elucidate the endocytic pathway of pneumococci.
T361 426829-427086 Sentence denotes Thirty minutes post infection pneumococci were co-localized with clathrin and 1 hour post infection pneumococcal trafficking was visualized to the various endosomal compartments including early, recycling and late endosomes by using GFPlabeled Rab proteins.
T362 427087-427283 Sentence denotes In conclusion, the results suggest that pneumococci are internalized via the clathrin-mediated endocytic pathway and then sorted into different endosomal compartments of the epithelial host cells.
T363 427284-427379 Sentence denotes However, the intracellular fate after reaching the phago-lysosomes is still a matter of debate.
T364 427380-427493 Sentence denotes Sustained eIF2 -phosphorylation antagonizes cell autonomous defense against S. aureus -toxin G. von Hoven* 1 , N.
T365 427494-427506 Sentence denotes Kloft 1 , C.
T366 427507-427522 Sentence denotes Neukirch 1 , W.
T367 427523-427540 Sentence denotes Bobkiewicz 1 , S.
T368 427541-427552 Sentence denotes Weis 1 , K.
T369 427553-427566 Sentence denotes Boller 2 , M.
T370 427567-427578 Sentence denotes Husmann 1 1
T1 427580-427723 Sentence denotes Previously we have shown that cells mount an autonomous defense against pore forming toxins, which allows them to survive membrane perforation.
T2 427724-427888 Sentence denotes For S. aureus -toxin we have demonstrated that clearance of pore complexes from the plasma membrane of HaCaT cells is but one requirement for cellular survival (1).
T3 427889-428144 Sentence denotes Because -toxin caused a state of energy crisis, starvation, and translational arrest, and because inhibition of PI3K prevented replenishment of cellular ATP, we further proposed that autophagy serves to maintain metabolic homeostasis in target cells (2) .
T4 428145-428280 Sentence denotes In keeping with this concept, autophagosome formation in -toxin-treated cells depended on eIF2 -phosphorylation, like in starved cells.
T5 428281-428390 Sentence denotes Because Huh7 cells fail to recover from attack by -toxin, we investigated eIF2phosphorylation in these cells.
T6 428391-428547 Sentence denotes Interestingly, eIF2 -phosphorylation was triggered by -toxin with similar kinetics as in HaCaT cells, but the response was markedly sustained in Huh7 cells.
T7 428548-428673 Sentence denotes Moreover, transmission electron microscopy analysis of toxin-treated Huh7 cells revealed stronger vesiculation in Huh7 cells.
T8 428674-428807 Sentence denotes When eIF2 -dephosphorylation in toxin-treated HaCaT cells was pharmacologically inhibited, cells failed to recover from toxin attack.
T9 428808-428971 Sentence denotes Since this effect was not due to a block of translation, sustained phosphorylation of eIF2 seems to promote killing of -toxintreated cells by an unusual mechanism.
T10 428972-429204 Sentence denotes Endothelial progenitor cells (EPCs) are a heterogeneous mixture of adult stem cells that play an essential role in revascularization after vascular damage and have shown promising results in regenerative therapy for ischemic injury.
T11 429205-429332 Sentence denotes In our work we investigated an unconventional method of improving the angiogenic potential of EPCs through bacterial infection.
T12 429333-429450 Sentence denotes Bartonella spp. are facultative intracellular pathogens and the only known bacteria to induce angiogenesis in humans.
T13 429451-429557 Sentence denotes Here we describe the course of a bacterial infection of EPCs with the vasculotropic bacterium B. henselae.
T14 429558-429715 Sentence denotes Our data demonstrate that EPCs are highly susceptible to B. henselae and that the infection does not disturb their differentiation towards endothelial cells.
T15 429716-429845 Sentence denotes Upon infection EPCs show a strong activation of hypoxia inducible factor-1 (HIF-1), the key transcription factor in angiogenesis.
T16 429846-430034 Sentence denotes This is followed by the signature HIF-1-dependent pro-angiogenic cell response including production of cytokines such as vascular endothelial growth factor (VEGF) and adrenomedullin (ADM).
T17 430035-430231 Sentence denotes Furthermore, B. henselae prevents apoptosis of EPCs and induces cell migration along a stromal cellderived factor (SDF)-1 gradient, both essential functional components of the angiogenic response.
T18 430232-430457 Sentence denotes Finally, when culture plates are coated with a basement membrane which simulates the extra-cellular matrix (Matrigel TM ), infected EPCs proliferate and assemble into complex, lumen containing vessel-like structures in vitro.
T19 430458-430788 Sentence denotes Experiments examining these structures reveal a strong up regulation of endothelial marker genes vascular endothelial growth factor receptor-2 (VEGF-2), endothelial nitric oxide synthase-3 (eNOS) and vascular endothelial cadherin (VE-Cadherin) indicating that the vessel-like structures are indeed be made up of endothelial cells.
T20 430789-430944 Sentence denotes Cumulatively, our data demonstrate that infection with B. henselae can improve the angiogenic capacity of EPCs and even induce vessel-like growth in vitro.
T21 430946-431147 Sentence denotes Former studies using cultured cells showed that an interaction of Yersinia via the adhesion factors YadA and Invasin with 1-integrins on the host cell site is a prerequisite for Yop translocation [1] .
T22 431148-431253 Sentence denotes In this study we want to show how YadA, Invasin and 1-integrins contribute to Yop translocation in vitro.
T23 431254-431364 Sentence denotes For this purpose a -lactamase reporter system was used to detect and quantify Yop injection in infected cells.
T24 431365-432022 Sentence denotes We will present evidence that (i) YadA and Invasin show striking differences how they contribute to Yop translocation. (ii) For Invasin mediated Yop translocation an intact 1-integrin cytoplasmic domain and its connection to the actin cytoskeleton seems to be important in terms of the integrity of the whole receptor and its ability to perform high affinity ligand binding. (iii) In contrast YadA does not require the 1-integrin cytoplasmic domain but might require additional factors beside integrins for effective Yop injection. (iiii) Depending on cell type YadA mediated Yop injection even occurs completely independent on the expression of 1integrins.
T25 432024-432138 Sentence denotes Streptococcus pneumoniae (pneumococci) are a Gram-positive bacteria highly adapted to the human respiratory tract.
T26 432139-432401 Sentence denotes S. pneumoniae, a microaerophilic pathogen and strictly fermentative, requires rapid and efficient response mechanisms against oxidative stress for the ability to grow and survive in the host and under reactive oxidant conditions like other facultative anaerobes.
T27 432402-432694 Sentence denotes The protection from cellular macromolecules from reactive oxygen species (ROS) produced endogenously during metabolism of dioxygen (O2) in the aerobic growth and exogenously by polymorphonuclear leukocytes (PMNs) or macrophages of host immune responses is essential for pneumococcal survival.
T28 432695-432812 Sentence denotes Beside other protection mechanisms, thioredoxin family proteins play an important role in oxidative stress tolerance.
T29 432813-432932 Sentence denotes This group of small ubiquitous proteins has a number of different important cellular functions in all living organisms.
T30 432933-433071 Sentence denotes Thioredoxins possess a highly conserved secondary structure with a CXXC active site motif and a common 3-D architecture thioredoxin motif.
T31 433072-433219 Sentence denotes Two putative surface-exposed lipoprotein genes were predicted in the pneumococcal genomes belonging to the family of the thioredoxin-like proteins.
T32 433220-433368 Sentence denotes These proteins are responsible of keeping the cellular reducing environment accepting electrons from NADPH and transferring them to other acceptors.
T33 433369-433578 Sentence denotes To investigate their role during pneumococcal growth, oxidative stress, phagocytosis and intracellular survival, the genes were mutated by allelic replacement in pneumococci with different genetic backgrounds.
T34 433579-433803 Sentence denotes In conclusion, the data will show the individual impact of the thioredoxin-like proteins and their additive effect on resistance of pneumococci against oxidative stress, which is essential for invasive pneumococcal diseases.
T35 433804-433996 Sentence denotes The cell surface proteins of Streptococcus pneumoniae (pneumococci) play a key role during the infection process and are therefore considered to be important virulence factors of the pathogen.
T36 433997-434166 Sentence denotes Surface proteins such as PspA and PspC are known to mediate bacterial adhesion to host cells, contribute to colonization and invasion and are involved in immune evasion.
T37 434167-434239 Sentence denotes Different clusters of proteins decorate the cell surface of pneumococci.
T38 434240-434350 Sentence denotes There are LPxTG cell wall-anchored proteins, choline-binding proteins such as PspC and PspA, and lipoproteins.
T39 434351-434512 Sentence denotes The surface-exposed lipoproteins of Grampositive bacteria fulfill important functions in cell signaling, protein export, substrate transport and protein folding.
T40 434513-434688 Sentence denotes Due to the pleiotropic effect upon inactivation of the lipoprotein components of ABC transporters they were initially identified as adhesins such as PsaA or virulence factors.
T41 434689-434852 Sentence denotes A direct contribution to virulence has yet only been shown for the lipoprotein SlrA, which belongs to the family of peptidyl-prolyl cis/trans isomerases, and PpmA.
T42 434853-435066 Sentence denotes The role of other lipoproteins for pneumococcal colonization, virulence and transmission has not been systematically investigated although functional genomic analysis points towards their implication in virulence.
T43 435067-435184 Sentence denotes Bioinformatic analysis of pneumococcal genomes revealed the presence of approximately 40 genes encoding lipoproteins.
T44 435185-435325 Sentence denotes Hypothetical lipoproteins and lipoproteins with unknown function were selected to explore their role for pneumococcal fitness and virulence.
T45 435326-435398 Sentence denotes Strikingly, all candidates were identified by surface proteome analysis.
T46 435399-435566 Sentence denotes Pneumococcal mutants of the selected ten lipoprotein encoding genes were generated by insertion deletion mutagenesis in pneumococci with different genetic backgrounds.
T47 435567-435716 Sentence denotes The initial comprehensive analysis included the growth behavior and stability of the mutants, which were unaltered even in chemically-defined medium.
T48 435717-435881 Sentence denotes To assess the effect of the individual lipoproteins on phagocytosis and intracellular survival, phagocytosis assays were conducted using the murine phagocytes J774.
T49 435882-436083 Sentence denotes The role of lipoproteins in pneumococcal adherence to and invasion into A549 lung alveolar epithelial cells was elucidated by the antibiotic protection assay and immunofluorescence microscopic studies.
T50 436084-436162 Sentence denotes The results revealed a various impact on intracellular survival and adherence.
T51 436163-436302 Sentence denotes A more detailed functional analysis will be performed using in vivo infection models and binding experiments with the recombinant proteins.
T52 436303-436462 Sentence denotes The Gram positive bacterium Staphylococcus aureus is a human pathogen causing several diseases including nosocomial infections such as sepsis and endocarditis.
T53 436463-436629 Sentence denotes One very important factor for colonising, resistance to antimicrobial peptides and antibiotics is a cell wall anchored glycopolymer known as wall teichoic acid (WTA).
T54 436630-436946 Sentence denotes While most of the S. aureus strains produce a ribitolphosphate (RboP) WTA polymer decorated with Nacetylglucusamine or D-alanine (D-ala), S. aureus strain L produces an extraordinary kind of WTA consisting of repetitive units of polyglycerolphosphate (GroP) substituted with by D-ala or Nacetylgalatosamine (GalNAc).
T55 436947-436981 Sentence denotes We renamed this WTA as L-type WTA.
T56 436982-437066 Sentence denotes So far staphylococcal phages can be classified into 4 major sergroups A, B, D and F.
T57 437067-437169 Sentence denotes They also differ in morphology e.g. in tail length, flexibility, contractibility, head shape and size.
T58 437170-437320 Sentence denotes Interestingly electron micrograph images revealed that serogroup L phage 187, a phage only infecting strain L is a doubleganger of serogroup B phages.
T59 437321-437418 Sentence denotes Although serogroup B phages have almost the same morphology they are not able to infect strain L.
T60 437419-437515 Sentence denotes So it was our intention to analyse if the L-type WTA serves as adsorption receptor of phage 187.
T61 437516-437696 Sentence denotes Therefore we created for the first time an L-type WTA deficient mutant in this strain background via deletion of the tagO gene responsible for the initial step of WTA biosynthesis.
T62 437697-437781 Sentence denotes We confirmed complete loss of WTA in S. aureus strain L tagO with a phosphate assay.
T63 437782-437911 Sentence denotes Moreover S. aureus strain L tagO is resistant to serogroup L and serogroup D phages shown in phage susceptibility and EOP assays.
T64 437912-438066 Sentence denotes To show that phage resistance is due to the loss of WTA phage susceptibility could be restored by complementation using a vector expressing the tagO gene.
T65 438067-438241 Sentence denotes Furthermore phage adsorption assays were performed to analyse if the observed phage resistance is due to the loss of the receptor or due to unknown post adsorption processes.
T66 438242-438341 Sentence denotes Our data clearly demonstrate that Ltype WTA serves as adsorption receptor of serogroup L phage 187.
T67 438342-438415 Sentence denotes Beside phage adsorption studies we analysed the mutant in further detail.
T68 438416-438504 Sentence denotes Growth kinetics demonstrates that loss of L-type WTA is dispensable for growth in vitro.
T69 438505-438573 Sentence denotes In contrast L-type WTA is essential for growth at high temperatures.
T70 438574-438712 Sentence denotes Furthermore the mutant produces lower amounts of biofilm so a possible role of L-type WTA in biofilm formation can not be excluded so far.
T71 438713-438845 Sentence denotes Taken together and compared to known functions of RboP/GlcNAc WTA type our data suggesting a similar role of this unique L-type WTA.
T72 438846-438976 Sentence denotes Small membrane-active peptides (PSM ) are required for phagosomal escape by Staphylococcus aureus in nonprofessional phagocytes M.
T73 438977-438990 Sentence denotes Grosz* 1 , D.
T74 438991-439007 Sentence denotes Schäfer 1 , S.S.
T75 439008-439025 Sentence denotes Chatterjee 2 , C.
T76 439026-439039 Sentence denotes Goerke 3 , C.
T77 439040-439051 Sentence denotes Background:
T78 439052-439127 Sentence denotes Staphylococcus aureus is one of the most common bacterial pathogens in men.
T79 439128-439240 Sentence denotes S. aureus is able to invade human host cells nonprofessional phagocytes (e.g. epithelial and endothelial cells).
T80 439241-439302 Sentence denotes We and others have reported a phagosomal escape of S. aureus.
T81 439303-439422 Sentence denotes However, there has been a controversy regarding the mechanism of phagosomal escape, which is only partially understood.
T82 439423-439431 Sentence denotes Methods:
T83 439432-439549 Sentence denotes Phagosomal integrity in human host cells (293 and HeLa) was monitored by pH determination with FITC-labeled bacteria.
T84 439550-439632 Sentence denotes Intraphagosomal pH was determined using FACS analysis 2 h and 6 h after infection.
T85 439633-439731 Sentence denotes As marker for acidification quenching of FITC fluorescence by low pH was used (Lâm et al., 2010) .
T86 439732-439859 Sentence denotes Additionally, a microscopic reporter recruitment technique was employed to demonstrate phagosomal escape (Giese et al., 2011) .
T87 439860-439941 Sentence denotes Isogenic mutants deficient in specific toxins were compared to wild type strains.
T88 439942-439950 Sentence denotes Results:
T89 439951-440168 Sentence denotes In both cell lines a loss of the intra-phagosomal pH 6 h after infection demonstrated an escape of S. aureus strains 6850 (strong -toxin producer), as well as the -toxin-negative strains LAC (USA300) and MW2 (USA400).
T90 440169-440265 Sentence denotes In all three strains, the PSM -deficient mutants were unable to reduce the acidification signal.
T91 440266-440325 Sentence denotes Complementation partially restored the wild type phenotype.
T92 440326-440444 Sentence denotes By contrast, other mutants tested (PSM , -toxin, PVL) behaved identical to the parental wild type strains LAC and MW2.
T93 440445-440573 Sentence denotes Atoxin-deficient mutant in strain 6850 did not differ from wild type, as determined by pH-monitoring and YFP-Fc co-localization.
T94 440574-440701 Sentence denotes Time course analysis using rifampicin inhibition demonstrated a susceptible window of roughly 3-4 hrs for strains LAC and 6850.
T95 440702-440801 Sentence denotes Furthermore, we have identified two additional strains with a moderate phagosomal escape phenotype.
T96 440802-440813 Sentence denotes Discussion:
T97 440814-440978 Sentence denotes Our data suggest that PSM , but not PSM , -toxin, or PVL, are required for phagosomal escape of the staphylococcal strains 6850, LAC, and MW2 in 293 and HeLa cells.
T98 440979-441041 Sentence denotes Surprisingly, -toxin appears not to be required in this model.
T99 441042-441170 Sentence denotes We were able to identify several escape-positive strains with different, distantly genetic backgrounds using a common mechanism.
T100 441171-441295 Sentence denotes growth inhibition assay and the deposition of activated complement components was analyzed by immunofluorescence microscopy.
T101 441296-441304 Sentence denotes Results:
T102 441305-441459 Sentence denotes Binding of complement regulators CFHR1, CFHR2, and CFHR5 to immobilized CRASP-4 could be detected by Western blotting and ELISA using specific antibodies.
T103 441460-441562 Sentence denotes Unexpectedly, CFH was not present in the eluate fraction as assessed by SDS-PAGE and Western blotting.
T104 441563-441672 Sentence denotes However, binding of serum-purified CFH to recombinant CRASP-4 was demonstrated by ELISA and Far Western blot.
T105 441673-441822 Sentence denotes Serum adsorption assays revealed that CRASP-4 producing borrelial cells were capable of binding CFHR1 and CFHR2 and to some extent CFHR5 but not CFH.
T106 441823-442071 Sentence denotes Functional analysis also shown that upon incubation with active human serum, CRASP-4 expressing spirochetes were killed by complement and large amounts of complement components C3, C6 and the membrane attack complex were deposited on their surface.
T107 442072-442083 Sentence denotes Discussion:
T108 442084-442230 Sentence denotes Taken together, we identified complement proteins CFHR2 and CFHR5 as novel ligands for the infection-associated CRASP-4 protein of B. burgdorferi.
T109 442231-442337 Sentence denotes Seemingly, binding of different CFHRs is not sufficient to protect cells from complement-mediated killing.
T110 442338-442466 Sentence denotes Thus, our data strongly suggest that CRASP-4 plays a subordinate but supporting role in complement resistance of B. burgdorferi.
T111 442467-442480 Sentence denotes Introduction:
T112 442481-442664 Sentence denotes Information on the genomes of S. dysgalactiae subsp. equisimilis (SDSE) is highly desirable since this pathogen causes a spectrum of infections similar to those caused by S. pyogenes.
T113 442665-442731 Sentence denotes Two genomes of SDSE (serological group G) were recently published.
T114 442732-442913 Sentence denotes Here we report on the complete genome sequence of SDSE strain AC-2713 possessing a group A cell wall carbohydrate (Lancefield) antigen and compare it to other streptococcal genomes.
T115 442914-443015 Sentence denotes Material and methods: SDSE strain AC-2713 originated from a blood culture of an adult (without STSS).
T116 443016-443130 Sentence denotes Pyrosequencing (shot-gun and an 8 kb paired-end library, combined 59-fold coverage -Roche/454 method) was applied.
T117 443131-443193 Sentence denotes Gap closure was achieved by Sanger sequencing of PCR products.
T118 443194-443273 Sentence denotes Assembly and finishing was obtained by applying different bioinformatics tools.
T119 443274-443384 Sentence denotes Annotation was performed by combining results of the "GenDB", "RAST", and "BASys" servers and manual curation.
T120 443385-443393 Sentence denotes Results:
T121 443394-443578 Sentence denotes The AC-2713 genome consists of one circular chromosome of 2,179,445 bp; 2,181 predicted protein coding sequences (CDSs) were identified; 15 rRNAs (5 rRNA loci) and 57 tRNAs were found.
T122 443579-443604 Sentence denotes The G+C content is 39.5%.
T123 443605-443782 Sentence denotes Three phage-related chromosomal islands (PRCIs) -but no complete prophages -were identified that did not contain any of the known phage-encoded virulence factors of S. pyogenes.
T124 443783-444090 Sentence denotes Two "clustered regularly interspaced short palindromic repeats" (CRISPRs) were identified having up to 19 spacers, some with homologies to S. pyogenes phages; this may explain the lack of complete phages in the AC-2713 genome and the low degree of rearrangements in SDSE genomes, in contrast to S. pyogenes.
T125 444091-444355 Sentence denotes Numerous orthologs of (non-phage-associated) virulence factor genes of S. pyogenes including determinants for an M-protein (emm stG485.0), pili, streptokinase, hemolysins (SLO, SLS), NADase, extracellular matrixprotein binding surface proteins, etc. were detected.
T126 444356-444403 Sentence denotes Spegg4 was found as the only superantigen gene.
T127 444404-444497 Sentence denotes Some unique features are found in the AC-2713 genome, including a salivaricin A gene cluster.
T128 444498-444580 Sentence denotes The putative genes for group A cell wall polysaccharide synthesis were identified.
T129 444581-444862 Sentence denotes Comparative analyses revealed a high degree of homology and colinearity with the published S. dysgalactiae genomes and closer relationships to S. pyogenes (based on available genomes) than to other beta-hemolytic streptococci including groups C and G streptococci of animal origin.
T130 444863-445122 Sentence denotes Conclusion/Discussion: AC-2713, an SDSE possessing Lancefield's group A antigen, is highly homologous to other S. dysgalactiae and more closely related to S. pyogenes than to other streptococcal species, including groups C and G streptococci of animal origin.
T131 445123-445226 Sentence denotes Surface proteins of Gram-positive bacteria contribute to the interaction with human Thrombospondin-1 T.
T132 445227-445241 Sentence denotes Kohler* 1 , N.
T133 445242-445254 Sentence denotes Gisch 2 , U.
T134 445255-445271 Sentence denotes Zähringer 2 , S.
T135 445272-445289 Sentence denotes Hammerschmidt 1 1
T136 445291-445389 Sentence denotes Adherence of bacteria to host cells is a multifactorial process and proceeds bacterial infections.
T137 445390-445544 Sentence denotes The versatile interplay between pathogenic bacteria and its host depend on numerous interactions of bacterial surface structures and host matrix proteins.
T138 445545-445676 Sentence denotes The matricellular glycoprotein Thrombospondin-1 (TSP-1) is mainly secreted by thrombocytes but also by many other human cell types.
T139 445677-445905 Sentence denotes TSP-1 is a multifunctional, multidomain 420 kDa homotrimer with a wide range of predicted functions in adherence and migration, cell morphology, proliferation and apoptosis as well as in interaction with extracellular proteases.
T140 445906-446134 Sentence denotes Furthermore, TSP-1 is part of the extracellular matrix and shows binding to different matrix proteins, including fibronectin, fibrinogen, heparin and furthermore to the surface receptors CD36, CD47 and integrin 5 1 (CD49e/CD29).
T141 446135-446250 Sentence denotes A recent study revealed a new role of TSP-1 for the interplay of different Gram-positive pathogens with host cells.
T142 446251-446445 Sentence denotes The TSP-1 was shown to act as a molecular bridge between host cells and Gram-positive bacteria, which facilitated adherence to and invasion into different human epithelial and endothelial cells.
T143 446446-446540 Sentence denotes Nevertheless, the receptor on the bacterial side as well as on the host side is still unknown.
T144 446541-446649 Sentence denotes In this work, TSP-1 was isolated from human buffy coats and purified via fast protein liquid chromatography.
T145 446650-446819 Sentence denotes The bioactivity of purified TSP-1 was verified in binding assays with human epithelial cells (A549) and different Gram-positive pathogenic bacteria using flow cytometry.
T146 446820-447051 Sentence denotes Surface plasmon resonance (SPR) studies with TSP-1 immobilized on CM5 biosensors and ligand overlay assays revealed a potential proteinous binding partner on the bacterial side in Staphylococcus aureus and Streptococcus pneumoniae.
T147 447052-447216 Sentence denotes To identify the protein of interest, 2Dgelelectrophoresis of surface protein fractions from S. aureus was performed and peptides were analyzed by mass spectrometry.
T148 447217-447304 Sentence denotes The results indicated binding candidates whose specific binding is under investigation.
T149 447305-447575 Sentence denotes The interaction of TSP-1 with different Gram-positive bacteria via surface exposed proteins is most likely important during colonization and bacterial dissemination and could be beneficial in the development of severe infections like infective endocarditis or pneumonia.
T150 447576-447711 Sentence denotes The discovery of antibiotics in 1929 was one of the most important milestones in medicine and in the fight against infectious diseases.
T151 447712-447860 Sentence denotes Today, the rapid evolution and the emergence of multidrug-resistant bacteria necessitate a further search for new antibiotic substances and sources.
T152 447861-448082 Sentence denotes Lantibiotics (lanthionine containing antibiotics) are ribosomally produced bacterial peptide antibiotics that show interesting antibacterial activities even in the nanomolar range against (multiresistant) human pathogens.
T153 448083-448234 Sentence denotes The characteristic thioether amino acids lanthionine and methyllanthionine are introduced by extensive enzyme-mediated posttranslational modifications.
T154 448235-448451 Sentence denotes These rare amino acids form intramolecular rings that are essential for the three-dimensional structure of lantibiotics, their enhanced stability against proteases and oxidation and convey the antimicrobial activity.
T155 448452-448619 Sentence denotes These features make lantibiotics interesting candidates or lead structures for novel antimicrobial applications in medical and food industry (Ross and Vanderas, 2011).
T156 448620-448847 Sentence denotes Blast searches employing the characteristic lantibiotic modification enzymes (LanM,B,C) in the NCBI database showed that ORFs coding for proteins involved in lantibiotic production are widespread in bacteria of different phyla.
T157 448848-449068 Sentence denotes Based on these genomic data, we identified putative lantibiotic gene clusters also in bacterial phyla for which production of lantibiotics had never been described before, like a cyanobacterial Nostoc punctiforme strain.
T158 449069-449190 Sentence denotes The focus of our project is the homologous and/or heterologous expression of those, so far uncharacterized, lantibiotics.
T159 449191-449346 Sentence denotes In this context, we were able to identify and characterize the novel two-peptide lantibiotic lichenicidin that is produced by Bacillus licheniformis DSM13.
T160 449347-449571 Sentence denotes Additionally, we revealed the presence of a partial lantibiotic gene cluster coding for the proteins involved in the producer self-protection against the well-known lantibiotic mersacidin in Bacillus amyloliquefaciens FZB42.
T161 449572-449762 Sentence denotes Transfer of the biosynthetic part of the mersacidin gene cluster into B. amyloliquefaciens FZB42 resulted in the successful expression of fully modified and active mersacidin in this strain.
T162 449763-449927 Sentence denotes Other putative lantibiotic producers, including an Anaerococcus thermophilum strain, were identified in this project and are still in the focus of the ongoing work.
T163 449928-449972 Sentence denotes Ross AC and Vederas JC., J Antibiot (Tokyo).
T164 449973-449994 Sentence denotes 2011 Jan; 64(1):27-34
T165 449995-449997 Sentence denotes M.
T166 449998-450011 Sentence denotes Türck* 1 , C.
T167 450012-450026 Sentence denotes Seifert 1 , L.
T168 450027-450046 Sentence denotes Mildenberger 1 , A.
T169 450047-450063 Sentence denotes Berscheid 1 , P.
T170 450064-450075 Sentence denotes Sass 1 , G.
T171 450076-450088 Sentence denotes Bierbaum 1 1
T172 450090-450198 Sentence denotes Gram-positive bacteria are characterized by a massive cell wall, formed by multiple layers of peptidoglycan.
T173 450199-450359 Sentence denotes The structural integrity of the cell wall directly affects the viability of the bacterium itself and biosynthesis of peptidoglycan has to be tightly controlled.
T174 450360-450481 Sentence denotes This also emphasizes the importance of cell wall targeting antibiotics in the clinical treatment of bacterial infections.
T175 450482-450665 Sentence denotes Despite that, much less is known in detail about how bacteria control the interplay between degradation and new synthesis of peptidoglycan to coordinate cell division and cell growth.
T176 450666-450892 Sentence denotes Two-component regulatory systems (TCSs), composed of a sensor histidine-kinase and a cognate response regulator protein, play a predominant role in the way how bacteria respond and adapt to environmental and stress conditions.
T177 450893-451044 Sentence denotes In S. aureus the essential YycFG (WalRK/VicRK) TCS plays a major role in the homeostasis of cell wall by controlling the expression of autolysins [1] .
T178 451045-451268 Sentence denotes In context with antiinfective treatment the VraSR TCS is involved in resistance to vancomycin and -lactam antibiotics [3, 4, 5] , and moreover confers mechanisms to sense inhibition / damage of the bacterial cell wall [6] .
T179 451269-451394 Sentence denotes In general, research on TC systems has been restricted to the use of truncated proteins, missing parts of the entire kinases.
T180 451395-451606 Sentence denotes Here, we want to show the in vitro phosphorylation activities of the full-length VraS and YycG kinases, of mutated versions of YycG and phosphotransfer to their cognate response regulator proteins VraR and YycF.
T181 451607-451609 Sentence denotes C.
T182 451610-451623 Sentence denotes Weber* 1 , K.
T183 451624-451637 Sentence denotes Krauel 2 , A.
T184 451638-451655 Sentence denotes Greinacher 2 , S.
T185 451656-451673 Sentence denotes Hammerschmidt 1 1
T186 451675-451871 Sentence denotes Gram-positive and Gram-negative bacteria are apart from being lifethreatening infectious pathogens also the supposed etiologic agents for the development of heparin-induced thrombocytopenia (HIT).
T187 451872-452084 Sentence denotes HIT is an IgGantibody-mediated adverse drug reaction against complexes of the positively-charged chemokine platelet factor 4 (PF4) and the most frequently used anionic anticoagulant in clinical medicine, heparin.
T188 452085-452393 Sentence denotes Interestingly, even heparin-naïve patients can generate IgG-antibodies specific for PF4/heparin complexes as soon as 4 days after exposure to heparin, presumably because these patients have encountered complexes similar to PF4/heparin before such as PF4 bound to anionic bacterial surfaces during infections.
T189 452394-452541 Sentence denotes Likely candidates for negatively charged molecules on bacterial surfaces are lipopolysaccharides (LPS) and teichoic acids (TA) as well as proteins.
T190 452542-452807 Sentence denotes In this study, we were able to pinpoint the bacterial binding partners of PF4 by showing that PF4 interacts with LPS of the Gram-negative model organisms Escherichia coli and Salmonella typhimurium and with teichoic acids of the Gram-positive Staphylococcus aureus.
T191 452808-452891 Sentence denotes This was shown in ELISA and competitive inhibition binding experiments experiments.
T192 452892-453024 Sentence denotes Surprisingly, E. coli and S. typhimurium mutants with successively shortened LPS-backbone displayed increasing PF4 binding capacity.
T193 453025-453091 Sentence denotes The highest binding was detected in E. coli waaC and E. coli waaA.
T194 453092-453148 Sentence denotes These mutants both lack the O-antigens and the core LPS.
T195 453149-453323 Sentence denotes However, the E. coli mutant waaA is additionally deficient in the 3deoxy-D-manno-octulosonic acids (KDO) suggesting that Lipid A is the structure contributing to PF4-binding.
T196 453324-453477 Sentence denotes Similarly, a S. aureus mutant with severely truncated wall teichoic acids (SA113 tagO) showed increased PF4 recruitment compared to the wild-type strain.
T197 453478-453685 Sentence denotes Additionally, pre-treatment of bacteria with pronase E and trypsin impaired PF4 binding to S. aureus but not to E. coli, indicating proteins as another major binding partner of PF4 on Gram-positive bacteria.
T198 453686-453804 Sentence denotes The anti-PF4/heparin antibodies generated upon binding of PF4 to bacteria enhanced phagocytosis of PF4coated bacteria.
T199 453805-454058 Sentence denotes Thus, by acting as a bacteria-opsonizing protein, PF4 is able to induce a humoral immune response specific for a wide variety of bacterial species and might therefore be an ancient host defense mechanism at the interface of innate and adaptive immunity.
T200 454059-454209 Sentence denotes HIF-1 is the key transcriptional factor involved in the adaption process of mammals to hypoxia and plays a crucial role in, e.g., cancer angiogenesis.
T201 454210-454299 Sentence denotes Recent evidence suggests a leading role of HIF-1 in inflammatory and infectious diseases.
T202 454300-454496 Sentence denotes We demonstrated previously that human pathogenic S. aureus leads to a HIF-1 activation in vitro, ex vivo and in vivo and that this HIF-1 activation is detrimental in a S. aureus peritonitis model.
T203 454497-454648 Sentence denotes To analyze the role of HIF-1 in S. aureus infections in more detail, we investigated the HIF-1 dependent host cell response upon a S. aureus infection.
T204 454649-454815 Sentence denotes For this purpose, control and HIF-1 -/-HepG2 cells were infected with S. aureus and gene expression was analyzed four hours later using Affymetrix WT 1.0 gene arrays.
T205 454816-454932 Sentence denotes Data revealed that expression of in total 253 genes was influenced of which 22 turned out to be regulated via HIF-1.
T206 454933-455030 Sentence denotes These 22 genes include genes involved in cell metabolism and genes known to affect tumour growth.
T207 455031-455297 Sentence denotes One of the upregulated genes was lysyl oxidase, (lox), a copper-dependent amine oxidase, which catalyzes the cross linking of collagen and elastin molecules in the extracellular matrix and might be involved in the development of a fibrosis after S. aureus infection.
T208 455298-455557 Sentence denotes Further experiments revealed that lox is upregulated in vitro and in vivo using HepG2 infection models, abscess-harbouring kidneys of intravenously infected NMRI-mice and tissue samples from patients suffering from microbiological proven S. aureus infections.
T209 455558-455710 Sentence denotes The role of lox in abscess formation and, moreover, in chronic infections known to result in fibrous scarring of tissues needs to be further elucidated.
T210 455711-455795 Sentence denotes Characterizing the anti-apoptotic activity of the Coxiella burnetii effector AnkG R.
T211 455796-455810 Sentence denotes Eckart* 1 , L.
T212 455811-455829 Sentence denotes Klingenbeck 1 , I.
T213 455830-455845 Sentence denotes Wittmann 1 , A.
T214 455846-456058 Sentence denotes Lührmann 1 1 Universitätsklinik Erlangen, Mikrobiologisches Institut, Erlangen, Deutschland Coxiella burnetii is a Gram-negative, obligate intracellular pathogen that causes Q-fever, a worldwide zoonotic disease.
T215 456059-456153 Sentence denotes Q-fever is a mild flu-like illness, but can be associated with chronic or even fatal outcomes.
T216 456154-456239 Sentence denotes C. burnetii are typically transmitted to humans by inhalation of infectious material.
T217 456240-456422 Sentence denotes For intracellular pathogens such as C. burnetii, manipulation of host cell pathways is important to ensure a productive infection and may play a role in establishing chronic disease.
T218 456423-456533 Sentence denotes C. burnetii employs a type IV secretion system (T4SS) and several substrates of the T4SS have been identified.
T219 456534-456671 Sentence denotes One of these substrate proteins, AnkG, inhibits pathogeninduced apoptosis, probably through binding to the host cell protein p32 (gC1qR).
T220 456672-456779 Sentence denotes The mitochondrial matrix protein p32 seems to be a pro-apoptotic protein, however the mechanism is unknown.
T221 456780-457020 Sentence denotes Studies with different truncation derivatives showed that the amino terminal AnkG1-69 region is sufficient to interact with p32 and to inhibit host cell apoptosis whereas AnkG70-339 did not interact with p32 nor inhibit host cell apoptosis.
T222 457021-457110 Sentence denotes Ectopically expressed AnkG shows a vesicular pattern with some mitochondrial association.
T223 457111-457235 Sentence denotes The p32 protein seems to be involved in bridging a signaling pathway that extends from the mitochondria to the cell nucleus.
T224 457236-457348 Sentence denotes Here, we aim to investigate how the interaction of AnkG with p32 determines the anti-apoptotic activity of AnkG.
T225 457349-457433 Sentence denotes Therefore, we tested whether the interaction of AnkG with p32 is direct or indirect.
T226 457434-457511 Sentence denotes Using GSTas well as His-pull-down experiments we demonstrated direct binding.
T227 457512-457699 Sentence denotes As p32 is a mitochondrial protein we hypothesized that if the anti-apoptotic activity of AnkG is mediated by p32, AnkG only inhibits the intrinsic and not the extrinsic apoptosis pathway.
T228 457700-457767 Sentence denotes This hypothesis was confirmed using ectopic expression experiments.
T229 457768-457895 Sentence denotes Furthermore, using localization studies we showed that AnkG shuttles from the cytosol to the nucleus after apoptosis induction.
T230 457896-458010 Sentence denotes Interestingly, ectopic expression showed that the N-terminus (AnkG1-69) is nuclear, while AnkG70-339 is cytosolic.
T231 458011-458083 Sentence denotes In both cases the localization did not change after apoptosis induction.
T232 458084-458246 Sentence denotes These data suggest that nuclear localisation as well as binding to p32 is necessary for the antiapoptotic activity of the Coxiella burnetii effector protein AnkG.
T233 458247-458341 Sentence denotes Staphylococcus saprophyticus is an important cause of urinary tract infections in young women.
T234 458342-458498 Sentence denotes The amino acid D-serine occurs in relatively high concentrations in human urine and has a bacteriostatic or toxic effect on many non-uropathogenic bacteria.
T235 458499-458612 Sentence denotes S. saprophyticus harbors the cytoplasmatic enzyme D-serine deaminase (DsdA), which is found in many uropathogens.
T236 458613-458704 Sentence denotes It is shown that DsdA enables S. saprophyticus to cleave Dserine into pyruvate and ammonia.
T237 458705-458778 Sentence denotes This is probably a factor that enables colonization of the urinary tract.
T238 458779-458993 Sentence denotes In contrast to E. coli the dsdA-operon of S. saprophyticus does not encode a specific D-serine-transporter, but there are three genes in the genome of S. saprophyticus that encode for putative Dserine-transporters.
T239 458994-459148 Sentence denotes SSP0286, SSP1070 and SSP2171 show a identity of 50 % and a similarity of 72 % to CycA an unspecific D-serine transporter of the uropathogen E.coli CFT073.
T240 459149-459266 Sentence denotes We created knock-out mutants of these genes in order to characterize the D-serine-transport of S. saprophyticus 7108.
T241 459267-459382 Sentence denotes In a defined medium D-serine elongates the lag-Phase of S. saprophyticus but not of the transporter mutant SSP1070.
T242 459383-459551 Sentence denotes We measured Dserine transport in an uptake essay by analyzing the remaining concentrations of D-serine in the supernatant of a bacterial suspension by NMR-spectroscopy.
T243 459552-459674 Sentence denotes In keeping with the observation of growth in the defined medium, the D-serine uptake of SSP1070 is lower in the NMR assay.
T244 459675-459790 Sentence denotes The reduction of D-serine uptake has therefore a positive effect on growth in defined medium under this conditions.
T245 459791-459906 Sentence denotes To take account of this results SSP1070 appears to be the major player in D-serine uptake in S. saprophyticus 7108.
T246 459907-460051 Sentence denotes The knowledge of D-serine transport and metabolism will presumably offer us more insights in to the physiology of virulence of S. saprophyticus.
T247 460053-460154 Sentence denotes A large group of bacterial surface proteins is represented by the family of autotransporter proteins.
T248 460155-460264 Sentence denotes Representatives belong to the type V secretion system (1) and are found in almost all Gram-negative bacteria.
T249 460265-460435 Sentence denotes Autotransporter proteins frequently represent important virulence factors, e.g. the trimeric autotransporter adhesin YadA of the enteropathogenic Yersinia enterocolitica.
T250 460436-460599 Sentence denotes Typically, a autotransporter consist of three functional domains: a N-terminal signal sequence, a C-terminal translocator domain and a passenger domain in between.
T251 460600-460825 Sentence denotes After synthesis in the cytosol and Sec-mediated transport across the inner membrane into the periplasm, the translocator domain forms a beta-barrel pore in the outer membrane, presumably with the help of the Bam complex (2) .
T252 460826-460918 Sentence denotes Through this pore, the passenger domain is then translocated to the surface of the cell (3).
T253 460919-461218 Sentence denotes A second, unrelated family of outer membrane proteins that expose passenger domains on the bacterial outer surface are the intimins and invasins, nonfimbrial adhesins from pathogenic bacteria, which specifically interact with host cell surface receptors and mediate bacterial attachment or invasion.
T254 461219-461401 Sentence denotes They are integrated into the bacterial outer membrane with the amino-terminal region, while the carboxy-terminal region of the polypeptide is exposed on the bacterial outer membrane.
T255 461402-461601 Sentence denotes Whereas the surface-localized parts of the protein are functionally well described, the topology and insertion of the N-terminal membrane domain and the translocation process have not been described.
T256 461602-461788 Sentence denotes To investigate the topology and the mechanism of translocation in more detail, we had a closer look at the amino acid sequence of invasin and intimin using SignalP, PsiBLAST and HHAlign.
T257 461789-461930 Sentence denotes Herefrom we got a prediction of the signal peptide and twelve different -strands, which might build up the -barrel within the outer membrane.
T258 461931-462032 Sentence denotes Out of these predictions, we developed topology models of the membrane anchor of invasin and intimin.
T259 462033-462215 Sentence denotes By introduction of HA-Tags defining the orientation of the translocator domain within the outer membrane and extensive immunofluorescence studies, we were able to confirm our models.
T260 462216-462438 Sentence denotes Moreover, we came to believe that Intimin (Int) and Invasin (Inv), two major pathogenicity factors of E.coli and Yersinia, are monomeric autotransporters, with the remarkable difference that their domain order is reversed.
T261 462439-462551 Sentence denotes Streptococcus anginosus is a commensal of the oral cavity, the gastrointestinal and the female urogenital tract.
T262 462552-462693 Sentence denotes It has clinical significance in abscess formation and has recently been suggested to play a pathogenic role in patients with cystic fibrosis.
T263 462694-462826 Sentence denotes An interesting feature of these bacteria is the inconsistent phenotype regarding Lancefield antigens as well as haemolytic activity.
T264 462827-462980 Sentence denotes While a considerable percentage of S. anginosus strains display a prominent ß-haemolytic phenotype, the corresponding genes have not been identified yet.
T265 462981-463107 Sentence denotes In different streptococcal species the ß-haemolysin is a modified short peptide (SLS) that is related to Class I bacteriocins.
T266 463108-463239 Sentence denotes It is encoded in the sag gene cluster including genes for the corresponding posttranslational modification and transport machinery.
T267 463240-463419 Sentence denotes By random chromosomal integration of the pGhost9:ISS1 transposition vector we generated a plasmid-based mutant library of the haemolytic Streptococcus anginosus strain ATCC 12395.
T268 463420-463591 Sentence denotes This library was screened for mutants showing a loss of the ß-haemolytic phenotype on blood agar plates and non-haemolytic mutants were selected for further investigation.
T269 463592-463913 Sentence denotes By sequencing the insertion sites of these mutants we identified thus far 10 different mutations sites in a gene cluster of 9 kb harbouring 9 open reading frames (ORFs), with significant similarities to the sag (SLS associated gene) gene cluster of Streptococcus pyogenes that encodes the haemolysin Streptolysin S (SLS).
T270 463914-463994 Sentence denotes ORFs corresponding to all of the 9 sag genes (sagA to sagI) could be identified.
T271 463995-464154 Sentence denotes Similarities of the deduced amino acids of the putative S. anginosus sag gene cluster to the Sag-proteins of S. pyogenes range from 37 % (sagF) to 81 % (sagD).
T272 464155-464298 Sentence denotes To further investigate the S. anginosus haemolysin, a functional haemolysin assay with culture supernatants and whole bacteria was carried out.
T273 464299-464510 Sentence denotes Haemolytic activity was only observed with whole cells, but not in the supernatant, indicating that like SLS of S. pyogenes, the S. anginosus haemolysin is able to lyse erythrocytes only in cell-associated form.
T274 464511-464705 Sentence denotes In summary we were able to successfully establish a pGhost9:ISS1 mutant library in S. anginosus and identify an SLS-like gene cluster as the genetic basis of S. anginosus ßhaemolysin production.
T275 464706-464846 Sentence denotes A magnetic isolation method to analyze differential Chlamydia pneumoniae interacting protein profiles in persistence infection Introduction:
T276 464847-464975 Sentence denotes Chlamydia pneumoniae (Cpn) is an obligate intracellular bacterial pathogen and causes acute infections of the respiratory tract.
T277 464976-465238 Sentence denotes Further, Cpn can also cause prolonged or chronic infections which may be due to persistent infections in respiratory and non-respiratory tissues and have been implicated in the development of a number of chronic diseases such as asthma, COPD and atherosclerosis.
T278 465239-465357 Sentence denotes Persistent chlamydiae exhibit characteristic protein expression profiles and are refractory to killing by antibiotics.
T279 465358-465464 Sentence denotes However, host-pathogen interacting proteins during persistent infection have not been investigated so far.
T280 465465-465609 Sentence denotes Recently, we developed a novel magnetic isolation technique for Chlamydiae using chlamydial lipid biotinylation and magnetic nanobeads labeling.
T281 465610-465847 Sentence denotes Key characteristics of this technique are gentle mechanical sonication of the plasma membrane, followed by a magnetic separation of labelled targets, which subsequently allows the biochemical detection of compartment-containing proteins.
T282 465848-465937 Sentence denotes In this study, we applied this method to investigate persistent Cpn interacting proteins.
T283 465938-466093 Sentence denotes Material and methods: FACS, confocal microscopy and electron microscopy were used to check labelling efficacy of biotinylated lipid and magnetic nanobeads.
T284 466094-466183 Sentence denotes Fractions containing magnetically labelled Cpn were isolated by using a magnetic chamber.
T285 466184-466249 Sentence denotes Interferon (IFN)-was used to induce persistent infections of Cpn.
T286 466250-466322 Sentence denotes Persistent Cpn interacting protein profile was investigated by SDS-PAGE.
T287 466323-466331 Sentence denotes Results:
T288 466332-466473 Sentence denotes Fractions containing magnetically labelled Cpn were efficiently separated from non-labeled cytoplasmic fractions by using a magnetic chamber.
T289 466474-466793 Sentence denotes In early infection (1 h p.i.), not only Cpn proteins like heat shock protein (HSP) 60, inclusion membrane protein (Inc) A and macrophage infectivity potentiator (MIP) but also early endosome marker proteins such as Rab5 and Rab4 were significantly detected in the magnetic "chlamydial" fraction compared to mock sample.
T290 466794-466889 Sentence denotes GAPDH served as a control and was strongly enriched in the non-magnetic "cytoplasmic" fraction.
T291 466890-467039 Sentence denotes Expression of HSP60, IncA and MIP was reduced in the magnetic "chlamydial" fraction in IFN-treated cells in comparison to non-treated cells 24 h p.i.
T292 467040-467204 Sentence denotes In contrast, 11 protein bands were enhanced detected in the magnetic "chlamydial" fraction of the IFN-treated cells by SDS-PAGE that needs further characterization.
T293 467205-467227 Sentence denotes Conclusion/Discussion:
T294 467228-467389 Sentence denotes Our results indicate the suitability of the novel magnetic isolation method to identify pathogen-host interacting proteins in a targeted and untargeted approach.
T295 467390-467565 Sentence denotes Escherichia coli strains are the most prevalent source of urinary tract infections (UTI), one of the most common bacterial infections in humans and a major cause of morbidity.
T296 467566-467771 Sentence denotes Uropathogenic E. coli (UPEC) causing UTI can be divided into pathogenic strains causing symptomatic UTI and nonpathogenic strains causing an asyptomatic bacteriuria (ABU) without provoking a host response.
T297 467772-467885 Sentence denotes The reason why ABU-associated E. coli strains do not evoke symptoms in the host is not completely understood yet.
T298 467886-468012 Sentence denotes Many ABU E. coli isolates seem to lack virulence associated phenotypes, although they carry many typical UPEC virulence genes.
T299 468013-468187 Sentence denotes It has been shown that such isolates express less virulence factors and that there is an evolution towards commensalism because of virulence attenuation and genome reduction.
T300 468188-468448 Sentence denotes Alterations in essential virulence genes including single base mutations, deletions or DNA rearrangements might be a general mechanism for long-term colonizing mucosal pathogens to evolve towards commensalism, consistent with a low immune response of the host.
T301 468449-468570 Sentence denotes In this study, the importance of the two-component system (TCS) BarA/UvrY for urovirulence of E. coli should be analysed.
T302 468571-468657 Sentence denotes The BarA/UvrY TCS is involved in bacterial adaption and survival in the urinary tract.
T303 468658-468893 Sentence denotes It plays a significant role in controlling a common regulatory network affecting a multitude of cellular functions, e.g. regulation of carbon metabolism, biofilm formation or motility and adhesion, and thus contributes to urovirulence.
T304 468894-469074 Sentence denotes In previous studies, genome sequence analysis revealed the frequent occurrence of single nucleotide polymorphisms in the uvrY gene upon prolonged colonization of the urinary tract.
T305 469075-469235 Sentence denotes This may indicate that uvrY is (i) a mutation hot spot and (ii) under positive selection during bacterial adaption during long-term growth in the urinary tract.
T306 469236-469476 Sentence denotes To correlate uvrY nucleotide sequence variability with prolonged colonization of the urinary tract, the uvrY gene was sequenced in 216 E. coli strains representing ABU and acute UTI isolates as well as a control group of commensal variants.
T307 469477-469575 Sentence denotes The sequence data obtained was compared with DNA sequences of housekeeping genes in these strains.
T308 469576-469713 Sentence denotes The results will be discussed in the light of genome plasticity and bacterial adaptation strategies to changing environmental conditions.
T309 469714-469923 Sentence denotes Despite being key for Staphylococcus epidermidis pathogenicity in implantassociated infections, large proportions of invasive Staphylococcus epidermidis isolates do not form biofilms under in vitro conditions.
T310 469924-470092 Sentence denotes We here tested the hypothesis that this apparent paradox is related to the existence of superimposed regulatory systems suppressing a multi-cellular biofilm life style.
T311 470093-470319 Sentence denotes Indeed, by transposon mutagenesis of clinical significant but biofilm negative S. epidermidis 1585, sarA, chief regulator of staphylococcal virulence, was identified as a negative regulator of S. epidermidis biofilm formation.
T312 470320-470505 Sentence denotes Genetic analysis found that inactivation of sarA induced expression of giant 1 MDa extracellular matrix binding protein (Embp) promoting intercellular adhesion and biofilm accumulation.
T313 470506-470626 Sentence denotes Furthermore, related to significant autolytic behaviour, sarA mutants extensively released eDNA into the biofilm matrix.
T314 470627-470813 Sentence denotes Increased autolysis, eDNA release and biofilm formation were directly linked with boosted processing of major autolysin AtlE by metallo protease SepA being overexpressed in sarA mutants.
T315 470814-471107 Sentence denotes Hence, our results provide first evidence that key regulator sarA, while promoting polysaccharide intercellular adhesin dependent biofilm formation on one hand, negatively controls S. epidermidis biofilm formation by suppressing Embp production and autolysis-related eDNA release on the other.
T316 471108-471270 Sentence denotes Hence, this report for the first time identifies a regulator enabling S. epidermidis to switch between polysaccharide-dependent and independent biofilm formation.
T317 471271-471465 Sentence denotes Furthermore, the novel function of sarA links SepA mediated escape from defensin dermicidine with biofilm related protection from phagocytosis and decreased proinflammatory macrophage responses.
T318 471466-471577 Sentence denotes Apparently, sarA plays a central role as regulator ensuring S. epidermidis adaptation to a hostile environment.
T319 471578-471837 Sentence denotes Streptococcus pneumoniae has evolved versatile strategies to adhere directly to human host cells or indirectly by interacting with components of the extracellular matrix (ECM), which is a prerequisite for pneumococci to colonize the upper respiratory airways.
T320 471838-472084 Sentence denotes Pneumococcal attachment to host components is mediated by bacterial surface molecules including cell wall components, choline-binding proteins, LPxTG proteins, lipoproteins and non-classical adhesins such as surface-associated glycolytic enzymes.
T321 472085-472236 Sentence denotes The major adhesin of pneumococci is PspC, the Pneumococcal surface protein C, which binds to the ectodomain of the human polymeric Ig receptor (hpIgR).
T322 472237-472284 Sentence denotes In addition, PspC recruits complement factor H.
T323 472285-472466 Sentence denotes Consequently, PspC is directly involved in pneumococcal adhesion to human host cells via its binding to the SC of hpIgR, and indirectly via the factor H-mediated adhesion mechanism.
T324 472467-472769 Sentence denotes Recently, we have shown that multimeric host-cellbound vitronectin (Vn), an adhesive glycoprotein present in plasma and the ECM, is exploited by pneumococci as a molecular bridge facilitating their adherence to and invasion into host cells by inducing proteins of the host signal transduction cascades.
T325 472770-472933 Sentence denotes Although the interaction of pneumococci with vitronectin and its cellular consequences were demonstrated, the pneumococcal adhesin for vitronectin remains unknown.
T326 472934-473050 Sentence denotes Here we provide first evidences that the multifunctional PspC protein is capable to interact with human vitronectin.
T327 473051-473196 Sentence denotes Surface plasmon resonance studies were conducted with vitronectin immobilized on a CM5 biosensor chip and different PspC derivatives as analytes.
T328 473197-473279 Sentence denotes The sensorgrams demonstrated binding of Nterminal PspC derivatives to vitronectin.
T329 473280-473383 Sentence denotes In addition, PspC and PspC-like Hic were expressed on the surface of non-pathogenic Lactococcus lactis.
T330 473384-473502 Sentence denotes Similar to pneumococci, the heterologous L. lactis but not the control strain interacted with immobilized vitronectin.
T331 473503-473618 Sentence denotes Remarkably, PspC-deficient pneumococci showed a lower capability to bind vitronectin as analyzed by flow cytometry.
T332 473619-473722 Sentence denotes Currently, proteomic approaches are being employed to confirm the interaction of PspC with vitronectin.
T333 473723-473881 Sentence denotes In order to localize the binding site for PspC in the vitronectin molecule different Vn derivatives were cloned into the pET28-TEV His6-tag expression vector.
T334 473882-473990 Sentence denotes The expressed and purified recombinant Vn fragments will be applied in biochemical and immunological assays.
T335 473991-474148 Sentence denotes In conclusion, PspC possesses vitronectin-binding activity, however, its role in pneumococcal pathogenesis via vitronectin has to be assessed in more detail.
T336 474149-474151 Sentence denotes K.
T337 474152-474167 Sentence denotes Blank* 1 , R.G.
T338 474168-474275 Sentence denotes Gerlach 1 1 Robert Koch-Institut, NG3, Wernigerode, Deutschland Salmonella enterica serovar Typhimurium (S.
T339 474276-474429 Sentence denotes Typhimurium) is a facultative intracellular pathogen able to cause acute gastroenteritis and diarrhea in humans and a typhoid fever-like disease in mice.
T340 474430-474473 Sentence denotes Many genes encoding virulence factors of S.
T341 474474-474545 Sentence denotes Typhimurium cluster together on Salmonella pathogenicity islands (SPI).
T342 474546-474782 Sentence denotes Salmonella pathogenicity island 4 (SPI-4) encodes a type I secretion system (SiiCDF) which secretes SiiE, a giant non-fimbrial adhesin shown to mediate intimate contact of Salmonella to the apical membrane of polarized epithelial cells.
T343 474783-474899 Sentence denotes SiiA and SiiB are two additional SPI-4-encoded inner membrane-associated proteins that might regulate SiiE function.
T344 474900-474983 Sentence denotes However, expression of SPI-4 is controlled by regulators encoded outside the locus.
T345 474984-475166 Sentence denotes Besides the central regulator HilA, the transcriptional activator SirA acts as a global regulator of Salmonella enteropathogenesis, controlling expression of SPI-1, as well as SPI-4.
T346 475167-475330 Sentence denotes SirA is part of the two-component regulatory system BarA/SirA sensing environmental signals and activating genes, among others important for motility and invasion.
T347 475331-475495 Sentence denotes Both, deletion of the entire SPI-4 locus, as well as deletion of sirA were shown to result in the same dramatic reduction in adhesion to polarized epithelial cells.
T348 475496-475668 Sentence denotes To gain more insight into SPI-4 function we want to investigate proteinprotein interactions of SPI-4 encoded proteins using a generic protein complex purification strategy.
T349 475669-475905 Sentence denotes The method includes (i) addition of a histidine-biotin-histidine (HBH) tag to the bait proteins (e.g. SiiB or SirA) via recombinant DNA techniques, (ii) in vivo biotinylation and (iii) in vivo cross-linking with para-formaldehyde (PFA).
T350 475906-476098 Sentence denotes The bacterially derived in vivo biotinylation signal peptide, inducing efficient biotin attachment to the HBH-tag, functions in eukaryotes like Saccharomyces cerevisiae as well as in bacteria.
T351 476099-476209 Sentence denotes The membrane-permeable cross-linking reagent PFA is used to stabilize transient and weak protein interactions.
T352 476210-476462 Sentence denotes Cross-linked complexes are enriched by tandem affinity purification (TAP) under fully denaturing conditions, followed by identification of proteins bound to the bait protein by liquid-chromatography in conjunction with tandem mass-spectrometry (LC-MS).
T353 476463-476625 Sentence denotes To verify the efficiency of this purification approach, we used the membrane-associated protein SiiB and the cytosolic protein SirA as bait protein, respectively.
T354 476626-476826 Sentence denotes We could immunodetect distinct sets of protein complexes for C-terminal HBH-tagged SiiB and SirA using an antibody against the RGS-H6 peptide implying low cross-reactivity against non-tagged proteins.
T355 476827-476993 Sentence denotes For MS analyses, we included two negative controls to eliminate the possibility of identifying background and non-specifically bound proteins as interacting partners.
T356 476994-477128 Sentence denotes Our results will provide new insights into the function of SPI-4 for successful adhesion to and subsequent colonization of host cells.
T357 477129-477131 Sentence denotes Y.
T358 477132-477147 Sentence denotes Novotny* 1 , L.
T359 477148-477166 Sentence denotes Marlinghaus 1 , M.
T360 477167-477179 Sentence denotes Korte 1 , S.
T361 477180-477194 Sentence denotes Neumann 1 , S.
T362 477195-477211 Sentence denotes Gatermann 1 , F.
T363 477212-477224 Sentence denotes Szabados 1 1
T1 477226-477453 Sentence denotes Staphylococcus lugdunensis is a coagulase-negative staphylococcus (CNS) that can cause severe infections, like skin and soft tissue infections, sepsis and potentially fatal endocarditis, similar to diseases caused by S. aureus.
T2 477454-477544 Sentence denotes In contrast to S. aureus, only very few virulence associated proteins have been described.
T3 477545-477709 Sentence denotes A fibrinogen binding protein and von Willebrand binding adhesin, and synergistic hemolysins have been discussed as putative pathogenicity factors in S. lugdunensis.
T4 477710-477965 Sentence denotes In this study we designed primer for the detection of two annotated putative fibrinogen binding adhesins and three annotated hemolysins in a collection of more than 100 different Staphylococcus lugdunensis strains, to detect the occurrence of these genes.
T5 477966-478084 Sentence denotes The prevalence of these fibrinogen binding protein and hemolysin genes were compared with their phenotypic correlates.
T6 478085-478194 Sentence denotes All strains were tested for their binding to fibrinogen and also on Columbia blood agar plates for hemolysis.
T7 478195-478302 Sentence denotes Except for the SLUSH gene cluster all the other alleged hemolysins and adhesins had a very high prevalence.
T8 478303-478397 Sentence denotes SLGD_00847 (slu_hemolysinIII) had a prevalence of 97 % and SLGD_00006 (slu_hemolysin) of 95 %.
T9 478398-478444 Sentence denotes The SLUSH gene cluster was only found in 45 %.
T10 478445-478550 Sentence denotes This indicates that the SLUSH gene cluster was absent or the gene cluster was different in these strains.
T11 478551-478673 Sentence denotes After 48 h of incubation 93.1 % of the S. lugdunensis strains showed hemolysis and 12 of them even had a strong hemolysis.
T12 478674-478766 Sentence denotes As already shown in a previously report, hemolysis after 48 h is typical for S. lugdunensis.
T13 478767-479029 Sentence denotes The high occurrence of the hemolysin genes also indicate that the hemolysis attribute is less likely determined by a single hemolysin gene but rather a supposed combined effect of several hemolysin genes or differences in their expression. (Sinha et al., 1999) .
T14 479030-479168 Sentence denotes In polarized cells 5 1 integrins are primarily located to the basolateral side, only a minor fraction is localized to the apical membrane.
T15 479169-479367 Sentence denotes We hypothesize that initial binding of FnBPs to apically located receptors increases the permeability of the endothelial barrier and thus facilitates access to the basolateral pool of 5 1 integrins.
T16 479368-479376 Sentence denotes Methods:
T17 479377-479531 Sentence denotes Human endothelial cells (EA.hy 926) were infected with different wild type and specific mutant / complemented staphylococcal strains on transwell filters.
T18 479532-479624 Sentence denotes Permeability was measured by transendothelial flux of FITC-dextran at different time points.
T19 479625-479732 Sentence denotes Inhibition experiments were performed with Src family protein-tyrosine kinase inhibitors genistein and PP2.
T20 479733-479793 Sentence denotes PP3, an EGFR inhibitor, served as a control for specificity.
T21 479794-479870 Sentence denotes Effects mediated by cytotoxicity were quantified by several viability tests.
T22 479871-480011 Sentence denotes Changes in impedance which reflect changes in cell morphology of the endothelial layer have been determined by real-time impedance analysis.
T23 480012-480020 Sentence denotes Results:
T24 480021-480207 Sentence denotes Infection with wild type S. aureus and S. carnosus heterologously expressing FnBPs, but not Clumping factor A (Clf A), induced an increase in permeability compared to non-infected cells.
T25 480208-480284 Sentence denotes By contrast, the nonpathogenic S. carnosus wild type had an opposite effect.
T26 480285-480372 Sentence denotes An FnBP double knock out mutant of wild type S. aureus failed to increase permeability.
T27 480373-480470 Sentence denotes Genistein and PP2 reduced loss of integrity by roughly 50%, whereas PP3 had no protective effect.
T28 480471-480694 Sentence denotes FnBP expression per se did not lead to substantial cell death, as determined by cell mass (crystal violet staining), mitochondrial respiratory activity (WST-1 conversion) and hypo-diploid DNA (modified cell cycle analysis).
T29 480695-480851 Sentence denotes Real-time impedance analysis yielded similar results as permeability data demonstrating that impedance declines upon infection with FnBP-expressing strains.
T30 480852-480863 Sentence denotes Discussion:
T31 480864-480964 Sentence denotes Our data show that FnBPs are required for an increase in permeability of the endothelial cell layer.
T32 480965-481042 Sentence denotes Src family kinases are crucial components of the integrin signaling pathways.
T33 481043-481220 Sentence denotes Partial protection by genistein and PP2 suggests that Src family kinases mediate an increased permeability of endothelial cells, supposedly by affecting tight junction proteins.
T34 481221-481318 Sentence denotes This effect might be mediated by phosphorylation of either tight junction or regulatory proteins.
T35 481319-481534 Sentence denotes Further studies will include scanning electron microscopy to determine changes in cell morphology, immunodetection and localization of tight junction proteins and siRNA transfection to knock down Src family kinases.
T36 481535-481700 Sentence denotes Streptococcus pneumoniae cell surface proteins play a central role during the infection process and are considered to be important virulence factors of the pathogen.
T37 481701-481842 Sentence denotes They are thought to be implicated in colonization, bacterial adhesion to and invasion into host cells, and immune evasion mechanisms as well.
T38 481843-481999 Sentence denotes Different clusters of proteins such as LPxTG cell wall-anchored proteins, choline-binding proteins, and lipoproteins decorate the pneumococcal cell surface.
T39 482000-482052 Sentence denotes Among them lipoproteins represent the largest group.
T40 482053-482173 Sentence denotes They are anchored to the membrane via a lipid moiety which is covalently attached by the diacylglyceryl transferase Lgt.
T41 482174-482253 Sentence denotes The signal peptide is cut off by the lipoprotein-specific signal peptidase Lsp.
T42 482254-482400 Sentence denotes A global analysis of pneumococcal surface-associated and extracellular proteins was performed using a combination of biotinylation and GeLC-MS/MS.
T43 482401-482632 Sentence denotes To study the fate of lipoproteins in pneumococci the surface-and exoproteome of non-encapsulated D39 cps was compared to the protein profiles of isogenic lgt, lsp, and lgt lsp mutants, which are deficient in lipoprotein maturation.
T44 482633-482765 Sentence denotes Based on the genome data 37 lipoproteins, 16 LPxTG-anchored proteins, and 12 choline-binding proteins were predicted for strain D39.
T45 482766-482823 Sentence denotes Out of them 33, 12, and 2, respectively, were identified.
T46 482824-482930 Sentence denotes The abundance of lipoproteins found to be surface-associated is significantly decreased in the lgt mutant.
T47 482931-483005 Sentence denotes Accordingly, their abundance in the exoproteome is considerably increased.
T48 483006-483138 Sentence denotes In contrast, the amount of surfaceexposed lipoproteins in the lsp mutant was unaffected for the majority of identified lipoproteins.
T49 483139-483269 Sentence denotes Only a few lipoproteins are less abundant, however, the corresponding exoproteome of the lsp mutant showed no significant changes.
T50 483270-483409 Sentence denotes The double knockout lgt lsp showed for the majority of lipoproteins similar amounts on the surface, while only a minority is less abundant.
T51 483410-483515 Sentence denotes Remarkably, the amount of lipoproteins in the exoproteome is significantly increased for D39 cps lgt lsp.
T52 483516-483599 Sentence denotes Streptococcus agalactiae colonizes the urogenital tract of 10-30% of healthy women.
T53 483600-483712 Sentence denotes During delivery it can be transmitted from the mother to the newborn and cause sepsis, pneumonia and meningitis.
T54 483713-483820 Sentence denotes Adhesion to extracellular matrix protein is an important pathogenicity mechanism of gram positive bacteria.
T55 483821-483993 Sentence denotes The laminin receptor on brain endothelial cells has recently been shown to be crucial for the initiation of contact between meningeal pathogens and the blood brain barrier.
T56 483994-484179 Sentence denotes In S. agalactiae the surface protein Lmb mediates the attachment to human placental laminin and is located on a pathogenicity islet that is absent in many bovine S. agalactiae isolates.
T57 484180-484326 Sentence denotes The crystal structure of Lmb was recently solved and shows that it belongs to the cluster 9 family of ABC (ATP binding cassette) transport system.
T58 484327-484544 Sentence denotes The structural organization of Lmb closely resembles ABC type solute binding proteins (SBPs) in which two structurally related globular domains interact with each other to form a metal binding cavity at the interface.
T59 484545-484655 Sentence denotes The bound zinc in Lmb is tetrahedrally coordinated to residues H66, H142, H206 and E281 from both the domains.
T60 484656-484830 Sentence denotes A highly disordered loop spanning residues 123-138 has been observed and it has been shown to play a role in metal uptake and release of the protein in homologous structures.
T61 484831-485094 Sentence denotes To analyze the role of individual amino acid residues for the binding properties of Lmb to immobilized laminin targeted mutations of the Zn binding motif were introduced into an lmb vector construct that allows the expression of lmb as a histidine fusion protein.
T62 485095-485202 Sentence denotes Binding of recombinant Lmb to laminin was investigated in ELISA plates coated with human placental laminin.
T63 485203-485384 Sentence denotes Analysis of the mutated and wild type versions of Lmb showed that a deletion of the metal binding loop and the mutation of His66 led to a severe reduction in the binding to laminin.
T64 485385-485660 Sentence denotes Being ubiquitous in all the serotypes of Group B Streptococcus (GBS), a detailed knowledge of the amino acid residues critical for laminin binding may allow the future development of targeted antimicrobial strategies interfering with the adhesion properties of S. agalactiae.
T65 485661-485674 Sentence denotes Introduction:
T66 485675-485817 Sentence denotes The immune effector molecule interferon (IFN)-arrests the intracellular growth of Chlamydia pneumoniae (Cpn) and induces persistent infection.
T67 485818-485953 Sentence denotes This implies the induction of the JAK-STAT signaling pathway and the tryptophan catabolising enzyme, indoleamine 2,3-dioxygenase (IDO).
T68 485954-486152 Sentence denotes Persistent infection is suggested to promote disease progression in chronic obstructive pulmonary disease (COPD) in which epithelial to mesenchymal cell transition (EMT) is known to be a key factor.
T69 486153-486294 Sentence denotes The disease is characterized by regional declines of the available oxygen content that are fostered by the underlying inflammatory processes.
T70 486295-486359 Sentence denotes Chlamydiae are well adapted to a low oxygen environment (<3%O2).
T71 486360-486492 Sentence denotes We therefore analyzed the effects of acute and persistent Cpn infections on EMT under different environmental oxygen concentrations.
T72 486493-486606 Sentence denotes Methods: HEp-2 and A549 cells were seeded in 6-well plates and treated with IFN-to induce chlamydial persistence.
T73 486607-486719 Sentence denotes After 24h incubation under normoxic (20%O2) or hypoxic conditions (2%O2), cells were infected with Cpn (CWL029).
T74 486720-486987 Sentence denotes Recovery assays were performed 72h p.i. and IFUs/μl were calculated by using immunofluorescence microscopy. p-STAT1 (Tyr701) and IDO expression were analyzed by western blot. -smooth muscle actin ( -SMA) expression level was analyzed by immunofluorescence microscopy.
T75 486988-487066 Sentence denotes Statistical analysis was performed with a one tailed, unpaired Student t-test.
T76 487067-487248 Sentence denotes Results: IFN-treatment was not effective to limit growth of Cpn at low oxygen concentrations (2% O2, 3x10 5 ±1x10 5 IFUs/μl) compared to normoxic conditions (20% O2, 0±0.2 IFUs/μl).
T77 487249-487385 Sentence denotes This was accompanied by diminished phosphorylation of STAT-1 (Tyr701) and subsequent reduced expression of IDO under hypoxic conditions.
T78 487386-487476 Sentence denotes Interestingly, Cpn could be rescued from IFN-induced persistence under hypoxic conditions.
T79 487477-487721 Sentence denotes The expression of the epithelial marker protein E-cadherin was significantly downregulated in acute and persistently infected A549 cells (p<0.05) whereas the mesenchymal marker protein -SMA was upregulated under normoxic and hypoxic conditions.
T80 487722-487808 Sentence denotes This effect was more pronounced when the cells were cultured under hypoxic conditions.
T81 487809-487905 Sentence denotes Conclusion: IFN-mediated immune control of Cpn depends on the environmental oxygen availability.
T82 487906-487968 Sentence denotes Thus, Cpn escapes host immune control in a hypoxic atmosphere.
T83 487969-488070 Sentence denotes Acute and persistent Cpn induce EMT in lung cells that is even more pronounced when oxygen is scarce.
T84 488071-488322 Sentence denotes Carbon catabolite repression (CCR) in bacteria is a widespread, global regulatory phenomenon that allows modulation of the expression of genes and operons involved in carbon utilization and metabolization in the presence of preferred carbon source(s).
T85 488323-488467 Sentence denotes In low-GC gram-positive bacteria, CCR is mediated mainly by the catabolite control protein A (CcpA), a member of the GalR-LacI repressor family.
T86 488468-488610 Sentence denotes In Staphylococcus aureus, CcpA has been shown to modulate the expression of metabolic genes and virulence determinants in response to glucose.
T87 488611-488853 Sentence denotes A second regulator that links carbon metabolism and virulence factor production in this organism is CodY, a sensor of carbon and nitrogen availability that responds to intracellular concentrations of branched-chain amino acids (BCAA) and GTP.
T88 488854-489082 Sentence denotes We recently reported that S. aureus produces a third regulatory molecule, NWMN_0641 (a member of the LysR family of transcriptional regulators with homology to CcpC of B. subtilis) that affects the expression of TCA cycle genes.
T89 489083-489287 Sentence denotes Here we show that this regulatory molecule also links carbon metabolism with virulence in S. aureus by altering the expression of important virulence factors, and affecting pathogenicity of this organism.
T90 489288-489437 Sentence denotes We found that inactivation of NWMN_0641 significantly decreased the transcription of the cap operon, and reduced the capsule formation of the mutant.
T91 489438-489699 Sentence denotes However, transcription of hla (encoding a-hemolysin), and of RNAIII, the effector molecule of the agr locus, were significantly increased by the mutation, suggesting that NWMN_0641 acts as a repressor of hla and RNAIII, and as an activator of cap transcription.
T92 489700-489931 Sentence denotes In a murine footpad infection model, the deletion mutant caused an increased footpad swelling, if compared to the wild-type and the complemented derivate, indicating a higher infectivity of the mutant in this systemic animal model.
T93 489932-490036 Sentence denotes The Hsp100/ClpATPases belong to a group of closely related proteins that operate as molecular chaperons.
T94 490037-490187 Sentence denotes These chaperons are regarded as an important element of bacteria to respond to various stress conditions by refolding or degrading misfolded proteins.
T95 490188-490320 Sentence denotes The major human pathogen Staphylococcus aureus expresses a number of such Hsp100/ClpATPases, namely ClpB, ClpC, ClpL, ClpX and ClpY.
T96 490321-490463 Sentence denotes In S. aureus, ClpB and ClpC are both regulated by the repressor CtsR, with ClpC probably supporting the active conformation of this repressor.
T97 490464-490534 Sentence denotes Under conditions of stress, ClpC shifts from CtsR to damaged proteins.
T98 490535-490693 Sentence denotes As a consequence CtsR looses its active conformation which results in a transcriptional induction of CtsR repressed genes, such as clpB, clpC and ctsR itself.
T99 490694-490822 Sentence denotes ClpB, and to a lesser extent, ClpC have been shown to promote the intracellular multiplication of S. aureus in epithelial cells.
T100 490823-491050 Sentence denotes Additionally, ClpC was found to stimulate the expression of citB (encoding the tricarboxylic acid (TCA) cycle enzyme aconitase) and to support the metabolization of acetate, thereby affecting the post-stationary-phase recovery.
T101 491051-491137 Sentence denotes Inactivation of clpC, on the other hand, led to an enhanced stationary-phase survival.
T102 491138-491274 Sentence denotes Based on these findings, we wondered about the impact of ClpB and ClpC on the intracellular persistence of S. aureus within human cells.
T103 491275-491508 Sentence denotes Our in vitro viability studies confirmed previous findings by showing that S. aureus DSM20231 clpC and clpBC mutant cultures possessed an increased stationary phase survival, if compared to the wild-type and the clpB mutant cultures.
T104 491509-491769 Sentence denotes In line with this, we detected an enhanced survival of the clpC and clpBC double mutants in keratinocytes 96 hours after the internalisation, while this was not the case with the clpB mutant, which showed a persistence rate comparable to that of the wild-type.
T105 491770-491970 Sentence denotes In terms of invasiveness, however, the clpB and clpBC mutants possessed significantly decreased internalization capacities, probably due to lowered productions of fibronectin binding proteins A and B.
T106 491971-492312 Sentence denotes These findings suggest that ClpB contributes to the invasiveness of this pathogen, but does not influence the persistence capacity of this organism, while a functional ClpC seems to decrease the intracellular survival capacity of S. aureus but does not affect the invasiveness of this organism, irrespective of its impact on clpB expression.
T107 492313-492485 Sentence denotes Aggregatibacter actinomycetemcomitans is a Gram-negative commensal bacterium of the oral cavity which has been associated with the pathogenesis of aggressive periodontitis.
T108 492486-492622 Sentence denotes However, genetic mechanisms underlying host susceptibility to periodontal infection and to periodontitis progress are still ill-defined.
T109 492623-492770 Sentence denotes This study aimed to identify genetic host factors which confer resistance against A. actinomycetemcomitans-induced periodontitis in a murine model.
T110 492771-492953 Sentence denotes NADPH oxidase-deficient mice (gp91phox KO), inducible nitric oxide synthase-deficient mice (iNOS KO), and C57BL/6 wild-type mice were orally inoculated with A. actinomycetemcomitans.
T111 492954-493121 Sentence denotes To analyse bacterial colonization in the different mouse strains at various time points after infection A. actinomycetemcomitans cultures of oral swabs were performed.
T112 493122-493306 Sentence denotes Furthermore, linear measurements and three-dimensional micro-computed tomography were used to quantify the degree of alveolar bone destruction as well as alveolar bone mineral density.
T113 493307-493463 Sentence denotes Persistence of A. actinomycetemcomitans in the murine oral cavity varied among the three mice strains showing the highest rate in infected gp91phox KO mice.
T114 493464-493727 Sentence denotes In accordance with A. actinomycetemcomitans persistence, gp91phox KO mice developed more severe periodontitis compared to iNOS KO and C57BL/6 wild-type mice and uninfected controls, characterized by significant higher alveolar bone loss and inflammatory reaction.
T115 493728-493913 Sentence denotes Additionally, a significantly lower alveolar bone mineral density was observed in all three groups of mice after infection with A. actinomycetemcomitans compared to uninfected controls.
T116 493914-494178 Sentence denotes Our results indicate that NADPH oxidase is important for resistance of the murine oral cavity against A. actinomycetemcomitans colonization which is clearly associated with alveolar bone destruction and influx of inflammatory cells in NADPH oxidase-deficient mice.
T117 494179-494327 Sentence denotes This model should give further insights into the molecular and genetic mechanisms of the A. actinomycetemcomitans induced periodontitis development.
T118 494328-494330 Sentence denotes N.
T119 494331-494348 Sentence denotes Ackermann* 1 , S.
T120 494349-494362 Sentence denotes Untiet 1 , G.
T121 494363-494376 Sentence denotes Anding 1 , J.
T122 494377-494601 Sentence denotes Heesemann 1 1 Max von Pettenkofer Institut, Bakteriologie, München, Deutschland Yersinia enterocolitica and Y. pseudotuberculosis are enteropathogenic bacteria causing diseases ranging from self-limiting diarrhoea to sepsis.
T123 494602-494720 Sentence denotes In the mouse model yersiniae pass the gut via M cells and invade Peyer´s patches with subsequent septic dissemination.
T124 494721-494877 Sentence denotes The trimeric autotransporter Yersinia adhesin A (YadA) is an essential mouse virulence (MV) factor for Y. enterocolitica, but not for Y. pseudotuberculosis.
T125 494878-494963 Sentence denotes It forms a lollipop-like structure with a head, connector, stalk and membrane anchor.
T126 494964-495047 Sentence denotes YadA in different Yersinia species shows differences in its head and stalk regions.
T127 495048-495263 Sentence denotes To compare the role of these differences in regard to autoagglutination, extracellular matrix or cell adherence, serum resistance and MV, different YadA variants were introduced into Y. enterocolitica strain WA-314.
T128 495264-495463 Sentence denotes Our results show that even though in vitro differences in collagen or fibronectin autoagglutination exist between those variants, there is no difference in their capability to induce mouse virulence.
T129 495464-495619 Sentence denotes These data show that all tested Y. enterocolitica and Y. pseudotuberculosis YadA variants are capable to mediate comparable MV in Y. enterocolitica WA-314.
T130 495620-495667 Sentence denotes Mycobacterium tuberculosis eats what it gets L.
T131 495668-495682 Sentence denotes Denkel* 1 , A.
T132 495683-495696 Sentence denotes Hampel 1 , R.
T133 495697-495710 Sentence denotes Bücker 2 , C.
T134 495711-495724 Sentence denotes Bolten 2 , C.
T135 495725-495743 Sentence denotes Wittmann 2 , F.-C.
T136 495744-495987 Sentence denotes Bange 1 1 Medizinische Hochschule Hannover, Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Hannover, Deutschland 2 Technische Universität Braunschweig, Institut für Bioverfahrenstechnik, Braunschweig, Deutschland Introduction:
T137 495988-496113 Sentence denotes New technologies such as fluxome analysis, metabolomics and transcriptomics allow an insight into the metabolism of bacteria.
T138 496114-496188 Sentence denotes These techniques often require a medium defined to a single carbon source.
T139 496189-496323 Sentence denotes Media, however, used for culture of Mycobacterium (M.) tuberculosis are quite complex and contain several carbon and nitrogen sources.
T140 496324-496551 Sentence denotes Here we show that M. tuberculosis growing in minimal medium with glucose as carbon source also metabolizes other supplemented ingredients like the fatty acids contamination of albumine and surprisingly also the detergent tween.
T141 496552-496560 Sentence denotes Methods:
T142 496561-496763 Sentence denotes For this study M. tuberculosis was cultivated in defined minimal medium with U-13 C labelled glucose as sole carbon source supplemented with albumine and the detergents tween or tyloxapol, respectively.
T143 496764-496919 Sentence denotes Methods used here involve isotopic tracer studies and GC-MS (gas chromatography mass spectrometry) for measurement of the labelling pattern of metabolites.
T144 496920-496928 Sentence denotes Results:
T145 496929-497057 Sentence denotes Using minimal medium with fully 13 C-labelled glucose, we expected to measure 1 % at the most non labelled amino acids by GC-MS.
T146 497058-497122 Sentence denotes However it turned out that 20 % of metabolites are not labelled.
T147 497123-497159 Sentence denotes Thus it leads us to two suggestions:
T148 497160-497276 Sentence denotes Either M. tuberculosis uses another carbon source beside the labelled glucose or the washing steps are insufficient.
T149 497277-497586 Sentence denotes Improvement of the washing steps did not change the fraction of non labelled amino acids, however the substitution of albumine (contaminated with 1 % fatty acids) against highly purified albumine (contaminated with 0.02 % fatty acids) could decrease the fraction of non labelled amino acids to less than 10 %.
T150 497587-497693 Sentence denotes When we replaced tween with tyloxapol we could further diminish the non labelled amino acids to about 5 %.
T151 497694-497929 Sentence denotes Further improvement of the protocol for preparation of M. tuberculosis cell extracts by using cellulose nitrate filter instead of harvesting the cells by centrifugation could diminish the part of non labelled amino acids to almost 0 %.
T152 497930-497941 Sentence denotes Conclusion:
T153 497942-498101 Sentence denotes In summary our studies showed that M. tuberculosis uses the minimal medium supplementations tween and fatty acids that adhere to albumine for their metabolism.
T154 498102-498273 Sentence denotes Thus new challenging approaches like fluxome analysis require a defined medium with carefully selected ingredients and well established protocols for harvesting the cells.
T155 498274-498276 Sentence denotes T.
T156 498277-498292 Sentence denotes Wille* 1 , R.G.
T157 498293-498364 Sentence denotes Gerlach Junior Research Group 3, Wernigerode, Deutschland Introduction:
T158 498365-498450 Sentence denotes Protein-protein interactions are important regulatory layers in all kingdoms of life.
T159 498451-498545 Sentence denotes They are involved in a vast set of cellular processes including virulence-associated pathways.
T160 498546-498718 Sentence denotes Identification and characterization of these interactions is one challenging task of the post-genomic era and crucial for understanding of the molecular bases of virulence.
T161 498719-498840 Sentence denotes Several methods have been successfully employed in the past decades to identify protein-protein interactions in bacteria.
T162 498841-498917 Sentence denotes These methods often include tedious and time-consuming manipulations of DNA.
T163 498918-499025 Sentence denotes The Gateway system is a fast and convenient tool, for efficient transfer of DNA-fragments between plasmids.
T164 499026-499152 Sentence denotes It uses a set of proprietary recombination sequences derived from the bacteriophage lambda site-specific recombination system.
T165 499153-499291 Sentence denotes Using a combination of different att-sites, MultiSite Gateway allows for site directed cloning of up to four fragments into one construct.
T166 499292-499299 Sentence denotes Method:
T167 499300-499423 Sentence denotes Here we developed a new set of Gateway vectors to facilitate the investigation of protein-protein interactions in bacteria.
T168 499424-499562 Sentence denotes The set includes custom made entry vectors for cloning of a gene of interest in front of an optimized RBS and modular destination vectors.
T169 499563-499771 Sentence denotes To test our approach, we designed a destination vector including two recombination loci in frame with the fragments of split Gaussia luciferase (Gluc), each under control of a tetracycline-inducible promoter.
T170 499772-499779 Sentence denotes Result:
T171 499780-499928 Sentence denotes Salmonella pathogenicity Island 1 (SPI-1) encodes for a type three secretion system (TTSS) together with effectors and cognate secretion chaperones.
T172 499929-500058 Sentence denotes The chaperone InvB binds the effector SipA via its N-terminal chaperone binding domain (CBD) comprising the first 47 amino acids.
T173 500059-500203 Sentence denotes As proof of principle, we wanted to characterize the interaction between the effector SipA and its chaperone InvB using the split Gluc approach.
T174 500204-500336 Sentence denotes A single Gateway recombination reaction resulted in the simultaneous integration of both genes in the customized destination vector.
T175 500337-500461 Sentence denotes In the final expression vector, C-and N-terminal portions of Gluc were fused to the Ctermini of InvB and SipA, respectively.
T176 500462-500517 Sentence denotes After transfer of constructs in Salmonella enterica sv.
T177 500518-500533 Sentence denotes Typhimurium (S.
T178 500534-500681 Sentence denotes Typhimurium), luminescence measurements revealed significant higher signal with full length SipA compared to its truncated version lacking the CBD.
T179 500682-500693 Sentence denotes Discussion:
T180 500694-500803 Sentence denotes In conclusion our system provides a valuable tool for investigating protein-protein interactions in bacteria.
T181 500804-500919 Sentence denotes The approach simplifies cloning steps and its modular nature enables fast adaption to all kinds of fusion proteins.
T182 500920-501089 Sentence denotes Streptococcus agalactiae (Group B Streptococci, GBS) is an important cause of human invasive infections in newborns, pregnant women and immunocompromised adult patients.
T183 501090-501194 Sentence denotes The ß-hemolysin of S. agalactiae is a surface associated toxin and regarded as a major virulence factor.
T184 501195-501304 Sentence denotes It is regulated by the cov two-component regulatory system, which controls numerous virulence factors of GBS.
T185 501305-501446 Sentence denotes Mutations of the cov gene have been associated with increased hemolytic activity, but impaired intracellular survival of GBS in granulocytes.
T186 501447-501726 Sentence denotes To determine the role of the ß-hemolysin for intracellular survival and to rule out the effect of other virulence factors controlled by cov, we investigated hemolytic and nonhemolytic GBS mutants for intracellular survival in primary human granulocytes and THP-1 monocytic cells.
T187 501727-501932 Sentence denotes We examined the role of ß-hemolysin for interaction with the monocytic and granulocytic cells using a serotype Ia S. agalactiae wild type strain and an isogenic nonhemolytic deletion mutant of this strain.
T188 501933-502005 Sentence denotes Both strains were fluorescently labeled with an EGFP expressing plasmid.
T189 502006-502212 Sentence denotes Following infection of eukaryotic cells with GBS at a multiplicity of infection (MOI) ranging from 5 to 25 the intracellular bacteria were evaluated by FACS analysis and culturing of intracellular bacteria.
T190 502213-502365 Sentence denotes Interestingly, the nonhemolytic mutants were able to survive in the intracellular environment in significantly higher numbers than the hemolytic strain.
T191 502366-502446 Sentence denotes A finding that was observed for primary granulocytes as well as for THP-1 cells.
T192 502447-502725 Sentence denotes To exclude the possibility that the observed differences in survival were due to host cell death induced by the hemolytic but not the non-hemolytic strain, Lactate Dehydrogenase (LDH) assays were carried out and confirmed the better survival capacity of the nonhemolytic strain.
T193 502726-502822 Sentence denotes The ß-hemolysin of GBS has previously been described as a potent inducer of IL-8 in macrophages.
T194 502823-502921 Sentence denotes To assess the induction of IL-8 following infection with GBS, ELISA determinations were performed.
T195 502922-503105 Sentence denotes While a considerable release of IL-8 could be observed, however we could not find a significant difference in the ability of hemolytic or nonhemolytic strains to induce the chemokine.
T196 503106-503249 Sentence denotes To determine the bacterial mediators of IL-8 release in this setting, cell wall preparations from both strains were incubated with THP-1 cells.
T197 503250-503337 Sentence denotes Both preparations were found to exert a potent proinflammatory stimulus on THP-1 cells.
T198 503338-503612 Sentence denotes In conclusion, our results indicate that the S. agalactiae ßhemolysin has a strong influence on the intracellular survival of GBS and that a tightly controlled regulation of ß-hemolysin expression is required for the successful establishment of GBS in different host niches.
T199 503613-503773 Sentence denotes In conclusion, the ability to express ZnuD upon zinc starvation was well conserved among meningococci, which argues for an essential role of meningococcal ZnuD.
T200 503774-503880 Sentence denotes It is intriguing that inactivating ZnuD mutation events seem to be restricted to only two clonal lineages.
T201 503881-503949 Sentence denotes In ST-23 cc the mutations are inserted by slipped strand mispairing.
T202 503950-504013 Sentence denotes Ex vivo events induced by culture must therefore be considered.
T203 504014-504155 Sentence denotes In our work, we firstly describe that 1-L-MT, but not 1-D-MT, can act as a nutritive substrate that replaces tryptophan in in vitro cultures.
T204 504156-504365 Sentence denotes In initial experiments we showed that 1-L-MT, but not 1-D-MT, allows the growth of tryptophan-auxotroph bacteria and parasites (Staphylococcus aureus ; Toxoplasma gondii) in the complete absence of tryptophan.
T205 504366-504497 Sentence denotes In line with that we describe that 1-L-MT in contrast to 1-D-MT counteracts IDO-dependent inhibition of human T-cell proliferation.
T206 504498-504651 Sentence denotes A detailed analysis showed that 1-L-MT, in contrast to 1-D-MT, is incorporated in human T-cell proteins when tryptophan is lacking in the culture medium.
T207 504652-504776 Sentence denotes In summary our data indicate that 1-L-MT inhibits the IDO-mediated antiproliferative activities via two independent effects:
T208 504777-504891 Sentence denotes On the one hand 1-L-MT acts as a competitive enzyme inhibitor and therefore reduces tryptophan degradation by IDO.
T209 504892-505027 Sentence denotes On the other hand 1-L-MT can replace tryptophan in protein biosynthesis and therefore allows bacterial, parasitical or cellular growth.
T210 505028-505181 Sentence denotes Further mass spectroscopy analysis is necessary to analyse whether 1-L-MT is incorporated into proteins either directly or after a demethylation process.
T211 505182-505299 Sentence denotes The hypothetical type VI secretion protein BPSS1504 is involved in virulence of Burkholderia pseudomallei Background:
T212 505300-505578 Sentence denotes Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains, especially the Panton-Valentine-leukocidin (PVL) producing strain USA300, have been associated with severe necrotizing infections, such as necrotizing pneumonia, which have high mortality rates.
T213 505579-505713 Sentence denotes Epidemiological studies revealed that necrotizing pneumonia is often preceded by influenza-like symptoms or by an influenza infection.
T214 505714-505831 Sentence denotes Nevertheless, the combined action of influenza virus and the pore-forming S. aureus toxin PVL is hardly investigated.
T215 505832-505840 Sentence denotes Methods:
T216 505841-506051 Sentence denotes In our study, we examined the effect of PVL on different cell types (granulocytes and epithelial cell line A549) with or without anteceded infection of the cells with the influenza virus Puerto Rico/8/34 (PR8).
T217 506052-506224 Sentence denotes The pro-inflammatory activation of the cells was measured by chemokine expression via RT-PCR and CD11b expression and cell death induction was determined by flow cytometry.
T218 506225-506296 Sentence denotes The integrity of the cell monolayers was evaluated by light microscopy.
T219 506297-506305 Sentence denotes Results:
T220 506306-506476 Sentence denotes We found that PVL induced cell activation and rapid cell death in granulocytes, but did not affect cell viability in non-professional phagocytes such as epithelial cells.
T221 506477-506692 Sentence denotes Influenza virus PR8 infection alone did not enhanced granulocyte cell death induction, but a previous infection with PR8 significantly enhanced the pro-inflammatory and cytotoxic effect of PVL on human granulocytes.
T222 506693-506931 Sentence denotes By contrast, cell viability of epithelial cells was not affected by single or combined stimulation with PVL and/or PR8, but the influenza virus had a highly activating impact on the chemokine expression (RANTES, IP-10, IL8) of A549 cells.
T223 506932-507180 Sentence denotes However, when we treated the epithelial cell monolayer with supernatants of human granulocytes, which were lysed by PVL and PR8, we found a dosedependent destruction of the epithelial cell monolayer, mainly caused by detachment of epithelial cells.
T224 507181-507371 Sentence denotes Fetal bovine serum, human serum and a Protease Inhibitor Cocktail interfered with the destructive impact of the granulocytes supernatant on A549 and preserved the integrity of the monolayer.
T225 507372-507383 Sentence denotes Conclusion:
T226 507384-507447 Sentence denotes Our findings show that PVL has a lysing effect on granulocytes.
T227 507448-507610 Sentence denotes The influenza virus has an activating impact on epithelial cells that could result in massive recruitment of granulocytes, which can be subsequently lysed by PVL.
T228 507611-507690 Sentence denotes PVL and influenza virus together showed even an enhanced impact on neutrophils.
T229 507691-507839 Sentence denotes The substances released by the lysed neutrophils, e.g. granule proteins and defensins, are able, in turn, to destruct the epithelial cell monolayer.
T230 507840-507992 Sentence denotes These results from our cell culture models suggest that PVL and influenza virus can act together and are possible agents to cause necrotizing pneumonia.
T231 507993-507995 Sentence denotes J.
T232 507996-508008 Sentence denotes Popp* 1 , T.
T233 508009-508024 Sentence denotes Dandekar 2 , W.
T234 508025-508372 Sentence denotes Eisenreich 3 1 Universität Osnabrück, Mikrobiologie, Osnabrück, Deutschland 2 Universität, Bioinformatik, Würzburg, Deutschland 3 Technische Universität, Biochemie, München, Deutschland The intracellular lifestyle of Salmonella enterica relies on the adaption to nutritional conditions within the Salmonella-containing vacuole (SCV) in host cells.
T235 508373-508480 Sentence denotes Based on previous results, metabolic requirements for Salmonella in the SCV shall be characterized further.
T236 508481-508762 Sentence denotes This includes (i) prediction of intracellular phenotypes of mutant strains based on metabolic modeling and experimental tests, (ii) isotopologue profiling using 13C compounds in intracellular Salmonella, and (iii) complementation of metabolic defects for attenuated mutant strains.
T237 508763-508945 Sentence denotes The collaborative work between the Dandekar and Hensel groups should lead to a comprehensive understanding of the metabolic requirements of the intracellular lifestyle of Salmonella.
T238 508946-509039 Sentence denotes Further analyses of intracellular phenotypes will be based on improved metabolic predictions.
T239 509040-509176 Sentence denotes Vice versa, based on analyses of mutant phenotypes and isotopologue analyses, iterative refinement of metabolic fluxes will be achieved.
T240 509177-509319 Sentence denotes Our vision is a comprehensive understanding of the adaption of Salmonella to its lifestyle in a membrane-bound compartment in mammalian cells.
T241 509320-509597 Sentence denotes Towards an integrated model and profiting from SPP1316, we will do comparisons of the Salmonella lifestyle with other pathogens and their metabolic requirements during infection such as cytoplasmic lifestyle found in Listeria or within a membrane compartment as for Legionella.
T242 509598-509600 Sentence denotes M.
T243 509601-509614 Sentence denotes Korte* 1 , L.
T244 509615-509633 Sentence denotes Marlinghaus 1 , S.
T245 509634-509648 Sentence denotes Neumann 1 , F.
T246 509649-509664 Sentence denotes Szabados 1 , N.
T247 509665-509684 Sentence denotes Pfennigwerth 1 , S.
T248 509685-509975 Sentence denotes Gatermann 1 1 Ruhr-Universität Bochum, Abteilung für Medizinische Mikrobiologie, Bochum, Deutschland S. saprophyticus is the only species of the staphylococci that is typically uropathogenic and the genome is the only one of all sequenced staphylococci which possesses a D-serine deaminase.
T249 509976-510030 Sentence denotes This enzyme converts D-serine to pyruvate and ammonia.
T250 510031-510117 Sentence denotes Interestingly, it is also present in other uropathogenic bacteria like E. coli (UPEC).
T251 510118-510271 Sentence denotes The amino acid D-serine is present in relatively high concentrations in human urine and is toxic or bacteriostatic to several non-uropathogenic bacteria.
T252 510272-510384 Sentence denotes Therefore the uncommon ability to degrade D-serine may play an important role for the virulence of uropathogens.
T253 510385-510498 Sentence denotes In addition, the presence of D-serine may be used as a cue by uropathogens for the presence in the urinary tract.
T254 510499-510639 Sentence denotes To analyze the metabolism and to understand the significance of D-serine catabolism of S. saprophyticus for virulence, we conducted 2D-gels.
T255 510640-510806 Sentence denotes To this end S. saprophyticus was grown with either glucose or D-serine as sole carbon and energy source in a chemically defined medium, which we developed previously.
T256 510807-510941 Sentence denotes Protein extracts of S. saprophyticus adapted to glucose or to D-serine were applied to 2D-gels and the protein patterns were analyzed.
T257 510942-511062 Sentence denotes Proteins which were reproducibly more than 2-fold upregulated were excised for a following identification via MALDI-TOF.
T258 511063-511304 Sentence denotes Most of the proteins upregulated under D-serine conditions affect the carbohydrate metabolism, the amino acid metabolism and the energy metabolism, but we also found the accessory protein of the urease, a virulence factor, to be upregulated.
T259 511305-511501 Sentence denotes These results lead us to the hypothesis that D-serine indeed is used as a cue for the presence within the urinary tract and regulates pathways that are important for viability in this environment.
T260 511502-511647 Sentence denotes Staphylococcus saprophyticus is a gram-positive and coagulase-negative pathogenic staphylococcus causing urinary tract infections in young women.
T261 511648-511753 Sentence denotes It is hydrophobic, able to bind fibronectin, laminin and collagen and hemagglutinates sheep erythrocytes.
T262 511754-511825 Sentence denotes Some of its surface proteins have been characterised in the early past.
T263 511826-511978 Sentence denotes The serine-aspartate repeat protein I (SdrI), contains the longest SD repeat region described so far (854 aa) and a LPXTG-motif for cell wall anchoring.
T264 511979-512065 Sentence denotes It is a member of the MSCRAMM protein family and shows a typical ABB domain structure.
T265 512066-512138 Sentence denotes Within the A domain a specific amino acid sequence (TYTFTNYVD) is found.
T266 512139-512316 Sentence denotes This motif or it variants is also found in many other fibronectin or collagen binding proteins from staphylococci, even in another surface protein of S. saprophyticus: the UafA.
T267 512317-512417 Sentence denotes Previous experiments have indicated that a SdrI mutant showed decreased fibronectin binding in vivo.
T268 512418-512548 Sentence denotes Following this hint isolated SdrI domains were tested for fibronectin binding in vitro using fibronectin coated microtiter plates.
T269 512549-512622 Sentence denotes These tests showed that fibronectin binding was mediated by the A domain.
T270 512623-512723 Sentence denotes To localise the area of binding within the A domain, it was divided into three parts: N1, N2 and N3.
T271 512724-512839 Sentence denotes N2, N3 and N2+3 were cloned into the pQE30Xa vector for overexpression and purification using a N-terminal His-Tag.
T272 512840-512963 Sentence denotes Binding assays were carried out with N2, N3, N2+3 and the whole A domain as a positive control in coated microtiter plates.
T273 512964-513007 Sentence denotes The bound protein was detected by an ELISA.
T274 513008-513160 Sentence denotes As expected the results showed a high binding activity for N2, a slightly reduced binding for N2+3 compared to N2 and N3 alone showed no binding at all.
T275 513161-513248 Sentence denotes N2 was further subjected to xray structure analysis but data analysis is still ongoing.
T276 513249-513343 Sentence denotes Suprisingly, the UafA knock out mutant was also associated with decreased fibronectin binding.
T277 513344-513455 Sentence denotes This protein has also been separated into its domains which will be used for the binding test mentioned before.
T278 513456-513621 Sentence denotes We expect to gain new insights into the A domain conformation of SdrI, the role of UafA and their implications for the uro-adherence of Staphylococcus saprophyticus.
T279 513622-513872 Sentence denotes Staphylococcus aureus, the cause of a wide spectrum of severe communityacquired and nosocomial infections, is recently acknowledged as an intracellular pathogen, as it can be internalized and persist in nonprofessional phagocytic cells in vitro [1] .
T280 513873-514012 Sentence denotes During the internalization process, S. aureus has to adapt to the new intracellular environment to survive or even persist within the host.
T281 514013-514247 Sentence denotes Recently, we were able to analyze this adaptive process of internalized S. aureus RN1HG using an infection assay which combines a pulse-chase labeling approach (SILAC [2] ), high capacity cell sorting, and gel-free proteomics [3, 4] .
T282 514248-514448 Sentence denotes The aim of this study is to comparatively investigate the adaptive response of S. aureus RN1GH wild type and isogenic agr mutant cells upon co-internalization by human bronchial epithelial cells (S9).
T283 514449-514614 Sentence denotes The accessory gene regulator (agr) is one of the major global regulators of S. aureus virulence, which positively influences the synthesis ofhemolysin and proteases.
T284 514615-514670 Sentence denotes In contrast adhesions are negatively influenced by agr.
T285 514671-514784 Sentence denotes There is striking evidence that this regulatory system plays an important role in the establishment of infection.
T286 514785-514914 Sentence denotes For instance, expression of agr is initially increased in the acute phase of infection in non-professional phagocytic cells [5] .
T287 514915-515077 Sentence denotes Furthermore, agr mutants are attenuated in a variety of animal models and it was also shown that the escape from the endosome requires agr-dependent factors [6] .
T288 515078-515246 Sentence denotes The striking advantage of a co-infection assay would be that both strains are being internalized simultaneously and adapt to the host under exactly the same conditions.
T289 515247-515572 Sentence denotes This allows us to study the competitive intracellular behavior of the two strains on the proteome level in a time-resolved fashion. (C. trachomatis) are the most frequent transmitted sexually disease in women that may result in severe clinical sequelae like pelvic inflammatory disease (PID), tubal occlusion and infertility.
T290 515573-515791 Sentence denotes It is suggested that once the acute infection is not efficiently cleared by the immune system intracellular persistence of chlamydiae eventually occur, leading to chronic inflammatory processes within infected tissues.
T291 515792-515969 Sentence denotes We could recently show that IFN-induced persistence of C. trachomatis in human fallopian tube cells can be overcome by decreasing the environmental oxygen concentration (<3%O2).
T292 515970-516079 Sentence denotes Under hypoxic conditions, chlamydiae start to replicate and re-differentiate to infectious elementary bodies.
T293 516080-516259 Sentence denotes The aim of our study was to characterize hypoxia induced reactivation of intracellular chlamydial growth with respect to pro-inflammatory immune responses and MAPkinase signaling.
T294 516260-516281 Sentence denotes Material and Methods:
T295 516282-516410 Sentence denotes Persistence was induced by treating C. trachomatis serovar D infected HeLa-229 cells with IFN-for 24h under normoxic conditions.
T296 516411-516517 Sentence denotes Persistent C. trachomatis were subsequently incubated under normoxic (20%O2) or hypoxic (2%O2) conditions.
T297 516518-516602 Sentence denotes Phosphorylation of ERK1/2 and p38 MAPkinases were detected by western blot analysis.
T298 516603-516688 Sentence denotes Expression levels of IL-1ß, IL-8, MCP-1, and TNF-mRNA were measured by real-time PCR.
T299 516689-516697 Sentence denotes Results:
T300 516698-516810 Sentence denotes Reactivation of persistent C. trachomatis induced by IFN-was observed under hypoxic but not normoxic conditions.
T301 516811-516930 Sentence denotes Preliminary experiments suggest that ERK1/2 and p38 MAPkinases are activated under normoxic but not hypoxic conditions.
T302 516931-517107 Sentence denotes In contrast, mRNA expression levels of the pro-inflammatory and chemotactic cytokines IL-1ß, IL-8, MCP-1, and TNF-were enhanced under hypoxic but not under normoxic conditions.
T303 517108-517120 Sentence denotes Discussions:
T304 517121-517300 Sentence denotes Our data indicate for the first time that persistent chlamydiae are recovered in a hypoxic environment that is found under physiological conditions in the female urogenital tract.
T305 517301-517454 Sentence denotes Intracellular reactivation of formerly persistent chlamydiae might perpetuate inflammatory processes that result in chronic sequelea in diseased tissues.
T306 517455-517457 Sentence denotes M.
T307 517458-517471 Sentence denotes Hopfe* 1 , B.
T308 517472-517488 Sentence denotes Henrich 1 1 Med.
T309 517489-517591 Sentence denotes Mikrobiologie und Krankenhaushygiene, Mycoplasmen Arbeitsgruppe, Düsseldorf, Deutschland Introduction:
T310 517592-517669 Sentence denotes Adherence to host tissue is one of the first steps of bacterial colonization.
T311 517670-517835 Sentence denotes In progress of infection, extracellular matrix (ECM) components became accessible at the host's surface as receptors for bacterial adhesion and bridges for invasion.
T312 517836-518058 Sentence denotes The cytoadhesion of Mycoplasma hominis, a facultative-pathogen of the human urogenital tract, depends on its ectoATPase activity which is uniquely mediated by OppA, the substrate-binding domain of an oligopeptide permease.
T313 518059-518279 Sentence denotes To characterize the binding of OppA to host cell structures, OppA mutants were generated with mutations of the Walker A and B motif of the ATPase domain and regions, which are conserved in OppA proteins of other species.
T314 518280-518519 Sentence denotes The attachment of the histidine-tagged mutants to extracellular matrix (ECM) components was analyzed in microplate format and compared to their adherence to HeLa cells in the presence of increasing amounts of ECM components as competitors.
T315 518520-518528 Sentence denotes Results:
T316 518529-518677 Sentence denotes Wild type OppA was shown to bind to sulfated components, like heparin, fucoidan and dextran sulfate, as well as to plasminogen, plasmin and laminin.
T317 518678-518711 Sentence denotes No binding was observed to mucin.
T318 518712-518803 Sentence denotes By the addition of plasminogen (1.2 μM) adhesion of OppA to HeLa cells was reduced to 45 %.
T319 518804-518901 Sentence denotes In the presence of fucoidan (5 μM) or dextran sulfate (0.2μM) adhesion was nearly abolished (6%).
T320 518902-519092 Sentence denotes Interestingly, mutation of the Walker A motif as well as deletion of the entire ATPase domain lead to a twofold enhanced binding to sulfated ECM structures, like heparin and dextran sulfate.
T321 519093-519392 Sentence denotes Deletion of CS1, an N-terminal region which is conserved in bacterial OppA proteins, resulted in a reduced binding of this OppA mutant to dextran sulfate (to 38 %) and heparin (17%) demonstrating that this region is involved in binding to sulfated components which are abundant in human endometrium.
T322 519393-519670 Sentence denotes The findings that the ecto-ATPase activity of OppA is essential for cytoadhesion of M. hominis whereas the CS1 region is more involved in ECM-binding suggest that these two functional regions play a different role in patho-physiological processes of host-pathogen interactions.
T323 519671-519673 Sentence denotes S.
T324 519674-519692 Sentence denotes Trüpschuch* 1 , R.
T325 519693-519706 Sentence denotes Prager 1 , A.
T326 519707-519719 Sentence denotes Fruth 1 , A.
T327 519720-519734 Sentence denotes Flieger 1 , W.
T328 519735-519987 Sentence denotes Rabsch FG 11 Bakterielle Infektionen, Wernigerode, Deutschland A monophasic variant of Salmonella Typhimurium not expressing the second phase flagellar antigene has become one of the predominant agents causing foodborne infections in humans in Germany.
T329 519988-520050 Sentence denotes In 2010 the monophasic variant represented 48% of all human S.
T330 520051-520143 Sentence denotes Typhimurium isolates sent to the National Reference Centre and was involved in 13 outbreaks.
T331 520144-520459 Sentence denotes In spite of variations in phage type, PFGE and resistance pattern within the emerging monophasic strains one type characterised by phage type DT193, PFGE pattern STYMXB.0131 and tetra-drugresistance towards antibiotics including ampicillin, streptomycin, sulfamerazine and tetracycline clearly dominates in Germany.
T332 520460-520550 Sentence denotes Meanwhile the same kind of strains is of increasing concern in various European countries.
T333 520551-520664 Sentence denotes Genome-based analyses displayed an 18.4 kb fragment adjacent to the thrW tRNA locus in this dominant strain type.
T334 520665-520806 Sentence denotes Sequencing of the detected insertion revealed 27 open reading frames and a significantly lower G+C content compared to the closely related S.
T335 520807-520905 Sentence denotes Typhimurium LT2 genome (47.4% vs. 52.2%), indicating its acquisition via horizontal gene transfer.
T336 520906-520946 Sentence denotes Transcripts have been shown for 24 ORFs.
T337 520947-521073 Sentence denotes On nucleotide level, homologous sequences for most of the open reading frames have been found in E. coli and Shigella genomes.
T338 521074-521190 Sentence denotes Protein BLAST analyses revealed several phage-related genes, indicating that the insertion might be of phage-origin.
T339 521191-521245 Sentence denotes ORF 10 had been predicted to code for a T3SS effector.
T340 521246-521325 Sentence denotes Our experiments suggest that its product is secreted but probably not via T3SS.
T341 521326-521453 Sentence denotes Infection experiments showed a reduced replication capacity of the island-deleted mutant in the mouse macrophage cell line RAW.
T342 521454-521598 Sentence denotes Further, it had been shown that the island is able to form a circular intermediate and therefore seems to be mobilised under certain conditions.
T343 521599-521756 Sentence denotes Aim of our studies is to uncover the function of the different gene products and the role of the whole island in terms of bacterial fitness or pathogenicity.
T344 521757-521827 Sentence denotes In Yersinia two pyruvate kinase (PK) isoenzymes (PykA and PykF) exist.
T345 521828-522015 Sentence denotes This glycolytic enzyme catalyses the irreversible conversion of phosphoenolpyruvate (PEP) into pyruvate coupled with the synthesis of ATP, and requires monovalent K + and divalent Mg 2+ .
T346 522016-522141 Sentence denotes Most bacterial PKs are allosterically regulated via fructose-1,6-bishosphate (FBP), ATP/AMP and/or monophosphorylated sugars.
T347 522142-522278 Sentence denotes We recombinantly produced Y.enterocolitica PykA and PykF in milligram amounts to characterise their structural and enzymatic properties.
T348 522279-522458 Sentence denotes PykA and PykF were overproduced in E. coli without affinity tags and combined anion exchange chromatography and size exclusion chromatography to yield highly homogeneous proteins.
T349 522459-522524 Sentence denotes We compared their enzymatic activities and allosteric regulation.
T350 522525-522750 Sentence denotes Further, we studied the conformational state of both isoenzymes in the absence and presence of the allosteric effector FBP applying small angle X-ray scattering (SAXS) and native polyacrylamide gel electrophoresis techniques.
T351 522751-522859 Sentence denotes Next, the spatial and temporal distribution of the two isoenzymes under various conditions shall be studied.
T352 522860-523087 Sentence denotes As expected, most high-pathogenicity-associated determinants of Y. enterocolitica subsp. enterocolitica (e.g. the High-Pathogenicity Island (HPI), Type 2 and Type 3 secretion systems) are absent from the O:3/4 genomes analysed.
T353 523088-523230 Sentence denotes However, the latter acquired putative virulence and fitness associated factors, which may substitute the classical yersinial virulence traits.
T354 523231-523548 Sentence denotes Gene clusters encoding a large 350 kDa RTxA-like toxin, an alternative chromosomal ysp T3SS, an insecticidal toxin, a pertactin,fimbriae, a putative hemolysin, an invasin, several toxin-antitoxin systems and a large 21 kbp flagellar cluster, similar to the flag-2 gene cluster of O:9/2, are evident O:3/4 acqusitions.
T355 523549-523689 Sentence denotes A filamentous PhiYep-1 prophage, which is highly homologous to CUS-1 of E. coli and Ypf prophage of Y. pestis, is present in the Y11 genome.
T356 523690-523763 Sentence denotes However, it has suffered successive deletions in strains Y8265 and Y5307.
T357 523764-523886 Sentence denotes A P2-like prophage designated PhiYep-3 is integrated in tRNA-Leu in the Y11 genome, but it is absent from Y8265 and Y5307.
T358 523887-523984 Sentence denotes We have demonstrated the precise excision of PhiYep-3 and the restoration of its attachment site.
T359 523985-524097 Sentence denotes The tRNA-Asn gene is occupied by HPI in O:8/1B and by an unidentified 14.9 kbp GIYep-01 genomic island in O:3/4.
T360 524098-524180 Sentence denotes The island has a functional integrase and can leave its tRNA-Asn integration site.
T361 524181-524313 Sentence denotes Conclusions Y. enterocolitica subsp. palearctica O:3/4 has an open genome that carries the traces of ancient and fresh acquisitions.
T362 524314-524486 Sentence denotes The acquired putative virulence and fitness-associated determinants may play an important role in host pathoadaptation and support dissemination of this group of pathogens.
T363 524487-524657 Sentence denotes Staphylococcus aureus adherence to host structures, which is a critical first step in S. aureus pathogenesis, is mediated by multiple adhesins with overlapping functions.
T364 524658-524981 Sentence denotes The role of some adhesins, particularly the MSCRAMs (microbial surface component recognizing adhesive matrix molecule) is already well established, whereas the function of the secreted and rebinding SERAMs (secretable expanded repertoire adhesive molecule), such as Emp (extracellular matrix binding protein) is less clear.
T365 524982-525147 Sentence denotes In this study, we focussed on the binding mechanism of Emp by elucidating the rebinding to the bacterial cell wall and by exploring binding sites to host structures.
T366 525148-525282 Sentence denotes For this, the Emp gene was devided into different subdomains (Emp1, Emp2, Emp3, Emp1+2, Emp2+3) , which were cloned and overexpressed.
T367 525283-525501 Sentence denotes To investigate Emp rebinding to the bacterial surface, adhesion assays were performed by adding bacterial suspensions of S. aureus strains Newman, Cowan I or 6850 to microtiter plates coated with Emp or its subdomains.
T368 525502-525604 Sentence denotes The C-terminus of the Emp protein (Emp2, Emp3) appears to be responsible for the binding to S. aureus.
T369 525605-525926 Sentence denotes Regarding the interaction between Emp and different extracellular matrix proteins (vitronectin, fibronectin, fibrinogen, collagen) as well as extracellular suprastructures from human skin and cartilage we found, that the Emp Nterminal part (Emp1, Emp2) is more important to mediate S. aureus adherence to host structures.
T370 525927-526055 Sentence denotes This could also be confirmed by Emp binding assays to different eukaryotic cell types, such as endothelial and epithelial cells.
T371 526056-526262 Sentence denotes As an alternative method, a peptide library comprised of 15 amino acids with 3 amino acids overlap, spanning full length Emp was used to identify clusters of amino acids showing binding activity to ligands.
T372 526263-526392 Sentence denotes However, Emp binding could not be broken down to short amino acid sequences, in fact, binding is mediated by the whole fragments.
T373 526393-526494 Sentence denotes Structural analysis (CD and 1 H-NMR) of Emp revealed that the protein is mainly forming random coils.
T374 526495-526589 Sentence denotes This is in line with the calculated values using the self-optimized prediction method (SOPMA).
T375 526590-526685 Sentence denotes We hypothesize, that the contact to the binding partner is essential for the protein structure.
T376 526686-526891 Sentence denotes Taken together, our results indicate that the N-terminal part of Emp mediates the binding to host structures, whereas the C-terminal part of Emp is responsible for the rebinding to the bacterial cell wall.
T1 526892-526894 Sentence denotes S.
T2 526895-526911 Sentence denotes Kositska* 1 , G.
T3 526912-526926 Sentence denotes Valenza 2 , M.
T4 526927-526944 Sentence denotes Abele-Horn 2 , H.
T5 526945-526962 Sentence denotes Hebestreit 3 , K.
T6 526963-527177 Sentence denotes Ohlsen 1 1 Institut für Molekulare Infektionsbiologie, Würzburg, Deutschland 2 Institut für Hygiene und Mikrobiologie, Würzburg, Deutschland 3 Kinderklinik, Universität Würzburg, Würzburg, Deutschland Introduction:
T7 527178-527283 Sentence denotes Cystic fibrosis (CF) is one of the most common lethal autosomal recessive disorders in white individuals.
T8 527284-527407 Sentence denotes The disease is caused by a mutation in the gene for the protein cystic fibrosis transmembrane conductance regulator (CFTR).
T9 527408-527492 Sentence denotes The most common death in CF is respiratory failure secondary to pulmonary infection.
T10 527493-527619 Sentence denotes Pseudomonas aeruginosa and Staphylococcus aureus are often co-isolated from CF respiratory cultures in 20-50% of all patients.
T11 527620-527729 Sentence denotes Methods: CF patients were surveyed for colonization with S. aureus and P. aeruginosa during the last 7 years.
T12 527730-527828 Sentence denotes Sputum cultures were collected whereas their routine clinical visits or admission to the hospital.
T13 527829-527975 Sentence denotes Pulsed -field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to determine the clonal relationship between the strains.
T14 527976-528063 Sentence denotes Microarray experiments allowed comparison of expression profiles of S. aureus isolates.
T15 528064-528087 Sentence denotes Results and Discussion:
T16 528088-528258 Sentence denotes In our study we characterised the phenotypical and genotypical features of S. aureus isolates in terms of antibiotic resistance, biofilm formation and mutation frequency.
T17 528259-528400 Sentence denotes PFGE revealed that some patients were infected with various S. aureus strains including MRSA or with the same strain during the whole period.
T18 528401-528573 Sentence denotes Ongoing studies are aimed to identify the gene expression profiles of S. aureus strains persistent over a period of several years in the presence of P. aeruginosa isolates.
T19 528574-528576 Sentence denotes D.
T20 528577-528597 Sentence denotes Chikkaballi * 1 , M.
T21 528598-528808 Sentence denotes Hensel 1 1 Universitaet Osnabrueck, Mikrobiologie, Osnabrueck, Deutschland Salmonella, an intracellular pathogen, lives and replicates in a membrane bound compartment called Salmonella containing vacuole (SCV).
T22 528809-529022 Sentence denotes While the entry of the pathogen into host cells is facilitated by the Salmonella pathogenicity island 1 (SPI1) encoded type three secretion system (T3SS), successful intracellular lifestyle requires the SPI2-T3SS.
T23 529023-529166 Sentence denotes A hallmark of intracellular Salmonella is the formation of extended tubular network from the SCV called as Salmonella-induced filaments (SIFs).
T24 529167-529279 Sentence denotes A recent study by our group has identified SIFs to be highly dynamic structures that extend, retract and branch.
T25 529280-529441 Sentence denotes Although SIFs were identified long back, their role in the intracellular life of Salmonella and the molecular mechanism underlying the dynamics is not yet known.
T26 529442-529593 Sentence denotes However, work by others and our group has identified that SIFs lie next to the microtubule tracks and are often associated or linked to motor proteins.
T27 529594-529756 Sentence denotes Because of these observations we hypothesized that the SIFs move along the microtubule tracks and require the activities of the motor proteins kinesin and dynein.
T28 529757-529938 Sentence denotes The identification of the roles of motor proteins in this context requires inhibiting their activities either by siRNA-mediated silencing or expression of dominant negative alleles.
T29 529939-530030 Sentence denotes However, these approaches render the cells compromised even before the bacterial infection.
T30 530031-530296 Sentence denotes Hence, in the present investigation, initial attempt was made to microinject specific antibodies against motor proteins at a specific time after the infection and characterize the roles of motor proteins in the intracellular life of Salmonella and dynamics of SIFs.
T31 530297-530394 Sentence denotes Streptococcus gallolyticus subsp. gallolyticus, an emerging pathogen of infective endocarditis D.
T32 530395-530408 Sentence denotes Hinse* 1 , T.
T33 530409-530423 Sentence denotes Vollmer 1 , C.
T34 530424-530437 Sentence denotes Knabbe 1 , J.
T35 530438-530484 Sentence denotes Dreier Bad Oeynhausen, Deutschland Background:
T36 530485-530594 Sentence denotes Infectious endocarditis (IE) is a disease with rising incidence and a high degree of mortality and morbidity.
T37 530595-530787 Sentence denotes 16s rDNA PCR analysis of explanted human heart valves revealed a significant relevance of Streptococcus gallolyticus subsp. gallolyticus (Sgg) (former S. bovis biotype I) as a causative agent.
T38 530788-530899 Sentence denotes Furthermore, the knowledge of virulence factors that contribute to the pathogenesis in Sgg IE is still limited.
T39 530900-530921 Sentence denotes Material and Methods:
T40 530922-531064 Sentence denotes To elucidate potential virulence genes of this bacterium, we performed ultra-high-throughput DNA sequencing using the 454 sequencing platform.
T41 531065-531260 Sentence denotes The selected strain was isolated in 2003 at the Herz-und Diabeteszentrum Nordrhein-Westfalen from an endocarditisderived blood culture of a 68-year-old woman with aortic heart valve endocarditis.
T42 531261-531336 Sentence denotes The GenDB and EDGAR software were used for genomic analysis and comparison.
T43 531337-531554 Sentence denotes Detection of microbial surface component recognizing matrix molecules (MSCRAMMs) in 41 isolates from different origins (clinical isolates IE patients, animal isolates, reference strains) was performed by PCR analysis.
T44 531555-531653 Sentence denotes Nick translation DIG labeling of plasmid DNA and Southern blotting was used for plasmid screening.
T45 531654-531662 Sentence denotes Results:
T46 531663-531816 Sentence denotes The genome of Sgg strain BAA-2069 contains a 2,356,444 bp circular DNA molecule with a G+C-content of 37.65% and a 20,765 bp plasmid designated as pSGG1.
T47 531817-531902 Sentence denotes Bioinformatic analysis predicted 2309 ORFs and the presence of 80 tRNAs and 21 rRNAs.
T48 531903-532015 Sentence denotes Additionally, 21 ORFs were detected on the plasmid pSGG1, including tetracycline resistance genes telL and tetM.
T49 532016-532094 Sentence denotes Screening of 41 Sgg isolates revealed one plasmid (pSGG2) homologous to pSGG1.
T50 532095-532255 Sentence denotes Furthermore, 21 adhesion proteins containing the cell wall-sorting motif LPxTG, which were shown to play a functional role in the pathogenesis, were identified.
T51 532256-532400 Sentence denotes Screening of 41 Sgg strains for these proteins revealed a heterogeneous distribution, correlating with binding characteristics to ECM-molecules.
T52 532401-532571 Sentence denotes The whole genome comparison to the recently sequenced Sgg strain UCN34 revealed significant differences, including habitant and host-pathogen interacting characteristics.
T53 532572-532707 Sentence denotes In order to detect a larger amount of putative virulence affecting genes in different strains, we designed a DNA microarray experiment.
T54 532708-532720 Sentence denotes Conclusions:
T55 532721-532881 Sentence denotes The analysis of the whole genome sequence of Sgg advanced the understanding of genetic factors concerning pathogenesis and adhesion to the ECM of this pathogen.
T56 532882-533032 Sentence denotes Furthermore we showed for the first time the occurrence of a resistance plasmid (pSGG1) with characteristics as a vector for lateral genetic transfer.
T57 533033-533275 Sentence denotes The alignment of microarray data and already evaluated phenotypic data, including ECM-binding, adhesion and invasion to host cells and biofilm formation capability of 41 strains, will substantiate the underlying genomic based pathomechanisms.
T58 533276-533278 Sentence denotes E.
T59 533279-533295 Sentence denotes Becker* 1 , J.H.
T60 533296-533544 Sentence denotes Hegemann Institut für Funktionelle Genomforschung der Mikroorganismen, Düsseldorf, Deutschland Chlamydiae are obligate intracellular bacteria that are responsible for a wide range of diseases of significant importance to clinical and public health.
T61 533545-533633 Sentence denotes C. trachomatis is a leading cause of sexually transmitted disease and blinding trachoma.
T62 533634-533746 Sentence denotes C. pneumoniae is an important respiratory pathogen recognized as a common cause of community acquired pneumonia.
T63 533747-533981 Sentence denotes Bioinformatical studies have led to the identification of the large family of putatively autotransported polymorphic membrane proteins (Pmps) with 9 members in C. trachomatis and 21 in C. pneumoniae, which are divided into 6 subtypes.
T64 533982-534137 Sentence denotes In previous studies, we characterized the cell surface localized protein Pmp21 as a bacterial adhesin mediating attachment of C. pneumoniae to human cells.
T65 534138-534300 Sentence denotes Furthermore, domain and mutational analysis identified the two Pmp-characteristic repetitive motifs GGA(I,L,V) and FxxN to be essential for the adhesion process .
T66 534301-534526 Sentence denotes We have now characterized the other subtypes of the Pmp protein family in the adhesion and infection process by selecting representative candidates of each Pmp subtype from C. pneumoniae and 2 orthologues from C. trachomatis.
T67 534527-534660 Sentence denotes Using the yeast display system as well as latex beads coated with recombinant protein, all tested Pmps were found to act as adhesins.
T68 534661-534834 Sentence denotes The Pmp adhesins are important for the C. pneumoniae infection process, as preincubation of human cells with the different recombinant proteins reduced subsequent infection.
T69 534835-535079 Sentence denotes Interestingly, using far western experiments we identified an interaction of Pmp21 with itself as with all other Pmps tested possibly suggesting that homo-and/or heteromeric Pmp adhesion complexes might be formed on the chlamydial cell surface.
T70 535080-535234 Sentence denotes Moreover, we examined the expression and post-translational processing of the proteins Pmp6, Pmp20, and Pmp21 during the infection cycle of C. pneumoniae.
T71 535235-535397 Sentence denotes The processing was not uniform but a Pmp-specific set of cleaved forms was detected in agreement with previous findings about Pmps being autotransporter proteins.
T72 535398-535646 Sentence denotes Cell fractionation experiments revealed that cleaved Pmp-fragments are partially released from the chlamydial surface and may exist in the inclusion lumen and possibly in the host cytosol perhaps serving an additional function as effector proteins.
T73 535647-535936 Sentence denotes Most interestingly, using immunofluorescence microscopy we detected that individual Pmp proteins were expressed only in a fraction of the inclusions analyzed possibly indicating a form of antigenic variation similar to what recently has been described for Pmp proteins from C. trachomatis.
T74 535937-536050 Sentence denotes [This work is supported by the DFG (SFB590 project C4) and the Manchot Graduate School "Molecules of Infection".]
T75 536051-536186 Sentence denotes The human pathogen Chlamydia pneumoniae nucleates and stabilizes F-actin via the secreted Cpn0572 protein to support host cell entry R.
T76 536187-536202 Sentence denotes Zrieq* 1 , J.H.
T77 536203-536392 Sentence denotes Hegemann Institut für Funktionelle Genomforschung der Mikroorganismen, Düsseldorf, Deutschland Chlamydiae are gram-negative obligate intracellular bacterial pathogens of humans and animals.
T78 536393-536466 Sentence denotes Among Chlamydiae species, C. pneumoniae is a common respiratory pathogen.
T79 536467-536642 Sentence denotes Like other pathogenic bacteria, It is believed that Chlamydiae deliver effector proteins to recruit F-actin at the site of entry to trigger their own entry into the host cell.
T80 536643-536883 Sentence denotes Among the few chlamydial effector proteins identified thus far, the C. trachomatis Tarp is translocated into the host cell where it colocalizes with F-actin underneath the attached bacteria and nucleates actin polymerization (Clifton et al.
T81 536884-536909 Sentence denotes PNAS 2004 , Jewett et al.
T82 536910-536921 Sentence denotes PNAS 2006 .
T83 536922-536999 Sentence denotes In this study, we characterized the C. pneumoniae homologous protein Cpn0572.
T84 537000-537058 Sentence denotes Cpn0572 is present in the infectious form of the bacteria.
T85 537059-537164 Sentence denotes About 20 minutes after infection Cpn0572 is secreted and could be found associated with the host F-actin.
T86 537165-537294 Sentence denotes To analyze the role of Cpn0572 in modulating the actin cytoskeleton, we used the yeast Saccaromyces cerevisiae as a model system.
T87 537295-537372 Sentence denotes Cpn0572-expressing yeast cells exhibited a strongly reduced growth phenotype.
T88 537373-537480 Sentence denotes The analysis revealed that Cpn0572 transformed the yeast actin cytoskeleton into 1-2 actin clumps per cell.
T89 537481-537562 Sentence denotes The GFP-Cpn0572 fusion protein exclusively colocalized with these F-actin clumps.
T90 537563-537635 Sentence denotes Similar observations were obtained in Cpn0572 transfected HEK-239 cells.
T91 537636-537826 Sentence denotes This actin phenotype is reminiscent of actin clumps formed in yeast and human cells treated with Jasplakinolide, a drug that induces actin polymerization and inhibits actin depolymerization.
T92 537827-537995 Sentence denotes Indeed, we could show that in yeast Cpn0572 stabilizes F-actin against the actin destabilizing drug Latrunculin A implying that Cpn0572 inhibits actin depolymerization.
T93 537996-538149 Sentence denotes Furthermore, we found evidence in vivo that Cpn0572 competes with and/or displaces the actin depolymerizing factor cofilin from binding F-actin in yeast.
T94 538150-538333 Sentence denotes These results were extended to in vitro experiments using mammalian F-actin and cofilin proteins, in which we could show that presence of Cpn0572 abrogates cofilin binding to F-actin.
T95 538334-538563 Sentence denotes In line with these data the actin clumps in Cpn0572 transfected HEK-293 cells were resistant to the actin depolymerizing drug cytochalasin D suggesting that Cpn0572 is indeed able to inhibit actin depolymerization in human cells.
T96 538564-538699 Sentence denotes Additionally, we showed also that recombinant Cpn0572 alone induces actin nucleation in vitro independent from other cellular proteins.
T97 538700-538828 Sentence denotes Finally, a deletion analysis identified a central domain within Cpn0572 (DUF1547) which is crucial for interaction with F-actin.
T98 538829-538938 Sentence denotes Conserved hydrophobic amino acids within this domain were identified as being essential for this interaction.
T99 538939-539166 Sentence denotes To our knowledge this is the first bacterial protein that exerts a dual function by inhibiting actin depolymerization and nucleating F-actin with the overall goal to maximize F-actin accumulation at the site of bacterial entry.
T100 539167-539214 Sentence denotes (The work was supported by SFB590, project C5).
T101 539215-539217 Sentence denotes A.
T102 539218-539412 Sentence denotes Sedlag* 1 1 Universität Ulm, Institut für Mikrobiologie und Biotechnologie, Ulm, Deutschland Listeria monocytogenes is the causative agent of listeriosis, a common and highly infectious disease.
T103 539413-539577 Sentence denotes Clinical manifestations are gastroenteritis, diarrhoea, and meningitis which in immune compromised, elderly or pregnant people frequently results in fatal outcomes.
T104 539578-539688 Sentence denotes During the first step of infection the intracellular pathogen Listeria monocytogenes is located in phagosomes.
T105 539689-539862 Sentence denotes Before maturing to fully acidified phagolysosomes, L. monocytogenes is able to actively lyse the membrane of this compartment and escapes to the cytosol where it replicates.
T106 539863-540114 Sentence denotes L. monocytogenes is equipped with a peptide sensing system which is homologous to the Staphylococcus aureus agr system and has been shown to be involved in the regulation of biofilm formation, in vitro and in vivo virulence and global gene expression.
T107 540115-540387 Sentence denotes In S. aureus, expression of the agr system consists of four genes encoding for an export-and processing protein (AgrB), a putative auto-inducing peptide (AgrD), a response regulator (AgrA), and a receptor kinase (AgrC) and its expression is controlled by the PII promoter.
T108 540388-540599 Sentence denotes Preliminary experiments with a PII-lux-fusion showed a lack of measurable luciferase activity in a deletion mutant of L. monocytogenes EGDe which lacks agrD, the gene encoding the putative auto inducing peptide.
T109 540600-540803 Sentence denotes To test, if the listerial AgrA acts as a transcription factor which, like the homolog in S.aureus, binds to the PII promoter a 6xHis-AgrA fusion was expressed in E.coli BL21 (DE3), isolated and purified.
T110 540804-540911 Sentence denotes The recombinant protein was tested for its binding capacity by electrophoretic mobility shift assay (EMSA).
T111 540912-540979 Sentence denotes 6xHis-AgrA was able to shift the full length PII promoter sequence.
T112 540980-541084 Sentence denotes This strongly suggests that the agr system of L. monocytogenes is subject to an auto-regulatory circuit.
T113 541085-541258 Sentence denotes A consensus AgrA-binding motif was established by in silico analysis of promoter sequences of the available peptide sensing systems from a variety of Gram-positive bacteria.
T114 541259-541376 Sentence denotes Using this consensus motif a number of putative binding sites were identified in the listerial PII promoter sequence.
T115 541377-541522 Sentence denotes These putative binding sites were analysed by EMSA using different fragments of the PII promoter and mutational alteration of the binding motifs.
T116 541523-541758 Sentence denotes Interestingly, a putative AgrA-binding motif was identifies in promoter of -prfA, the major virulence regulator of L. monocytogenes which could explain the strong regulation of this gene in the agrD-deletion mutant observed previously.
T117 541759-541825 Sentence denotes Indeed, 6xHis-AgrA was able to shift a PCRamplified prfA promoter.
T118 541826-542039 Sentence denotes This further confirms the involvement of agr peptide sensing in virulence regulation of L. monocytogenes and is the first indication of a direct regulation of virulence genes in L. monocytogenes by the agr system.
T119 542040-542241 Sentence denotes A novel class of antibacterial acyldepsipeptides (ADEPs) exerts prominent antibacterial activity against Gram-positive bacteria including multiresistant Staphylococcus aureus in vitro and in vivo [1] .
T120 542242-542334 Sentence denotes ADEPs act via a yet unprecedented mechanism by dysregulating the caseinolytic protease ClpP.
T121 542335-542439 Sentence denotes Usually, the activity of ClpP is tightly controlled by ATP-dependent Clp-ATPases and accessory proteins.
T122 542440-542690 Sentence denotes ADEPs overcome these tight control mechanisms, switching ClpP from a regulated to an uncontrolled protease that predominantly targets unfolded or flexible proteins as well as nascent polypeptides at the ribosome in the absence of Clp-ATPases [1, 2] .
T123 542691-542867 Sentence denotes Although the activity of ADEPs can be explained on the molecular level of its target ClpP, the specific events that finally lead to bacterial cell death still remained unknown.
T124 542868-542995 Sentence denotes In our study, we investigated the effect of ADEP treatment on different Gram-positive species using high-resolution microscopy.
T125 542996-543352 Sentence denotes In the presence of low inhibitory ADEP concentrations, the coccoid cells of S. aureus and Streptococcus pneumoniae swelled to more than 3-fold the volume of wild type cells, and the rod-shaped cells of B. subtilis grew into very long filaments, which reached 60-to 100-fold the length of untreated cells, clearly indicating stalled bacterial cell division.
T126 543353-543486 Sentence denotes To gain further insights into the underlying molecular mechanism, we followed the events that led to the inhibition of cell division.
T127 543487-543748 Sentence denotes Cell division in Gram-positive bacteria is concerted by the divisome, which is highly dynamic and is characterized by a time-dependent two-step assembly of specific cell division proteins like FtsZ, FtsA, ZapA and EzrA as well as PBP2B, FtsW, DivIVA, etc. [3] .
T128 543749-543904 Sentence denotes We observed that ADEP treatment led to the inhibition of septum formation in S. aureus and B. subtilis, while chromosome segregation was rather unaffected.
T129 543905-544105 Sentence denotes Localization studies with GFP-labeled cell division proteins revealed that the ADEP-ClpP complex interferes with key components of early cell division and therefore perturbs normal divisome formation.
T130 544106-544349 Sentence denotes By analyzing cell extracts of ADEP-treated S. aureus and B. subtilis cells, immunoblotting revealed that the ADEP-ClpP proteolytic complex degrades essential components of the cell division machinery and, hence, destines the bacteria to death.
T131 544350-544544 Sentence denotes ADEPs demonstrate that beside their interesting antibacterial potency they are excellent tools to examine central mechanism of bacterial physiology, like cell division and regulated proteolysis.
T132 544545-544547 Sentence denotes A.
T133 544548-544565 Sentence denotes Berscheid* 1 , P.
T134 544566-544579 Sentence denotes Sass 1,2 , A.
T135 544580-544593 Sentence denotes Jansen 1 , M.
T136 544594-544611 Sentence denotes Oedenkoven 1 , C.
T137 544612-544625 Sentence denotes Szekat 1 , A.
T138 544626-544645 Sentence denotes Strittmatter 3 , G.
T139 544646-544663 Sentence denotes Gottschalk 3 , G.
T140 544664-544676 Sentence denotes Bierbaum 1 1
T141 544678-544804 Sentence denotes Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are commonly treated with the glycopeptide vancomycin.
T142 544805-544905 Sentence denotes However, MRSA strains with reduced susceptibility to vancomycin have emerged during the last decade.
T143 544906-545024 Sentence denotes Due to increased antibiotic treatment failure, there is an obvious need to understand how bacteria develop resistance.
T144 545025-545218 Sentence denotes In an approach to study the impact of an elevated mutation frequency on vancomycin resistance development, we had previously isolated the highly vancomycin resistant S. aureus strain VC40 (MIC:
T145 545219-545277 Sentence denotes 64 μg/ml) after serial passage of strain RN4220 mutS (MIC:
T146 545278-545413 Sentence denotes 2 μg/ml), a mutS gene deletion mutant of the laboratory strain RN4220, in the presence of increasing concentrations of vancomycin (1) .
T147 545414-545700 Sentence denotes In the current study, antimicrobial susceptibility testing of strain VC40 additionally revealed cross resistance against the lipopeptide antibiotic daptomycin, which has recently been introduced for the treatment of complicated skin and skin structure infections (cSSSI) caused by MRSA.
T148 545701-545908 Sentence denotes To gain further insights into the resistance phenotype of S. aureus VC40, the full genome sequences of this strain and its parent strain S. aureus RN4220 mutS were determined using 454 sequencing technology.
T149 545909-546100 Sentence denotes In strain VC40, a total of 79 mutations in genes related to cell wall metabolism, transport and gene regulation, including the two-component regulatory systems VraSR and WalKR, were detected.
T150 546101-546313 Sentence denotes Since the latter regulatory proteins significantly affect the biosynthesis and turnover rates of the bacterial cell wall (2, 3) , we performed further morphological studies using transmission electron microscopy.
T151 546314-546489 Sentence denotes Here, strain VC40 was characterized by an abnormal cell envelope morphology including an increased cell wall thickness that may result from deregulated VraRS or WalKR systems.
T152 546490-546630 Sentence denotes Cell wall thickening has previously been linked to vancomycin resistance and even has been correlated with daptomycin cross resistance (4) .
T153 546631-546727 Sentence denotes However, these findings alone cannot explain the high-level resistance phenotype of strain VC40.
T154 546728-546960 Sentence denotes Further characterization of strain VC40 is necessary and will provide important information on how the accumulation of mutations resulted in the autonomous development of vancomycin and daptomycin high-level resistance in S. aureus.
T155 546961-546963 Sentence denotes S.
T156 546964-546980 Sentence denotes Schielke* 1 , G.
T157 546981-547188 Sentence denotes Häcker 1 1 Universität Freiburg, Institut für Medizinische Mikrobiologie und Hygiene, Freiburg, Deutschland Chlamydia trachomatis is specialized for survival and growth within the human epithelial host cell.
T158 547189-547339 Sentence denotes In vivo, the bacteria very likely face adverse replication conditions, in particular when the immune response is activated and cytokines are secreted.
T159 547340-547466 Sentence denotes Such conditions can be mimicked in a number of ways, for instance by the addition of -lactam antibiotics to cultures in vitro.
T160 547467-547535 Sentence denotes In these conditions Chlamydiae enter a so-called "persistent" state.
T161 547536-547839 Sentence denotes The persistent phenotype is characterized by enlarged, aberrant reticulate bodies (RBs; the replicative form of Chlamydia), which are neither able to divide nor to re-differentiate into infectious elementary bodies (EBs) but which can be re-activated upon withdrawal of the persistence-inducing reagent.
T162 547840-548059 Sentence denotes As persistent infections are thought to be relevant for the outcome of chlamydial disease, especially in chronic infections, we are investigating the development of the chlamydial inclusion during this persistent state.
T163 548060-548313 Sentence denotes Infection of human cervical epithelial cells (HeLas), mouse oviduct epithelial cells (C57epi.1) and mouse embryonic fibroblasts (MEFs) with C. trachomatis L2 in the presence of penicillin G resulted in the development of aberrant, non-proliferating RBs.
T164 548314-548410 Sentence denotes Although there were only few of these RBs per inclusion, the vacuole grew substantially in size.
T165 548411-548532 Sentence denotes We have proposed that growth of the inclusion during productive infection requires the secreted chlamydial protease CPAF.
T166 548533-548645 Sentence denotes During persistent infection, expression and activity of CPAF was also detectable over a time course of 64 hours.
T167 548646-548785 Sentence denotes Maturation of C. trachomatis is dependent on host cell lipids, which are acquired upon CPAF-dependent fragmentation of the Golgi apparatus.
T168 548786-548931 Sentence denotes During penicillin-induced persistence Golgi fragmentation likewise occurred, resulting in the formation of Golgi ministacks around the inclusion.
T169 548932-549185 Sentence denotes These results suggest that even upon entry into the 'persistent' state during treatment with penicillin G, C. trachomatis can maintain the machinery for lipid acquisition and inclusion growth, very likely by maintaining production and secretion of CPAF.
T170 549186-549277 Sentence denotes Understanding this chlamydial activity may help understanding persistent infection in vivo.
T171 549278-549291 Sentence denotes Introduction:
T172 549292-549396 Sentence denotes Staphylococcus aureus small colony variants (SCV) are frequently isolated from cystic fibrosis patients.
T173 549397-549489 Sentence denotes Defective airway clearance and prolonged antibiotic treatment predisposes for SCV formation.
T174 549490-549501 Sentence denotes Objectives:
T175 549502-549674 Sentence denotes To assess the relevance of intra-strain diversity on the hostpathogen interaction in the present study we analyzed consecutive S. aureus SCV isolates from the same patient.
T176 549675-549695 Sentence denotes Methods and Results:
T177 549696-549801 Sentence denotes Pulsed-field gel electrophoresis and MALDI biotyping demonstrated strain relatedness of all SCV isolates.
T178 549802-549915 Sentence denotes Nevertheless, SCV isolates could be distinguished by spa-typing, e.g. one isolate displayed a truncated spa-type.
T179 549916-550146 Sentence denotes Interestingly, Western blot analysis of protein A expression revealed that the truncated spa gene was associated with a protein of decreased size, and that two SCV isolates could be distinguished by increased protein A expression.
T180 550147-550237 Sentence denotes For functional comparison of SCV isolates we compared TLR2 activity and biofilm formation.
T181 550238-550368 Sentence denotes The results obtained indicated that increased SpA expression correlated with increased TLR2 activity and biofilm forming capacity.
T182 550369-550558 Sentence denotes An enhancing role for protein A was subsequently confirmed by comparison of a spa-deficient S. aureus mutant SA113 Dspa to the isogenic wild type strain SA113 WT in TLR2 and biofilm assays.
T183 550559-550571 Sentence denotes Conclusions:
T184 550572-550752 Sentence denotes We conclude that quantitative differences in protein A expression reflect intra-strain variability and translate to functional differences modulating the host-pathogen interaction.
T185 550753-551072 Sentence denotes However, we can only speculate that concomitant changes observed in the microbial environment (e.g. infection and colonization with other microbial species), in the pulmonary and clinical status of the patient and in the antibiotic regimes may be responsible for the functional and genetic changes in the SCV phenotype.
T186 551073-551218 Sentence denotes While surface-associated motility has long been described in Acinetobacter lacking flagella, the mechanism of motility has never been elucidated.
T187 551219-551384 Sentence denotes Acinetobacter baumannii, an often multidrug-resistant nosocomial pathogen, may profit from motility during infection or while persisting in the hospital environment.
T188 551385-551551 Sentence denotes To identify determinants of Acinetobacter motility, a library of transposon mutants of strain ATCC 17978 was generated and screened for mutants with motility defects.
T189 551552-551660 Sentence denotes We could identify two akinetic mutants carrying transposon insertions in the dat and ddc gene, respectively.
T190 551661-551912 Sentence denotes These neighbouring genes encode L-2,4-diaminobutyrate:2ketoglutarate 4-aminotransferase and L-2,4-diaminobutyrate decarboxylase, respectively, and contribute to synthesis of 1,3-diaminopropane (DAP), a polyamine ubiquitously produced in Acinetobacter.
T191 551913-552007 Sentence denotes The motility defect of both mutants could be cured by supplementing semi-solid media with DAP.
T192 552008-552191 Sentence denotes Making use of naturally competent clinical Acinetobacter isolates, we verified by transformation of the mutants' DNA that ddc and dat were required for motility in different isolates.
T193 552192-552274 Sentence denotes Collectively, surface-associated motility of Acinetobacter baumannii requires DAP.
T194 552275-552340 Sentence denotes Current work aims at elucidating how DAP contributes to motility.
T195 552341-552343 Sentence denotes A.
T196 552344-552357 Sentence denotes Weiss* 1 , J.
T197 552358-552374 Sentence denotes Soltwisch 2 , J.
T198 552375-552638 Sentence denotes Brockmeyer 1 1 Institut für Lebensmittelchemie, Forschung, Münster, Deutschland 2 Institut für Medizinische Physik und Biophysik, Münster, Deutschland Enterohemorrhagic Escherichia coli (EHEC) naturally occur in the intestinal tract of cattle and other ruminants.
T199 552639-552695 Sentence denotes Humans are mainly infected via contaminated animal food.
T200 552696-552793 Sentence denotes In addition, infections via vegetables are possible, as shown by the current outbreak in Germany.
T201 552794-552917 Sentence denotes The clinical outcome of EHEC infection range from intestinal dysfunctions to the lifethreatening hemolytic-uremic syndrome.
T202 552918-553045 Sentence denotes To date, systematic therapy is hampered by the fact that the detailed mechanism of pathogenicity is still not fully understood.
T203 553046-553114 Sentence denotes EHEC express a number of factors that might contribute to virulence.
T204 553115-553185 Sentence denotes One of them is the plasmid-encoded extracellular serine protease EspP.
T205 553186-553338 Sentence denotes It has been shown that EspP might interact with hemostasis and immune response (1,2) but a systematic screening of relevant substrates is still lacking.
T206 553339-553423 Sentence denotes We therefore screened for novel substrates of EspP using human plasma and platelets.
T207 553424-553546 Sentence denotes Plasma was first depleted from albumins and IgG and incubated with purified recombinant EspP or the inactive mutant S263A.
T208 553547-553673 Sentence denotes Fragmentation of plasma proteins was assessed by SDS-PAGE or 2D gel electrophoresis and compared to the nonincubated proteins.
T209 553674-553846 Sentence denotes Analysis of EspP substrates in platelets was performed with platelet lysates to obtain whole cell extracts or collagen-activated platelets to analyze the secreted proteins.
T210 553847-553966 Sentence denotes Following incubation with EspP or S263A protein pattern were investigated by aforementioned electrophoresis techniques.
T211 553967-554201 Sentence denotes To indentify novel substrates, protein spots that shifted in gel electrophoretic mobility compared to the negative control were digested in-gel and subjected to mass spectrometrical analysis using MALDI-TOF-MS as well as nano-ESI-MS².
T212 554202-554387 Sentence denotes Altogether, it has been shown that EspP cleaves several physiologically relevant proteins in plasma as well as in platelets including protease inhibitors and surface adhesion molecules.
T213 554388-554530 Sentence denotes Further studies will elucidate the contribution of the newly identified substrates to the role of EspP as a possible virulence factor of EHEC.
T214 554531-554653 Sentence denotes HlyA from E. coli is secreted in an unfolded state and requires the C-terminal located amino acids for its lytic function.
T215 554654-554656 Sentence denotes S.
T216 554657-554670 Sentence denotes Thomas 1 , T.
T217 554671-554686 Sentence denotes Jumpertz 1 , P.
T218 554687-554700 Sentence denotes Bakkes 1 , S.
T219 554701-554714 Sentence denotes Smits* 1 , L.
T220 554715-554996 Sentence denotes Schmitt 1 1 Heinrich Heine University, Institut für Biochemie, Duesseldorf, Deutschland Escherichia coli haemolysin A (HlyA), an RTX toxin, is secreted probably as an unfolded intermediate, by the type I (ABC transporter-dependent) pathway, utilizing a C-terminal secretion signal.
T221 554997-555086 Sentence denotes However, the mechanism of translocation and post-translocation folding is not understood.
T222 555087-555266 Sentence denotes We identified a mutation (hlyA99) at the extreme C terminus, which is dominant in competition experiments, blocking secretion of the wild-type toxin co-expressed in the same cell.
T223 555267-555435 Sentence denotes This suggests that unlike recessive mutations which affect recognition of the translocation machinery, the hlyA99 mutation interferes with some later step in secretion.
T224 555436-555627 Sentence denotes Indeed, the mutation reduced haemolytic activity of the toxin and the activity of betalactamase when the latter was fused to a C-terminal 23 kDa fragment of HlyA carrying the hlyA99 mutation.
T225 555628-555848 Sentence denotes A second mutant (hlyAdel6), lacking the six C-terminal residues of HlyA, also showed reduced haemolytic activity and neither mutant protein regained normal haemolytic activity in in vitro unfolding/refolding experiments.
T226 555849-555991 Sentence denotes Tryptophan fluorescence spectroscopy indicated differences in structure between the secreted forms of wild-type HlyA and the HlyA Del6 mutant.
T227 555992-556106 Sentence denotes These results suggested that the mutations affected the correct folding of both HlyA and the betalactamase fusion.
T228 556107-556262 Sentence denotes Thus, we propose a dual function for the HlyA C terminus involving an important role in post-translocation folding as well as targeting HlyA for secretion.
T229 556263-556381 Sentence denotes The Hly translocator complex exports a variety of passenger proteins when fused N-terminal to this secretion sequence.
T230 556382-556432 Sentence denotes However, not all fusions are secreted efficiently.
T231 556433-556604 Sentence denotes Here, we demonstrate that the maltose binding protein (MalE) lacking its natural export signal and fused to the HlyA secretion signal is poorly secreted by the Hly system.
T232 556605-556782 Sentence denotes Anticipating that folding kinetics might be limiting, we introduced the 'folding' mutation Y283D into MalE and indeed the fusion protein was now secreted at a much higher level.
T233 556783-556884 Sentence denotes This level was further enhanced by the introduction of a second MalE folding mutation (V8G or A276G).
T234 556885-557017 Sentence denotes Folding analysis revealed that all mutations reduced the refolding rate of the substrate, whereas the unfolding rate was unaffected.
T235 557018-557141 Sentence denotes Thus, HlyA is secreted in a unfolded state and folds into its lytic conformation upon binding of calcium ion in the medium.
T236 557142-557144 Sentence denotes H.
T237 557145-557159 Sentence denotes Funken* 1 , A.
T238 557160-557172 Sentence denotes Knapp 1 , M.
T239 557173-557187 Sentence denotes Brocker 2 , S.
T240 557188-557202 Sentence denotes Wilhelm 1 , F.
T241 557203-557222 Sentence denotes Rosenau 1,3 , K.-E.
T242 557223-557448 Sentence denotes Jaeger 1 1 University Duesseldorf, IMET, Jülich, Deutschland 2 Forschungszentrum Jülich, Jülich, Deutschland 3 University Ulm, Biotechnology, Ulm, Deutschland One of the most essential elements for living of organism is iron.
T243 557449-557667 Sentence denotes Iron acts as a cofactor for a range of redox enzymes which are involved in many central cellular processes like electron transport or DNA synthesis, it is also essential for amino acid and pyrimidine biosynthesis [1] .
T244 557668-557830 Sentence denotes One characteristic feature of fluorescent pseudomonads is their ability to produce the yellowgreen fluorescent pigment pyoverdine as their major siderophore [2] .
T245 557831-558000 Sentence denotes The best studied pyoverdine is that of the opportunistic human pathogen Pseudomonas aeruginosa, where it is required for growth and plays a major role in virulence [3] .
T246 558001-558127 Sentence denotes Among other factors, pyoverdine biosynthesis in P. aeruginosa is regulated by the iron starvation (IS) sigma factor PvdS [4] .
T247 558128-558300 Sentence denotes We have applied a proteomic approach using the 2D DIGE system and transcriptional analysis by qPCR to study the role of P. aeruginosa lipase LipA for pyoverdine production.
T248 558301-558391 Sentence denotes We clearly demonstrated that the lipase has a regulatory effect on the pvdS transcription.
T249 558392-558541 Sentence denotes A lipA negative mutant of P. aeruginosa ( lipA) showed a significant lower pyoverdine concentration in culture supernatant compared to the wild type.
T250 558542-558677 Sentence denotes In addition, the secretome of P. aeruginosa lipA revealed differences in PvdS regulated proteins including the proteases PrpL and AprA.
T251 558678-558787 Sentence denotes These proteins are only present in the lipA deficient strain when lipase expression is complemented in trans.
T252 558788-558993 Sentence denotes Results obtained by qPCR revealed a significantly decreased level of pvdS m-RNA in P. aeruginosa lipA and, as a consequence, the transcription of several other PvdS regulated genes was also down-regulated.
T253 558994-559207 Sentence denotes These results indicate that the LipA plays an important role in the regulation of the iron starvation sigma factor PvdS in P. aeruginosa thereby influencing the synthesis of pyoverdine and other virulence factors.
T254 559208-559210 Sentence denotes A.
T255 559211-559225 Sentence denotes Pelzer* 1 , S.
T256 559226-559238 Sentence denotes Serci 1 , F.
T257 559239-559258 Sentence denotes Rosenau 1,2 , K.-E.
T258 559259-559272 Sentence denotes Jaeger 1 , S.
T259 559273-559647 Sentence denotes Wilhelm 1 1 Heinrich-Heine-University Duesseldorf, Institute for Molecular Enzyme Technology, Juelich, Deutschland 2 Ulm University, Institute of Pharmaceutical Biotechnology, Ulm, Deutschland Pseudomonas aeruginosa is ubiquitously distributed, living in wet or humid surroundings ranging from soil to the human body and it produces a huge variety of extracellular proteins.
T260 559648-559726 Sentence denotes An important part of the arsenal of bacterial virulence factors are proteases.
T261 559727-559826 Sentence denotes Proteases are enzymes that hydrolyze peptide bonds and can therefore degrade proteins and peptides.
T262 559827-559981 Sentence denotes As proteins are one of the basic building blocks in nature, proteases can influence a broad range of biological functions including the infection process.
T263 559982-560237 Sentence denotes The opportunistic human pathogen P. aeruginosa has an arsenal of efficient proteases that helps establishing and maintaining an infection and thereby controlling and modifying the environment according to the needs of the bacterium within the host tissue.
T264 560238-560382 Sentence denotes Some of these proteases, like Elastase and Protease IV, are well characterized but others exist, of which nothing is known (Hoge et al., 2010) .
T265 560383-560466 Sentence denotes We focused on serine proteases that belong to the peptidase S8 family (subtilases).
T266 560467-560512 Sentence denotes These subtilases are encoded as preproenzyme.
T267 560513-560715 Sentence denotes They carry a signal peptide that drives their translocation through the cytoplasmic membrane and a propeptide that acts as a folding mediator required to give the protease its final native conformation.
T268 560716-560849 Sentence denotes By homology, we have identified the open reading frames PA1242 (sprP) and PA3535 (sprS) in the genome sequence of P. aeruginosa PAO1.
T269 560850-560976 Sentence denotes The gene sprP encodes for a so far hypothetical protein as a putative member of the subtilisin-like serine protease family S8.
T270 560977-561065 Sentence denotes The gene product of sprP contains a predicted signal sequence and a peptidase S8 domain.
T271 561066-561174 Sentence denotes Sequence analysis revealed the presence of an additional element in the domain organization of the protease.
T272 561175-561299 Sentence denotes SprP carries, in addition to its signal peptide and the S8 domain, a domain of unknown function (DUF) between both elements.
T273 561300-561413 Sentence denotes After the protease activity of SprP was demonstrated, we characterized different virulence associated phenotypes.
T274 561414-561647 Sentence denotes We were able to illustrate an eminent role of SprP for the virulence of P. aeruginosa because the deletion of sprP resulted in a loss of motility, an increased biofilm formation, and the accumulation of cell aggregates during growth.
T275 561648-561739 Sentence denotes The second subtilase SprS belongs to the autotransporter protein family (type V secretion).
T276 561740-561824 Sentence denotes In the case of SprS, the N-terminal passenger domain contains the protease function.
T277 561825-561947 Sentence denotes Similar to the sprP deletion strain, the P. aeruginosa DsprS showed a loss of motility and an increased biofilm formation.
T278 561948-562097 Sentence denotes Additionally the secretome showed that the sprS-negative strain produces less virulence associated extracellular proteases compared to the wild type.
T279 562098-562179 Sentence denotes Initial results suggest a regulatory relationship in between these two proteases.
T280 562180-562296 Sentence denotes Finally, these results indicate that the two novel proteases, SprP and SprS, play a role in P. aeruginosa virulence.
T281 562297-562475 Sentence denotes The iron-regulated gene A (IrgA) homologue adhesin (Iha) is encoded in O island (OI) 43 and 48 in the genome of prototypic enterohemorrhagic E. coli (EHEC) O157:H7 strain EDL933.
T282 562476-562559 Sentence denotes The role of this molecule in the virulence of EHEC O157 is incompletely understood.
T283 562560-562625 Sentence denotes In uropathogenic E. coli Iha functions as a siderophore receptor.
T284 562626-562769 Sentence denotes We identified several EHEC O157:H7 strains that underwent spontaneous internal or complete deletions of OI-43 and/or OI-48 leading to iha loss.
T285 562770-562984 Sentence denotes The iha-negative derivatives displayed, under iron-limited conditions that promote iha transcription, significantly lower adherence to human intestinal epithelial cells than did their iha-positive parental strains.
T286 562985-563101 Sentence denotes Moreover, iha loss reduced the ability of strains to grow in liquid as well as on solid media under iron limitation.
T287 563102-563443 Sentence denotes Isogenic iha deletion mutant of the parental strain displayed adherence capacity and growth ability comparable to that of the spontaneous ihanegative derivative, whereas iha complementation of the iha deletion mutant and the iha-negative spontaneous derivative restored the adherence and the growth speed to the level of the parental strain.
T288 563444-563579 Sentence denotes These data suggest that Iha plays dual roles in the virulence and/or fitness of EHEC O157:H7, as an adhesin and a siderophore receptor.
T289 563580-563700 Sentence denotes The S. epidermidis accumulation associated protein Aap: proteolytic processing and consequences for biofilm formation M.
T290 563701-563719 Sentence denotes Kotasinska* 1 , M.
T291 563720-563732 Sentence denotes Busch 1 , M.
T292 563733-563746 Sentence denotes Michel 1 , N.
T293 563747-563762 Sentence denotes Schommer 1 , C.
T294 563763-563779 Sentence denotes Burdelski 1 , G.
T295 563780-563792 Sentence denotes Kroll 1 , H.
T296 563793-563808 Sentence denotes Schlüter 1 , H.
T297 563809-563987 Sentence denotes Rohde 1 1 Universitätsklinikum Eppendorf, Klinische Chemie, Hamburg, Deutschland Implant associated Staphylococcus epidermidis infections essentially depend on biofilm formation.
T298 563988-564093 Sentence denotes S. epidermidis biofilm formation depends on polysaccharide PIA and accumulation associated protein (Aap).
T299 564094-564205 Sentence denotes 160 kDa Aap is essentially consists of an N-terminal domain A (aa 1 -609) and a repetitive C-terminal domain B.
T300 564206-564322 Sentence denotes Importantly, Aap mediates biofilm formation only after proteolytic processing, during which the A domain is removed.
T301 564323-564392 Sentence denotes In turn, the repetitive B domain then induces cell cluster formation.
T302 564393-564560 Sentence denotes Therefore, proteolytic processing is of fundamental importance for Aap function, however, the underlying mechanisms and proteases involved have not been characterized.
T303 564561-564700 Sentence denotes Here we used specific S. epidermidis mutants, recombinant Aap sub-domains and N-terminal sequencing to characterize dynamic Aap processing.
T304 564701-564744 Sentence denotes Aap is cleaved at least at three positions.
T305 564745-564827 Sentence denotes Two cleavage sites were identified within the A domain at position aa 283 and 335.
T306 564828-564922 Sentence denotes A third cleavage site is located at the junction between domain A and B at position of aa 596.
T307 564923-565093 Sentence denotes By using supernantants from protease overexpressing S. epidermidis mutant 1457-M10sarA and mutant 1457-M10sarAsepA processing of Aap was assigned to metalloprotease SepA.
T308 565094-565204 Sentence denotes Experiments using protease inhibitors suggest that Aap cleavage could also occur through additional proteases.
T309 565205-565349 Sentence denotes Our results indicate that S. epidermidis uses specific proteases for the controlled and coordinated processing and functional activation of Aap.
T310 565350-565677 Sentence denotes Given the importance of Aap-mediated S. epidermidis biofilm formation for pathogenesis of foreign-material associated infections our results could open new ways for the future development of therapeutic approaches, e.g. by interference with protease dependent dynamics of pro-aggregative S. epidermidis cell surface properties.
T311 565678-565763 Sentence denotes Tryptophan is an essential amino acid for human beings as well as for microorganisms.
T312 565764-565911 Sentence denotes Therefore, the reduction of local tryptophan concentrations in tissues has an influence on the growth of microorganisms that have invaded the body.
T313 565912-566063 Sentence denotes In human cells the IFN-inducible enzyme indoleamine 2,3-dioxygenase (IDO) degrades tryptophan and is therefore able to mediate an antimicrobial effect.
T314 566064-566178 Sentence denotes It could be shown, that in in vitro systems an IDO activity inhibits the growth of bacteria, parasites or viruses.
T315 566179-566304 Sentence denotes Additionally, it was observed that IDO has immunoregulatory functions, as it controls the activation and survival of T-cells.
T316 566305-566378 Sentence denotes IDO is a dioxygenase that catalyses the oxidative cleavage of tryptophan.
T317 566379-566509 Sentence denotes Up to now, however, it remained unclear under which oxygen conditions IDO promotes its antimicrobial and immunoregulatory effects.
T318 566510-566622 Sentence denotes Therefore, the enzymatic activity of IDO in different tissues with variable oxygen supply needs to be clarified.
T319 566623-566735 Sentence denotes In recent studies we could show that the activity of IDO is inhibited under hypoxic conditions in vitro (1% O2).
T320 566736-566856 Sentence denotes In comparison to normoxic conditions, IDO produces a reduced amount of kynurenine, the downstream product of tryptophan.
T321 566857-566940 Sentence denotes In line with this observation, the antibacterial effect is abrogated under hypoxia.
T322 566941-567138 Sentence denotes We could find that the IDO-mediated inhibition of Staphylococcus aureus as well as group B Streptococcus growth was abrogated in IFN-stimulated human glioblastoma cells or HeLa-cells, respectively.
T323 567139-567262 Sentence denotes Hypoxia not only impaired IDO enzyme activity but also resulted in reduced IFN-mediated signaling via the JAK-Stat pathway.
T324 567263-567376 Sentence denotes IFN-induced IDO enzyme activity and JAK-Stat signaling were not inhibited when oxygen concentrations were >5% O2.
T325 567377-567573 Sentence denotes These findings indicate that the influence of the oxygen concentration on IDO function could be important for the defense against several pathogens that affect organs with different oxygen supply.
T326 567574-567716 Sentence denotes Phospholipases A (PLA) represent a group of bacterial virulence factors with the ability to interact with host membranes and lipid signalling.
T327 567717-567913 Sentence denotes Only a few PLA are proven virulence factors 1,2 but the total number of known PLA is still limited and the details about host-pathogen interaction mechanisms involving PLA remain to be elucidated.
T328 567914-568056 Sentence denotes The opportunistic human pathogen Pseudomonas aeruginosa produces a variety of virulence factors; however, a PLA has not been described so far.
T329 568057-568186 Sentence denotes A comprehensive in silico analyses of the P. aeruginosa PA01 genome revealed number of genes encoding putative lipolytic enzymes.
T330 568187-568304 Sentence denotes Here, we describe the identification and characterization of PlbF, a novel phospholipase B produced by P. aeruginosa.
T331 568305-568446 Sentence denotes PlbF displaying sequence homology to prokaryotic and eukaryotic PLA, lipases and esterases involved in lipid metabolism and lipid signalling.
T332 568447-568545 Sentence denotes We succeeded to purify this enzyme and to demonstrate its esterase and phospholipase B activities.
T333 568546-568652 Sentence denotes Subcellular localisation experiments revealed its association with the membrane fraction of P. aeruginosa.
T334 568653-568797 Sentence denotes More detailed localisation studies suggested that PlbF is most likely anchored to the bacterial inner membrane, as an integral membrane protein.
T335 568798-568967 Sentence denotes Our experimental data are in agreement with in silico analyses which have predicted the presence of hydrophobic transmembrane helices in the N-terminal sequence of PlbF.
T336 568968-569088 Sentence denotes Additionally, we recognised within the PlbF amino acid sequence a signature motif present in integral membrane proteins.
T337 569089-569211 Sentence denotes This motif is required for helix-helix interactions of inner membrane proteins, suggesting that PlbF may exist as a dimer.
T338 569212-569286 Sentence denotes This assumption was experimentally confirmed by cross linking experiments.
T339 569287-569500 Sentence denotes The contribution of PlbF to P. aeruginosa virulence was investigated in a D. melanogaster virulence modelby using a plbF negative P. aeruginosa mutant which was compared to the wild type strain P. aeruginosa PA01.
T340 569501-569674 Sentence denotes Significant attenuation of the plbF negative mutant strain in killing of flies was observed confirming PlbF may represent a novel virulence factor produced by P. aeruginosa.
T341 569675-569898 Sentence denotes Further investigations of the cellular function of PlbF, particularly the relation of PlbF dimerisation and virulence of P. aeruginosa PAO1 might elucidate novel mechanisms of PLA-based pathogenicity mechanisms in bacteria.
T342 569899-570110 Sentence denotes Non-coding small RNAs (ncRNAs) have been recognised as important players in regulatory networks controlling gene expression in bacteria, and recent research has led to profound insights into their way of action.
T343 570111-570292 Sentence denotes Thus, while trans-encoded ncRNAs often act through direct base pair interaction with target-mRNAs, cis-encoded ncRNAs directly regulate the transcript within which they are encoded.
T344 570293-570500 Sentence denotes Many cis-encoded ncRNAs are located in the 5' untranslated region (5' UTR) of genes and operons and control downstream gene transcription by acting as riboswitches through the binding of various metabolites.
T345 570501-570693 Sentence denotes Here we describe the detection and characterisation of a novel cis-encoded ncRNA in the 5' UTR preceding the methionine biosynthesis genes in S. aureus, S. epidermidis and other staphylococci.
T346 570694-571033 Sentence denotes Bioinformatic analyses of the staphylococcal met leader RNA revealed homologies and predicted structural elements associated with Bacillus T-box systems which represent terminator/antiterminator transcription control elements of Gram-positive bacteria that undergo conformational change upon specific binding of charged or uncharged tRNAs.
T347 571034-571216 Sentence denotes In the staphylococcal met leader RNA conserved sequence stretches were identified such as a T-box motif and a transcription terminator, as well as, unexpectedly, a CodY binding site.
T348 571217-571454 Sentence denotes In vitro binding assays with the met leader RNA and tRNA Met demonstrated specific binding interaction between the two RNA species, and mutational studies of the T-box motif revealed the importance of single nucleotides for tRNA binding.
T349 571455-571661 Sentence denotes Transcript levels of the met leader RNA and the downstream methionine biosynthesis genes were shown to vary between different staphylococcal strains, but to be generally induced under methionine starvation.
T350 571662-571883 Sentence denotes The pleiotropic repressor CodY seems to regulate the overall transcription of the T-box system, but does not interfere with the tight methionine-dependent regulation of the biosynthesis genes by the met leader RNA itself.
T351 571884-572152 Sentence denotes Various S. aureus RNase mutants were investigated for their influence on met leader RNA and metoperon transcription and stability, and these experiments demonstrated the involvement of RNase Y, RNase J2 and RNase III in met leader RNA/ mRNA processing and degradation.
T352 572153-572298 Sentence denotes The combined data demonstrate that in staphylococci de novo methionine biosynthesis is obviously under the control of a T-box regulatory element.
T353 572299-572480 Sentence denotes This is unusual and in contrast to other Gram-positive bacteria which regulate this pathway mostly by S-box systems which represent S-adenosyl methionine (SAM)-binding riboswitches.
T354 572481-572483 Sentence denotes A.
T355 572484-572499 Sentence denotes Mehlitz* 1 , G.
T356 572500-572513 Sentence denotes Krohne 2 , T.
T357 572514-572742 Sentence denotes Rudel 1 1 Universität Würzburg, Mikrobiologie, Würzburg, Deutschland 2 Universität Würzburg, Elektronenmikrokopie, Würzburg, Deutschland Simkania negevensis is a Chlamydia-like emerging pathogen implicated in pulmonary diseases.
T358 572743-572819 Sentence denotes However, knowledge regarding its intracellular accommodation remains sparse.
T359 572820-573011 Sentence denotes Here, we demonstrated that expansion of the S. negevensis vacuole within epithelial cells is accompanied by massive spatial reorganization of host mitochondria and endoplasmic reticulum (ER).
T360 573012-573176 Sentence denotes Spatial reorganization was mitochondria-and ER-specific as the Golgi apparatus appeared intact and was positioned between the nucleus and the S. negevensis vacuole.
T361 573177-573304 Sentence denotes Ultrastructural analysis and 3D reconstruction revealed that S. negevensis forms one large vacuole located within the ER lumen.
T362 573305-573436 Sentence denotes Location of the vacuole within the ER led to the formation of a so far not described pathogen-containing triple membrane structure.
T363 573437-573513 Sentence denotes Expression of the non-fimbrial large adhesin BapA of Salmonella enterica sv.
T364 573514-573528 Sentence denotes Typhimurium A.
T365 573529-573549 Sentence denotes Felipe-López* 1 , B.
T366 573550-573563 Sentence denotes Haßing 1 , M.
T367 573564-573824 Sentence denotes Hensel 1 1 FB05 Universität Osnabrück, Mikrobiologie, Osnabrück, Deutschland Two adhesins were recently described at the locus of the Salmonella Pathogenicity Island 4 (SPI4) and SPI9 to be secreted by type 1 secretion systems (T1SSs) encoded at the same loci.
T368 573825-573949 Sentence denotes SiiE of the SPI4 is necessary for adhesion to polarized epithelial cells whereas BapA was associated with biofilm formation.
T369 573950-574081 Sentence denotes To overexpress BapA, its putative promoter was substituted with the SPI4 promoter by red recombination in a SPI4 background strain.
T370 574082-574211 Sentence denotes Mutant and luciferase reporter strains of the bap operon as well as csgDBA mutant strains were also created by red recombination.
T371 574212-574270 Sentence denotes All strains used were derived from NCTC 12023 (WT) strain.
T372 574271-574372 Sentence denotes Western Blot (WB) and immunostaining were performed by using BapA kindly provided by Dr. Lasa, Spain.
T373 574373-574532 Sentence denotes Psii promoter exchange clones, SPI4, bapA and bapD and WT strains were incubated either with rigorous shaking or in static conditions for 72 h at 37° or 30° C.
T374 574533-574722 Sentence denotes Bacterial cells and supernatant aliquots were taken from 2 to 72 h at several time intervals for WB analysis and expression kinetics, which were performed using the luciferase reporter gen.
T375 574723-574818 Sentence denotes Static cultures of BapA overexpressing strains incubated at 37° C developed biofilm after 72 h.
T376 574819-575053 Sentence denotes Further results from WB analyses revealed a band with the theoretical molecular weight of BapA in bacterial pellets after the first 24 h; then, as in bacterial pellets as in supernatants at 48 h; and finally, only supernatant at 72 h.
T377 575054-575284 Sentence denotes Expression kinetics were contrary to the observed phenotypes obtained in WB with the WT strain, since expression of the bapA or bapD in static conditions was around 10 fold higher at 30° C than at 37° C after 8 h in WT background.
T378 575285-575392 Sentence denotes One could conclude that BapA transcript underground degradation so that no signal can be observed using WB.
T379 575393-575476 Sentence denotes Finally, images from CLSM from bacterial biofilm revealed several cluster patterns.
T380 575477-575637 Sentence denotes In comparison to WT strain, bapA strains formed very large clusters but BapA overexpressing strains were found as small groups of bacteria containing 3 7 cells.
T381 575638-575730 Sentence denotes On the other hand, single isolated bacterial cells were only observed using the csgD strain.
T382 575731-575821 Sentence denotes Altogether, these results showed that BapA may play a role in the surface of the bacteria.
T383 575822-575946 Sentence denotes Its secretion pattern depending on the time and medium conditions could suggest a protective function to the bacterial cell.
T384 575947-576109 Sentence denotes Further analysis of regulation of the expression and its possible role of invasion/adhesion is currently in progress to understand the role of BapA in host cells.
T1 576110-576230 Sentence denotes Helicobacter pylori HtrA is a new secreted virulence factor that cleaves E-cadherin to disrupt intercellular adhesion B.
T2 576231-576244 Sentence denotes Hoy* 1,6 , M.
T3 576245-576257 Sentence denotes Löwer 2 , C.
T4 576258-576271 Sentence denotes Weydig 1 , G.
T5 576272-576286 Sentence denotes Carra 1,6 , N.
T6 576287-576303 Sentence denotes Tegtmeyer 3 , T.
T7 576304-576335 Sentence denotes Geppert 4 , P.r Schröder 5 , N.
T8 576336-576349 Sentence denotes Sewald 5 , S.
T9 576350-576364 Sentence denotes Backert 3 , G.
T10 576365-576592 Sentence denotes Schneider 2, 4 Helicobacter pylori (Hp) is a gram-negative bacterium that colonizes the gastric epithelium of approximately 50% of world`s population, leading to the development of inflammatory diseases and even gastric cancer.
T11 576593-576696 Sentence denotes To promote full virulence, Hp disrupts the gastric epithelial barrier to enter the intercellular space.
T12 576697-576808 Sentence denotes However, the underlying mechanism of Hp-induced epithelial barrier subversion is currently not well understood.
T13 576809-577046 Sentence denotes Here, we report an entirely new function of the serine protease and chaperone HtrA as a new secreted virulence factor from Hp, which cleaves the ectodomain of E-cadherin, an important molecule in cell-cell adhesions and tumor suppressor.
T14 577047-577194 Sentence denotes Upon infection, we detected the extracellular E-cadherin domain as a soluble fragment in the supernatants of Hp-colonized gastric epithelial cells.
T15 577195-577292 Sentence denotes Correspondingly the amount of full length E-cadherin in cellular lysates was drastically reduced.
T16 577293-577412 Sentence denotes Using pharmacological inhibitors and RNAi we excluded the involvement of host cell proteases in E-cadherin degradation.
T17 577413-577504 Sentence denotes However, we identified HtrA as a secreted bacterial factor exhibiting proteolytic activity.
T18 577505-577695 Sentence denotes In vitro studies using recombinant E-cadherin revealed that active HtrA specifically cleaved the N-terminal ectodomain of E-Cadherin, whereas an inactive, point-mutated HtrA variant did not.
T19 577696-577858 Sentence denotes Most importantly, using a computer assisted homology model of the E. coli HtrA homologue DegP we designed and synthesized the small-molecule HpHtrA inhibitor HHI.
T20 577859-578102 Sentence denotes HHI efficiently blocked HtrA activity in E-cadherin cleavage experiments and on Hp-colonized epithelial cells, thereby significantly reducing intercellular invasion of Hp through epithelial monolayers and without affecting bacterial viability.
T21 578103-578295 Sentence denotes Taken together, these data imply, that HtrA mediated E-Cadherin shedding during Hp infection disrupts epithelial barrier functions thus allowing the bacteria to access the intercellular space.
T22 578296-578413 Sentence denotes We assume that blocking of HtrA activity might help combat Hp persistence in the early phases of bacterial infection.
T23 578414-578427 Sentence denotes Introduction:
T24 578428-578553 Sentence denotes Oxidative stress converts methionine to a mixture of methionine-S-sulfoxide (Met-S-SO) and methionine-R-sulfoxide (Met-R-SO).
T25 578554-578636 Sentence denotes Methionine sulfoxide reductases protect bacteria by repairing oxidized methionine.
T26 578637-578751 Sentence denotes MsrA and MsrB reduce MetSO, the former being specific for the S-form and the latter being specific for the R-form.
T27 578752-578849 Sentence denotes In this study we characterize MsrA, MsrB and a third methionine sulfoxide reductase, fRMsr, in S.
T28 578850-578939 Sentence denotes Typhimurium. fRMsr has been previously shown to be specific for free Met-R-SO in E. coli.
T29 578940-578948 Sentence denotes Methods:
T30 578949-579021 Sentence denotes Methods used for this study are the generation of deletion mutants in S.
T31 579022-579274 Sentence denotes Typhimurium by one-step inactivation via homologous recombination and the transduction with the bacteriophage P22int, H2O2 susceptibility assays, infection experiments in the macrophage cell line RAW264.7, competitive infections of Balb/cJ mice with S.
T32 579275-579398 Sentence denotes Typhimurium, growth experiments of Salmonella on methionine and methionine sulfoxide, overexpression and purification of S.
T33 579399-579486 Sentence denotes Typhimurium His-tagged MsrA and MsrB, NADPH linked methionine reductase activity assay.
T34 579487-579495 Sentence denotes Results:
T35 579496-579536 Sentence denotes Here we show that deletion of msrA in S.
T36 579537-579667 Sentence denotes Typhimurium increased susceptibility to exogenous H2O2 and reduced bacterial replication inside activated macrophages and in mice.
T37 579668-579726 Sentence denotes In contrast, a msrB mutant showed the wild type phenotype.
T38 579727-579816 Sentence denotes We constructed fRMsr and msrB mutant strains in a methionine auxotrophic background of S.
T39 579817-579829 Sentence denotes Typhimurium.
T40 579830-579914 Sentence denotes The fRMsr mutant but not the msrB mutant failed to utilize free oxidized methionine.
T41 579915-580067 Sentence denotes Recombinant MsrA was active against both free and peptidyl Met-S-SO, whereas recombinant MsrB was only weakly active and specific for peptidyl Met-R-SO.
T42 580068-580235 Sentence denotes To examine the role of fRMsr in oxidative stress response we compared a fRMsr single mutant and a fRMsr/msrB double mutant, and found that fRMsr affects survival of S.
T43 580236-580310 Sentence denotes Typhimurium following exposure to H2O2, growth in macrophages and in mice.
T44 580311-580322 Sentence denotes Conclusion:
T45 580323-580368 Sentence denotes Thus in summary, we showed that mutants of S.
T46 580369-580533 Sentence denotes Typhimurium lacking components of the methionine sulfoxide reductase pathway are attenuated in vitro when exposed to H2O2, inside activated macrophages and in mice.
T47 580534-580668 Sentence denotes Previously, MsrA and MsrB were considered to be the principle enzymes of the msr-system that play a role in oxidative stress response.
T48 580669-580790 Sentence denotes Here we showed that in addition fRMsr contributes significantly to antagonize the damage caused by oxidative stress in S.
T49 580791-580803 Sentence denotes Typhimurium.
T50 580804-581043 Sentence denotes Upon Salmonella enterica serovar Typhimurium infection of the gut, an early line of defense is the gastrointestinal epithelium which senses the pathogen and intrusion along the epithelial barrier is one of the first events towards disease.
T51 581044-581393 Sentence denotes Recently, we showed that high intracellular amounts of the secondary messenger c-di-GMP in S. typhimurium abolished stimulation of a pro-inflammatory immune response and inhibition of invasion of the gastrointestinal epithelial cell line HT-29 suggesting regulation of transition between biofilm formation and virulence by c-di-GMP in the intestine.
T52 581394-581763 Sentence denotes Here we show that highly complex c-di-GMP signaling networks consisting of distinct groups of c-di-GMP synthesizing and degrading proteins modulate the virulence phenotypes IL-8 production, invasion and in vivo colonization in the streptomycin-treated mouse model implying a spatial and timely modulation of virulence properties in S. typhimurium by c-di-GMP signaling.
T53 581764-581913 Sentence denotes Inhibition of the invasion phenotype by c-di-GMP is associated with inhibition of secretion of the type three secretion system effector protein SipA.
T54 581914-582099 Sentence denotes Inhibition of the invasion and IL-8 phenotype by cdi-GMP (partially) requires the major biofilm activator CsgD and/or BcsA the synthase for the extracellular matrix component cellulose.
T55 582100-582286 Sentence denotes Our findings show that c-di-GMP signaling is at least equally important in the regulation of Salmonella-host interaction as in the regulation of biofilm formation at ambient temperature.
T56 582287-582289 Sentence denotes A.
T57 582290-582305 Sentence denotes Lüttich* 1 , S.
T58 582306-582321 Sentence denotes Brunke 1,2 , I.
T59 582322-582450 Sentence denotes Jacobsen 1 Candida albicans is both a successful commensal and a pathogen of humans that can infect a broad range of body sites.
T60 582451-582606 Sentence denotes During the infection process, C. albicans has the potential to adapt to different host niches and to survive the attack of phagocytes, such as macrophages.
T61 582607-582691 Sentence denotes Previous evidence suggests that microevolution plays a central role in this process.
T62 582692-582892 Sentence denotes For example, Forche et al. [1] found that C. albicans rapidly reacts to its environment by undergoing chromosome-level genetic variations, which are sufficient to generate new variants of C. albicans.
T63 582893-583060 Sentence denotes In this study, we used an in vitro experimental microevolution approach to identify factors and activities necessary for survival within, and escape from, macrophages.
T64 583061-583199 Sentence denotes Using this system, we also investigated whether C. albicans can utilise alternative signal transduction pathways under selective pressure.
T65 583200-583311 Sentence denotes First, we analysed a range of mutants with defects in hyphal-formation for capacity to escape from macrophages.
T66 583312-583408 Sentence denotes Normally, four hours post-infection, more than 50 % of wild type cells escaped from macrophages.
T67 583409-583522 Sentence denotes Although none of the tested mutants achieved such high escape rates, many were capable of lower levels of escape.
T68 583523-583600 Sentence denotes An efg1/cph1 double mutant, on the other hand, was entirely unable to escape.
T69 583601-583741 Sentence denotes We therefore chose this strain for further investigation and monitored its adaptation to macrophages during a series of co-culture passages.
T70 583742-583848 Sentence denotes After 23 passages, the efg1/cph1 mutant began to exhibit dramatic phenotypic changes and formed filaments.
T71 583849-583919 Sentence denotes These filaments enabled the evolved mutant to escape from macrophages.
T72 583920-584107 Sentence denotes Surprisingly, the strain was also now able to form filaments under other conditions and, in contrast to the original efg1/cph1 mutant, has regained the ability to invade epithelial cells.
T73 584108-584196 Sentence denotes Furthermore, it has significantly higher virulence in an alternative model of infection.
T74 584197-584363 Sentence denotes The phenotype was stable in the absence of selective pressure, indicating that microevolutionary events have bypassed the reliance on Cph1 and Efg1 for filamentation.
T75 584364-584459 Sentence denotes QRT-PCR, PFGE and high-throughput sequencing did not indicate major chromosomal rearrangements.
T76 584460-584668 Sentence denotes These and further investigations of the genetic cause of the adaptation will yield insights into the genomic and genetic plasticity of this important fungal pathogen during adaptation to specific host niches.
T77 584669-584671 Sentence denotes A.
T78 584672-584687 Sentence denotes Koenigs* 1 , C.
T79 584688-584708 Sentence denotes Hammerschmidt 1 , D.
T80 584709-584730 Sentence denotes Barthel Introduction:
T81 584731-584896 Sentence denotes Borrelia burgdorferi, the etiological agent of Lyme disease exploits an array of strategies in order to overcome the host immune response and establish an infection.
T82 584897-585057 Sentence denotes Those strategies involve interactions of spirochetes with diverse host proteins, including plasminogen, extracellular matrix proteins and complement regulators.
T83 585058-585232 Sentence denotes A number of genetically unrelated outer surface proteins, i.e. OspA, OspC, CspA, ErpA, ErpC, and ErpP have previously been identified as ligands for host-derived plasminogen.
T84 585233-585321 Sentence denotes Here we report the identification of a novel Borrelia plasminogen binding protein, BopA.
T85 585322-585343 Sentence denotes Material and Methods:
T86 585344-585406 Sentence denotes Binding of plasminogen to BopA was first assessed using ELISA.
T87 585407-585655 Sentence denotes To verify whether bound plasminogen/plasmin is proteolytically active, we performed substrate degradation assays utilizing both a chromogenic substrate, D-valyl-leucyl-lysine-pnitroanilidedihydrocholride as well as the natural substrate fibrinogen.
T88 585656-585839 Sentence denotes For further characterization of the BopA-plasminogen interacting region we created a number of truncated versions of BopA and investigated binding with various plasminogen constructs.
T89 585840-585909 Sentence denotes Finally, the bopA gene expression was assessed using RT-PCR analysis.
T90 585910-585918 Sentence denotes Results:
T91 585919-586024 Sentence denotes When compared to other borrelial proteins, BopA displayed the strongest binding capacity for plasminogen.
T92 586025-586294 Sentence denotes Bound plasminogen was converted to proteolytically active plasmin in the presence of urokinase type plasminogen activator and subsequently cleaved the chromogenic substrate D-valyl-leucyl-lysine-p-nitroanilidedihydrochloride as well as the natural substrate fibrinogen.
T93 586295-586422 Sentence denotes Employing diverse truncated borrelial proteins, the plasminogen interacting site has been mapped within the C-terminus of BopA.
T94 586423-586625 Sentence denotes Lysine residues located within this particular -helical C-terminal domain appeared to be crucial for binding plasminogen, as the lysine analog tranexamic acid significantly inhibited these interactions.
T95 586626-586816 Sentence denotes By utilizing several plasminogen constructs containing single or multiple Kringle domains, it was determined that the Kringle domain 4 most likely represents the major BopA interaction site.
T96 586817-586921 Sentence denotes Finally, RT-PCR analysis revealed that in vitro cultivated spirochetes expressed the BopA encoding gene.
T97 586922-586933 Sentence denotes Discussion:
T98 586934-587061 Sentence denotes Here we present data on the identification and characterization of a novel plasminogen binding protein, BopA of B. burgdorferi.
T99 587062-587172 Sentence denotes BopA binds host plasminogen and upon activation surface-bound plasmin proteolytically cleaved host fibrinogen.
T100 587173-587328 Sentence denotes Binding of plasminogen/plasmin might endow B. burgdorferi with the capability to resist opsonization and to degrade components of the extracellular matrix.
T101 587329-587549 Sentence denotes Furthermore, interaction with the plasminogen activation system represents an important virulence factor and enables spirochaetes to successfully establish an infection and disseminate more efficiently in the human host.
T102 587550-587714 Sentence denotes In vitro infection studies are usually based on cell culture monolayers (two dimensional systems), which are infected with the respective pathogenic microorganisms.
T103 587715-587838 Sentence denotes A limitation of this model is the restricted dimensionality which contrasts three dimensional (3D) host tissue environment.
T104 587839-588118 Sentence denotes Using a 3D collagen gel setup (3D-CoG), it has been shown that Yersinia enterocolitica as a prototype of an extracellularly replicating pathogen exhibits unusual growth behaviour in this system, resembling in vivo microcolony formation in infected tissues (Freund et al., 2008) .
T105 588119-588275 Sentence denotes Here we present first results of the growth behaviour of Staphylococcus aureus and its interaction with neutrophils in this model using confocal microscopy.
T106 588276-588398 Sentence denotes S. aureus is known to multiply extracellularly in tissue, terminating in abscess formation due to neutrophil infiltration.
T107 588399-588506 Sentence denotes In a first approach we compared the growth behavior of strain Newman and the derived mutants saeRS and ica.
T108 588507-588658 Sentence denotes Surprisingly, we found that strain Newman and both mutants formed bacterial clusters similar to those observed in cell culture medium without collagen.
T109 588659-588769 Sentence denotes In a second approach we supplemented the 3D-CoG with defined plasma proteins to create tissue-like conditions.
T110 588770-588928 Sentence denotes Strikingly, strain Newman and the ica mutant formed microcolonies of uniform size which were surrounded by concentric structures of different optical density.
T111 588929-589007 Sentence denotes In contrast, the saeRS mutant formed microcolonies devoid of these structures.
T112 589008-589097 Sentence denotes Finally, we studied the interaction of murine neutrophils with S. aureus grown in 3D-CoG.
T113 589098-589197 Sentence denotes In 3D-CoG alone neutrophils migrated towards S. aureus microcolonies and phagocytosed the microbes.
T114 589198-589383 Sentence denotes In contrast, the concentric structures formed in 3D-CoG supplemented with plasma proteins were able to prevent neutrophils from infiltration and phagocytosis of S. aureus microcolonies.
T115 589384-589560 Sentence denotes By including further mutants in our study we were able to identify several S. aureus pathogenicity factors which are involved in S. aureus protection against neutrophil attack.
T116 589561-589636 Sentence denotes Pathogen vacuole proteomics of Legionella pneumophilainfected phagocytes C.
T117 589637-589653 Sentence denotes Hoffmann* 1 , I.
T118 589654-589667 Sentence denotes Finsel 1 , D.
T119 589668-589681 Sentence denotes Becher 2 , M.
T120 589682-589695 Sentence denotes Hecker 2 , H.
T121 589696-589724 Sentence denotes Hilbi München, Deutschland 2
T122 589726-589854 Sentence denotes Legionella pneumophila is an opportunistic human pathogen and the causative agent of the severe pneumonia Legionnaires´ disease.
T123 589855-590030 Sentence denotes Upon internalization of the Gram-negative bacterium into phagocytic cells, a unique "Legionella-containing vacuole" (LCV) is formed, the prerequisite for intracellular growth.
T124 590031-590188 Sentence denotes Fusion with lysosomes is avoided, but LCVs acquire endosomal markers and associate with mitochondria, early secretory vesicles and the endoplasmic reticulum.
T125 590189-590417 Sentence denotes Internalization, LCV formation and intracellular growth of L. pneumophila are dependent on the bacterial Icm/Dot type IV secretion system (T4SS), which translocates more than 250 different "effector" proteins into the host cell.
T126 590418-590640 Sentence denotes The identification of the biochemical and cellular function of some of these Icm/Dot substrates is challenging, as the deletion of single or multiple effectors in many cases does not show an intracellular growth phenotype.
T127 590641-590773 Sentence denotes We established a simple and fast method to isolate and purify intact LCVs from the social soil amoeba Dictyostelium discoideum (1) .
T128 590774-591059 Sentence denotes The straightforward two-step protocol includes enrichment of LCVs by immunomagnetic separation using an antibody against an effector that selectively binds to phosphatidylinositol-4-phosphate exclusively on LCVs, followed by density centrifugation through a Histodenz gradient cushion.
T129 591060-591172 Sentence denotes The proteome of the isolated intact LCVs was analysed by LC-MS/MS and revealed more than 500 host cell proteins.
T130 591173-591325 Sentence denotes Recently, we successfully adapted this protocol to isolate intact LCVs from macrophages, allowing comparative proteomic studies of different host cells.
T131 591326-591515 Sentence denotes First results indicate a high overlap of identified host proteins, yet also reveal a number of small GTPases and other host proteins that previously have not been identified as LCV markers.
T132 591516-591673 Sentence denotes We currently confirm the relevance for LCV formation of novel host cell proteins on the LCV by fluorescence microscopy, pulldown assays and functional tests.
T133 591674-591897 Sentence denotes The opportunistic pathogens Legionella pneumophila and Legionella longbeachae employ a conserved mechanism to replicate in amoebae and macrophages within a unique compartment called the "Legionellacontaining vacuole" (LCV).
T134 591898-592072 Sentence denotes Formation of LCVs requires the bacterial Icm/Dot type IV secretion system which, for L. pneumophila, translocates more than 250 "effector proteins" into the target host cell.
T135 592073-592294 Sentence denotes L. pneumophila SidM and SidC are effector proteins that anchor to the phosphoinositide (PI) lipid PI(4)P on the cytosolic face of LCVs, where they interfere with host cell vesicle trafficking and signal transduction (1) .
T136 592295-592424 Sentence denotes Pulldown experiments with PI lipids coupled to agarose beads revealed that L. longbeachae SidC also specifically binds to PI(4)P.
T137 592425-592539 Sentence denotes L. pneumophila modifies the PI pattern of LCVs in an Icm/Dot dependent manner, yet the mechanism remained elusive.
T138 592540-592718 Sentence denotes We recently discovered a bacterial PI phosphatase, which is translocated into the host via the Icm/Dot T4SS and preferentially hydrolyses poly-phosphorylated PIs yielding PI(4)P.
T139 592719-592839 Sentence denotes This PI phosphatase, termed LppA, might shape the LCV PI pattern to promote binding of L. pneumophila effector proteins.
T140 592840-592985 Sentence denotes The genetically tractable social amoeba Dictyostelium discoideum has been used in a number of studies to analyze LCV formation of L. pneumophila.
T141 592986-593159 Sentence denotes The PI 5-phosphatase OCRL1 and its Dictyostelium homologue Dd5P4 localize to LCVs, restrict intracellular bacterial growth and are implicated in retrograde trafficking (1) .
T142 593160-593271 Sentence denotes Similarly, intracellular replication of L. longbeachae was found to be enhanced in D. discoideum lacking Dd5P4.
T143 593272-593408 Sentence denotes Current efforts aim at a detailed characterization of LCV formation and intracellular replication of L. longbeachae using D. discoideum.
T144 593409-593506 Sentence denotes The epidermal growth factor receptor is a cellular target for the C. pneumoniae adhesion Pmp21 K.
T145 593507-593525 Sentence denotes Mölleken* 1 , J.H.
T146 593526-593631 Sentence denotes Hegemann Funktionelle Genomforschung, Düsseldorf, Deutschland Chlamydiae are significant human pathogens.
T147 593632-593793 Sentence denotes C. pneumoniae is an important respiratory pathogen associated with 5 to 10 % of communityacquired cases of pneumonia worldwide and a seroprevalence of up to 70%.
T148 593794-593948 Sentence denotes Our group is concentrating on the first and initial step of the infection, the adhesion to and subsequent internalization of chlamydiae by the human cell.
T149 593949-594025 Sentence denotes Recently we characterized the first chlamydial adhesin -human receptor pair.
T150 594026-594282 Sentence denotes We could demonstrate that glycosaminoglycans (GAGs) on host cell surfaces are involved in attachment and subsequent infection by C. pneumoniae and that the bacterial surface-associated OmcB protein is the GAG-binding adhesin (Moelleken and Hegemann 2008) .
T151 594283-594446 Sentence denotes Using our Yeast Surface Display system in combination with a functional adhesion test procedure we subsequently identified as a second C. pneumoniae adhesin Pmp21.
T152 594447-594565 Sentence denotes In adhesion studies, using purified recombinant Pmp21 protein, we were able to show that it could bind to human cells.
T153 594566-594742 Sentence denotes By generating deletion variants and single amino acid mutations within the protein, we mapped the adhesion properties to short repetitive amino acid motives GGA(I, V) and FXXN.
T154 594743-594958 Sentence denotes Inhibition of the interaction between Pmp21 and the host cell by an anti-Pmp21 antibody or soluble recombinant Pmp21 protein let to a reduced infection as well as a reduced binding of the bacteria to the human cell.
T155 594959-595076 Sentence denotes In a biochemical approach we tried to identify a human cellular interaction partner for the chlamydial Pmp21 protein.
T156 595077-595272 Sentence denotes We generated a recombinant Pmp21 protein fragment that is analogous to a natural processed fragment found on EBs and used this protein to isolate interaction partners from the human cell surface.
T157 595273-595390 Sentence denotes Through mass spectrometry we identified the epidermal growth factor receptor (EGFR1) as a potential target for Pmp21.
T158 595391-595580 Sentence denotes Depletion of the EGF receptor by siRNA knock down showed a 60% decreased infection with C. pneumoniae in comparison to non-transfected cells or cells transfected with a non targeting siRNA.
T159 595581-595753 Sentence denotes Most importantly a knockdown of EGFR resulted in a diminished binding of fluorescently labeled bacteria, indicating that EGFR indeed serves as a receptor for C. pneumoniae.
T160 595754-595909 Sentence denotes Vice versa we could enhance the infection approximately by the factor two, when we overexpressed EGFR in cells deficient for this receptor tyrosine kinase.
T161 595910-596053 Sentence denotes Deletion of the ligand-binding domain within the receptor significantly reduced the infection due to a reduced internalization of the bacteria.
T162 596054-596191 Sentence denotes These findings imply that C. pneumoniae uses EGFR as a receptor for infection with Pmp21 on the bacterial surface as interaction partner.
T163 596192-596246 Sentence denotes The work was supported by the DFG (FOR729, project 3).
T164 596247-596359 Sentence denotes Reactive oxygen species (ROS) are conventionally considered to be part of the immune response to kill pathogens.
T165 596360-596497 Sentence denotes The obligate intracellular bacteria, Chlamydia trachomatis (C. trachomatis), however evolved mechanisms to use ROS for its own advantage.
T166 596498-596616 Sentence denotes C. trachomatis-induced, increased ROS levels activate caspase-1 by an inflammasome protein, NLRP3 dependent mechanism.
T167 596617-596717 Sentence denotes Proteolytic activity of capase-1 is required for chlamydial inclusion formation in epithelial cells.
T168 596718-596905 Sentence denotes Despite the positive effect of ROS on chlamydial development, it is incompletely understood how different cellular conditions that lead to altered ROS production affect chlamydial growth.
T169 596906-597064 Sentence denotes Therefore we aimed to decipher the molecular mechanisms underlying the beneficial effect of ROS on chlamydial inclusion formation and inflammasome activation.
T170 597065-597158 Sentence denotes Host cell mitochondrial and cytoplasmic ROS formations were modified by different approaches.
T171 597159-597345 Sentence denotes Hypoxia and inhibition of mitochondrial complex III by Antimycin A that increased the concentrations of mitochondrial ROS supported and decreased C. trachomatis development respectively.
T172 597346-597501 Sentence denotes Induction of cytoplasmic ROS by addition of small amount of exogenous tert-butyl hydrogen peroxide or cobalt-chloride increased C. trachomatis development.
T173 597502-597675 Sentence denotes The role of cytoplasmic NADPH that is essential for regeneration of host cell oxidative defense system was studied in cells permanently overexpressing the NAD-kinase enzyme.
T174 597676-597941 Sentence denotes Consequent host cell signaling changes were followed by detection of hypoxia inducible factor, HIF-1 stabilization and phosphorylation of the acetyl-coenzym Acarboxylase (ACC) enzyme that might mediate the host cell metabolic changes that support chlamydial growth.
T175 597942-598150 Sentence denotes Fluorescence lifetime imaging of the metabolic coenzyme, NAD(P)H by two-photon microscopy was used to precisely characterize the effect of different sources of ROS on the metabolic activity of C. trachomatis.
T176 598151-598307 Sentence denotes Changes in chlamydial metabolism were detected by measuring the relative amount of protein-bound NAD(P)H and its fluorescence lifetime inside the inclusion.
T177 598308-598454 Sentence denotes We also observed the trafficking of Golgi and lysosomes into close proximity to the nucleus in C. trachomatis-infected cells by live cell imaging.
T178 598455-598589 Sentence denotes This organelle trafficking might escort the perinuclear clustering of inflammasomes that is required for their ROS-induced activation.
T179 598590-598732 Sentence denotes Our studies about the role of ROS in chlamydial development contribute to the understanding of the fundamentals of host-pathogen interactions.
T180 598733-598942 Sentence denotes Chlamydial growth and progeny formation showed different regulation depending on the concentrations and origin of ROS generated in the host cell and on the consequent HIF-1 stabilization and metabolic changes.
T181 598943-598945 Sentence denotes B.
T182 598946-598960 Sentence denotes Prusty* 1 , L.
T183 598961-598973 Sentence denotes Böhme 1 , T.
T184 598974-599071 Sentence denotes Rudel 1 1 University Of Würzburg, Department of Microbiology, Würzburg, Deutschland Introduction:
T185 599072-599300 Sentence denotes Both human herpes virus 6 (HHV6) and Chlamydia (C. trachomatis or C. pneumoniae) are detected together in different human disorders including chronic fatigues syndrome (CFS) and autoimmune disorders like multiple sclerosis (MS).
T186 599301-599429 Sentence denotes The possible interaction between these two types of pathogens and its influence on their fate inside host cell is still unknown.
T187 599430-599451 Sentence denotes Material and methods:
T188 599452-599583 Sentence denotes We used an in vitro epithelial cell culture model to study the cross talk between HHV6 and C. trachomatis during their coinfection.
T189 599584-599592 Sentence denotes Results:
T190 599593-599682 Sentence denotes Under in vitro growth conditions acute HHV6 infection reduced the Chlamydial infectivity.
T191 599683-599804 Sentence denotes On contrary, the bacterial replication and infectivity was strongly up regulated in latent HHV6 containing HeLa229 cells.
T192 599805-599879 Sentence denotes Interestingly, C. trachomatis favored enhanced entry and survival of HHV6.
T193 599880-600052 Sentence denotes In a co-infection model, both pathogens affected each other's entry, survival as well as infectivity and influenced the replication and transcription pattern of each other.
T194 600053-600064 Sentence denotes Conclusion:
T195 600065-600174 Sentence denotes Our findings strongly argue for an increased biological significance of co-infection of both these pathogens.
T196 600175-600376 Sentence denotes Transposon mutants of Mycoplasma agalactiae: exploiting a modified QTD PCR methodology as a novel strategy for sequencing Tn insertion sites and its optimization for selective quantitative screening S.
T197 600377-600390 Sentence denotes Hegde* 1 , M.
T198 600391-600408 Sentence denotes Zimmermann 1 , R.
T199 600409-600427 Sentence denotes Rosengarten 1 , R.
T200 600428-600461 Sentence denotes Chopra-Dewasthaly* 1 1 IBMH, Vet.
T201 600462-600466 Sentence denotes Med.
T202 600467-600471 Sentence denotes Uni.
T203 600472-600608 Sentence denotes Vienna, Department of pathobiology, Vienna, Oesterreich As mycoplasmas are poorly understood pathogens, they are difficult to eradicate.
T204 600609-600825 Sentence denotes Despite considerable advances in the area of mycoplasma molecular genetics in the past decade, progress towards understanding their pathogenicity mechanisms is hindered by the paucity of available genetic tools (1) .
T205 600826-601045 Sentence denotes Mycoplasma agalactiae (MA) is an important ruminant pathogen for which the pathogenicity mechanisms are largely unknown, mainly because of the previous incapability to genetically manipulate it until the year 2005 (2) .
T206 601046-601276 Sentence denotes We initiated studies in this direction (2) and have now constructed a transposon (Tn) mutant library with insertions in almost all important categories including hypothetical, lipoprotein, transporter and metabolism related genes.
T207 601277-601604 Sentence denotes For the functional analysis of MA genes, the Tn mutants were subjected to several in vitro screening protocols, especially to those which could have a direct role in its pathogenicity, such as biofilm formation, growth, adherence and antibiotic sensitivity, in order to shortlist them for an experimental sheep infection study.
T208 601605-601821 Sentence denotes This resulted in the identification of many important mutants in known and hypothetical genes displaying significant phenotypes, such as pdhB, uhpT and MAG0110 in growth and many lipoprotein genes in adhesion assays.
T209 601822-602002 Sentence denotes Although reported for some mycoplasmas, direct genomic sequencing and other methods to sequence the Tn insertion sites are not always successful, or are rather costly or laborious.
T210 602003-602127 Sentence denotes We have exploited and developed a unique PCR strategy that is an integral part of the QTD method (3) for sequencing mutants.
T211 602128-602338 Sentence denotes This involves selective and specific amplification of the Tninsertion sites using the same pair of primers for all mutants in a ligationmediated PCR, and offers an easy and fast alternative sequencing strategy.
T212 602339-602648 Sentence denotes Since QTD is a novel, efficient, high-throughput method capable of also identifying quantitative phenotypes during mixed population screening (3), we have adapted it to MA and optimized various parameters to successfully reproduce specific amplification of individual as well as a pool of selected Tn-mutants.
T213 602649-602901 Sentence denotes The study not only highlights the biological significance of some specific mutated genes but also paves the way for the successful application of a novel QTD method in mycoplasmas to facilitate a better understanding of their biology and pathogenicity.
T214 602902-602913 Sentence denotes Background:
T215 602914-603007 Sentence denotes Inflammatory activation of the endothelium leads to a release of von Willebrand factor (VWF).
T216 603008-603158 Sentence denotes Exposed to blood flow, VWF gets activated by a conformational change from a globular protein to stretched mulitmerized ultra large VWF (ULVWF) fibers.
T217 603159-603259 Sentence denotes These ULVWF fibers bind platelets at sites of vascular injury even under high shear rate conditions.
T218 603260-603345 Sentence denotes Moreover, ULVWF is considered as a potential binding partner for pathogenic bacteria.
T219 603346-603471 Sentence denotes It is worth to emphasize that ULVWF fibers can be formed on an intact endothelial cell layer, independent of vascular damage.
T220 603472-603573 Sentence denotes Previous studies indicate that Staphylococcus aureus binds to VWF via staphylococcal protein A (SpA).
T221 603574-603633 Sentence denotes However, the influence of ULVWF fibers is not known so far.
T222 603634-603755 Sentence denotes Therefore, the present study focused on the impact of ULVWF for the adhesion of S. aureus to an intact endothelial layer.
T223 603756-603764 Sentence denotes Methods:
T224 603765-603861 Sentence denotes To simulate blood flow conditions at different shear rates we used various microfluidic devices.
T225 603862-604044 Sentence denotes Human umbilical vein endothelial cells (HUVECs) were perfused with fluorescence-labelled S. aureus (Cowan I) enabling quantification of bacterial adhesion by fluorescence microscopy.
T226 604045-604204 Sentence denotes To investigate the role of SpA, we compared the adhesion of S. aureus wild type to SpA-deficient isogenic mutant and to the non pathogenic S. carnosus (TM300).
T227 604205-604442 Sentence denotes To prove whether the binding of S. aureus to the endothelium is specifically mediated by ULVWF we used in some experiments the specific VWF degrading protease ADAMTS-13 (a disintegrin and metalloproteinase with thrombospondin motifs 13).
T228 604443-604451 Sentence denotes Results:
T229 604452-604554 Sentence denotes The overall capacity of ULVWF to bind S. aureus was dramatically higher in comparison to globular VWF.
T230 604555-604673 Sentence denotes SpA supported the binding of S. aureus mainly to the globular form of VWF which is in agreement with previous studies.
T231 604674-604755 Sentence denotes By contrast, binding of S. aureus to ULVWF fibers was largely independent of SpA.
T232 604756-604903 Sentence denotes Adhesion of S. carnosus to ULVWF was comparable to S. aureus indicating an adhesion mechanism independent of S. aureus-specific surface structures.
T233 604904-604983 Sentence denotes In addition, fixed and live staphylococci adhered to a similar extent to ULVWF.
T234 604984-605115 Sentence denotes In experiments using ADAMTS-13 an instantaneously cleavage of ULVWF was documented and thereby an inhibition of bacterial adhesion.
T235 605116-605127 Sentence denotes Discussion:
T236 605128-605273 Sentence denotes Within the context of clinical studies a significant contribution of VWF and ADAMTS-13 to vascular disease progression has been shown previously.
T237 605274-605416 Sentence denotes Here, we show for the first time that S. aureus is able to bind to an intact endothelial cell layer under physiological blood flow conditions.
T238 605417-605506 Sentence denotes Binding was mainly due to ULVWF fibers and independent of SpA but dependent of ADAMTS-13.
T239 605507-605609 Sentence denotes Our data suggest an adhesion mechanism independent of secreted factors or surface associated adhesins.
T240 605610-605752 Sentence denotes However, the acting molecular mechanism that triggers strong binding of S. aureus to ULVWF is currently unknown and requires further research.
T241 605753-605869 Sentence denotes Streptococcus pneumoniae induces release of von Willebrand factor from primary human endothelial cells Introduction:
T242 605870-605941 Sentence denotes Streptococcus pneumoniae is colonizing the human nasopharyngeal cavity.
T243 605942-606075 Sentence denotes Invasive pneumococcal infections cause meningitis, pneumonia and septic dissemination within the vascular system (Cartwright, 2002) .
T244 606076-606202 Sentence denotes The pore-forming cytotoxin pneumolysin is described as a major virulence factor of pneumococci (Mitchell and Mitchell, 2010) .
T245 606203-606371 Sentence denotes Pulmonary microvascular endothelial cells (HPMEC) form the inner lining of blood microvessels and produce specialized storage granules named Weibel-Palade Bodies (WPB).
T246 606372-606598 Sentence denotes WPB contain procoagulant factors like von Willebrand factor (vWF) and proinflammatory interleukin 8 (IL-8) and are released in response to inflammatory or stress-related stimuli (Weibel and Palade 1964; Rondaij et al., 2004) .
T247 606599-606620 Sentence denotes Material and Methods:
T248 606621-606826 Sentence denotes Confluently grown primary HPMEC were cultured on glass cover slips or in two chamber transwell systems and were infected with serotype 35A and serotype 2 pneumococci and with pneumolysin-deficient mutants.
T249 606827-606915 Sentence denotes Bacterial adherence and WPB exocytosis was visualized by immune fluorescence microscopy.
T250 606916-607023 Sentence denotes Cell confluency was controlled by measurement of biological impedance using the CellZscope (Nanoanalytics).
T251 607024-607110 Sentence denotes Quantification of vWF and IL-8 within cell culture supernatant was performed by ELISA.
T252 607111-607119 Sentence denotes Results:
T253 607120-607264 Sentence denotes Microscopic quantification demonstrated that pneumococcal adherence to confluent HPMEC correlates with a decreasing amount of intracellular WPB.
T254 607265-607398 Sentence denotes Following time course of infection with pneumococci, increasing amounts of vWF and IL-8 were determined in cell culture supernatants.
T255 607399-607702 Sentence denotes Release of vWF and of IL8 was slightly lower after infection with pneumolysin-deficient pneumococcal mutants compared to wild-type bacteria, but sublytical amounts of recombinant pneumolysin significantly stimulated WPB exocytosis in similar levels as physiological agonists like histamine and thrombin.
T256 607703-607855 Sentence denotes Moreover, the vWF release was determined in cell culture supernatant after infection of HPMEC from apical and basolateral cell surface in equal amounts.
T257 607856-607867 Sentence denotes Discussion:
T258 607868-608036 Sentence denotes This report describes for the first time an induction of WPB exocytosis by pneumococcal adherence and by the cytotoxin pneumolysin using primary lung endothelial cells.
T259 608037-608302 Sentence denotes The stimulation of vWF and IL8 release after apical and basolateral infection implies a stimulation of WPB exocytosis in both directions: from inside the vasculature and also following invasive pneumococcal transmigration from pulmonary tissue into the bloodstream.
T260 608303-608458 Sentence denotes These data describe a new endothelial-specific response to severe pneumococcal infection and provide new insights into the pathophysiological consequences.
T261 608459-608461 Sentence denotes H.
T262 608462-608476 Sentence denotes Büttner 1 , M.
T263 608477-608493 Sentence denotes Christner 1 , H.
T264 608494-608657 Sentence denotes Rohde* Medizinische Mikrobiologie, Virologie und Hygiene, Hamburg, Deutschland Staphylococcus epidermidis is a leading cause of foreign body associated infections.
T265 608658-608739 Sentence denotes Pathogenicity of the species is mainly related to its biofilm forming capability.
T266 608740-608862 Sentence denotes Biofilm formation crucially depends on bacterial interactions with surface organized host extracellular matrix components.
T267 608863-609003 Sentence denotes Here we analyzed the role of giant 1 MDa extracellular matrix binding protein Embp in S. epidermidis adherence to surface bound fibronectin.
T268 609004-609343 Sentence denotes Analysis of fibronectin interactions with recombinant Embp sub-domains, either comprising Found In Various Architectures-(FIVAR-) domains alone or in combination with G-related albumin binding-(GA-) domains, in ELSA-based assays and by surface plasmon resonance produced evidence that FIVAR domains are sufficient for Embp-Fn interactions.
T269 609344-609812 Sentence denotes In addition, Far Western blotting experiments mapped Embp fibronectin interactions to FnIII 12 -13 domains By making use of Embp expressing S. epidermidis strain 1585v and isogenic, Embp-negative mutant 1585v-M135 the functional relevance of Embp-Fn interactions for S. epidermidis surface colonization was then tested in semi-quantitative adherence assays, showing that Embp-expression is sufficient and necessary for S. epidermidis binding to surface immobilized Fn.
T270 609813-610129 Sentence denotes The importance of Embp for bacterial adherence to fibronectin was further validated by testing surrogated host S. carnosus TM300 carrying a plasmid allowing for expression of a fusion protein comprising FIVAR domains, an N-terminal YSIRK-containing export signal and the putative C-terminal cell wall binding region.
T271 610130-610292 Sentence denotes Whereas wild type S. carnosus exhibited almost no Fnadherence capacity, in trans expression of this construct lead to a significant increase in bacterial binding.
T272 610293-610379 Sentence denotes These results establish the role of Embp as a bona fide bacterial fibronectin adhesin.
T273 610380-610575 Sentence denotes To test if S. epidermidis adherence to fibronectin also involves interactions with FnIII domains 12 -13, adherence of S. epidermidis 1585v to immobilized Fn was tested in the presence of heparin.
T274 610576-610651 Sentence denotes Here, a dose dependent inhibition of S. epidermidis adherence was recorded.
T275 610652-610808 Sentence denotes Therefore, as heparin specifically binds to FnIII 12, this particular domain is obviously crucial for Embp mediated S. epidermidis adherence to fibronectin.
T276 610809-610997 Sentence denotes Our findings establish the role of Embp as an important S. epidermidis adhesin mediating adherence to immobilized fibronectin, thereby contributing to the first phase of biofilm formation.
T277 610998-611110 Sentence denotes The underlying molecular basis of Embp -fibronectin interactions involves a novel, so far undescribed mechanism.
T278 611111-611113 Sentence denotes R.
T279 611114-611128 Sentence denotes Ziobro* 1 , B.
T280 611129-611141 Sentence denotes Henry 1 , E.
T281 611142-611153 Sentence denotes Gulbins 1 1
T282 611155-611281 Sentence denotes Pseudomonas aeruginosa is the leading respiratory pathogen and eventual cause of death for patients with Cystic Fibrosis (CF).
T283 611282-611422 Sentence denotes Ceramide, a bioactive sphingolipid involved in cell signalling, has been shown to assemble along the cell membrane of the airway epithelium.
T284 611423-611533 Sentence denotes Patients with CF have a relative abundance of ceramide enriched domains, termed platforms if of a larger size.
T285 611534-611642 Sentence denotes Accumulation of domains has been directly correlated with patient maturation and vulnerability to infection.
T286 611643-611773 Sentence denotes Increased assemblages of ceramide are a molecular outlier of patients with CF and act as potential stages for aberrant signalling.
T287 611774-611993 Sentence denotes In this study we show that although commonly found along the basal lateral membrane, Beta 1 integrin maintains a permanent position on ceramide enriched domains on the luminal side of airway epithelial cells in CF mice.
T288 611994-612173 Sentence denotes Visualization of ceramide, Beta 1 integrin and P. aeruginosa via immunohistochemistry indicates co-localization at the point of infection on trachea and bronchi epithelial tissue.
T289 612174-612280 Sentence denotes Beta 1 integrin is known to have a RGD binding domain which facilitates binding of various other microbes.
T290 612281-612564 Sentence denotes P. aeruginosa's adherence via its RGD sequence was tested both in vitro by treating Chang cells with and in vivo by inhalation of GRGDN peptide prior to infection; both treatments diminished the virulence of the bacteria as measured by bacterial adherence and mucocilliary clearance.
T291 612565-612770 Sentence denotes Our data indicates that ceramide enriched domains, and the Beta 1 integrin molecules they host, play an important role in the virulence of P. aeruginosa within the airways of patients with Cystic Fibrosis.
T292 612771-612905 Sentence denotes Additionally, we observed that competitive inhibition of an RGD sequence receptor decreased the pathogenicity of the bacteria in vivo.
T293 612906-613169 Sentence denotes These two findings suggest the RGD binding domain of Beta 1 integrin may act as an adherence mechanism for P. aeruginosa and that increased clustering of this receptor in patients with CF may play an important role in the infectious manifestations of the disease.
T294 613170-613269 Sentence denotes Preliminary data indicates inhalation of RGD peptides may be a novel treatment option for patients.
T295 613270-613280 Sentence denotes Figure 1 :
T296 613281-613380 Sentence denotes Confocal microscopy of immunofluorescence staining for Beta 1 Integrin in the bronchial epithelium.
T297 613381-613464 Sentence denotes A shift is observed from basal lateral to apical side of the tissue in CFTR-/-mice.
T298 613465-613664 Sentence denotes Paraffin embeded tissue (A,B,C) and in situ stained paraffin embedded tissue (D,E) The type three secretion system (TTSS) of Yersinia enterocolitica (Ye) is crucial to evade the host immune response.
T299 613665-613807 Sentence denotes After adhesion of the bacteria via YadA or Invasin (Inv) to 1 integrins, the TTSS injects Yersinia outer proteins (Yops) into host cells [1] .
T300 613808-613981 Sentence denotes To dissect the role of YadA, Invasin and 1 integrins for Yop injection a -lactamase reporter system was used to measure Yop injection into leukocytes by flow cytometry [2] .
T301 613982-614234 Sentence denotes In vitro infection of splenic leukocytes with a low infection dose of a Ye YadA Inv strain, revealed that Yop injection into DCs, macrophages and PMNs occurs to some extent and to a much lower extent in B and T cells in a YadA-and Inv-dependent manner.
T302 614235-614347 Sentence denotes Yop injection after infection with a YadA mutant occurred with similar frequency in all tested cell populations.
T303 614348-614471 Sentence denotes After infection of a Inv strain PMNs were targeted for Yop injection with the highest and T cell with the lowest frequency.
T304 614472-614642 Sentence denotes Detection of Yop injection was performed in mouse infection models which resulted in similar bacterial load in the spleen when wildtype and mutant Yersinia were compared.
T305 614643-614891 Sentence denotes After infection with a YadA mutant no Yop injection could be detected while a Inv mutant showed nearly similar Yop injection as a wildtype strain, indicating that YadA but not Inv is crucial for Yop injection into leukocytes during mouse infection.
T306 614892-615065 Sentence denotes To investigate the role of 1 integrins Yop injection into PMNs, macrophages and B cells was investigated using 1 depleted splenocytes derived from conditional knockout mice.
T307 615066-615239 Sentence denotes 1 integrin depletion had no impact on Yop injection by a Inv mutant but reduced significantly Yop injection by a YadA mutant after infection of B cells, PMNs or macrophages.
T308 615240-615351 Sentence denotes Our data provide evidence that during systemic mouse infection YadA but not Inv is essential for Yop injection.
T309 615352-615483 Sentence denotes Inv mediated Yop injection into leukocytes in contrast to YadA mediated Yop injection can occur in a 1 integrin independent manner.
T310 615484-615862 Sentence denotes Trimeric autotransporter adhesin-dependent adherence of Bartonella henselae, Bartonella quintana and Yersinia enterocolitica to matrixcomponents and endothelial cells under static and dynamic flow conditions Trimeric autotransporter adhesins (TAAs) are important virulence factors of Gram-negative bacteria responsible for adherence to extracellular matrix (ECM) and host cells.
T311 615863-616147 Sentence denotes Here, we analyzed three different TAAs [Bartonella adhesin A (BadA) of Bartonella henselae, variably expressed outer membrane proteins (Vomps) of Bartonella quintana, Yersinia adhesin A (YadA) of Yersinia enterocolitica] for mediating bacterial adherence to ECM and endothelial cells.
T312 616148-616327 Sentence denotes Using static (cell culture vials) and dynamic (capillary flow chambers) experimental settings, adherence of wildtype bacteria and the respective TAA-negative strains was analyzed.
T313 616328-616483 Sentence denotes Under static conditions, ECM adherence of B. henselae, B. quintana and Y. enterocolitica was strongly dependent on the expression of their particular TAAs.
T314 616484-616622 Sentence denotes YadA of Y. enterocolitica did not mediate bacterial binding to plasma or cellular fibronectin neither under static nor dynamic conditions.
T315 616623-616797 Sentence denotes TAA-dependent host cell adherence appeared more significant under dynamic conditions although the total number of bound bacteria was diminished compared to static conditions.
T316 616798-617045 Sentence denotes Dynamic models expand the methodology to perform bacterial adherence experiments under more realistic blood-stream like conditions and allow to dissect the biological role of TAAs in ECM and host cell adherence under static and dynamic conditions.
T317 617046-617048 Sentence denotes A.
T318 617049-617071 Sentence denotes Felipe-López* 1 , C.A.
T319 617072-617085 Sentence denotes Danzer 2 , M.
T320 617086-617513 Sentence denotes Hensel 1 1 FB05 Universität Osnabrück, Mikrobiologie, Osnabrück, Deutschland 2 Universitätsklinikum FAU Erlangen Nürnberg, Mikrobiologisches Institut, Erlangen, Deutschland In order to adhere and invade polarized epithelial cells at the intestine, Salmonella injects virulence proteins by the type 3 secretion system-1 (T3SS-1) into the host cell which re-organize the actin cytoskeleton allowing the bacterial internalization.
T321 617514-617680 Sentence denotes Recently, the microvilli effacement of polarized epithelial cells caused by Salmonella was described as consequence of cooperation of the adhesin SiiE and the T3SS-1.
T322 617681-618128 Sentence denotes To investigate the disruption of the brush border of polarized epithelial cells, SL1344 wild-type (WT), sipA, sopA, sopB, sopE, sopE2mutant strains, M712 strain, which lacks the five mentioned effector proteins, and their complemented derivates were used for invasion experiments in MDCK Pf and C2bbe1 cell lines, the later were also used to evaluate the disruption of the trans-epithelial electrical resistance (TEER) by the Salmonella infection.
T323 618129-618329 Sentence denotes Results compared to the WT-strain showed that whereas the single sopE-mutant strain was two third less invasive, one third of the invasiveness was recovered by the single introduction of sopE in M712.
T324 618330-618411 Sentence denotes On the other hand, the M712 + sopE abrogated the TEER similarly to the WT strain.
T325 618412-618538 Sentence denotes No microvilli effacement and small ruffles were observed in actinstained MDCK cells infected by the sopE-single mutant strain.
T326 618539-618626 Sentence denotes Microvilli in these infected cells seemed to be fused to the Salmonella invasion point.
T327 618627-618778 Sentence denotes Further results demonstrated that cells infected by the WT strain presented reticular actin around the entry point at the apical side of the host cell.
T328 618779-619100 Sentence denotes Additionally, M712 + sipA strain intimately anchored to the microvilli; M712 + sopA or + sopB enhanced the adherence compared to the background mutant strain; M712 + sopE2 induced small accumulation of actin; and finally, M712 + sopE effaced the microvilli and also induced reticular actin as observed with the WT strain.
T329 619101-619238 Sentence denotes On the other hand, chain-like structures at the former tight-junction were observed in ZO-1 stained C2bbe1 cells infected by M712 + sopE.
T330 619239-619584 Sentence denotes To follow the invasion process, time lapse imaging using EGFP-LifeAct MDCK cells revealed that bacteria intimately adhered to the microvillus during the first 5 min.; thereafter, microvilli effacement simultaneously occurred with the membrane ruffling from 10 to 15 min.; finally, after 20 min. ruffles were fused promoting bacterial clustering.
T331 619585-619645 Sentence denotes A delayed invasion of sopE-strain was observed after 25 min.
T332 619646-619790 Sentence denotes During this time Salmonella migrated until the microvilli basement, there then induced moderate membrane ruffling without microvilli effacement.
T333 619791-619940 Sentence denotes Altogether, these results showed that microvilli effacement, reticular actin and disruption of tight junctions only depended on the presence of SopE.
T334 619941-620096 Sentence denotes Deletion of other effector proteins also caused changes in the ruffle morphology and spatial distribution of bacteria at the apical side of the host cells.
T335 620097-620210 Sentence denotes Further work is in progress to understand the molecular basis of the microvilli destruction caused by Salmonella.
T336 620211-620213 Sentence denotes M.
T337 620214-620227 Sentence denotes Fulde* 1 , M.
T338 620228-620240 Sentence denotes Rohde 1 , A.
T339 620241-620258 Sentence denotes Hitzmann 1 , K.T.
T340 620259-620277 Sentence denotes Preissner 2 , D.P.
T341 620278-620300 Sentence denotes Nitsche-Schmitz 1 , A.
T342 620301-620319 Sentence denotes Nerlich 1,3 , G.S.
T343 620320-620335 Sentence denotes Chhatwal 1 , S.
T344 620336-620645 Sentence denotes Bergmann 1, 4 1 Helmholtz Zentrum für Infektionsforschung, Medizinische Mikrobiologie, Braunschweig, Biochemie, Giessen, Deutschland 3 Stiftung Tierärztliche Hochschule Hannover, Mikrobiologie, Hannover, Deutschland 4 Technische Universität Braunschweig, Mikrobiologie, Braunschweig, Deutschland Introduction:
T345 620646-620732 Sentence denotes Streptococcus canis is able to cause severe invasive diseases mainly in cats and dogs.
T346 620733-620920 Sentence denotes Increased incidences of streptococcal infections in humans have also been attributed to S. canis and have thus prompted an investigation into the mechanisms underlying their pathogenesis.
T347 620921-620941 Sentence denotes Methods and Results:
T348 620942-621043 Sentence denotes Binding of plasmin(-ogen) is a common virulence trait for establishing invasive bacterial infections.
T349 621044-621144 Sentence denotes Plasma-absorption analysis demonstrated a strong interaction between S. canis and human plasminogen.
T350 621145-621358 Sentence denotes Furthermore, in comparative radioactive binding assays using human or canine plasminogen, no differences in quality of interaction could be observed, a prerequisite for establishing infections across host species.
T351 621359-621486 Sentence denotes Binding of plasminogen depends on the presence of the S. canis M-like protein (SCM) genetically organized in the mga-scm locus.
T352 621487-621639 Sentence denotes Surface plasmon resonance technique revealed dissociation constants in the nanomolar range indicating a specific and biologically important interaction.
T353 621640-621765 Sentence denotes The plasminogen-binding site is located in the N-terminal part of SCM ranging from aminoacid 90 to 112 of the mature protein.
T354 621766-621982 Sentence denotes Furthermore, Western blot analysis using truncated fragments of plasminogen revealed an interaction with the C-terminal part comprising kringle domain 5 and the catalytic domain, also known as miniplasminogen (mPlg).
T355 621983-622088 Sentence denotes To the best of our knowledge, SCM represents the first bacterial receptor directly interacting with mPLG.
T356 622089-622267 Sentence denotes Even though S. canis possess no streptokinase activity, the use of eukaryotic plasminogen activator urokinase enables the bacterium to immobilize plasmin activity on its surface.
T357 622268-622364 Sentence denotes This led to the degradation of purified fibrinogen and of synthetically aggregated fibrin clots.
T358 622365-622599 Sentence denotes Furthermore, electron microscopic illustration and time lapse imaging demonstrated plasmin-mediated transmigration of fibrinous thrombi by S. canis and, therefore, may represent a mechanism for bacterial dissemination within the host.
T359 622600-622611 Sentence denotes Discussion:
T360 622612-622717 Sentence denotes Although S. canis is a zoonotic pathogen, very little is known about its virulence factors or mechanisms.
T361 622718-622881 Sentence denotes Indeed, as described for other streptococcal pathogens (1, 2), S. canis utilizes similar strategies for tissue destruction and bacterial dissemination in the host.
T362 622882-622985 Sentence denotes However, binding of SCM to mPlg presents a mechanistically different bacterial-plasminogen interaction.
T363 622986-623091 Sentence denotes The role of such interactions in the etiopathogenesis of these infections requires further investigation.
T364 623092-623284 Sentence denotes Burkholderia pseudomallei, the etiological agent of melioidosis, and Burkholderia thailandensis are closely related organisms mainly found in the soil in Southeast Asia and Northern Australia.
T365 623285-623489 Sentence denotes The genome of B. pseudomallei encodes six type VI secretion systems (T6SSs) that contain the complete repertoire of T6S core components; orthologs of five of these systems are present in B. thailandensis.
T366 623490-623619 Sentence denotes The type VI secretion system is a widespread and multicomponent apparatus involved in bacterial and eukaryotic cell interactions.
T367 623620-623774 Sentence denotes The Burkholderia T6SSs present in a genome are evolutionarily distinct, leading us to hypothesize that each secretion system performs a discrete function.
T368 623775-623890 Sentence denotes To test this, we inactivated each T6SS individually and investigated their role in virulence and bacterial fitness.
T369 623891-624002 Sentence denotes We found that only one T6SS mutant, T6SS-5, lost virulence in an aerosol delivered murine model of melioidosis.
T370 624003-624177 Sentence denotes This is likely due to a specific role T6SS-5 plays in the subversion of or resistance against MyD88 dependent immune response since T6SS-5 is highly virulent in MyD88 -/mice.
T371 624178-624330 Sentence denotes Next, we screened B. thailandensis T6SS mutants against a diverse array of 31 Gram positive and negative bacterial species in growth competition assays.
T372 624331-624479 Sentence denotes The proliferation and survival of B. thailandensis in the presence of several competitor species requires T6SS-1 in a cell contact dependent manner.
T373 624480-624605 Sentence denotes Intriguingly, two of these species, Pseudomonas fluorescens and Pseudomonas putida, are also soil inhabitants in the tropics.
T374 624606-624718 Sentence denotes In addition, T6SS-1 is essential for the persistence of B. thailandensis in mixed biofilm assays with P. putida.
T375 624719-624816 Sentence denotes Data on T6S substrates are scarce and no Burkholderia T6S effectors have been identified to date.
T376 624817-624926 Sentence denotes To shed light on the mechanism of action of T6SS-5, we aimed at identifying proteins secreted by this system.
T377 624927-625056 Sentence denotes For this, we utilized quantitative Orbitrap mass spectrometry to compare the secretomes of B. thailandensis wild type and T6SS-5.
T378 625057-625126 Sentence denotes Two proteins, Hcp-5 and VgrG-5, were identified as T6SS-5 substrates.
T379 625127-625294 Sentence denotes Hcp-5 and VgrG-5 are extracellular structural components of the T6S apparatus with high structural similarity to phage tail tube and tail spike proteins, respectively.
T380 625295-625450 Sentence denotes Interestingly, VgrG-5 contains a putative C terminal effector domain that does not display significant sequence similarity to proteins with known function.
T381 625451-625624 Sentence denotes We showed that the C terminal domain of VgrG-5 is required for B. thailandensis to induce the formation of multinucleated giant cells in phagocytic and non-phagocytic cells.
T382 625625-625834 Sentence denotes Together the data suggest an important role of Burkholderia T6S in determining the outcome of an infection by directly targeting eukaryotic host cells or by allowing the bacteria to persist in the environment.
T1 625835-626098 Sentence denotes Amino acid metabolism in Streptococcus pneumoniae as analyzed by 13 C-isotopologue profiling Streptococcus pneumoniae (pneumococci) produce a large fraction of ABC transporter systems involved in uptake and metabolism of sugars, soluble compounds and amino acids.
T2 626099-626325 Sentence denotes In this study the carbon metabolism was assessed by 13 C isotopologue perturbation after growth of pneumococci in chemically defined media (CDM) in presence of [U-13 C6]glucose or [1,2-13 C2] glucose as the sole carbon source.
T3 626326-626397 Sentence denotes GC/MS analysis showed a pattern of labeled and non-labeled amino acids.
T4 626398-626566 Sentence denotes Lack of isotopologue labeling was indicated for His, Glu, Pro, Gly and the branched amino acids (BCCA) Ile, Leu, Val, which were all also required as supplement in CDM.
T5 626567-626648 Sentence denotes Asp is not synthesized via the general pathway including pyruvate and acetyl-CoA.
T6 626649-626748 Sentence denotes Instead, oxaloacetate is synthesized from phosphoenolpyruvate (PEP) and CO2 by the PEP carboxylase.
T7 626749-626878 Sentence denotes The oxaloacetate is then converted into Asp, while in a lateral reaction the essential glutamate is converted into 2oxoglutarate.
T8 626879-626939 Sentence denotes These reactions are catalyzed by the aspartate transaminase.
T9 626940-627013 Sentence denotes Furthermore, the shikimate pathway could be determined by GC/MS analysis.
T10 627014-627160 Sentence denotes Complete labeled aromatic amino acids are de novo synthesized from erythrose-4-phosphate and two molecules of PEP via the intermediate chorismate.
T11 627161-627354 Sentence denotes Strikingly, unlabeled Gly [M+0] in [U-13 C6]glucose supplemented CDM indicated the inability of pneumococci to synthesize Ser from complete labeled 3-P-D-glycerate and in a subsequent step Gly.
T12 627355-627492 Sentence denotes In contrast, pneumococci synthesize Ser by glycine hydroxymethylation from 5,10-methylenetetrahydrofolate as part of the One Carbon Pool.
T13 627493-627597 Sentence denotes This pathway was verified by using [U-13 C2]glycine supplemented CDM, resulting in labeled [M+2] serine.
T14 627598-627734 Sentence denotes Finally, the general carbon and amino acid metabolism was also investigated for pneumococcal mutants deficient in PavA, CodY, and G6PDH.
T15 627735-627817 Sentence denotes However, the isotopologue profiles of these mutants were similar to the wild-type.
T16 627818-628046 Sentence denotes In conclusion, these data show for the first time the variety of amino acid synthesis pathway in pneumococci and the essential role of pneumococcal amino acid transporter systems for bacterial fitness und host milieu adaptation.
T17 628047-628238 Sentence denotes The human innate immune system counteracts bacterial invaders by multiple antimicrobial mechanisms including polymorphonuclear leukocytes (PMN), which represent the most efficient phagocytes.
T18 628239-628355 Sentence denotes The human formyl peptide receptor 2 (FPR2) belongs to the family of seven-transmembrane G-protein coupled receptors.
T19 628356-628575 Sentence denotes Recently, we could show that FPR2 is crucial for recruiting and activating PMN in staphylococcal infections, because it senses concentrations of the major staphylococcal cytolysins phenol-soluble modulin (PSM) peptides.
T20 628576-628685 Sentence denotes Moreover, FPR2 adjusts PMN responses with respect to PSM release and pathogenicity of staphylococcal species.
T21 628686-628756 Sentence denotes Enterococci represent another group of important nosocomial pathogens.
T22 628757-628966 Sentence denotes In this study, we show that not only staphylococci but also certain enterococci are capable of producing ligands for FPR2 thereby underscoring the important role of this receptor in antibacterial host defense.
T23 628967-629120 Sentence denotes PMN chemotaxis and intracellular calcium influx were induced in a dosedependent manner by supernatants of Enterococcus faecalis and Enterococcus faecium.
T24 629121-629179 Sentence denotes This effect could be inhibited by the FPR2antagonist WRW4.
T25 629180-629349 Sentence denotes For both, Enterococcus faecalis and Enterococcus faecium vancomycin-resistant isolates elicited considerably stronger PMN responses than vancomycin-susceptible isolates.
T26 629350-629490 Sentence denotes Calcium flux in receptor-transfected HL-60 cells showed that only E. faecium eliciting a FPR2-specific response whereas E. faecalis did not.
T27 629491-629589 Sentence denotes However, both species activated the FPR2 paralog FPR1, which senses bacterial formylated peptides.
T28 629590-629817 Sentence denotes The enterococcal genomes do not encode peptides with apparent similarity to PSM peptides and no peptides with PSM-like properties could be identified in enterococcal culture supernatants in sufficient quantitites for detection.
T29 629818-630095 Sentence denotes To further characterize the unknown FPR2 ligands produced by E. faecium supernatants were treated with proteases, which completely abolished the ability to stimulate FPR2 transfected HL-60 cells thereby indicating that the unknown FPR2 ligands of E. faecium represent peptides.
T30 630096-630259 Sentence denotes In conclusion, we were able to demonstrate that certain enterococci produce peptide-derived microbial associated molecular patterns, which are sensed by human PMN.
T31 630260-630367 Sentence denotes Structure-function relationship of zwitterionic cell wall polymer dependent immune modulatory mechanisms C.
T32 630368-630387 Sentence denotes Weidenmaier* 1 , S.
T33 630388-630401 Sentence denotes Wanner 1 , M.
T34 630402-630419 Sentence denotes Rautenberg 1 , S.
T35 630420-630431 Sentence denotes Baur 1 , L.
T36 630432-630668 Sentence denotes Kull 1 1 Universität Tübingen, IMIT, Tübingen, Deutschland Recent advances on the field of glycobiology led to discovery of novel mechanisms by which bacterial cell wall glycopolymers can directly modulate the host immune response (1) .
T37 630669-630888 Sentence denotes It had been demonstrated that the structural prerequisite of the T cell stimulatory activity of bacterial capsular polysaccharides (CP) is the presence of positive and negative charge centres within the repeating units.
T38 630889-631101 Sentence denotes We could recently show that the important pathogenicity factor (2) cell wall teichoic acid (WTA) of S. aureus has a similar activity and is able to activate CD4-T cells in an MHCII presentation depended way (3) .
T39 631102-631189 Sentence denotes This mechanism modulates the development of skin abscess formation in an in vivo model.
T40 631190-631538 Sentence denotes Interestingly mostly pathogenic Gram-positive bacterial species seem to exhibit the ribitol-Pi repeating units with D-alanine (however WTA structures differ in side chain sugar composition or contain additional repeating unit sugars), whereas non pathogenic species often have shorter glycerol-Pi repeating units or no D-alanine modifications (1) .
T41 631539-631774 Sentence denotes Our data demonstrates that the glycerol-pi containing Stapyhlococcus epidermis WTA is not able to efficiently stimulate T cell proliferation whereas ribitol-Pi WTA of S. aureus is an efficient MHCII dependent T cell activating polymer.
T42 631775-631928 Sentence denotes We therefore postulate that the WTA structure and expression profile play a role in defining the pathogenic potential of Gram-positive bacterial species.
T43 631929-632081 Sentence denotes In addition it is also of great interest to understand the regulatory events of WTA biosynthesis in pathogenic Gram-positive bacteria such as S. aureus.
T44 632082-632223 Sentence denotes It is not known so far whether the amount of teichoic acids differs during the development of an infection or even between bacterial strains.
T45 632224-632339 Sentence denotes Interestingly, we find that highly pathogenic S. aureus strains exhibit an elevated WTA amount in their cell walls.
T46 632340-632631 Sentence denotes Purified protein-free cell wall fractions from these strains induce T cell proliferation and cytokine production more efficiently than cell wall from less pathogenic strains In addition we could demonstrate that cell wall fractions of these strains are more active in skin abscess formation.
T47 632632-632721 Sentence denotes Thus, up regulation of WTA biosynthesis contributes to pathogenic potential of S. aureus.
T48 632722-632830 Sentence denotes Characterization of highly divergent African Staphylococcus aureus isolates from wild and captive animals F.
T49 632831-632870 Sentence denotes Schaumburg* 1 1 Uniklinik Münster, Med.
T50 632871-632920 Sentence denotes Mikrobiologie, Münster, Deutschland Introduction:
T51 632921-632976 Sentence denotes Staphylococcus aureus is colonizing humans and animals.
T52 632977-633172 Sentence denotes Recent reports from industrialized countries have described the impact of zoonotic S. aureus transmissions from livestock, companion animals and meat for the colonization and infection in humans.
T53 633173-633308 Sentence denotes S. aureus colonization of undomesticated sub-Saharan animals could pose a risk for inter-species transmission and infections in humans.
T54 633309-633506 Sentence denotes We therefore characterize S. aureus isolates from wild and captive animals from sub-Saharan Africa to obtain data for further investigations of cross-species transmission of S. aureus in this area.
T55 633507-633515 Sentence denotes Methods:
T56 633516-633673 Sentence denotes We screened 204 living captive (n = 39) or human-habituated (n = 59) animals and bushmeat samples (n = 106) from Ivory Coast, Gabon, Zambia and South Africa.
T57 633674-633776 Sentence denotes S. aureus isolates were confirmed by nuc gene detection and sequencing of the ribosomal 16S rRNA gene.
T58 633777-633845 Sentence denotes Antibiotic susceptibility was tested using VITEK2 automated systems.
T59 633846-634030 Sentence denotes Isolates were genotyped using S. aureus protein A (spa) typing and multilocus sequence typing (MLST) and were characterized by their possession of genes encoding for virulence factors.
T60 634031-634250 Sentence denotes Results: S. aureus was found in 35% of all samples including 35 isolates from human primates (gorilla and chimpanzees, 48.6%), 34 from nonhuman primates (47.2%), two from ruminants (2.8%) and one from a wild cat (1.4%).
T61 634251-634297 Sentence denotes Noteworthy, 26 isolates were nuc PCR negative.
T62 634298-634486 Sentence denotes Resistance to penicillin, tetracycline and cotrimoxazole was 12%, 6% and 1%, respectively and was significantly associated with captive animals (OR = inf., 95% CI = 32.48-inf., p < 0.001).
T63 634487-634509 Sentence denotes MRSA was not detected.
T64 634510-634587 Sentence denotes The most prevalent toxin genes were sei (23.6%), seh (16.7%) and seg (15.3%).
T65 634588-634667 Sentence denotes The prevalence of classical superantigen-encoding genes (sea-see, tst) was low.
T66 634668-634736 Sentence denotes Three isolates contained Panton-Valentine leukocidin encoding genes.
T67 634737-634847 Sentence denotes Among 72 isolates, we found 42 different spa types and so far 28 confirmed different MLST sequence types (ST).
T68 634848-634936 Sentence denotes The most prevalent spa types were t7821 (11%), t6963 (6%) and t6962 and t7723 (6% each).
T69 634937-635000 Sentence denotes The predominant STs were ST1 and ST182 (11% each) and ST9 (6%).
T70 635001-635193 Sentence denotes The STs 2058 STs , 2059 STs , 1873 STs , 2022 STs , 2067 STs , 1857 STs , 1872 and 1874 formed a highly divergent group when using their concatenated sequences and the neighbor-joining method.
T71 635194-635205 Sentence denotes Conclusion:
T72 635206-635302 Sentence denotes Our results suggest that colonization of human and nonhuman primates with S. aureus is frequent.
T73 635303-635431 Sentence denotes One third of S. aureus isolates belonged to STs, which were highly divergent from those described in isolates colonizing humans.
T74 635432-635530 Sentence denotes This will provide an excellent tool to investigate potential cross-species transmission in future.
T75 635531-635533 Sentence denotes A.
T76 635534-635549 Sentence denotes Grisold* 1 , B.
T77 635550-635563 Sentence denotes Würstl 2 , M.
T78 635564-635578 Sentence denotes Hoenigl 3 , T.
T79 635579-635594 Sentence denotes Valentin 3 , H.
T80 635595-635608 Sentence denotes Salzer 3 , E.
T81 635609-635623 Sentence denotes Leitner 1 , G.
T82 635624-635637 Sentence denotes Feierl 1 , J.
T83 635638-635650 Sentence denotes Posch 1 , R.
T84 635651-635664 Sentence denotes Krause 3 , G.
T85 635665-635904 Sentence denotes Zarfel 1 1 Medical University , Institute of Hygiene, Graz, Austria, Oesterreich 2 Max von Pettenkofer Institute, Munich, Germany, Deutschland 3 Medical University , Department of Infectious Diseases, Graz, Austria, Oesterreich Objectives:
T86 635905-636012 Sentence denotes Carbapenem-resistant Enterobacteriaceae have emerged as a major public health problem throughout the world.
T87 636013-636142 Sentence denotes Namely the newly described resistance mechanism conferred by New Delhi metallo-ßlactamase 1 (NDM-1) activated worldwide interest.
T88 636143-636265 Sentence denotes We report the emergence of carbapenem-resistant Enterobacteriaceae and involved carbapenem hydrolyzing enzymes in Austria.
T89 636266-636274 Sentence denotes Methods:
T90 636275-636459 Sentence denotes In a 10-year retrospective analysis (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) Enterobacteriaceae resistance data were checked for resistance to carbapenems.
T91 636460-636630 Sentence denotes Detected strains were further analyzed for involved carbapenem hydrolyzing enzymes using PCR methods for the detection of eg MBL/IMP, SPM, VIM, GIM or NDM-1 respectively.
T92 636631-636723 Sentence denotes Rep-PCR on the DiversiLab instrument (BioMerieux) was used to determine clonal relationship.
T93 636724-636732 Sentence denotes Results:
T94 636733-636937 Sentence denotes Over a ten-year-period carbapenem-resistant Enterobacteriaceae were isolated from 13 hospitalized patients with the first isolation in the year 2005 and a remarkable increase of involved patients in 2010.
T95 636938-637065 Sentence denotes Carbapenem-resistant Enterobacteriaceae comprise 8 Klebsiella pneumoniae, 4 Klebsiella oxytoca and one Escherichia coli strain.
T96 637066-637146 Sentence denotes Detected carbapenemases belonged to the metallo-ß-lactamases NDM-1, VIM and IMP.
T97 637147-637208 Sentence denotes K. pneumoniae carbapenemase (KPC) was first detected in 2010.
T98 637209-637322 Sentence denotes NDM-1 was detected in three patients, in two cases in K. pneumoniae, and one patient with NDM-1 positive E. coli.
T99 637323-637386 Sentence denotes All three cases occurred between December 2009 and August 2010.
T100 637387-637603 Sentence denotes All detected carbapenem-resistant Enterobacteriaceae strains were multiresistant, and exhibited susceptibility to tigecycline (13/13), colistin (10/13), amikacin (10/13), fosfomycin (5/13) and gentamicin (3/13) only.
T101 637604-637691 Sentence denotes Carbapenem-resistant Enterobacteriaceae has become a substantial global health problem.
T102 637692-637743 Sentence denotes We report the emergence of such strains in Austria.
T103 637744-637851 Sentence denotes Carbapenem-resistant Enterobacteriaceae were detected in a comparable low number of 13 patients since 2001.
T104 637852-637975 Sentence denotes But regarding international data there is an urgent need of active surveillance on carbapenem-resistant Enterobacteriaceae.
T105 637976-638154 Sentence denotes Both local reports and linked data of national and international surveillance networks may be successful approaches to an effective containment of these multi-resistant bacteria.
T106 638155-638223 Sentence denotes Analysis of the population structure of Anaplasma phagocytophilum C.
T107 638224-638237 Sentence denotes Winter 1 , T.
T108 638238-638256 Sentence denotes Wolfsperger 1 , N.
T109 638257-638590 Sentence denotes Wüppenhorst 1 , F. von Loewenich* 1 1 Institut für Medizinische Mikrobiologie, Universität Freiburg, Freiburg, Deutschland Anaplasma phagocytophilum is a Gram-negative, tick-transmitted, obligate intracellular bacterium that elicits acute febrile diseases in humans and domestic animals such as sheep, cattle, horses, dogs, and cats.
T110 638591-638655 Sentence denotes Roe deer and red deer are regarded as potential reservoir hosts.
T111 638656-638831 Sentence denotes Considerable strain variation has been suggested to occur within this species, because isolates from humans and animals differed in their pathogenicity for heterologous hosts.
T112 638832-639007 Sentence denotes In order to explain host preference and epidemiological diversity, we used multi locus sequence typing (MLST) for the molecular characterization of A. phagocytophilum strains.
T113 639008-639117 Sentence denotes 237 samples from humans, domestic animals, potential reservoir hosts, and Ixodes ricinus ticks were analyzed.
T114 639118-639269 Sentence denotes Because of double infections the typabilitiy was only 69% (164/237). eBURST analysis revealed one clonal complex with sequence type (ST) 55 as founder.
T115 639270-639416 Sentence denotes Strains belonging to this clonal complex originated from dogs (54/56), horses (10/10), humans (6/6), cats (2/2), sheep (3/42) , and ticks (14/20).
T116 639417-639496 Sentence denotes All samples from the US were ST 64, what might reflect their geographic origin.
T117 639497-639578 Sentence denotes The phylogenetic analysis of the concatenated sequences showed two main clusters.
T118 639579-639619 Sentence denotes Tick samples were found in both of them.
T119 639620-639686 Sentence denotes Apart from that, cluster 2 was restricted exclusively to roe deer.
T120 639687-639824 Sentence denotes As A. phagocytophilum strains infecting humans, dogs and horses belonged to one clonal complex, they seem to share the same epidemiology.
T121 639825-639922 Sentence denotes Their biological similarity is reflected in the results of previous cross-infections experiments.
T122 639923-640185 Sentence denotes Because the concatenated sequences from I. ricinus ticks are found in both clusters, they might be involved in the transmission cycle of A. phagocytophilum strains that infect humans and domestic animals (cluster 1) and of those that infect roe deer (cluster 2).
T123 640186-640380 Sentence denotes In contrast to red deer, roe deer are probably not the reservoir for granulocytic anaplasmosis in humans and domestic animals, as their concatenated sequences formed a clearly separated cluster.
T124 640381-640642 Sentence denotes The identification and description of disease outbreaks as well as the improvement of our knowledge on population genetics, epidemiology and ecology of bacterial pathogens begins with the ability to differentiate bacterial pathogens via reliable typing methods.
T125 640643-640868 Sentence denotes Especially for the delineation of extraintestinal pathogenic Escherichia coli (ExPEC) and commensal E.coli this is a challenging task, because certain ExPEC and commensal E.coli strains may be geno-and phenotypically similar.
T126 640869-641206 Sentence denotes We set our focus on E.coli clonal complex 73, a prominent ExPEC lineage, harboring phenotypically diverse ExPEC, asymptomatic bacteruria but also commensal E.coli variants, as a model to evaluate and improve two already established systems, i.e. "multi locus sequence typing" (MLST) and generic E.coli "multi locus VNTR analysis" (MLVA).
T127 641207-641319 Sentence denotes We tested these typing systems for their ability to differentiate between the three phenotypic E.coli subgroups.
T128 641320-641429 Sentence denotes Furthermore, we evaluated how additional markers improve the discriminatory potential of both typing methods.
T129 641430-641569 Sentence denotes Our results will be discussed regarding the advancement of typing and molecular epidemiology of extraintestinal pathogenic Escherichia coli
T130 641570-641669 Sentence denotes No evidence for transmissions of septic salmonellosis in children from Ghana by water from wells H.
T131 641670-641690 Sentence denotes Frickmann* 1, 2 , D.
T132 641691-641706 Sentence denotes Dekker 3 , N.G.
T133 641707-641721 Sentence denotes Schwarz 3 , N.
T134 641722-641736 Sentence denotes Sarpong 4 , Y.
T135 641737-641756 Sentence denotes Adu-Sarkodie 5 , S.
T136 641757-641773 Sentence denotes Poppert 6 , R.M.
T137 641774-641786 Sentence denotes Hagen 1 , J.
T138 641787-642178 Sentence denotes May 3 1 Bundeswehrkrankenhaus Hamburg, Hamburg, Deutschland 2 Universitätsklinikum Rostock, Institut für Medizinische Mikrobiologie, Virologie und Hygiene, Rostock, Epidemiologie, Hamburg, Deutschland 4 Kumasi Centre for Collaborative Research, Kumasi (Ghana) , Ghana 5 Kwame Nkrumah University of Science and Technology, Kumasi (Ghana) , Ghana Diagnostik, Hamburg, Deutschland Introduction:
T139 642179-642362 Sentence denotes Kumasi, Ghana, is one model region for epidemiological investigations on the transmission of infectious diseases in the tropical setting, where we analyzed bacterial causes of sepsis.
T140 642363-642539 Sentence denotes Salmonella enterica is one of the quantitatively most important blood culture isolates from children in the rural village Agogo and the surrounding area, Ashanti Region, Ghana.
T141 642540-642673 Sentence denotes However, the exact mode of transmission of this typically food-borne pathogen in that particular part of the world is widely unknown.
T142 642674-642748 Sentence denotes Most children in this region do not have access to bottled drinking water.
T143 642749-642824 Sentence denotes We therefore investigated contamination of drinking water from local wells.
T144 642825-642846 Sentence denotes Material and methods:
T145 642847-643026 Sentence denotes Within a study period of 2 years, 192 Salmonella enterica blood culture isolates from children 15 years of age and 40 isolates from 13 wells were collected, partly longitudinally.
T146 643027-643093 Sentence denotes Salmonella strains were repetitively isolated from selected wells.
T147 643094-643319 Sentence denotes Species identity was biochemically determined by API-E (bioMeriéux) as well as H2S synthesis on triple-sugar-iron-agar ("Kligler"-agar) in Kumasi, Ghana, and confirmed by Fluorescence-in-situ-hybridization and PCR in Hamburg.
T148 643320-643507 Sentence denotes Serotyping was performed from all isolates and identical serotypes that were identified within a 1-month-range in one village were further typed by Pulsed-fieldgel-electrophoresis (PFGE).
T149 643508-643659 Sentence denotes PFGE patterns were compared with the software Gel Compar II (Applied Maths) using the settings DICE, UPGMA, 0.5% optimization, 1.0% position tolerance.
T150 643660-643705 Sentence denotes Clonal identity was accepted at 95% identity.
T151 643706-643714 Sentence denotes Results:
T152 643715-643772 Sentence denotes All analyzed wells were colonized by Salmonella enterica.
T153 643773-643916 Sentence denotes Serotyping demonstrated that the reaction patterns of the isolates from the wells were clearly distinguishable from the blood culture isolates.
T154 643917-644076 Sentence denotes PFGEanalysis of 76 clinical isolates in 8 villages showed a great diversity of different clones but also some endemic clones circulating in particular regions.
T155 644077-644353 Sentence denotes Eight S. enterica isolates that were isolated from one particular well were discriminated to 7 distinct clones by PFGE thus demonstrating that a single well can be colonized by different S. enterica strains and that individual clones can persist about several weeks in a well.
T156 644354-644475 Sentence denotes Discussion: PFGE data from the blood culture isolates indicate that endemic clones circulate within the local population.
T157 644476-644506 Sentence denotes The source was not identified.
T158 644507-644619 Sentence denotes Serotyping gave no evidence that the water from the wells plays a major role in the transmission of S. enterica.
T159 644620-644789 Sentence denotes The interpretation is limited because different distinct serotypes may colonize a well at the same time while only one individual clone was selected per isolation event.
T160 644790-644985 Sentence denotes The high number of isolates from the wells with no matches to blood culture isolates during the whole study interval is more likely to support the hypothesis of alternative modes of transmission.
T161 644986-645089 Sentence denotes Further studies addressing the role of food for the transmission of S. enterica in Ghana should follow.
T162 645090-645211 Sentence denotes Enterovirus surveillance in Germany -one system of two intentions: diagnostic of CNS-infections and polio surveillance K.
T163 645212-645228 Sentence denotes Neubauer* 1 , K.
T164 645229-645242 Sentence denotes Beyrer 2 , S.
T165 645243-645258 Sentence denotes Böttcher 1 , S.
T166 645259-645370 Sentence denotes Diedrich FG 15, Berlin, Deutschland 2 Niedersächsisches Landesgesundheitsamt, Hannover, Deutschland Background:
T167 645371-645647 Sentence denotes Since the Global Polio Eradication Initiative (GPEI) has been launched in 1988, the global incidence of polio has been reduced by over 99% and the transmission of indigenous wild poliovirus was interrupted in all but four countries (Afghanistan, Pakistan, India, and Nigeria).
T168 645648-645997 Sentence denotes Although the WHO region Europe has been certified polio-free in 2002, the risk of reintroduction remains (e.g. polio outbreak in Tajikistan with spread to Russian Federation, Turkmenistan, and Kazakhstan in 2010) and strategies such as vaccination and surveillance of polio have to be maintained until the global polio eradication has been achieved.
T169 645998-646228 Sentence denotes In 2010 the responsibilities for the German programme of polio surveillance -namely the enterovirus surveillance -was handed over from the Governmental Institute of Public Health of Lower Saxony to the Robert Koch Institute (RKI).
T170 646229-646445 Sentence denotes In this context the enterovirus surveillance system was extended and can now be used for both, the enterovirus diagnostics in patients with aseptic meningitis / encephalitis and patients with acute flaccid paralysis.
T171 646446-646578 Sentence denotes The examination of one faecal sample or CSF per CNS / AFP patient is offered all paediatric and neurological clinics free of charge.
T172 646579-646656 Sentence denotes Specimens are analyzed for enteroviruses using PCR, culture and virus typing.
T173 646657-646758 Sentence denotes A network of 14 labs was established, each of these participating regularly in the proficiency tests.
T174 646759-646908 Sentence denotes In 2010, about 47% of all paediatric hospitals and 11% of all neurology departments in Germany made use of the offer of free enterovirus diagnostics.
T175 646909-647018 Sentence denotes In the five years till end of 2010 a total of ~13,500 samples were tested; on average 2,500 samples annually.
T176 647019-647103 Sentence denotes In about 65% of cases stool samples were tested, the remaining 35% were CSF samples.
T177 647104-647309 Sentence denotes The age distribution of patients reflects the great interest of paediatric clinics on the enterovirus diagnostics: overall, about 83% of samples originate from children <15 years of age (median = 7 years).
T178 647310-647431 Sentence denotes The seasonal pattern of samples sent for testing shows the classical dominance of enteroviruses during the summer months.
T179 647432-647532 Sentence denotes The overall PCR-positivity rate amounts to ~30% (33.4% for stool samples and 19.6% for CSF samples).
T180 647533-647644 Sentence denotes So far, more than 2,800 samples were serotyped and >50 different serotypes were detected (ECHO 30 most common).
T181 647645-647711 Sentence denotes Polioviruses were not detected between 2006 and 2010, as expected.
T182 647712-647952 Sentence denotes The results demonstrate that the high-quality enterovirus surveillance is a) well accepted by the clinicians and b) allows Germany to meet its obligations to WHO in establishing a reliable surveillance system to prove the polio-free status.
T183 647953-648187 Sentence denotes Furthermore, the high rate of enterovirus positive results and the distribution of cases all over Germany give a good picture of the circulating enteroviruses in Germany, thus, contributing to outbreak detection and infection control.
T184 648188-648371 Sentence denotes Characterization of extended spectrum -lactamase producing Escherichia coli: comparison of phylogenetic grouping by the DiversiLab system, mononucleotide motifs and genetic markers T.
T185 648372-648387 Sentence denotes Vollmer* 1 , C.
T186 648388-648401 Sentence denotes Knabbe 1 , J.
T187 648402-648450 Sentence denotes Dreier Bad Oeynhausen, Deutschland Introduction:
T188 648451-648608 Sentence denotes Rapid and reliable methods to identify the molecular relatedness in clinical isolates are essentially needed to investigate potential routes of transmission.
T189 648609-648779 Sentence denotes Randomly amplified polymorphic DNA amplification (RAPD) techniques have been shown as a useful and comparatively simple tool for the discrimination of strain relatedness.
T190 648780-648908 Sentence denotes Recently, the DiversiLab (DL, bioMérieux) system was released as an automated system for the analysis of microbial fingerprints.
T191 648909-649059 Sentence denotes The main advantage, compared to RAPD, is the amplification of defined different sized repetitive fragments within the non-coding region of the genome.
T192 649060-649244 Sentence denotes In the present study, the performance of the DL system as a routine typing tool for extended-spectrum ß-lactamase (ESBL) producing E. coli was evaluated in comparison to other methods.
T193 649245-649266 Sentence denotes Material and Methods:
T194 649267-649622 Sentence denotes The phylogenetic relatedness of 107 clinical isolates of ESBL-producing E. coli were compared using the DiversiLab system (DL, bioMérieux), amplification of mononucleotide repeats (MNRs; yaiN, ycgW, b2345, ykgE) and three candidate gene markers (chuA, yjaA, TSPE4.C2), which were able to distinct between the main four phylogenetic groups A, B1, B2 and D.
T195 649623-649722 Sentence denotes Isolates were estimated to be related by the DL system at >95 % homology using Pearson correlation.
T196 649723-649731 Sentence denotes Results:
T197 649732-649909 Sentence denotes The DL system revealed two larger clusters of related isolates (cluster I, 18 strains; cluster II: six strains) as well as twelve clusters of two, three or four related strains.
T198 649910-649998 Sentence denotes Isolates of cluster I belong to the phylogenetic group B2 and had none of the four MNRs.
T199 649999-650081 Sentence denotes Isolates of cluster II were typed as B1 and showed amplification of all four MNRs.
T200 650082-650197 Sentence denotes Other DL-clusters also showed a concordance regarding their phylogenetic group and their MNR amplification pattern.
T201 650198-650272 Sentence denotes ESBL-typing by these three different methods showed excellent correlation.
T202 650273-650403 Sentence denotes Amplicons of MNRs were currently sequenced to evaluate the phylogenetic inference and confirm the discrimination of the DL system.
T203 650404-650552 Sentence denotes The discriminatory power of the DL system will be further evaluated in comparison to pulse-field-gelelectrophoresis and multi-locus sequence typing.
T204 650553-650717 Sentence denotes Further investigations include the evaluation of the DL with an in house automated RAPD-PCR using different established primers followed by chip-gelelectrophoresis.
T205 650718-650813 Sentence denotes Sequence typing and expression analysis of 4CMenB targets in capsule null locus meningococci H.
T206 650814-650827 Sentence denotes Claus* 1 , M.
T207 650828-650842 Sentence denotes Jördens 1 , P.
T208 650843-650854 Sentence denotes Kriz 2 , M.
T209 650855-650869 Sentence denotes Musilek 2 , A.
T210 650870-650884 Sentence denotes Tremmel 1 , U.
T211 650885-651126 Sentence denotes Vogel 1 1 Universität Würzburg, Institut für Hygiene und Mikrobiologie, Würzburg, Deutschland 2 National Institute of Public Health, National Reference Laboratory for Meningococcal Infections, Prag, Tschechische Republik Background and aims:
T212 651127-651454 Sentence denotes The multi-component vaccine against serogroup B meningococci (4CMenB) comprises three major recombinant antigens including Neisserial adhesion A (NadA), factor H binding protein (fHbp), and Neisserial heparin binding antigen (NHBA) as well as outer-membrane vesicles, which especially induce antibodies against P1.4 of porin A.
T213 651455-651559 Sentence denotes The purpose of this study was to estimate the effect of 4CMenB on capsule null locus (cnl) meningococci.
T214 651560-651581 Sentence denotes Material and methods:
T215 651582-651916 Sentence denotes Forty-one strains of cnl meningococci from three countries (Burkina Faso, Czech Republic, and Germany) belonging to nine sequence types (ST) and four clonal complexes were antigen sequence typed and analysed for antigen expression by genetic typing and the recently published Meningococcal Antigen Typing System (MATS, Donnelly et al.
T216 651917-651924 Sentence denotes 2010) .
T217 651925-651949 Sentence denotes Results and conclusions:
T218 651950-652150 Sentence denotes Strains of the ST-198 complex harboured an fHbp variant, which is included in the vaccine, and expressed the antigen above the positive bactericidal threshold (PBT) as defined for serogroup B strains.
T219 652151-652212 Sentence denotes Expression of NHBA above the PBT was seen in strains of the .
T220 652213-652320 Sentence denotes Only one strain harboured the nadA gene and P1.4 of porin A was not present among all analysed cnl strains.
T221 652321-652422 Sentence denotes The data suggest that bactericidal antibodies will be effective only on a subset of cnl meningococci.
T222 652423-652618 Sentence denotes A biological effect on the ST-198 complex and on ST-845 will depend on antibody levels at the mucosal surface and their capacity to block transmission, for which data are currently not available.
T223 652619-652621 Sentence denotes C.
T224 652622-652635 Sentence denotes Rohde* 1 , K.
T225 652636-652651 Sentence denotes Selezska 2 , M.
T226 652652-652672 Sentence denotes Kazmierczak 1,3 , J.
T227 652673-653287 Sentence denotes Sikorski 1, 3, 4 1 DSMZ, Mikrobiologie, Braunschweig, Deutschland 2 HZI, Braunschweig, Deutschland 3 DSMZ, Braunschweig, Deutschland 4 Deutschland Introduction:The lytic potential of novel Pseudomonas aeruginosa (PA) phages against a large number of novel environmental and clinical PA isolates was challenging to be studied 1.) to find new potential therapy phages, a medical approach in the fight against PA and 2.) to find a possible relationship between lysis phenomena in the phylogenetic tree of these hosts and the allocation of clusters of lysed strains: a phylogenetic approach using lysis as "phenotype".
T228 653288-653434 Sentence denotes The latter addresses the potential role of phages in the microevolutionary diversification of PA strains into different genetic lineages (clades).
T229 653435-653492 Sentence denotes Are phages relevant drivers of major phylogenetic groups?
T230 653493-653584 Sentence denotes The large pool of new hosts and phages offered a test system to respond to these questions.
T231 653585-653606 Sentence denotes Material and Methods:
T232 653607-653698 Sentence denotes Environmental PA strains were isolated from German rivers across different water qualities.
T233 653699-653772 Sentence denotes Clinical isolates came from different sources, mostly chronic infections.
T234 653773-653845 Sentence denotes All strains were genetically typed using the Pseudomonas ArrayTube chip.
T235 653846-653908 Sentence denotes Phages ("JG series") were isolated from sewage (courtesy of J.
T236 653909-653921 Sentence denotes Garbe and M.
T237 653922-653949 Sentence denotes Schobert, TU Braunschweig).
T238 653950-654109 Sentence denotes Phage assay: overnight cultures were adjusted to 10 9 cfu/mL, drops of 5μL phage suspension were added in 4x4 patterns per agar plate in the overlay technique.
T239 654110-654118 Sentence denotes Results:
T240 654119-654222 Sentence denotes Serotypes of the strains were determined, motility phenotypes, virulence factors and biofilm formation.
T241 654223-654307 Sentence denotes All strains were grouped acc. their core genomic similarity into four main clusters.
T242 654308-654372 Sentence denotes The phages were known to infect PA via an LPS membrane receptor.
T243 654373-654511 Sentence denotes We analysed the lysis phenotype of seven highly lytic phages of this set of 28 phages on 190 strains of PA, 99 environmental, 91 clinical.
T244 654512-654602 Sentence denotes The phages showed substantial differences in the lysis phenotypes on the set of 190 hosts.
T245 654603-654658 Sentence denotes The proportion of lysed strains ranged from 70% to 35%.
T246 654659-654743 Sentence denotes Pairwise phage comparison indicated high or low phenotypic similarity of the phages.
T247 654744-654799 Sentence denotes All results finally suggest two unrelated phage groups.
T248 654800-654841 Sentence denotes One phage seemed phenotypically distinct.
T249 654842-654909 Sentence denotes Clinical PA were more sensitive to phage attack than environmental.
T250 654910-654962 Sentence denotes Lysis phenotypes appeared to depend on the serotype.
T251 654963-655101 Sentence denotes Some serotypes showed large proportions of full lysis, others appeared to confer phage resistance, some were sensitive only to few phages.
T252 655102-655210 Sentence denotes On the level of intraspecies host diversity, some genetic lineages with different phage sensitivity emerged.
T253 655211-655295 Sentence denotes The host lineages differed also in the presence and abundance of specific serotypes.
T254 655296-655418 Sentence denotes Two phages were far more effective in lysing clinical than environmental strains, independent of the genetic host lineage.
T255 655419-655430 Sentence denotes Conclusion:
T256 655431-655569 Sentence denotes From the phage lysis patterns we conclude there is therapeutic potential of some of these highly lytic phages with broad PA host spectrum.
T257 655570-655756 Sentence denotes From the lysis phenotypes we conclude that phages, via the diversity of host receptor types, have potential to contribute to the intraspecies microevolutionary diversification within PA.
T258 655757-655930 Sentence denotes From additional own studies (not shown) we have reasons to believe that the population dynamics of PA were probably shaped by environmental forces, not by clinical habitats.
T259 655931-656023 Sentence denotes The results contribute to the knowledge on the evolutionary dynamics of PA phage resistance.
T260 656024-656026 Sentence denotes A.
T261 656027-656040 Sentence denotes Sabat* 1 , M.
T262 656041-656058 Sentence denotes Chlebowicz 1 , H.
T263 656059-656095 Sentence denotes Grundmann 1 , J.M. van Dijl 1 , A.W.
T264 656096-656203 Sentence denotes Friedrich 1 1 University Medical Center Groningen, Medical Microbiology, Groningen, Niederlande Background:
T265 656204-656337 Sentence denotes Methicillin-resistant Staphylococcus aureus (MRSA) is an increasing health problem in both hospital and community settings worldwide.
T266 656338-656495 Sentence denotes MRSA are characterized by the carriage of diverse Staphylococcal cassette chromosome mec (SCCmec) elements, conferring resistance to beta-lactam antibiotics.
T267 656496-656600 Sentence denotes To date, ten major SCCmec types were described on the basis of the combination of ccr and mec complexes.
T268 656601-656770 Sentence denotes Although, several PCR-based procedures were designed for typing the SCCmec cassettes, they were often found to be limited in their ability to characterize MRSA isolates.
T269 656771-656934 Sentence denotes The goal of our study was to compare the SCCmec typing results produced by well-established multiplex-PCR protocol with those obtained by a microarray-based assay.
T270 656935-656943 Sentence denotes Methods:
T271 656944-657195 Sentence denotes Two hundred-six non-repetitive MRSA isolates were obtained from the collection previously characterized by spa typing, Multiple-Locus Variable-Number Tandem Repeat Analysis (MLVA) and Multiple-Locus Variable-Number Tandem Repeat Fingerprinting (MLVF).
T272 657196-657342 Sentence denotes All isolates were subjected to SCCmec-typing using a multiplex-PCR protocol described by Kondo et al., (Kondo et al., Antimicrob Agents Chemother.
T273 657343-657361 Sentence denotes 51:264-74, 2007) .
T274 657362-657476 Sentence denotes Microarrays (StaphyType by Alere Technologies GmbH, Jena, Germany) were implemented in 117 selected MRSA isolates.
T275 657477-657485 Sentence denotes Results:
T276 657486-657623 Sentence denotes Using the multiplex-PCR protocol, 179 MRSA isolates were typeable whereas from 27 MRSA isolates, the SCCmec type could not be determined.
T277 657624-657832 Sentence denotes The most prevalent SCCmec type was SCCmec type IV (n=108), followed by SCCmec type V (n=36), SCCmec type I (n=20), SCCmec type II (n=8), SCCmec type III (n=4), SCCmec type VI (n=2) and SCCmec type VIII (n=1).
T278 657833-657980 Sentence denotes We further analyzed by the microarray-based approach all MRSA isolates of SSCmec types I-III,VI and VIII as well as all non-typeable MRSA isolates.
T279 657981-658172 Sentence denotes Based on MLVA, MLVF and spa typing characteristics, representatives of the most prevalent SCCmec types, IV and V (n=30 and n=25, respectively), were selected for microarrays characterization.
T280 658173-658302 Sentence denotes For all isolates typeable by the multiplex-PCR method, there was perfect agreement of the results between the two methods tested.
T281 658303-658409 Sentence denotes Among 27 isolates non-typeable by multiplex-PCR approaches almost all (n=26) were typeable by microarrays.
T282 658410-658528 Sentence denotes Among them the SCCmec type V was the most prevalent (n=21), followed by SCCmec type IV (n=4), and SCCmec type I (n=1).
T283 658529-658593 Sentence denotes A single MRSA was non-typeable based on the microarrays results.
T284 658594-658606 Sentence denotes Conclusions:
T285 658607-658763 Sentence denotes The microarray approach appeared to be more efficient than the PCR-based protocol in identification of SCCmec types among genetically diverse MRSA isolates.
T286 658764-658922 Sentence denotes Moreover, the microarray-based method is an inexpensive and fast procedure, and can be easily implemented for investigations in routine clinical laboratories.
T287 658923-658966 Sentence denotes North branch, Korea, Republik Introduction:
T288 658967-659048 Sentence denotes Rotavirus is the major cause of severe diarrhea among infants and young children.
T289 659049-659264 Sentence denotes Rotaviruses, belonging to a genus of doublestranded RNA viruses in the family Reoviridae, infect the mature villus epithelial cells of the small intestine, often leading to fever, vomiting, and diarrhea in children.
T290 659265-659273 Sentence denotes Methods:
T291 659274-659569 Sentence denotes A total of 5,872 stool specimens collected from patient with acute gastroenteritis in northern Gyeonggi province of Korea during 3 years surveilence (August 2007 through July 2010) were analyzed by using antigen-capture enzyme-linkes immunosorbent assay(ELISA) and RT-PCR followed by sequencing.
T292 659570-659578 Sentence denotes Results:
T293 659579-659917 Sentence denotes Of the 5,872 specimens, rotavirus were selected 347 specimens as rotavirus-positive specimens by means of antigen-capture enzyme-linkes immunosorbent assay(ELISA Streptococcus agalactiae (group B streptococci, GBS) is a common commensal of the female urogenital tract and is well known for causing life-threatening infections in neonates.
T294 659918-660178 Sentence denotes For several years now increasing infection rates of immunocompromized adult patients have been reported In rare cases an infection in neonates, pregnant women and immunocompromized elderly patients can lead to a toxic shock syndrome with necrotizing fasciitis.
T295 660179-660307 Sentence denotes Streptococcal toxic shock syndrome (STSS) is a highly invasive bacterial infection with a mortality rate ranging between 30-70%.
T296 660308-660460 Sentence denotes It is typically caused by Streptococcus pyogenes and mutations of a general virulence regulator (cov) have been reported in association with STSS cases.
T297 660461-660771 Sentence denotes To investigate if similar mutations are found in S. agalactiae strains or if a certain clone is responsible for this uncommon course of S. agalactiae infections, we performed a detailed molecular analysis and DNA sequencing of the cov virulence regulator genes in a collection of 30 S. agalactiae STSS strains.
T298 660772-660879 Sentence denotes The strains originated from 28 adults and 2 children and were collected in the US, Germany and Switzerland.
T299 660880-661008 Sentence denotes Molecular typing was performed by multi locus sequence typing (MLST), capsular typing, and surface protein determination by PCR.
T300 661009-661122 Sentence denotes For analysis of the cov gene locus the genes covS and covR were amplified by PCR and subjected to DNA sequencing.
T301 661123-661288 Sentence denotes Molecular subtyping by MLST showed that 7 of the 30 S. agalactiae STSS strains were ST1 and capsular serotype type V and 5 belonged to ST23 and capsular serotype Ia.
T302 661289-661350 Sentence denotes The rest of the strains harbored different common MLST types.
T303 661351-661446 Sentence denotes Genetic analysis showed a 1 bp deletion leading to a frame shift in covS in one of the strains.
T304 661447-661598 Sentence denotes The two component cov regulator represses hemolysis and the mutation was associated with an increased hemolysis and pigmentation in the mutated strain.
T305 661599-661673 Sentence denotes Another strain harbored a 3 bp deletion that had previously been reported.
T306 661674-661847 Sentence denotes The rest of the strains had cov alleles matching the genes sequence of completely sequenced S. agalactiae strains in the Genbank database or were harboring silent mutations.
T307 661848-662030 Sentence denotes In conclusion virulence regulator gene mutations in strains causing streptococcal toxic shock syndrome as they have been described for S. pyogenes can also be found in S. agalactiae.
T308 662031-662259 Sentence denotes A detailed molecular epidemiologic characterization of STSS S. agalactiae strains by MLST, capsular typing and surface protein determination showed them to be heterogenous, despite a certain predominance of ST1 and ST23 strains.
T309 662260-662262 Sentence denotes M.
T310 662263-662279 Sentence denotes Kalitzky* 1 , T.
T311 662280-662292 Sentence denotes Flohr 1 , D.
T312 662293-662307 Sentence denotes Meißner 1 , L.
T313 662308-662406 Sentence denotes Grüter 1 1 Partnerschaftspraxis für Labormedizin, Mikrobiologie, Hameln, Deutschland Introduction:
T314 662407-662501 Sentence denotes Pathogenic E. coli species represent a major cause for diarrhea in various groups of patients.
T315 662502-662753 Sentence denotes Whereas Enteropathogenic E. coli (EPEC) is a well known cause of diarrhea in babies, Enterohaemorrhagic E. coli (EHEC) and Shiga-toxin-producing E. coli (STEC) are known to cause infections especially by intake of food contaminated with bovine faeces.
T316 662754-662913 Sentence denotes Recently, an outbreak of a novel clone of E. coli bearing the rare serotype O104:H4 has threatened Northern Germany and challenged the health care authorities.
T317 662914-663026 Sentence denotes We report data collected in our routine medical laboratory during the clinical investigation of stool specimens.
T318 663027-663048 Sentence denotes Material and Methods:
T319 663049-663298 Sentence denotes During a period of three years, more than 20,000 stool specimens from patients with diarrhea or enteritis originating from Lower Saxony and surrounding counties were screened by real-time multiplex PCR for the presence of pathogenic E. coli species.
T320 663299-663432 Sentence denotes Briefly, characteristic genes for EHEC, STEC, EAEC, EPEC, and EIAC, ETEC were detected by an in-house real-time multiplex PCR method.
T321 663433-663441 Sentence denotes Results:
T322 663442-663585 Sentence denotes Our PCR technique detects Enterohaemorrhagic E. coli and Shigatoxin producing E. coli with a higher sensitivity than verotoxin-ELISAtechniques.
T323 663586-663656 Sentence denotes Pathogenic E. coli species were found as causative agents at all ages.
T324 663657-663746 Sentence denotes Interestingly, EPEC were not only found in young children, but in adult patients as well.
T325 663747-663813 Sentence denotes EHEC and STEC do not show any preference for a certain age either.
T326 663814-663991 Sentence denotes The E. coli species responsible for the recent outbreak bears a combination of pathogenic factors that has not been found in our 20,000 stool probes during the past three years.
T327 663992-664003 Sentence denotes Discussion:
T328 664004-664230 Sentence denotes According the microbiologic-infectiologic quality standards (MIQ) of the German Society for Hygiene and Microbiology, screening of stool samples for pathogenic E. coli species should only be performed under certain conditions.
T329 664231-664335 Sentence denotes Following this standard, EHEC, STEC, EAEC and EPEC would not have been detected in most of our patients.
T330 664336-664506 Sentence denotes For example, the MIQ standard only considers EPEC a causative agent in adult patients if they are immunocompromised or have persisting diarrhea for more than three weeks.
T331 664507-664547 Sentence denotes Our data contradict this recommendation.
T332 664548-664661 Sentence denotes Autochthonous leishmaniasis in Germany: survey of the prevalence of sand flies and infected rodents in Bavaria C.
T333 664662-664676 Sentence denotes Bogdan* 1 , S.
T334 664677-664694 Sentence denotes Haeberlein 1 , K.
T335 664695-664707 Sentence denotes Knoll 1 , S.
T336 664708-664722 Sentence denotes Eßbauer 2 , U.
T337 664723-664937 Sentence denotes Schleicher 1 1 Universitätsklinikum Erlangen, Mikrobiologisches Institut, Erlangen, Deutschland 2 Institut für Mikrobiologie der Bundeswehr, Abteilung Virologie und Rickettsiologie, München, Deutschland Background:
T338 664938-665118 Sentence denotes Zoonotic cutaneous and visceral leishmaniasis are caused by protozoan parasites of the genus Leishmania, which are transmitted by sand fly vectors from reservoir animals to humans.
T339 665119-665217 Sentence denotes Within Europe the disease is highly prevalent in countries and islands of the Mediterranean basin.
T340 665218-665385 Sentence denotes In recent years evidence was presented that argues for a northward spread of leishmaniasis into previously non-endemic areas such as Switzerland, Slowenia and Austria.
T341 665386-665526 Sentence denotes Within Germany sand flies of the species Phlebotomus mascitii and P. pernicious were detected in the Rhine valley (Naucke et al., Parasitol.
T342 665527-665531 Sentence denotes Res.
T343 665532-665539 Sentence denotes 2008) .
T344 665540-665611 Sentence denotes Also, a case of autochthonous human leishmaniasis (Bogdan et al., Clin.
T345 665612-665619 Sentence denotes Infect.
T346 665620-665624 Sentence denotes Dis.
T347 665625-665795 Sentence denotes 2001 ) and three cases of autochthonous canine and equine leishmaniasis have recently been reported from Bavaria (Gothe et al., Kleintierpraxis 1999; Koehler et al., Vet.
T348 665796-665806 Sentence denotes Parasitol.
T349 665807-665814 Sentence denotes 2002) .
T350 665815-665996 Sentence denotes In the present study, we investigated whether sand flies are prevalent in Bavaria and whether wild mice carry Leishmania parasites and therefore might function as reservoir animals.
T351 665997-666006 Sentence denotes 1 1 Univ.
T352 666007-666065 Sentence denotes Hosp., Institute for Hygiene, Münster, Deutschland 2 Univ.
T353 666066-666078 Sentence denotes Hosp., Dept.
T354 666079-666448 Sentence denotes Periodontology, Münster, Deutschland 3 Ridom GmbH, Münster, Deutschland Enterohemorrhagic Escherichia coli (EHEC) O157:H7/Hbelong to the highly pathogenic subgroup of Shiga toxin-producing E. coli (STEC) and cause severe illness, including watery or bloody diarrhea and the hemolytic uremic syndrome, the most common cause for acute renal failure in children worldwide.
T355 666449-666659 Sentence denotes It is already widely accepted that EHEC O157 branched from O55:H7 to both Sorbitol fermenting (SF) O157:Hand three clusters of non-SF EHEC O157:H7 (Leopold et al., Proc Natl Acad Sci USA (2009) 106(21):8713-8).
T356 666660-666763 Sentence denotes However, until now the involved hypothetical intermediate SF O157:H7 clone had not yet been identified.
T357 666764-667046 Sentence denotes Using published genome sequence data of different O55:H7 and O157 strains, a DNA sequence-based approach including 94 coding regions of the core genome was established and applied to a strain collection of 50 strains (48 SF and non-SF O157 and 2 O55) to investigate their phylogeny.
T358 667047-667208 Sentence denotes Electopherogram data were analyzed and subsequent phylogenetic analyses were performed using the Ridom SeqSphere software version 0.9 beta (Ridom GmbH, Münster).
T359 667209-667334 Sentence denotes Moreover, the data was compared with previously performed multiple-locus variable-number tandem-repeats analysis (MLVA) data.
T360 667335-667505 Sentence denotes Sequence analysis of in total 51,229 nucleotides per strain resulted in 111 single nucleotide polymorphisms (SNPs) that were discovered in 76 of the 94 investigated loci.
T361 667506-667641 Sentence denotes With an overall variant ratio of 0.22%, 58 SNPs were non-synonymous (52.3%) indicating a certain evolutionary pressure on each subtype.
T362 667642-667701 Sentence denotes The combination of SNPs resulted in 27 different genotypes.
T363 667702-667804 Sentence denotes Of these, certain SNPs or SNP genotypes could be determined to differentiate each serotype or cluster.
T364 667805-667925 Sentence denotes Interestingly, SNP data enabled us to further differentiate amongst EHEC O157 subgroups, especially within EHEC O157:H7.
T365 667926-668033 Sentence denotes In contrast, SNP data confirmed the already determined high genotypic conservation within SF EHEC O157:H -.
T366 668034-668172 Sentence denotes The SNP data was further investigated to elaborate new typing techniques using fewer loci in order to get a comparable rate of resolution.
T367 668173-668296 Sentence denotes Therefore, characterization of only 25 of the 111 observed SNP localizations enabled a differentiation of all 27 genotypes.
T368 668297-668593 Sentence denotes In addition, a genotype suitable to fit the hypothetical intermediate clone was identified from an animal isolate with the SF O157:H7 phenotype, thereby complementing the EHEC stepwise evolutionary model and complementing the phylogenetic step from SF O55:H7 to either SF O157:Hor non-SF O157:H7.
T369 668594-668772 Sentence denotes In summary, SNP-based typing of EHEC O55 and O157enabled us to close the gap within the evolutionary model of EHEC O157 by identification of an isolate of the hypothetical clone.
T370 668773-669011 Sentence denotes Moreover, the identified SNPs and associated genotypes open up new prospects and challenges to develop alternative typing methods with less costs and comparable rates of resolution for epidemiological investigations and phylogeny studies.
T371 669012-669025 Sentence denotes Introduction:
T372 669026-669205 Sentence denotes Different studies revealed the emerging zoonotic behaviour of extended host spectrum genotypes (EHSGs) like livestock-associated (LA-) MRSA which belong to clonal complex (CC)398.
T373 669206-669370 Sentence denotes At present, MRSA is one of the major causes of wound infections in small animals and horses, and infected animals may provide a significant threat to public health.
T374 669371-669519 Sentence denotes Based on these findings, the aim of this study was to determine the proportion of LA-MRSA in wound swabs obtained from companion animals in Germany.
T375 669520-669541 Sentence denotes Material and Methods:
T376 669542-669691 Sentence denotes Between Nov 2010 and May 2011, 1.814 wound swabs originating from different geographic regions in Germany were investigated for the presence of MRSA.
T377 669692-669835 Sentence denotes All MRSA were further characterized by spa-typing, susceptibility testing and presence of genes encoding for Panton-Valentine leukocidin (PVL).
T378 669836-669844 Sentence denotes Results:
T379 669845-669953 Sentence denotes Overall, the general MRSA distribution was 5.6% among clinical samples of companion animal and horse origin.
T380 669954-670185 Sentence denotes Regarding distribution of the MRSA among the different animal species, canine samples (1196) showed a distribution of 4.9% (n= 59), feline samples (399) 5.8% (n= 23) and those of equine origin (219) 9.1% (n= 20) MRSA, respectively.
T381 670186-670419 Sentence denotes Interestingly, the proportion of spa-types frequently associated with livestock-associated MRSA (e.g. CC398, t011, t034) among the strain collection was 18,6% for canine MRSA, 4,3% for feline MRSA, and 90% for those of equine origin.
T382 670420-670632 Sentence denotes In contrast to the previously reported frequent occurrence of CC8associated MRSA among clinical isolates of equine origin, our current results revealed a predominance of LA-MRSA in MRSA positive wound infections.
T383 670633-670841 Sentence denotes A similar trend was seen in wound infections due to MRSA in dogs, which in the past had frequently been associated with CC22-and CC5-strains, while currently we identified several cases associated with CC398.
T384 670842-670853 Sentence denotes Conclusion:
T385 670854-671237 Sentence denotes The detected proportions of CC398-associated MRSA in horses and dogs suggests a rapid spread of LA-associated strains into animals other than livestock and raise the question whether these EHSG-MRSA may harbour certain factors or abilities (genetic and/ or functional) mediating the obvious successful spread among different mammalian species apart from their methicillin-resistance.
T386 671238-671345 Sentence denotes Genotyping of Pilus Islet-1 and Pilus Islet-2 in Streptococcus pneumoniae Clinical Isolates from Germany G.
T387 671346-671360 Sentence denotes Gámez 1,2 , A.
T388 671361-671376 Sentence denotes Gómez* 1,2 , A.
T389 671377-671390 Sentence denotes Zander 1 , N.
T390 671391-671426 Sentence denotes Schade 3 , M. van der Linden 3 , S.
T391 671427-671603 Sentence denotes Hammerschmidt 1 1 Universität Greifswald, Abteilung Genetik der Mikroorganismen, Interfakultäres Institut für Genetik und Funktionelle Genomforschung, Greifswald, Deutschland 2
T1 671605-671776 Sentence denotes The recent discovery of long filamentous pilus-like structures extending from the surface of the pneumococcus has opened a new window in the research field of pneumococci.
T2 671777-671973 Sentence denotes Two pilus regions have been identified in the pneumococcal genome: PI-1 (Pilus Islet-1) and PI-2 (Pilus Islet-2) and, at least for PI-1, their role in pneumococcal pathogenesis has been suggested.
T3 671974-672165 Sentence denotes We analyzed the distribution of both pili among 419 pneumococcal clinical isolates from Germany, divided in three different groups: invasive (n=233), non-invasive (n=73) and carriers (n=113).
T4 672166-672385 Sentence denotes Fortythree of the 91 pneumococcal serotypes were represented in this set of samples, including those used in the 7-valent, 10-valent and 13-valent pneumococcal conjugate vaccines (PCVs) and 23-valent PPSV23 (Pneumovax).
T5 672386-672461 Sentence denotes Overall, 135 (32,2%) pneumococcal clinical isolates were positive for pili:
T6 672462-672524 Sentence denotes 17,2% for PI-2, 13,4% for PI-1 and 1,7% for both pilus islets.
T7 672525-672658 Sentence denotes The frequency within the invasive group (45,5%) was higher than the frequency within non-invasive (13,7%) and carrier (16,8%) groups.
T8 672659-672807 Sentence denotes The 89% of the pilus-positive invasive isolates were positive for PI-2, whereas 2/3rd of the pilus-positive carrier isolates were positive for PI-1.
T9 672808-672928 Sentence denotes Surprisingly, the majority (71,4%) of the pilus-positive non-invasive isolates were positive for both pneumococcal pili.
T10 672929-672998 Sentence denotes The double-positive isolates belong to the serotypes 19F, 19A and 7F.
T11 672999-673108 Sentence denotes Serotypes 6A and 9V are strongly related to PI-1, whereas PI-2 was present mainly in serotypes 1, 7F and 11A.
T12 673109-673274 Sentence denotes Only two (9V and 19F) of the pneumococcal serotypes identified as pilus-positive in this study are present in all currently available vaccines against S. pneumoniae.
T13 673275-673359 Sentence denotes In contrast, serotype 6A and 11A are included only in PCV13 and PPSV23 respectively.
T14 673360-673543 Sentence denotes The clonality of the pilus-containing isolates identified here is being determined by MLST analysis and their correlation with antibiotic resistance profiles will also be established.
T15 673544-673746 Sentence denotes Finally, these data show the scenario of pneumococcal pili distribution among pneumococcal clinical isolates in Germany and may support the potential of PI-1 and PI-2 constituents as vaccine candidates.
T16 673747-673758 Sentence denotes Background:
T17 673759-673883 Sentence denotes Hospital-acquired methicillin resistant Staphylococcus aureus (HA-MRSA) is a growing public health concern across the world.
T18 673884-673944 Sentence denotes Early detection of clusters is vital for patient protection.
T19 673945-674210 Sentence denotes Pulsed-field gel electrophoresis (PFGE) is considered a 'gold standard' in detecting important outbreak events and widely used despite several limitations such as limited resolution and lack of digitized data, making intra-and interlaboratory comparisons difficult.
T20 674211-674484 Sentence denotes Spa typing, which relies on sequence variation within the S. aureus protein A gene (spa), has been proven to be a successful tool for investigating the relatedness of strains and the sequence data is easily stored in databases and communicated between researchers globally.
T21 674485-674571 Sentence denotes Optical Mapping (OM) creates linear restriction enzyme maps from single DNA molecules.
T22 674572-674813 Sentence denotes These maps can distinguish chromosomal events that are ~>1.5 kb in size, providing a higher resolution of data, as well as fragment organization within the genome, as compared to PFGE (~30 kb detection limit) and probably also to spa typing.
T23 674814-674896 Sentence denotes Here we compare these techniques in a retrospective analysis of a HA-MRSA cluster.
T24 674897-674905 Sentence denotes Methods:
T25 674906-675038 Sentence denotes Four epidemiologically related isolates, 5 unrelated clinical isolates and one isolate of indeterminable epidemiology were analyzed.
T26 675039-675211 Sentence denotes Strains were previously characterized using spa typing, and 3 of 5 unrelated isolates were chosen based on an identical spa type with the epidemiologically related strains.
T27 675212-675369 Sentence denotes The remaining two isolates were different spa types, yet one had the same sequence type (multilocus sequence typing), as the epidemiologically related group.
T28 675370-675473 Sentence denotes The relatedness of all studied strains was determined using PFGE and OM (OpGen, Gaithersburg, MD, USA).
T29 675474-675569 Sentence denotes Results: PFGE results confirmed the close relation of all 4 epidemiologically related isolates.
T30 675570-675738 Sentence denotes Three of these 4 strains, as well as the isolate of indeterminable epidemiology, had identical PFGE banding patterns while the fourth isolate differed by a single band.
T31 675739-675916 Sentence denotes The three unrelated isolates with identical spa types were more closely related to the epidemiologically-linked group (though not identical) than the 2 with different spa types.
T32 675917-676059 Sentence denotes The OM data validated the PFGE data, but with a higher resolution showing additional restriction sites in strains that were identical by PFGE.
T33 676060-676072 Sentence denotes Conclusions:
T34 676073-676175 Sentence denotes Our data illuminate the varying levels of data resolution that are offered by spa typing, PFGE and OM.
T35 676176-676304 Sentence denotes PFGE alone was able to differentiate epidemiologically unrelated strains from the cluster strains that shared the same spa type.
T36 676305-676448 Sentence denotes OM data corresponded with both the PFGE and spa typing results, yet with a higher resolution, providing more detailed phylogenetic information.
T37 676449-676643 Sentence denotes This increased resolution may be useful in tracking the progress of a cluster, though further investigations are needed to verify that OM alone is sufficient to identify the spread of pathogens.
T38 676644-676646 Sentence denotes M.
T39 676647-676661 Sentence denotes Merker* 1 , T.
T40 676662-676673 Sentence denotes Kohl 2 , J.
T41 676674-676691 Sentence denotes Kalinowski 2 , S.
T42 676692-676712 Sentence denotes Ruesch-Gerdes 3 , L.
T43 676713-676726 Sentence denotes Truebe 1 , A.
T44 676727-676741 Sentence denotes Roetzer 1 , S.
T45 676742-676996 Sentence denotes Niemann 1 1 Forschungszentrum Borstel, Molekulare Mykobakteriologie, Borstel, Deutschland 2 University of Bielefeld, CEBITEC, Bielefeld, Deutschland 3 Forschungszentrum Borstel, National Reference Center for Mycobacteria, Borstel, Deutschland Background:
T46 676997-677196 Sentence denotes Transmission and phylogenetic background of clinical Mycobacterium tuberculosis complex isolates can be analyzed by genotyping techniques (IS6110 DNA fingerprinting, spoligotyping, MIRU-VNTR typing).
T47 677197-677452 Sentence denotes Although these methods have a discriminatory power high enough to discriminate clinical isolates on the strain level, they are not able to visualize microevolution events e.g. within one patient in the case of treatment failure and resistance development.
T48 677453-677465 Sentence denotes Methodology:
T49 677466-677646 Sentence denotes To adress this question, we applied Next Generation Sequencing (NGS) to analyze the genomes of serial isolates obtained from two different patients with long duration of treatment.
T50 677647-677744 Sentence denotes The time period between the first and last isolate obtained from one patient was up to 36 months.
T51 677745-677926 Sentence denotes Both strains belong to the Beijing lineage; all isolates from one patient exhibited the same IS6110 RFLP and 24 loci MIRU-VNTR typing patterns, thus excluding exogenous reinfection.
T52 677927-677946 Sentence denotes Principal findings:
T53 677947-678041 Sentence denotes From all isolates high quality reads with a mean genome coverage of at least 30 were obtained.
T54 678042-678146 Sentence denotes The reads were mapped to the H37rv reference sequence to identify single nucleotide polymophisms (SNPs).
T55 678147-678429 Sentence denotes Demonstrating the high quality of the data obtained, we were able to identify all polymorphisms in classical resistance genes conferring resistance to first line (isoniazid, rifampicin, ethambutol, pyrazinamide and streptomycin) and second line antibiotics (ofloxacin, capreomycin).
T56 678430-678574 Sentence denotes Interestingly, the emergence of SNPs in resistance genes coincides with the development of phenotypic resistance within the course of treatment.
T57 678575-678721 Sentence denotes In addition, we identified more than 20 SNPs that were acquired by particular isolates in comparison to the parental (first) strain, respectively.
T58 678722-678781 Sentence denotes These are actually being validated by classical sequencing.
T59 678782-678794 Sentence denotes Conclusions:
T60 678795-678909 Sentence denotes Our results reveal a striking diversity in serial patient isolates that cannot be captured by standard genotyping.
T61 678910-679043 Sentence denotes Functional consequences e.g. as additional resistance mechanisms or compensatory evolution are currently under investigation. (PVL) .
T62 679044-679112 Sentence denotes Therefore the existence of two different pathovars was hypothesized.
T63 679113-679336 Sentence denotes We investigated the microevolution and population structure of S.aureus ST121.To elucidate the mechanisms of emergence and epidemic spread we investigated the microevolution by means of single nucleotide polymorphism (SNP).
T64 679337-679479 Sentence denotes SNP discovery was performed by using dHPLC for 117 genetic loci (46839 bp) within an international strain collection of 102 S.aureus isolates.
T65 679480-679571 Sentence denotes We identified 231 bi-allelic polymorphisms (BiP), corresponding to 1 BiP per 0.2 kilobases.
T66 679572-679718 Sentence denotes The 231 bi-allelic polymorphisms were used to construct a minimum spanning tree (MST) which was associated in 86 haplotypes among the 102 strains.
T67 679719-679850 Sentence denotes Results indicate that two subpopulations have evolved into different pathovars that are associated with distinct clinical pictures.
T68 679851-680200 Sentence denotes In addition we could show that much of the population structure of ST121 is local, largely endemic and spread to other countries or continents occur only on rare occasions.Furthermore, and in agreement to prior results in ST5, ST22 and ST239, mapping of spa types onto the MST revealed several spa types whose patterns seemed to reflect homoplasies.
T69 680201-680389 Sentence denotes It was recently demonstrated that MRSA accumulate genomic diversity sufficiently fast to document the pathogen's spatial spread over long distances, e. g., between countries or continents.
T70 680390-680585 Sentence denotes It is currently unclear, however, whether genomic variation may also enable the reconstruction of transmission chains between individual patients, e. g. during an outbreak of hospital infections.
T71 680586-680703 Sentence denotes We have applied Illumina sequencing to compare the full genomes from multiple MRSA isolates from a hospital outbreak.
T72 680704-680786 Sentence denotes Phylogenetic analyses indicated a marked temporal signal in the DNA sequence data.
T73 680787-680995 Sentence denotes An isolate from the index patient (i. e., the presumed starting point of the outbreak) had a basal position in the phylogenetic tree, and isolates from later time points were more distal from the tree's root.
T74 680996-681124 Sentence denotes In cases where multiple isolates from individual patients were available, we observed significant genomic diversity among these.
T75 681125-681320 Sentence denotes However, not each and every patient got infected by a unique MRSA, and MRSA genomes from several patients were indistinguishable or very closely related, suggesting potential transmission events.
T76 681321-681430 Sentence denotes It appears that genome sequencing holds promising potential for tracing MRSA spatial spread at a local scale.
T77 681431-681441 Sentence denotes Objective:
T78 681442-681561 Sentence denotes The identification of risk factors prior to hospitalization is a central element in the prevention and control of MRSA.
T79 681562-681721 Sentence denotes In Germany, screening is recommended based on the guidelines of the Robert-Kochinstitute; yet, the screening intensity is largely varying between institutions.
T80 681722-681832 Sentence denotes In order to introduce a regional program on MRSA control, a detailed knowledge of the MRSA burden is required.
T81 681833-682024 Sentence denotes To this effect, for the first time for an entire German State, we have a conducted a State-wide admission prevalence screening (APS) in 24 hospitals of Saarland during October-November, 2010.
T82 682025-682033 Sentence denotes Methods:
T83 682034-682212 Sentence denotes For screening, combined throat and nasal swabs were taken from patients immediately upon admission using a novel swab (ESwab, Copan, Italy) system for enhanced pathogen recovery.
T84 682213-682410 Sentence denotes Samples were processed by a novel, fully automated, roboter driven specimen processor (WASP, Copan, Italy) and plated on selective biplate media for MRSA and methicillin sensitive S. aureus (MSSA).
T85 682411-682510 Sentence denotes All analyses were performed in the State Hygiene laboratory at the University of Saarland Hospital.
T86 682511-682711 Sentence denotes Of 20,007 specimen tested, 436 (423 throat/nose swabs, 13 wound swabs) were positive (prevalence, 2.2/100 admissions; 13,9% of total S.aureus); prevalence varied between institutions (0/100 -9.0/100).
T87 682712-683009 Sentence denotes These figures are higher than those previously determined in other German APS programs, and they also differ from those previously reported from other central European regions or institutions. spa type 003 (ST225) was most frequent among MRSA followed by spa type 504 (previously rarely reported).
T88 683010-683202 Sentence denotes A detailed risk assessment (multivariate analysis) revealed that previous MRSA history, chronic care, and presence of a skin wound/ulcer were significantly associated with MRSA carrier status.
T89 683203-683356 Sentence denotes The conduct of a large regional APS study allowed for comparative determination of MRSA colonization/infection upon admission in an entire Federal State.
T90 683357-683691 Sentence denotes These results are important to estimate the burden of MRSA cost and disease for the hospitals, for designing a cost-effective continuous screening program for the region, and for developing a rational approach to combat the circle of MRSA introduction from the community and care facilities into hospitals and back into the community.
T91 683692-683882 Sentence denotes As a side effect, a general enhanced attention of general public, institution administrators, and policy makers towards the quality goals of MRSAarNetz was observed. (Schouls et al., 2006) .
T92 683883-683952 Sentence denotes It assesses the variation at 8 different VNTR loci within the genome.
T93 683953-684078 Sentence denotes To date, however, little is known about the rate of MLVA variation and hence its usefulness for the tracing of transmissions.
T94 684079-684388 Sentence denotes We attempted to describe the extent of diversity across epidemiological samples differing in the number of transmissions contained within them; to control for similar genetic background, we only compared strains sharing the same "finetype" (i.e. strains with the same serogroup, PorA, and FetA sequence type).
T95 684389-685167 Sentence denotes Following samples, ordered by the presumed number of transmissions within them, were included: strains from long time carriers (20 strains from 4 carriers); strains from cases with invasive disease and close contacts (11 strains in the proximity of 5 index cases); strains from a cross-sectional study of carriers (36 strains belonging to 9 finetypes); strains from clusters of invasive disease by ET-15 meningococci (26 strains from 6 clusters); strains from finetype-specific spatio-temporal clusters of invasive disease (36 strains from 10 finetypespecific clusters computed with SaTScan version 5.1.3); and strains from randomly distributed cases of invasive disease belonging to the same finetypes as spatio-temporal clusters (72 control strains belonging to 10 finetypes).
T96 685168-685275 Sentence denotes Categorical distances were computed between MLVA-profiles of strains sharing the same finetype and setting.
T97 685276-685397 Sentence denotes Variation of MLVA types within a finetype differed significantly across settings (p<0.001, Kruskal-Wallis test; Figure) .
T98 685398-685578 Sentence denotes No variation of types was observed within long-time carriers and in strains obtained from index cases and close contacts containing zero and one to few transmissions, respectively.
T99 685579-685729 Sentence denotes Interestingly. strains from finetype-specific clusters of disease by ET-15 showed less variation than strains from the cross-sectional carrier sample.
T100 685730-685898 Sentence denotes In contrast, mean distance within strains from spatio-temporal clusters and randomly distributed cases (controls) of invasive disease was considerably higher (Figure) .
T101 685899-686141 Sentence denotes Our preliminary analyses describe the diversity of MLVA types in strains belonging to the same finetype obtained from different epidemiological settings; specifically, variation seems to increase according to presumed number of transmissions.
T102 686142-686312 Sentence denotes To corroborate these findings and to estimate the rate of variation after one transmission, further analyses of long-time carrier-and Index-carrier-samples are under way.
T103 686313-686444 Sentence denotes The prudent use of antibiotics is a major preventive measure to forestall the selection and spread of multidrug-resistant bacteria.
T104 686445-686608 Sentence denotes Here we assessed the density of antibiotic use in hospitals cooperating within a quality network for the prevention and control of healthcare-associated infection.
T105 686609-686616 Sentence denotes Method:
T106 686617-686846 Sentence denotes Within the Dutch-German EurSafety Health-net project (www.eursafety.eu), German hospitals in the EUREGIO Gronau/Enschede provided data on the use of antibiotics in intensive care units (ICUs) and other (non-intensive care) wards.
T107 686847-686936 Sentence denotes Data were assessed separately for the second half-year 2009 and the first half-year 2010.
T108 686937-687170 Sentence denotes The use of antibiotics was reported by hospital pharmacies based on the Anatomical Therapeutic Chemical (ATC) code and in (milli-) gram and transformed into daily defined doses (DDD) as defined by the World Health Organization (WHO).
T109 687171-687239 Sentence denotes Comparability was achieved by calculating DDD/100 patient-days (pd).
T110 687240-687248 Sentence denotes Results:
T111 687249-687339 Sentence denotes Thirty-four regional hospitals within the network provided data on the use of antibiotics.
T112 687340-687463 Sentence denotes The median (mean) density of antibiotic use on non-ICU wards was 48 (56) DDD/100 pd in 2009 and 47 (53) DDD/100 pd in 2010.
T113 687464-687718 Sentence denotes Of note, in the non-ICU setting, extended-spectrum penicillins (J01CA), cephalosporins (J01DB-DE), carbapenems (J01DH) and quinolones (J01M) accounted (2009/2010) for 2.8%/2.4%, 39%/41.7%, 1.9%/2.2% and 11.7%/8% of the total antibiotic use, respectively.
T114 687719-687841 Sentence denotes On 25 ICUs, the median (mean) density of antibiotic use was 108 (110) DDD/100 pd in 2009 and 109 (114) DDD/100 pd in 2010.
T115 687842-688046 Sentence denotes On different ICUs the use of the broad-spectrum antibiotics vancomycin, teicoplanin, linezolid, carbapenems and tigecyclin together represented between 5% and 48% of the total use of antimicrobial agents.
T116 688047-688058 Sentence denotes Discussion:
T117 688059-688210 Sentence denotes The total regional use of antibiotics was comparable to data from other regions published in the German GERMAP 2008 and the Dutch NethMap 2010 reports.
T118 688211-688363 Sentence denotes However, between the hospitals, we observed a broad range of antibiotic use both with respect to the density and the types of antimicrobial agents used.
T119 688364-688535 Sentence denotes Together with regional data on bacterial antibiotic-resistance, this data can be used by the network hospitals for discussions aiming to improve local antibiotic policies.
T120 688536-688692 Sentence denotes Algorithm for empirical treatment for the prudent use of glycopeptides in hematologic patients: a tool in the control of vancomycin resistant enterococci X.
T121 688693-688705 Sentence denotes Zhou* 1 , M.
T122 688706-688720 Sentence denotes Kerkhof 2 , L.
T123 688721-688734 Sentence denotes Span 3 , A.W.
T124 688735-688753 Sentence denotes Friedrich 1 , J.P.
T125 688754-689016 Sentence denotes Arends 1 1 Universitary Medical Centre Groningen, Medical Microbiology, Groningen, Niederlande 2 Universitary Medical Centre Groningen, Epidemiology, Groningen, Niederlande 3 Universitary Medical Centre Groningen, Hematology, Groningen, Niederlande Introduction:
T126 689017-689088 Sentence denotes Enterococcus faecium has become a major cause of nosocomial infections.
T127 689089-689252 Sentence denotes Especially patients with hematologic malignancies are at high risk because of their immunocompromised status and therefore adequate antibiotic therapy is required.
T128 689253-689388 Sentence denotes However for the control of vancomycin resistant enterococci (VRE) antibiotic stewardship with a focus on prudent use becomes necessary.
T129 689389-689583 Sentence denotes The aim of this study was to determine risk factors in order to develop an algorithm for optimal empirical treatment in those patients who are at risk of E. faecium blood stream infection (BSI).
T130 689584-689592 Sentence denotes Methods:
T131 689593-689783 Sentence denotes Retrospectively demographic, clinical and microbiological data in 33 patients with an E. faecium BSI were compared to 66 control patients during a 5-year study period at the hematology unit.
T132 689784-689945 Sentence denotes Multivariate logistic regression was used to explore the independent risk factors in order to develop a prognostic model to determine the risk of E. faecium BSI.
T133 689946-689954 Sentence denotes Results:
T134 689955-690305 Sentence denotes Significant associations of E. faecium BSI were found with age, hospital stay prior to blood culture, duration of hospitalization 1 year before admission, fever prior to blood culture, severity and duration of neutropenia, CRP (C-reactive protein) at time of blood culture withdrawal, colonization with E. faecium prior to blood culture and diarrhea.
T135 690306-690396 Sentence denotes E. faecium BSIs were found associated with more severe disease and higher mortality rates.
T136 690397-690714 Sentence denotes Independent risk factors for E. faecium BSI were age > 59 years (5.47; 1.6-18.2) , hospital stay prior to blood culture > 14 days (4.78; 1.3 -18 .0), fever > 1 day (4.02; 1.3-12.8) , colonization with E. faecium 30 days prior to blood culture (OR 3.83; ) and abdominal pain, diarrhea or neutropenia (5.95; 1.1-31.4) .
T137 690715-690726 Sentence denotes Conclusion:
T138 690727-690853 Sentence denotes Empirical treatment should be considered in those hematologic patients who are at high risk for development of E. faecium BSI.
T139 690854-691030 Sentence denotes Using a prognostic model for risk stratification as designed in this study prudent use of glycopeptides becomes possible and might be helpful controlling further spread of VRE.
T140 691031-691042 Sentence denotes Objectives:
T141 691043-691198 Sentence denotes We report an outbreak of Klebsiella pneumoniae with carbapenem resistance or decreased susceptibility in five patients in a tertiary care hospital in 2010.
T142 691199-691281 Sentence denotes Molecular and phenotypical investigations of the clinical isolates were performed.
T143 691282-691290 Sentence denotes Methods:
T144 691291-691373 Sentence denotes Antibiotic susceptibilities of K. pneumoniae were determined by Vitek2 and E-test.
T145 691374-691494 Sentence denotes DNA from K. pneumoniae isolates were screened by PCR for the presence of KPC, VIM, IMP, NDM-1 and Oxa-48 betalactamases.
T146 691495-691633 Sentence denotes Genetic relatedness was investigated using Enterobacterial Repetitive Intergenic Consensus (ERIC)-and repetitive sequence based (REP)-PCR.
T147 691634-691642 Sentence denotes Results:
T148 691643-691765 Sentence denotes Ertapenem-resistant K. pneumoniae were isolated from rectal swabs and urine samples from five patients during an outbreak.
T149 691766-691968 Sentence denotes Of these five patients, one was only colonized, two exhibited urinary tract infections and two patients were severely ill suffering from sepsis and/or pneumonia due to ertapenem-resistant K. pneumoniae.
T150 691969-692118 Sentence denotes Infected patients were successfully treated with high dose meropenem and gentamicin, and, in case of pneumonia, in addition with inhalative colistin.
T151 692119-692230 Sentence denotes All K. pneumoniae were resistant to penicillins, cephalosporins, ertapenem, fluoroquinolones, and cotrimoxazol.
T152 692231-692348 Sentence denotes Some of the strains exhibited elevated MICs towards meropenem, doripenem or imipenem, but still remained susceptible.
T153 692349-692450 Sentence denotes Oxa-48 was identified by PCR and DNA sequencing in all K. pneumoniae isolates from the five patients.
T154 692451-692578 Sentence denotes ERIC-and REP-PCR analysis of the K. pneumoniae isolates of all patients showed an identical profile suggesting a clonal spread.
T155 692579-692720 Sentence denotes In one of the patients an Oxa-48 producing Escherichia coli was isolated from rectal swabs in addition to the Oxa-48 producing K. pneumoniae.
T156 692721-692875 Sentence denotes Prior to the emergence of the Oxa-48 expressing E. coli a less resistant E. coli with an ESBL phenotype could be isolated several times from this patient.
T157 692876-693030 Sentence denotes This E. coli was negative for the presence of the Oxa-48 gene but showed an identical DNA fingerprint pattern compared with the Oxa-48 expressing E. coli.
T158 693031-693130 Sentence denotes This strongly suggests the in vivo transfer of Oxa-48 from K. pneumoniae to E. coli in the patient.
T159 693131-693142 Sentence denotes Discussion:
T160 693143-693228 Sentence denotes We report the outbreak of Oxa-48 producing K. pneumoniae in a tertiary care hospital.
T161 693229-693369 Sentence denotes Epidemiological, molecular and phenotypical data suggest a horizontal interspecies transfer of Oxa-48 from K. pneumoniae to E. coli in vivo.
T162 693370-693383 Sentence denotes H Background:
T163 693384-693521 Sentence denotes Increasing prevalence of clinical isolates of Vancomycinresistant enterococci (VRE) in patients of an ICU was noticed during 6 to 9/2010.
T164 693522-693668 Sentence denotes Therefore a VRE screening was implemented to determine both the rate of colonisation at ICU admission and the rate of acquisition during ICU stay.
T165 693669-693677 Sentence denotes Methods:
T166 693678-693767 Sentence denotes From 10 to 12/2010 an universal VRE screening was introduced on an interdisciplinary ICU.
T167 693768-693909 Sentence denotes Perianal swabs were taken from all patients at admission and once weekly until ICU discharge and analysed by culture for the presence of VRE.
T168 693910-694122 Sentence denotes Pre-existing VRE carriage was assumed if samples taken within 72 hours after admission were positive; nosocomial acquisition was defined as a positive follow-up screening after a negative result at ICU admission.
T169 694123-694246 Sentence denotes VRE-positive patients were characterised in detail and were monitored for developing VRE infections during hospitalisation.
T170 694247-694255 Sentence denotes Results:
T171 694256-694348 Sentence denotes 737 patients were screened for VRE at least once and 111/737 patients (15.1%) were positive.
T172 694349-694417 Sentence denotes 126/737 (22%) patients could be included in the follow-up screening.
T173 694418-694651 Sentence denotes Because 42/111 (37.8%) of the VRE-positive patients had been hospitalised for more than 72 hours before the first screening swabs, the distinction community vs. nosocomial acquisition could be made only for the remaining 69 patients.
T174 694652-694781 Sentence denotes 32/69 (43.5%) of these patients were VRE carriers at ICU admission, 37/69 patients (56.5%) did acquire VRE during their ICU stay.
T175 694782-694853 Sentence denotes The mean time from admission to first VRE detection was 20 days (range:
T176 694854-694884 Sentence denotes 4-54) in the nosocomial cases.
T177 694885-694967 Sentence denotes Interestingly, none of the VREpositive patients was previously known to carry VRE.
T178 694968-695147 Sentence denotes A total of 37 (33.3%) patients had been treated with Vancomycin prior to VRE detection; 8 patients (7.2%) were known to be colonised/infected with Methicillin resistant S. aureus.
T179 695148-695313 Sentence denotes Only 6/111 of VRE positive patients (5.4%) developed a definite VRE infection (3 bloodstream and 3 abdominal infections); 4 patients had an asymptomatic bacteriuria.
T180 695314-695416 Sentence denotes Mortality in VRE patients was 14.4% (14/111) regardless to their VRE infection or colonisation status.
T181 695417-695429 Sentence denotes Conclusions:
T182 695430-695567 Sentence denotes Universal screening revealed a substantial VRE positivity rate of 15.1% including a high proportion of nosocomial acquired cases (56.5%).
T183 695568-695649 Sentence denotes Reasons for the high VRE prevalence may be the previous Vancomycin therapy (33%).
T184 695650-695708 Sentence denotes The clinical significance of VRE detection was low (5.4%).
T185 695709-695814 Sentence denotes Neither did colonisation predict consecutive infection, nor was the VRE status associated with mortality.
T186 695815-695905 Sentence denotes Therefore, we did not implement a universal VRE screening subsequent to this study period.
T187 695906-696037 Sentence denotes Moreover, concerning infection control standard contact isolation precautions without single room isolation seemed to be justified.
T188 696038-696185 Sentence denotes Further studies are warranted to determine the role of the environmental contamination especially in a setting of moderate hand hygiene compliance.
T189 696186-696188 Sentence denotes E.
T190 696189-696203 Sentence denotes Köster* 1 , E.
T191 696204-696214 Sentence denotes Ott 1 , D.
T192 696215-696228 Sentence denotes Sohr 2 , I.F.
T193 696229-696281 Sentence denotes Chaberny 1 1 Hannover Medical School, Inst. for Med.
T194 696282-696409 Sentence denotes Microbiology and Hospital Epidemiology, Hannover, Inst. for Hygiene and Environmental Medicine, Berlin, Deutschland Background:
T195 696410-696496 Sentence denotes A routine surveillance for MRSA was established since 2001 at Hannover Medical School.
T196 696497-696599 Sentence denotes In order to identify nosocomial transmission episodes molecular protein A (spa)-typing was introduced.
T197 696600-696757 Sentence denotes Method: MRSA infections of all inpatients were analysed according to CDC definitions and were classified into nosocomial or imported cases from 2004 to 2007.
T198 696758-696908 Sentence denotes Sequencing of the repeat region of the Staphylococcus protein A gene (spa typing) were performed from microbiological isolates of all MRSA infections.
T199 696909-696917 Sentence denotes Results:
T200 696918-696996 Sentence denotes In total 2,460 MRSA cases including 358 (14.6%) MRSA infections were detected.
T201 696997-697183 Sentence denotes Surgical site infections were the most frequently type of MRSA infection (64.8%) followed by primary bloodstream infection (12.0%), pneumonia (11.7%) and urinary tract infection (11.5%).
T202 697184-697252 Sentence denotes In addition, secondary bloodstream infections (10.6%) were detected.
T203 697253-697347 Sentence denotes A significant reduction of MRSA infections was observed from 2004 to 2006 (OR 0.363; p<0.001).
T204 697348-697394 Sentence denotes The spa typing revealed 49 distinct spa types.
T205 697395-697562 Sentence denotes The spa type t032 (so called "Barnimer" strain) was the predominant strain type (68.4%) followed by t003 (4.5%), t020 (2.8%), t456 (2.5%), t004 (2.2%) and t022 (2.2%).
T206 697563-697702 Sentence denotes The proportion of infections with secondary bloodstream infections were significantly associated with the spa type t003 (OR 6.75; p=0.004).
T207 697703-697801 Sentence denotes The spa type t032 was significantly less associated to secondary bloodstream infections (p=0.041).
T208 697802-697813 Sentence denotes Conclusion:
T209 697814-697940 Sentence denotes The molecular spa typing provided additional information to the epidemiological associations during routine MRSA surveillance.
T210 697941-698026 Sentence denotes The spa type t003 seemed to be a more invasive MRSA strain contrary to spa type t032.
T211 698027-698084 Sentence denotes Further studies were needed to verify these observations.
T212 698085-698087 Sentence denotes T.
T213 698088-698104 Sentence denotes Juretzek* 1 , M.
T214 698105-698126 Sentence denotes Armengol Porta 1 , A.
T215 698127-698141 Sentence denotes Bühling 1 , R.
T216 698142-698156 Sentence denotes Haubold 1 , M.
T217 698157-698169 Sentence denotes Pohle 1 , F.
T218 698170-698183 Sentence denotes Schwab 1 , W.
T219 698184-698206 Sentence denotes Bär 1 1 Inst. für med.
T220 698207-698399 Sentence denotes Mikrobiologie und Krankenhaushygiene, Cottbus, Deutschland During the last years the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infections in german hospitals has raised.
T221 698400-698644 Sentence denotes In order to know the impact of MRSA at the hospital admission and to develop strategies to reduce the risk of spread of MRSA inside and outside the hospitals an epidemiological study in seven german hospitals of South Brandenburg was conducted.
T222 698645-698781 Sentence denotes We evaluated the incidence of MRSA carriage at hospital admission and we investigated some risk factors associated to MRSA colonisation.
T223 698782-698876 Sentence denotes Moreover the epidemiological distribution and characterisation of MRSA genotypes was analyzed.
T224 698877-698885 Sentence denotes Methods:
T225 698886-699022 Sentence denotes Over a two month period onset patients who accepted the study from all clinical wards were asked for risk factors and screened for MRSA.
T226 699023-699120 Sentence denotes Nasal and wounds swabs were plated on MRSA chromogenic selective agar and incubated for 48 hours.
T227 699121-699252 Sentence denotes Suspected MRSA colonies were confirmed by rapid tests (catalase, coagulase) and an antimicrobial susceptibility test was performed.
T228 699253-699616 Sentence denotes The assignation of the MRSA isolates to the different clonal complexes was performed by analyzing their gene pattern including virulence, adhesion, resistance and other factors with DNA chip technology (Alere/Clondiag Jena represents the predominant multidrug-resistant organism and is a wellknown causative agent of healthcare-associated infections in hospitals.
T229 699617-699717 Sentence denotes However, only few data about its overall prevalence in certain geographical areas are yet available.
T230 699718-699729 Sentence denotes Objectives:
T231 699730-699903 Sentence denotes To determine the prevalence of MRSA including the proportion of Panton-Valentine Leukocidin (PVL) positive strains among inpatients in the rural district Hannover (Germany).
T232 699904-700000 Sentence denotes In addition we checked for patient subgroups that may be at an increased risk for MRSA carriage.
T233 700001-700008 Sentence denotes Method:
T234 700009-700209 Sentence denotes A point-prevalence study was conducted in 17 hospitals from October 1, 2009 through March 31, 2010 Inpatients were screened by cultures from nose, throat and broken skin and patient data was recorded.
T235 700210-700303 Sentence denotes MRSA isolates were tested for PVL and were analysed by staphylococcal protein A (spa) typing.
T236 700304-700378 Sentence denotes Results: MRSA was isolated from 118 (3.9%) of 3,013 consenting inpatients.
T237 700379-700447 Sentence denotes The MRSA prevalence n different hospitals ranged from 0.8% to 12.5%.
T238 700448-700550 Sentence denotes 42% of all identified MRSA carriers were detected on internal medicine wards and 30% on surgery wards.
T239 700551-700596 Sentence denotes 92 (78%) were newly identified MRSA carriers.
T240 700597-700647 Sentence denotes In 3 of 17 hospitals an alert system was in place.
T241 700648-700694 Sentence denotes The spa typing revealed 26 distinct spa types.
T242 700695-700920 Sentence denotes The spa type t032 (so called "Barnimer" strain) was the predominant strain type (67%) followed by t003 (3%), t011 (3%), t020 (3%) and t002 (2% As commonly known, MRSA is one of the most important hospital accquired pathogens.
T243 700921-701126 Sentence denotes Within healthcare institutions, Methicillin-resistant Staphylococcus aureus (MRSA) can be transmitted from patient to patient or via healthcare workers' (HCW) hands, clothes, equipment and the environment.
T244 701127-701289 Sentence denotes Few studies have evaluated the prevalence of MRSA colonization among HCW outside outbreak situations and the importance of staff as a real source of transmission.
T245 701290-701441 Sentence denotes Therefore, this investigation focuses on MRSA among hospital employees and evaluates the efficency of decolonisation therapy among health-care workers.
T246 701442-701645 Sentence denotes 726 staff members (83 physicans, 514 nurses and 109 persons without direct contact to patients) out of nine hospitals within the German part of the Dutch-German EUREGIO participated in the investigation.
T247 701646-701739 Sentence denotes Nasalpharyngeal swabs were taken from each staff member on Mondays and subsequent Wednesdays.
T248 701740-701820 Sentence denotes Risk factors for MRSA carriership were determined by standardized questionnaire.
T249 701821-701924 Sentence denotes Both swabs were microbiologically processed and spa-typed in order to exclude or determine carriership.
T250 701925-702007 Sentence denotes Only stable MRSA carriers with two positive swabs underwent eradication treatment.
T251 702008-702081 Sentence denotes Additionally, the skin of all stable MRSA positive personnel was swabbed.
T252 702082-702325 Sentence denotes In case of skin-colonization participants were released from work until successful MRSA-eradication (verified two days after the end of antimicrobial therapy); otherwise only the first two days of eradication therapy had to take place at home.
T253 702326-702406 Sentence denotes Long-term eradication success was controlled one or six month after eradication.
T254 702407-702483 Sentence denotes 33 out of 726 (4,5 %) staff members were MRSA-positive in al least one swab.
T255 702484-702531 Sentence denotes 23 of them (3,2 %) were positive in both swabs.
T256 702532-702674 Sentence denotes Among physicians, MRSA prevalence was lower (1,2%) than in nurses (3,7 %) and other staff members without direct contact to patients (2,8 %) .
T257 702675-702791 Sentence denotes A positive MRSA anamnesis and acne were identified to be statistical significant risk factors for MRSA colonisation.
T258 702792-702882 Sentence denotes 90 % of the stable MRSA carriers underwent decolonization therapy parallel to normal work.
T259 702883-703007 Sentence denotes The therapy was successful after the first eradication cycle in 100 % of the cases and could be verified in follow-up swaps.
T260 703008-703047 Sentence denotes Nine different spa-types were detected.
T261 703048-703203 Sentence denotes The most frequent ones were typical hospital-acquired ha-MRSA t003 and t032 whereas lifestockassociated la-MRSA were not found at all under staff carriers.
T262 703204-703384 Sentence denotes The study showed that hospital staff is frequently colonized with typical hospital-accquired MRSA outside outbreak situations and MRSA prevalence differs among professional groups.
T263 703385-703482 Sentence denotes Notable, livestock-associated MRSA were not found in the EUREGIO where a high pig density exists.
T264 703483-703596 Sentence denotes Importantly, all HCW were successfully decolonized no matter if they were eradicated at home or parallel to work.
T265 703597-703726 Sentence denotes Decolonisation parallel to work reduced the days of inability to work among HCW in comparison to complete decolonisation at home.
T266 703727-703737 Sentence denotes Objective:
T267 703738-703909 Sentence denotes Rates of VREF remain constant or are declining in Germany in recent years, but molecular analyses suggest a genotype switch from vanAtype to vanB-type among hospital VREF.
T268 703910-704082 Sentence denotes We investigated an outbreak with VanB type VREF involving 74 neonate patients from an intensive care unit of a German hospital to resolve emergence and spread of VanB VREF.
T269 704083-704093 Sentence denotes Materials:
T270 704094-704179 Sentence denotes All samples were taken at a single neonatal ICU during a 9months period in 2008/2009.
T271 704180-704239 Sentence denotes Enterococci were isolated non-selectively on Columbia Agar.
T272 704240-704277 Sentence denotes Species was identified biochemically.
T273 704278-704367 Sentence denotes Vancomycin MICs were determined by E-test and the genotype by a PCR-hybridization method.
T274 704368-704615 Sentence denotes Altogether 75 E. faecium were sent to the Focal Lab for Enterococci at the RKI for further analyses including confirmation of the resistance phenotypes and genotypes, molecular typing and characterization (PFGE, MLST, plasmid typing by rep genes).
T275 704616-704624 Sentence denotes Results.
T276 704625-704673 Sentence denotes We confirmed 75 VREF of which 73 possessed vanB.
T277 704674-704831 Sentence denotes Of the 73 vanB VREF 36 (49%) were vancomycin-resistant according to EUCAST criteria (MIC >4 mg/L) and only 5 (7%) when using the CLSI breakpoint (=>32 mg/L).
T278 704832-705033 Sentence denotes Additional resistances included ampicillin (n=73; 100%), high-level (HL) ciprofloxacin (67; 92%); HL gentamicin (61; 84%), HL streptomycin (63; 86%), erythromycin (68; 93%), and tetracycline (10; 14%).
T279 705034-705130 Sentence denotes All were susceptible to linezolid and daptomycin; one was resistant to tigecycline (MIC= 1mg/L).
T280 705131-705236 Sentence denotes PFGE analysis revealed one major clone prevalent over the entire study period represented by 56 isolates.
T281 705237-705298 Sentence denotes Another 17 vanB VREF and 2 vanA VREF were clonally different.
T282 705299-705429 Sentence denotes The major outbreak strain revealed MLST type ST192, a hospital-associated strain type widely prevalent in Germany in recent years.
T283 705430-705529 Sentence denotes Plasmid typing identified prevalence of pLG1-, pRE25-and pRUM-like plasmids in the outbreak strain.
T284 705530-705615 Sentence denotes Preliminary molecular characterization revealed a chromosomally located vanB cluster.
T285 705616-705627 Sentence denotes Discussion:
T286 705628-705754 Sentence denotes An increasing number of vanB VREF involved in outbreaks is also reported in other European countries (Sweden, France, Poland).
T287 705755-706111 Sentence denotes A molecular analysis of a recent outbreak strain from Sweden revealed vanBtype ST192 VREF associated with a distribution of a specific vanB2-Tn5382 cluster on pRUM-like plasmids; in our case the corresponding vanB cluster was not plasmid-located but chromosomally-borne although ST192 isolates contained several plasmids including also a pRUM-like variant.
T288 706112-706123 Sentence denotes Conclusion:
T289 706124-706238 Sentence denotes Reliable detection of low level vancomycin expression in vanB-type VREF is a challenge in the clinical laboratory.
T290 706239-706390 Sentence denotes Its delayed detection or unnoticed appearance may support further spread of vanB VREF and underestimate its true prevalence among the clinical setting.
T291 706391-706564 Sentence denotes Molecular (screening) methods to detect these strains are here superior to phenotypic assays including also screening agar plates with a comparably high vancomycin inoculum.
T292 706565-706715 Sentence denotes Emerging multiresistant Pseudomonas aeruginosa and its hidden reservoirs in hospitals -molecular studies of a nosocomial outbreak in a high risk area.
T293 706716-706815 Sentence denotes Multiresistant Pseudomonas (P.) aeruginosa is an increasing problem in health care facilities [1] .
T294 706816-706934 Sentence denotes As one of the most important inhabitants of biofilms this Nonfermenter can consist in different reservoirs in clinics.
T295 706935-707064 Sentence denotes The siphon traps of washing basins are colonized by different germs and may be an underestimated source of pathogen distribution.
T296 707065-707358 Sentence denotes Long-term surveillance from 2007 to 2009 within a hemato-oncological setting with 24 isolation units for patients with allogenic stem cell transplantation revealed the association of emerging multiresistant P. spp. infections with detection of suspicious strains in siphons of patients´ rooms.
T297 707359-707545 Sentence denotes All affected patients received same antibiotic prophylaxis and underwent same treatment with multiple daily throat rinsing against oropharyngeal mucositis in front of the washing basins.
T298 707546-707642 Sentence denotes During this procedure it was very possible that the patients inhaled aerosols from siphon water.
T299 707643-707764 Sentence denotes Responsible for this outbreak two strains could be detected by RAPD-PCR and specific P. aeruginosa microarray genotyping.
T300 707765-707893 Sentence denotes Comparison of outbreak strains with strains isolated from siphons of patients´ rooms showed same antibiotic resistance patterns.
T301 707894-708058 Sentence denotes Resistotype 1 had broad resistance to common antibiotics with susceptibility only to Polymyxin B, whereas resistotype 2 was susceptible to Polymyxin B and Amikacin.
T302 708059-708150 Sentence denotes All Carbapenem-resistant strains were screened positive for VIM-Metallobetalactamases [2] .
T303 708151-708286 Sentence denotes The outbreak could be stopped by implementation of so called Hygiene siphons at the washing basin in all of the 24 isolation units (Fa.
T304 708287-708298 Sentence denotes BIOREC -Dr.
T305 708299-708350 Sentence denotes SCHLUTTIG Medizinischer Apparatebau, Dresden) [3] .
T306 708351-708362 Sentence denotes Conclusion:
T307 708363-708524 Sentence denotes Here we report the molecular studies of an outbreak of multiresistant P. spp. in a hemato-oncological ward with optimal conditions concerning hygiene management.
T308 708525-708714 Sentence denotes Even with maximum barrier infection control measures undertaken to avoid nosocomial infections hidden reservoirs like siphon traps of washing basins can be the source of pathogen outbreaks.
T309 708715-708853 Sentence denotes The installation of Hygiene siphons in isolation units for high risk patients can help to reduce the distribution of multiresistant germs.
T310 708854-709024 Sentence denotes Early molecular-epidemiological analysis is urgently needed to verify outbreak situations and is essential to implement specific and effective infection control measures.
T311 709025-709138 Sentence denotes The gastrointestinal tract harbours a complex microbial ecosystem, engaged in continuous crosstalk with the host.
T312 709139-709374 Sentence denotes The balanced relationship between intestinal epithelial cells (IECs) and gut microbes can be disturbed, resulting in the activation of the mucosal immune system which contributes to the development of inflammatory bowel diseases (IBD).
T313 709375-709527 Sentence denotes We used a T84 cell culture model to analyze the cellular responses of IECs co-incubated with the Gram-negativ probiotic strain E.coli Nissle 1917 (EcN).
T314 709528-709730 Sentence denotes As IBD may develop after defects of barrier function and based on our microarray data obtained with co-incubated IECs we focussed on proteins (ZO-2, PAR-3, PAR-6) of the apical junctional complex (AJC).
T315 709731-709838 Sentence denotes We used transepithelial resistance (TER) of T84 cells as a read-out to monitor epithelial barrier function.
T316 709839-710005 Sentence denotes By microRNA-profiling of IECs after co-incubation with probiotics we identified microRNAs (miRNAs) that correspond to proteins of the AJC (qRT-PCR, Western-Blotting).
T317 710006-710102 Sentence denotes Barrier function was enforced by probiotic E.coli and disrupted after EPEC infection (E2348/69).
T318 710103-710245 Sentence denotes Even after barrier disruption by EPEC, barrier function could be restored by incubation with probiotic EcN as monitored by an increase in TER.
T319 710246-710410 Sentence denotes We confirmed the participation of microRNAs (miRNAs) in the regulation of the AJC proteins by transfecting T84 cells with miRNAs, miRNA-inhibitors and miRNA-mimics.
T320 710411-710532 Sentence denotes These studies show that miRNAs are involved in strengthening probiotic effects and the barrier disrupting effect of EPEC.
T321 710533-710660 Sentence denotes This study revealed molecular responses of IECs specifically induced by the Gram-negative probiotic E. coli strain Nissle 1917.
T322 710661-710813 Sentence denotes Further insight into the molecular targets of specific miRNAs might foster the development of new strategies for treatment of gastrointestinal diseases.
T323 710814-710909 Sentence denotes Effects of pathogenic, and probiotic E. coli on Immune Cells at the gastrointestinal barrier H.
T324 710910-710923 Sentence denotes Salia* 1 , N.
T325 710924-710939 Sentence denotes Kramko 1 , M.A.
T326 710940-711126 Sentence denotes Schmidt 1 1 Zentrum für Molekularbiologie der Entzündung, Infektiologie, Münster, Deutschland The gut immune system keeps the microbiota of the gastrointestinal tract under surveillance.
T327 711127-711330 Sentence denotes Besides commensal and probiotic microorganisms there are also bacterial pathogens such as enteropathogenic E.coli that can cause diarrhea by exploitation of host cells and subversion of defense measures.
T328 711331-711442 Sentence denotes How these pathogens interact with and subvert (immune) cells of the intestinal barrier is not fully understood.
T329 711443-711635 Sentence denotes In this study we address the effects of the enteropathogenic E. coli (EPEC) strain E2348/69 and of the probiotic E. coli Nissle 1917 (EcN) strain on human PMBC and human monocytic THP-1 cells.
T330 711636-711778 Sentence denotes Dendritic cells derived from human PBMC and monocytic THP-1 cells were infected with EPEC wt, EPEC mutant strains or the probiotic strain EcN.
T331 711779-711856 Sentence denotes The cytotoxicity of these bacteria was evaluated by analyzing cell viability.
T332 711857-712077 Sentence denotes In addition, morphological changes within the actin network were analyzed via confocal immunofluorescence microscopy and influences on the expression of proteins involved in inflammation via qRT-PCR and Western Blotting.
T333 712078-712216 Sentence denotes In contrast to infections of THP-1 cells with E. coli C600 or EcN, EPEC infection exhibit a dramatic increase in cytotoxicity (threefold).
T334 712217-712425 Sentence denotes Analysis of the actin cytoskeleton of THP-1 cells revealed that EPEC wt as well as effector protein deletion mutants are able to induce pedestals with classic actin accumulation at bacterial attachment sites.
T335 712426-712500 Sentence denotes This is not observed with an EPEC mutant lacking the effector protein Tir.
T336 712501-712629 Sentence denotes Interestingly, pedestal formation can be faintly detected also with mutants without functional type III secretion system (T3SS).
T337 712630-712830 Sentence denotes Concerning the expression of cytokines we observed an upregulation of the anti-inflammatory IL-10 upon infection of DC or THP-1 cells with EcN (two and fivefold) in contrast to infection with EPEC wt.
T338 712831-713037 Sentence denotes By analyzing the influence of EPEC and EcN on the NFkB signal cascade in THP-1 cells we observed an activation of the pathway upon EcN and E. coli C600 coincubation but an inhibition upon EPEC coincubation.
T339 713038-713245 Sentence denotes The results show that EPEC exhibit destructive activity on immune cells where virulence factors such as the T3SS-secreted effector proteins enable the bacteria to specifically target host cellular processes.
T340 713246-713433 Sentence denotes The comparison of effects of pathogenic and probiotic bacteria on immune cells might reveal further insights into the molecular mechanisms that are impaired in gastrointestinal disorders.
T341 713434-713535 Sentence denotes Role of dendritic cell activation as well as Toll-like receptor 2 and 4 expression while DSS colitis.
T342 713536-713628 Sentence denotes Hosts live normally in symbiosis with their intestinal microbiota due to a long coevolution.
T343 713629-713786 Sentence denotes Somehow this tolerance is abolished in IBD patients, which leads to a strong and long lasting inflammation irregularly interrupted by short remission phases.
T344 713787-713957 Sentence denotes In order to investigate the role of dendritic cells and their TLR2 and TLR4 expression while acute phase inflammation the Dextran sodium sulfate (DSS) model was employed.
T345 713958-714016 Sentence denotes Therefore DSS was administered to C57BL/6 mice for 7 days.
T346 714017-714097 Sentence denotes Mice displayed significant weight loss and an increasing disease activity index.
T347 714098-714207 Sentence denotes Lamina propria (LP) and mesenteric lymph node (MLN) DCs were isolated of untreated mock and DSS treated mice.
T348 714208-714439 Sentence denotes LP DCs of DSS treated mice owned significantly increased levels of MHCII, CD40, CD80 and CD86 representing a more activated DC state as compared to DCs of mock mice, whereas MLN DCs did not differ concerning their activation state.
T349 714440-714553 Sentence denotes Furthermore DSS mice featured higher levels of myeloid DCs in the LP as well as in the MLN compared to mock mice.
T350 714554-714719 Sentence denotes As well had DSS mice a significantly increased percentage of CD103 + DCs in the LP in comparison to mock mice, whereas amounts of CD103 + DCs in MLN were vice versa.
T351 714720-714843 Sentence denotes Surprisingly LP and MLN DCs of DSS mice expressed significantly higher amounts of TLR2 and TLR4 in comparison to mock mice.
T352 714844-715011 Sentence denotes Increased appearance of CD103 + DCs and higher amounts of TLR2 and TLR4 are likely to be a counter regulation of the host in order to suppress developing inflammation.
T353 715012-715017 Sentence denotes H.-P.
T354 715018-715030 Sentence denotes Horz* 1 , T.
T355 715031-715044 Sentence denotes Küpper 2 , M.
T356 715045-715060 Sentence denotes Hettlich 2 , A.
T357 715061-715074 Sentence denotes Introduction:
T358 715075-715231 Sentence denotes The bacterial inventory of the human oral cavity is already fairly comprehensive and composed of about 800 different species many of which are uncultivated.
T359 715232-715354 Sentence denotes However, the dynamics and variability of the oral microflora in relation to health or disease are still poorly understood.
T360 715355-715544 Sentence denotes Our aim was to test the hypothesis, that the development of oral symptoms after multiple stressing of the immunosystem is related to the composition and diversity of the initial microflora.
T361 715545-715778 Sentence denotes Material and Methods As a model system, we analyzed the oral microbiota from 58 healthy individuals during a challenging expedition in remote regions of the Himalayas (implying hygiene, diet, temperature, physical and mental stress).
T362 715779-715906 Sentence denotes Plaque samples were taken at begin (Bhulebhule) and destination (3,000 meter difference in altitude) seven days later (Manang).
T363 715907-716041 Sentence denotes Twentyeight individuals remained symptom-free (Group I) while 30 participants developed dental problems, mostly gingivitis (Group II).
T364 716042-716209 Sentence denotes The oral microbiota was monitored via terminal restriction fragment length polymorphism (T-RFLP) analysis of amplified 16S rRNA-genes directly from the plaque samples.
T365 716210-716392 Sentence denotes Variation in T-RFLP datasets were analyzed based on the Additive Main Effects and Multiplicative Interactions (AMMI) model using the T-Rex software package (http://trex.biohpc.org/).
T366 716393-716401 Sentence denotes Results:
T367 716402-716588 Sentence denotes Variation from T-RF main effects was at least 95% indicating that the majority of variation in T-RFLP profiles was due to inherent differences in microbial communities among individuals.
T368 716589-716858 Sentence denotes However, when comparing Group I and II an interaction signal of about 3% was consistently observed (under varying analytical parameters) while this was not the case when the two time points of sampling (i.e. sampling at start and end of the trekkingtour) were compared.
T369 716859-717083 Sentence denotes Likewise the Shannon-Weaver-index of diversity and the Jaccard-index of similarity indicated, that Group I and II differed significantly in the heterogeneity of their oral microbial communities at begin of the trekking tour.
T370 717084-717233 Sentence denotes Based on the Human Oral Microbiome Database (HOMD) distinct bacterial taxa could be elucidated as key-components explaining the observed differences.
T371 717234-717245 Sentence denotes Discussion:
T372 717246-717545 Sentence denotes The data indicate that there is a discernable relationship between susceptibility to clinical symptoms and the indigenous, pre-formed oral microbiota which means that the initial composition of the oral ecosystem determines whether or not symptoms develop upon exposure to various stress-parameters.
T373 717546-717645 Sentence denotes Role of I B in induction of semimature dentritic cells and maintenance of intestinal homeostasis C.
T374 717646-717661 Sentence denotes Klameth* 1 , K.
T375 717662-717684 Sentence denotes Schulze-Osthoff 2 , D.
T376 717685-717702 Sentence denotes Hildebrand 2 , K.
T377 717703-717820 Sentence denotes Bode 3 I B is a member of the I B protein family and a recently described regulator of the transcription factor NF B.
T378 717821-717918 Sentence denotes But unlike all other I Bs, I B seems not only to be an inhibitor but more of a modulator of NF B.
T379 717919-718076 Sentence denotes While the production of the proinflammatory cytokine TNF-is inhibited in presence of I B , other genes like IL-6, IL-12p40 and GMCSF are highly up regulated.
T380 718077-718236 Sentence denotes We hypothesize that I B at least partially contributes to DC semimaturation, as this process was shown to be dependent on presence of IL-6 and absence of TNF-.
T381 718237-718344 Sentence denotes Furthermore this DC differentiation type promotes rather tolerogenic than proinflammatory T-cell responses.
T382 718345-718433 Sentence denotes Due to this fact I B might play a crucial role in maintenance of intestinal homeostasis.
T383 718434-718715 Sentence denotes To clarify the role of which might lead to the development of new strategies in treatment of inflammatory bowl diseases we analysed and compared in a first step the I B mRNA and protein expression in naïve, semimature and mature DC using quantitative PCR and western blot analysis.
T384 718716-718802 Sentence denotes Additionally the expression of I B in restimulation of semimature DC was investigated.
T385 718803-718936 Sentence denotes Future experiment will involve I B deficient mice, for generation of BMDC and the analysis of the impact of I B on acute DSS colitis.
T1 718937-718939 Sentence denotes A.
T2 718940-718957 Sentence denotes Zschüttig* 1 , C.
T3 718958-718973 Sentence denotes Eichhorn 1 , S.
T4 718974-718987 Sentence denotes Meltke 1 , K.
T5 718988-719005 Sentence denotes Zimmermann 2 , T.
T6 719006-719205 Sentence denotes Wassenaar 3 The probiotic drug Symbioflor 2 (E. coli DSM 17252) contains six Escherichia coli genotypes named E. coli G1/2, G3/10, G4/9, G5, G6/7 and G8, all functionally representing one population.
T7 719206-719336 Sentence denotes Symbioflor 2 is a liquid preparation containing live E. coli cells, which were recently classified as biosafety level 1 organisms.
T8 719337-719492 Sentence denotes The product is well established as a probiotic drug for years and it is widely used especially for the treatment of Colon irritable in adults and children.
T9 719493-719657 Sentence denotes The genomes of all six genotypes have been sequenced by pyrosequencing and sequencing by synthesis technology and overall twelve different plasmids were identified.
T10 719658-719755 Sentence denotes Whole-genome comparison of the six genotypes allow a subdivision into four different genomotypes.
T11 719756-719858 Sentence denotes We could also show that all Symbioflor 2 E. coli clearly belong to the cluster of apathogenic E. coli.
T12 719859-720047 Sentence denotes However, it is largely unknown which E. coli genotype is responsible for the products probiotic character and which genes and proteins are involved in the development of a probiotic trait.
T13 720048-720184 Sentence denotes E. coli G3/10 has already attracted interest due to its ability of inducing defensin production in cell culture and in human volunteers.
T14 720185-720457 Sentence denotes Using an in vitro adherence assay with human intestinal epithelial cells we could further show that E. coli G3/10 significantly decreases the adhesion of enteropathogenic E. coli strain E2348/69 (EPEC) to host cells, comparable to the probiotic strain E. coli Nissle 1917.
T15 720458-720573 Sentence denotes The EPEC adherence assay is a suitable surrogate test system to evaluate probiotic action of a given microorganism.
T16 720574-720705 Sentence denotes The sequenced genomes together with in vitro data will help us to gain further insight into the probiotic behavior of Symbioflor 2.
T17 720706-720835 Sentence denotes Oral biofilms are supposed to be involved in a number of oral diseases such as plaque formation, dental caries and periodontitis.
T18 720836-720964 Sentence denotes Thus, a more detailed knowledge of oral microbiota in biofilm formation is important for a better understanding of pathogenesis.
T19 720965-721210 Sentence denotes In the present pilot study the composition of human microbiota during intraoral biofilm formation was analysed in 7 healthy volunteers (females, aged 24 to 26) using bovine dental enamel slabs harvested after 3 and 30 minutes, 2, 6 and 24 hours.
T20 721211-721277 Sentence denotes In parallel, interproximal dental plaque and saliva were analysed.
T21 721278-721480 Sentence denotes The microbial communities were characterized by T-RFLP (Terminal Restriction Fragment Length Polymorphism) analysis of 16S rRNA genes applying three different restriction enzymes, HaeIII, MspI and HhaI.
T22 721481-721685 Sentence denotes T-RFLP signals of the phylogenetic groups were assigned to sequence data obtained by DGGE (Denaturing Gradient Gel Electrophoresis) analysis or in silico analysis of published sequence data, respectively.
T23 721686-721918 Sentence denotes Initially, 70 samples were investigated by DGGE revealing that the variety of bands was restricted to few signals in all dental biofilm samples whereas a higher diversity of bands was found in saliva and proximal interdental plaque.
T24 721919-722046 Sentence denotes Semiquantitative analysis by T-RFLP confirmed lower microbial diversity in dental biofilms compared to other oral compartments.
T25 722047-722259 Sentence denotes 55 specific HaeIII T-RF signals could be assigned to various phylogenetic groups and we could show that microbial community structure was different between dental biofilms, saliva and interproximal dental plaque.
T26 722260-722378 Sentence denotes The abundance of streptococci was highest in dental biofilms whereas Bacteroidetes were predominantly found in saliva.
T27 722379-722498 Sentence denotes The relative abundance of Veillonella spp. varied during biofilm formation with lowest frequencies in 6 hours biofilms.
T28 722499-722617 Sentence denotes Other phylogenetic groups e.g. the group of Haemophilus/Neisseria were detected with equal frequencies in all samples.
T29 722618-722895 Sentence denotes The similarities in the community structure in different oral compartments were estimated by calculation of similarity index Chao-Jaccard revealing the lowest similarity between microbial communities in saliva and interproximal dental plaque compared to 2 hours dental biofilm.
T30 722896-723019 Sentence denotes The highest similarity was calculated for microbial communities in 6 hours dental biofilms versus 24 hours dental biofilms.
T31 723020-723168 Sentence denotes We conclude that characteristic signatures of microbiota can be found in saliva, interproximal dental plaque and also during oral biofilm formation.
T32 723169-723368 Sentence denotes Culture-independent analysis of microbiota may provide new insights into the microbial diversity in various communities which was shown here for different oral compartments and growing oral biofilms.
T33 723369-723380 Sentence denotes Background:
T34 723381-723485 Sentence denotes Candida albicans, a diploid asexual fungus is a part of the normal flora including the vaginal surfaces.
T35 723486-723554 Sentence denotes In healthy individuals, C. albicans normally does not cause disease.
T36 723555-723749 Sentence denotes However, when the balance of the normal flora is altered e. g. during antibiotic or hormonal therapy, C. albicans can cause painful cutaneous or subcutaneous infections including vulvovaginitis.
T37 723750-723823 Sentence denotes In germany around five million women suffer from candidal vulvovaginitis.
T38 723824-723885 Sentence denotes Lactobacilli belong to the physiological flora of the vagina.
T39 723886-723971 Sentence denotes While some studies show the beneficial effect of lactobacilli on bacterial vaginosis.
T40 723972-724046 Sentence denotes However, the role of lactobacilli in candidal vulvovaginitis is not known.
T41 724047-724145 Sentence denotes One well known pathogenicity factor of candida spp. is the transition from yeast to hyphal growth.
T42 724146-724300 Sentence denotes Therefore we analyzed how far lactobacilli may influence the yeast-hyphal transition has been described as a factor that facilitates the invasion process.
T43 724301-724309 Sentence denotes Methods:
T44 724310-724452 Sentence denotes In total we used five different C. albicans strains which were isolated from women presenting different microbial environment in their vagina.
T45 724453-724605 Sentence denotes The different Candida spp. were incubated with distinct Lactobacillus strains (L. gasseri, L.crispatus und L. jenseni) isolated from the vagina as well.
T46 724606-724797 Sentence denotes We analyzed how far the different Lactobacillus strains influence the viability (XTT_reduction assay) of Candida as well as the expression of hyphae-specific genes (ALS3, HWP1, HYR1) Results:
T47 724798-724927 Sentence denotes Only L. crispatus inhibited the growth of all Candida albicans isolates under study and diminished their viability significantly.
T48 724928-725026 Sentence denotes In contrast L. gasseri led to a significant increase in viability of all Candida albicans strains.
T49 725027-725079 Sentence denotes L. jensenii exhibited no influenec on fungal growth.
T50 725080-725234 Sentence denotes The effect of the different Lactobacillus spp. on yeast to hyphae transition was strongly dependent on the type of the respective Candida albicans strain.
T51 725235-725246 Sentence denotes Conclusion:
T52 725247-725368 Sentence denotes From our results one may conclude that lactobacillus spp. may either favour or inhibit pathogenicity of Candida albicans.
T53 725369-725522 Sentence denotes In addition to genetic predisposition, environmental factors such as commensal bacteria contribute to the development of inflammatory bowl disease (IBD).
T54 725523-725644 Sentence denotes The gut of mammalians is colonised by a complex flora of microorganisms containing 500 -1000 different bacterial species.
T55 725645-725813 Sentence denotes These bacterial populations contribute to the health of the host, among other things, by promoting proper immune systeme development and limiting pathogen colonization.
T56 725814-725983 Sentence denotes Bacteroides vulgatus mpk was shown to have the ability to prevent Colitis, whereas E. coli mpk induces intestinal inflammation in interleukin-2-defiient (IL-2 -/-) mice.
T57 725984-726022 Sentence denotes The mechanism however remains unclear.
T58 726023-726164 Sentence denotes In the current study we analyse the composition of the intestinal microbiota of T-cell transferred Rag1 -/by 454-Seqeuncing of 16S RNA genes.
T59 726165-726401 Sentence denotes With this method we want to reveal differences between (1) the gut microbiota of mice that develop colitis compared to mice that stay healthy and (2) the composition of the intestinal microbiota before and during development of colitis.
T60 726402-726493 Sentence denotes Cystic fibrosis (CF) is a recessive inherited disease caused by mutations of the CFTR gene.
T61 726494-726582 Sentence denotes In CF patients morbidity is severely influenced by chronic polymicrobial lung infection.
T62 726583-726699 Sentence denotes Routine microbiological testing is focused on the detection of few typical pathogens using culture-based techniques.
T63 726700-726784 Sentence denotes Thus, complexity of microbial community in cystic fibrosis lung cannot be displayed.
T64 726785-727031 Sentence denotes However, by the use of new culture-independent techniques unbiased semi-quantitative analysis of complex bacterial communities are now available which could help to better understand the role of microbiota in colonisation and pathogenicity of CF.
T65 727032-727245 Sentence denotes In the present pilot study 51 respiratory samples of 37 CF patients in three different age groups (0 to 10, 11 to 20 and >20 years) were analysed both by culture-based and new culture-independent methods (T-RFLP).
T66 727246-727541 Sentence denotes Corresponding to previous studies, an age-dependent infection rate was found by culture-based analysis for the classical CF related pathogens with highest rates of S. aureus infections in the 0 to 10 year old group and rising P. aeruginosa infection rate in older patients (11-20 and >20 years).
T67 727542-727990 Sentence denotes This finding was supported by the culture-independent analysis of bacterial nucleic acid in respiratory samples using T-RFLP, however, we also found new aspects of respiratory infections by the culture-independent method with increasing rates of anaerobic bacteria in CF patients at older age which potentially contribute to clinical progression of CF lung disease; however, prospective clinical studies are required to confirm this new assumption.
T68 727991-728142 Sentence denotes Culture-independent analysis also provides a quantitative view to the relative abundance of specific pathogens in bacterial communities of CF patients.
T69 728143-728406 Sentence denotes High relative abundance of Staphylococci (24%), Neisseria / Haemophilus (16%), Streptococci (9%), Bacteroidetes (6%) and Veillonella (5%) were found whereas the Pseudomonas species were detected with surprisingly low frequencies in samples (1%) of all age groups.
T70 728407-728668 Sentence denotes We conclude that culture-independent analysis (e.g. T-RFLP) will fundamentally change the current view of polymicrobial lung infection in CF patients due to the unbiased detection of potentially new respiratory pathogens and due to the new quantitative aspects.
T71 728669-728846 Sentence denotes We suppose that very soon culture-independent analysis of the respiratory microbiota could become a new diagnostic standard for optimized diagnostics and therapy of CF patients.
T72 728847-728860 Sentence denotes Introduction:
T73 728861-728938 Sentence denotes Commensals comprise an important component of the micro-ecology of epithelia.
T74 728939-729119 Sentence denotes Yet, opposite to the upper respiratory tract and the gut, the lower respiratory tract including lung has been considered free of foreign microorganisms, so it is vastly unexplored.
T75 729120-729237 Sentence denotes To understand how lung micro-ecology affects host defense and infection, it is essential to identify lung commensals.
T76 729238-729259 Sentence denotes Material and methods:
T77 729260-729428 Sentence denotes We identified bacterial species in sterilely excised murine lungs by combining various culture systems and 16S rDNA analysis using cloning or 454 pyrosequencing method.
T78 729429-729622 Sentence denotes We compared inbred mice maintained under specific-pathogen-free (SPF) or inbred and wild-derived mice of distinct geographical origin from a non-SPF-facility as well as mice caught in the wild.
T79 729623-729705 Sentence denotes To localize bacteria in lungs, Fluorescence in situ Hybridization (FISH) was used.
T80 729706-729833 Sentence denotes Finally, bacterial communities in each group of mice were compared by divergence-based diversity analyses on the species level.
T81 729834-729842 Sentence denotes Results:
T82 729843-729970 Sentence denotes We isolated aerobes and facultative anaerobes including Pasteurellaceae, Enterobacteria and Firmicutes from lungs of Plön mice.
T83 729971-730102 Sentence denotes Additional species including non-cultivatable Betaproteobacteria and Enteric bacteria were identified by 16S rDNA cloning analysis.
T84 730103-730247 Sentence denotes In addition, we confirmed that major species of lung commensal is Betaproteobacteria, or Ralstonia sp. by high-throughput pyrosequencing method.
T85 730248-730387 Sentence denotes Notably, we detected bacterial communities on alveolar epithelia by eubacteria specific FISH in the absence of inflammatory lung pathology.
T86 730388-730643 Sentence denotes Comparison of bacterial community between mouse groups shows that overall composition of Phyla or Family level was similar regardless of mouse origin, but rarefaction curve by diversity or richness measure shows significant differences between each group.
T87 730644-730666 Sentence denotes Conclusion/Discussion:
T88 730667-730759 Sentence denotes We screened mouse lung microbiome by traditional culture method and robust genetic analysis.
T89 730760-730955 Sentence denotes Ultimately, we expect that our lung isolates or major species by non-culture method can be introduced into mouse model experiment for lung commensalism as well as coinfection with lung pathogens.
T90 730956-731117 Sentence denotes Identification of commensal bacteria in murine lungs will pave the path to study influence of respiratory tract microbiome on infection and immunity of the lung.
T91 731118-731200 Sentence denotes The salivary microbiome as an indicator of human-microbe coevolution Introduction:
T92 731201-731324 Sentence denotes Human-associated bacteria hold great potential for shedding light on the history of our ancestors (i.e. migration pattern).
T93 731325-731462 Sentence denotes The concept of human-bacterial co-evolution has been demonstrated for Helicobacter pylori however its analysis requires stomach-biopsies.
T94 731463-731629 Sentence denotes We currently test the hypothesis that oral microbes may function as chronometer of human evolution thereby allowing a fine-scale differentiation of human populations.
T95 731630-731651 Sentence denotes Material and Methods:
T96 731652-731789 Sentence denotes Saliva samples from ten volunteers each from 12 areas world-wide, representing diverse ethnic groups are the initial focus of this study.
T97 731790-732058 Sentence denotes Variations in the 16S-23S rDNA internal transcribed spacer region of Fusobacterium nucleatum and the gdh, (encoding for the glucosedehydrogenase) and the gtf (encoding for the glycosyl-transferase) of the mitis-streptococci are analyzed by culture-independent methods.
T98 732059-732133 Sentence denotes In addition, T-RFLP for whole community genetic profiling will be applied.
T99 732134-732142 Sentence denotes Results:
T100 732143-732314 Sentence denotes The ITS region of F. nucleatum and the gdh of mitis-streptococci could be directly PCR-amplified from saliva DNA-extract using newly designed as well as published primers.
T101 732315-732436 Sentence denotes All ten individuals tested from the countries Germany, Turkey, Poland, Bolivia, and Congo were positive for F. nucleatum.
T102 732437-732522 Sentence denotes Nine of ten individuals from Germany and Congo were tested positive also for the gdh.
T103 732523-732636 Sentence denotes Sequence analysis of ten clones per individual showed a high intra-and inter-individual diversity for both genes.
T104 732637-732830 Sentence denotes Based on phylogenetic tree reconstruction most clusters consisted of sequence types which were representative for multiple countries, however, one country was typically dominating in a cluster.
T105 732831-732938 Sentence denotes In addition, we also found unique clusters explicitly consisting of sequence types specific to one country.
T106 732939-732950 Sentence denotes Discussion:
T107 732951-733072 Sentence denotes Oral microbes found in saliva may exhibit a clonal structure and thus be representative for a distinct geographic region.
T108 733073-733229 Sentence denotes Phylogenetic tools such as UniFrac are necessary to compute the distances within and between hosts in order to discern geographical and ethnic associations.
T109 733230-733575 Sentence denotes Given that saliva is increasingly preferred in sampling humans as a source of DNA for epidemiologic and population genetic studies, the salivary microbiome may hold additional valuable information for providing new perspectives on unsolved human migration patterns -an issue of great social, anthropological, and medical importance Introduction:
T110 733576-733700 Sentence denotes Clinical studies revealed that the probiotic mixture VSL#3 is protective in the context of inflammatory bowel disease (IBD).
T111 733701-733877 Sentence denotes We previously showed that reduced cecal inflammation in VSL#3-fed IL-10 -/mice correlates with reduced levels of interferon-inducible protein (IP)-10 in cecal epithelial cells.
T112 733878-734034 Sentence denotes Mechanistically, IP-10 degradation in intestinal epithelial cells (IEC) was mediated by cell surface proteins of VSL#3derived Lactobacillus paracasei (L.p).
T113 734035-734170 Sentence denotes The aim of the present study was to identify the active probiotic structure triggering the loss of the proinflammatory chemokine IP-10.
T114 734171-734401 Sentence denotes Methods: IEC stimulation experiments, cell-free incubation assays, ileal explant culture and in vivo experiments using a murine ileitis model were used to investigate the probiotic mechanism of L.p and its physiological relevance.
T115 734402-734507 Sentence denotes Chromatographic fractionation and LC-MS/MS analysis were used to identify the active probiotic structure.
T116 734508-734601 Sentence denotes Isogenic mutants were generated to proof the protective activity of the identified structure.
T117 734602-734610 Sentence denotes Results:
T118 734611-734714 Sentence denotes In addition to fixed bacteria also conditioned media of L.p (CM L.p) mediated the loss of IP-10 in IEC.
T119 734715-734938 Sentence denotes Further in vitro experiments including PMSF inhibitor studies revealed that L.p expresses a secreted and cell wall associated serine protease of L.p that selectively and directly degrades IP-10 in an IEC-independent manner.
T120 734939-735024 Sentence denotes Beside IP-10, CM L.p selectively targets an array of pro-inflammatory chemokines (eg.
T121 735025-735092 Sentence denotes I-TAC, Fractalkine), whereas all tested cytokines were not cleaved.
T122 735093-735311 Sentence denotes Furthermore, explant culture experiments using inflamed intestinal tissue from experimental IBD models revealed that established IP-10 gradients in the tissue are also destroyed by this host cell-independent mechanism.
T123 735312-735585 Sentence denotes The physiological relevance of this anti-inflammatory mechanism was proven by intraperitoneal injection of the active bacterial supernatant into an experimental ileitis model, resulting in selective degradation of ileal IP-10, lymphocyte recruitment and ileal inflammation.
T124 735586-735742 Sentence denotes Chromatographic fractionation of the probiotic supernatant and subsequent LC-MS/MS analysis indicated lactocepin of L.p to be the active probiotic protease.
T125 735743-736012 Sentence denotes The generation of a lactocepin-negative mutant from a similarly active, but transformable, human Lactobacillus casei isolate resulted in loss of its IP-10 degrading capacity, finally proving that probiotic lactocepin is the active anti-inflammatory bacterial structure.
T126 736013-736024 Sentence denotes Conclusion:
T127 736025-736234 Sentence denotes The present study reveals the selective degradation of proinflammatory chemokines including IP-10 by probiotic lactocepin as a specific structure-related probiotic mechanism of a clinically relevant probiotic.
T128 736235-736560 Sentence denotes The identification of lactocepin as a probiotic structure could enable a more structure-based evaluation of probiotic bacteria for therapeutical interventions in the context of chronic inflammation PWV05 Flagellin and tcpC are essential factors of the protective effect of E. coli Nissle strain 1917 in DSS-induced colitis S.
T129 736561-736573 Sentence denotes Menz* 1 , K.
T130 736574-736589 Sentence denotes Gronbach 1 , P.
T131 736590-736601 Sentence denotes Adam 2 , A.
T132 736602-736615 Sentence denotes Wieser 3 , S.
T133 736616-736631 Sentence denotes Schubert 3 , U.
T134 736632-736790 Sentence denotes Dobrindt 4 , T. (EcN) is as effective as mesalazine in maintenance of remission in ulcerative colitis and shortens the duration of diarrhea in young children.
T135 736791-736955 Sentence denotes We studied in a preclinical model of acute colitis whether EcN protects from disease and analysed the bacterial mechanism underlying the anti-inflammatory capacity.
T136 736956-737065 Sentence denotes Methods: C57BL/6 and TLR5 -/mice were fed with either EcN or EcN fliC or EcN tcpC and treated with 3, 5% DSS.
T137 737066-737125 Sentence denotes Body weight and disease activity index were assessed daily.
T138 737126-737266 Sentence denotes At the end of the experiment the colon length and weight was measured and inflammation was determined by histological analyses of the colon.
T139 737267-737390 Sentence denotes Furthermore activation and maturation of lamina propria and mesenteric lymph node dendritic cells and T cells was analysed.
T140 737391-737399 Sentence denotes Results:
T141 737400-737516 Sentence denotes In wild type mice E. coli Nissle protects from DSS induced colitis whereas the protection is reduced in TLR5 -/mice.
T142 737517-737653 Sentence denotes In line with this the fliC mutant strain was less effective in protecting the host from disease as compared to the EcN wild type strain.
T143 737654-737805 Sentence denotes However a second bacterial factor tcpC also contributes to the protective effect of EcN as the tcpC mutant strain was not able to protect from disease.
T144 737806-737885 Sentence denotes Administration of the double mutant fliC tcpC of EcN evidences this conclusion.
T145 737886-737898 Sentence denotes Conclusions:
T146 737899-737987 Sentence denotes EcN ameliorates a DSS induced acute colitis via flagellin and the secreted protein tcpC.
T147 737988-738097 Sentence denotes However contribution of further bacterial factors to the anti-inflammatory effect of EcN can not be excluded.
T148 738098-738248 Sentence denotes More than a marine propeller -the flagella of the probiotic Escherichia coli strain Nissle 1917 is also the major adhesin mediating binding to mucin2.
T149 738249-738426 Sentence denotes The flagella of the probiotic E. coli strain Nissle 1917 (EcN) is not just responsible for motility but also for EcN's ability to induce the production of human beta-defensin 2.
T150 738427-738484 Sentence denotes Here, we report a third function of this organell in EcN.
T151 738485-738681 Sentence denotes Ex vivo studies with EcN, its isogenic non-flagellated mutant EcN fliCand the complemented mutant proved the flagella to be essential for efficient adherence to cryosections of human gut biopsies.
T152 738682-738774 Sentence denotes A hyperflagellated EcN variant adhered even more efficient to this cryosections than EcN WT.
T153 738775-738956 Sentence denotes In contrast, in vitro studies showed that neither overexpression nor the lack of flagella has a significant effect on the adhesion to the human epithelial cell lines Caco-2 and T24.
T154 738957-739100 Sentence denotes The observed discrepancy is most likely due to the lack of mucin on these cell lines, whereas mucus might still be present in the cryosections.
T155 739101-739266 Sentence denotes This interpretation is supported by the finding, showing flagella-dependent adhesion of EcN to the human epithelial cell line LS174T, which produces mucins in vitro.
T156 739267-739464 Sentence denotes Binding of the EcN flagella to mucin was further demonstrated by the inhibition of EcN adhesion to cryosections after preincubation of flagellated EcN with mucin2 in a mucin2-dose dependent manner.
T157 739465-739650 Sentence denotes In addition, we were able to show the hyperflagellated EcN variant to adhere much more efficient to mucin2 compared to EcN WT, whereas the non-flagellated mutant EcN fliCadhered barely.
T158 739651-739805 Sentence denotes In contrast, the E. coli strain CFT073 harboring and expressing an identical fliC gene adhered as inefficient to mucin2 as EcN fliC for unknown reason(s).
T159 739806-739930 Sentence denotes Finally, electron microscopic examination of EcN revealed this E. coli strain to express only one flagella at one cell pole.
T160 739931-740031 Sentence denotes This is in contrast to the general assumption, that E. coli are peritrichously flagellated bacteria.
T161 740032-740179 Sentence denotes We conclude, the flagella of EcN is its major adhesin for binding to mucin2, the major constituent of mucus in the small and large human intestine.
T162 740180-740284 Sentence denotes Therefore, this organelle might be of importance for efficient, transient colonization of the human gut.
T163 740285-740380 Sentence denotes Natural ligands of the aryl hydrocarbon receptor control intestinal lymphoid organogenesis E.A.
T164 740381-740392 Sentence denotes Kiss 1 , C.
T165 740393-740410 Sentence denotes Vonarbourg 1 , S.
T166 740411-740426 Sentence denotes Kopfmann 1 , C.
T167 740427-740439 Sentence denotes Esser 1 , A.
T168 740440-740677 Sentence denotes Diefenbach* 1 1 University of Freiburg, IMMH, Freiburg, Deutschland Intestinal homeostasis is a prerequisite for the active extraction of metabolites by the absortive intestinal epithelium and for protection against intestinal pathogens.
T169 740678-740737 Sentence denotes However, the underlying molecular networks are ill defined.
T170 740738-740847 Sentence denotes ROR t-expressing innate lymphocytes (ROR t + ILC) play an important role in promoting intestinal homeostasis.
T171 740848-741073 Sentence denotes ROR t + ILC constitutively produce interleukin (IL-)22, a cytokine instructing epithelial expression of antimicrobial genes required for the protection against intestinal infections with attaching and effacing (A/E) bacteria.
T172 741074-741276 Sentence denotes Furthermore, subsets of ROR t + ILC have lymphoid tissue-inducing (LTi) function required for the postnatal development of intestinal lymphoid tissue harboring microflora-reactive IgA-producing B cells.
T173 741277-741512 Sentence denotes While bacteria control the generation of B cell-containing isolated lymphoid follicles (ILF) from B cell-free cryptopatches, it is unknown whether the development of cryptopatches is programmed or also instructed by environmental cues.
T174 741513-741756 Sentence denotes The family of Per-Arnt-Sim (PAS) domain-containing proteins is conserved throughout all eukaryotic species and it is believed that this group of proteins has evolved to control adaptation of multicellular organisms to environmental challenges.
T175 741757-741998 Sentence denotes One of its members, the aryl hydrocarbon receptor (AhR) functions as a ligand-inducible transcription factor that, upon binding of its ligands, instructs transcriptional programs that include the induction of xenobiotic metabolizing enzymes.
T176 741999-742142 Sentence denotes Although the role of the AhR in the response to environmental toxins is appreciated, its role in the mucosal immune system is not well defined.
T177 742143-742281 Sentence denotes We addressed the role of the AhR for development and function of ROR t + ILC by generating mice lacking the AhR in ROR t + ILC (Ahr LTi ).
T178 742282-742345 Sentence denotes Interestingly, Ahr LTi lacked genesis of cryptopatches and ILF.
T179 742346-742518 Sentence denotes While ROR t + ILC developed independent of the AhR before birth, they required AhR signals for postnatal expansion which was a prerequisite for the formation of CP and ILF.
T180 742519-742685 Sentence denotes Postnatal maintenance of ROR t + ILC was controlled by dietary phytochemicals (e.g., plant-derived glucosinolates and polyphenols) that served as natural AhR ligands.
T181 742686-742818 Sentence denotes AhR-mediated signals stabilized Kit expression by ROR t + ILC and mice with defective Kit signaling had crippled development of ILF.
T182 742819-742992 Sentence denotes AhR-deficient mice were highly susceptible to infections with Citrobacter rodentium, a mouse model of A/E infections such as those with enterohemorrhagic strains of E. coli.
T183 742993-743183 Sentence denotes Our results demonstrate that plant-derived, dietary AhR ligands dynamically regulate postnatal development of lymphoid follicles by controling the pool size of ROR t + ILC with LTi function.
T184 743184-743368 Sentence denotes Thus, sensing of dietary compounds allows dynamic adaptation of immune cells to environmental cues required to establish intestinal homeostasis and resistance to intestinal infections.
T185 743369-743380 Sentence denotes Background:
T186 743381-743508 Sentence denotes Proteolytic degradation of extracellular matrix by matrix metalloproteases is a serious consequence of intestinal inflammation.
T187 743509-743640 Sentence denotes The aim of our study was to investigate whether proteases produced by commensal gut bacteria contribute to the pathogenesis of IBD.
T188 743641-743766 Sentence denotes For this purpose we focused on the zinc dependent metalloprotease Gelatinase (GelE) from Enterococcus faecalis (E. faecalis).
T189 743767-743775 Sentence denotes Results:
T190 743776-744133 Sentence denotes In order to investigate the impact of bacterial GelE on the development of chronic intestinal inflammation we monoassociated gnotobiotc wild type (Wt) and Interleukin 10 deficient (IL-10 -/-) mice with GelE expressing E. faecalis strain OG1RF and isogenic mutant strains that lack GelE expression including TX5264 (gelE deletion) and TX5266 (fsrB deletion).
T191 744134-744276 Sentence denotes Histological analysis revealed a significant reduction of distal and proximal colonic tissue pathology of IL-10 -/mice in the absence of GelE.
T192 744277-744374 Sentence denotes Wt mice did not develop colitis demonstrating the commensal character of the E. faecalis strains.
T193 744375-744534 Sentence denotes Epithelial barrier function seemed to be affected by GelE as the presence of extracellular E-Cadherin protein domain was significantly reduced in IL-10 -/mice.
T194 744535-744731 Sentence denotes Furthermore, using LC-MS/MS analysis, we could indentify cleavage sites for GelE in the sequence of recombinant murine E-Cadherin suggesting the possibility for a direct GelE-mediated degradation.
T195 744732-744960 Sentence denotes Stimulation of distal colon segments with proteolytically active GelE revealed the loss of barrier function in mice susceptible to intestinal inflammation (IL-10 -/-, TNF ARE/Wt , NOD2 -/-) before tissue pathology has developed.
T196 744961-745054 Sentence denotes The loss of barrier integrity was associated with reduced levels of extracellular E-Cadherin.
T197 745055-745235 Sentence denotes Finally, Ptk6 cells exhibited decreased transepithelial electrical resistance and increased translocation of permeability markers after stimulation with GelE from OG1RF background.
T198 745236-745437 Sentence denotes We could show specificity for the GelE-mediated loss of barrier function by using an E. faecalis lipoprotein as unrelated control and the broad spectrum MMP inhibitor Marimastat to block GelE activity.
T199 745438-745586 Sentence denotes Conclusion: E. faecalis GelE impairs intestinal mucosal barrier function associated with the degradation of extracellular E-Cadherin protein domain.
T200 745587-745745 Sentence denotes Our data clearly demonstrates the role of GelE, as a commensal-derived protease, in the development of intestinal inflammation in the diseasesusceptible host.
T201 745746-745801 Sentence denotes Dendritic cells (DCs) can provide different phenotypes.
T202 745802-745892 Sentence denotes They can display an immature DC (iDC) phenotype or an activated mature DC (mDC) phenotype.
T203 745893-745967 Sentence denotes Recently a third phenotype has been discovered, termed semimature (smDCs).
T204 745968-746150 Sentence denotes These smDCs are able to take up antigen, but not to process it and they show reduced expression of T cell activating costimulatory molecules and a reduced expression of MHC class II.
T205 746151-746181 Sentence denotes SmDC fail to polarize T cells.
T206 746182-746382 Sentence denotes Sm BMDCs show reduced cleaving of the invariant chain (Ii) compared to mDCs, a major regulator of the MHC class II transport to the cell surface, so leading to reduced MHC class II surface expression.
T207 746383-746505 Sentence denotes The cleaving of Ii is catalyzed by the endosomal protease CatS, which is regulated by the endogenous inhibitor Cystatin C.
T208 746506-746610 Sentence denotes Indeed, mice lacking Cystatin C provide a significant higher susceptibility towards DSS induced colitis.
T209 746611-746825 Sentence denotes Therefore we suggest that semi-maturation plays an important role in maintaining the intestinal homeostasis and that regulation of Cathepsin S could be a potential target for the treatment of colitis. respectively.
T210 746826-746971 Sentence denotes An average number of 4202 sequences were obtained per tissue sample (mean length, 234 bp), and 98.5% of sequences could be classified into phyla.
T211 746972-747033 Sentence denotes Flora composition differed markedly between MIT and MHH mice.
T212 747034-747184 Sentence denotes MHH mice shared 600 (20%) of detected species with the MIT mice, and showed an overall lower richness of the bacterial flora compared to the MIT mice.
T213 747185-747261 Sentence denotes Several bacterial phyla were exclusively detected in either MHH or MIT mice.
T214 747262-747438 Sentence denotes Most notably, all caecum samples from MIT mice yielded large numbers of sequences assigned to the Deferribacteres phylum, while this phylum was completely absent from MHH mice.
T215 747439-747619 Sentence denotes Conclusion: C57/BL6J IL-10 ko mice housed under SPF conditions in two different institutions differed markedly in their susceptibility to H. hepaticus-induced intestinal pathology.
T216 747620-747830 Sentence denotes These differences were associated with highly significant differences of microbiota composition, which are likely responsible for attenuating responses to H. hepaticus in mice housed at one of the institutions.
T217 747831-747965 Sentence denotes The data highlight the importance of characterizing the intestinal microbiome when interpreting the results from murine models of IBD.
T218 747966-748060 Sentence denotes Usefulness of automatisation in a university diagnostic laboratory: more than meets the eye C.
T219 748061-748205 Sentence denotes MacKenzie* 1 1 Heinrich-Heine-Universität, Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Düsseldorf, Deutschland Introduction:
T220 748206-748556 Sentence denotes Laboratrory automatisation has made rapid and important advances in the past few years with increasing technology, standardisation of specimen collection and integration of multiple processes from simple inoculation of media to imaging and identification of microorganisms using sophisticated technology such as MALDI-TOF in a semi-automated fashion.
T221 748557-748761 Sentence denotes University laboratories are traditionally not very high throughput diagnostic services and often have personell structuring which is markedly different to that of their counterparts in the private sector.
T222 748762-748860 Sentence denotes Thus they have not been the target for many commercial companies manufacturing these technologies.
T223 748861-749007 Sentence denotes There is however a need for efficiency and improved quality provided by standardised procedures that are optimally furnished by automated systems.
T224 749008-749016 Sentence denotes Methods:
T225 749017-749391 Sentence denotes A university diagnostic laboratory providing a service to approximately 1800 in-patient beds, most of which (1200) are in a tertiary care / university setting and the remainder in a satellite teaching hospital also providing tertiary care, underwent a trial of an automated inoculation apparatus, with a view to further automatisation and integration of advanced technology.
T226 749392-749400 Sentence denotes Results:
T227 749401-749594 Sentence denotes The implementation of the system proceeded in two main phases; firstly the introduction of a standardised specimen collection system essential for the full implementation of any automatisation.
T228 749595-749873 Sentence denotes Thus in the primary phase the eSwab (Copan-Italia) was introduced universally approximately 18 months before the second phase in which the automatic inoculator or specimen processor (WASP -walk away specimen processor -MAST/Copan-Italia) was set up in the diagnostic laboratory.
T229 749874-750158 Sentence denotes After initiation, which included a unidirectional connection to the LIS, testing of the media plates for compatiility and training of the technical staff, the system was run in parallel with the manual inoculation for a standard period in order to run comparisons for quality control.
T230 750159-750265 Sentence denotes Thereafter the system was put "on-line" and run in the diagnostic services as the sole specimen processor.
T231 750266-750366 Sentence denotes Staff re-allocation and processng tme, specimen quality and errors were all documented and analysed.
T232 750367-750378 Sentence denotes Discussion:
T233 750379-750607 Sentence denotes The introduction and full implementation of an automated specimen processor in the microbiology diagnostic service for a typical large university hospital has far reaching consequences to the work-flow and staff time-allocation.
T234 750608-750726 Sentence denotes Especially in this setting, the use of highly qualified staff for repetitive tasks demanding low skill is inefficient.
T235 750727-750899 Sentence denotes University laboratories are generally unable to employ staff commensurate for low-skill tasks and furthermore are not in the position to cut costs through staff reductions.
T236 750900-751098 Sentence denotes In this setting automatisation can be of great benefit by freeing technical staff for more tasks more appropriate to their training and also to the demands of a tertiary care / reference laboratory.
T237 751099-751239 Sentence denotes Introduction: T. pallidum spp. pallidum, is the causative agent of syphilis, a sexual transmitted disease of variable clinical presentation.
T238 751240-751414 Sentence denotes As the spirochete resists conventional cultivation and to overcome test specific limitations, laboratory diagnosis is based on a stepwise approach using immunological assays.
T239 751415-751590 Sentence denotes However, independent clinical evaluation of such tests is not mandatory within the EU and currently, a large number of assays of variable analytical quality are on the market.
T240 751591-751742 Sentence denotes Here we present a summary report of the results obtained by the German proficiency testing program for syphilis serology between 2007 and 2010 Methods:
T241 751743-752217 Sentence denotes From 2007 to 2010 eight proficiency testing surveys were conducted in Germany by the central reference laboratory for bacteriological serodiagnostics now situated at the Institute for Laboratory Medicine at the Northwest Medical Centre in Frankfurt, Germany, in cooperation with the Institute of Standardisation in the Medical Laboratory e.V. (INSTAND e.V.), Düsseldorf and with reference laboratories of the Bacteriologic Infection Serology Study Group of Germany (BISSGG).
T242 752218-752380 Sentence denotes In each testing, two samples obtained from infected patients or healthy donors were sent out to the participants for analysis under routine laboratory conditions.
T243 752381-752590 Sentence denotes Assessment of reference test results was performed according to standard operating procedures of INSTAND and current guidelines of the German chamber of physicians as published recently (Müller et al., 2010) .
T244 752591-752740 Sentence denotes Moreover, 13.5% of laboratories wrongly reported evidence for active infection in cases with past syphilis or in sera from seronegative blood donors.
T245 752741-752752 Sentence denotes Discussion:
T246 752753-752962 Sentence denotes With the liberalisation of the in vitro diagnostics market in the EU a wide range of serological assays for syphilis diagnostics of different diagnostic quality was promoted for use in diagnostic laboratories.
T247 752963-753252 Sentence denotes In this study we confirm previous investigations that, although positive trends are obvious, the sensitivity and specificity of currently applied assays for syphilis diagnostics can still vary substantially, depending on the stage of disease, the type of test and the kind of manufacturer.
T248 753253-753508 Sentence denotes Our results demonstrate that ongoing external quality control of routine diagnostic laboratories and in depth clinical evaluation of diagnostic assays is clearly needed to monitor test quality and to ensure a high level of quality in syphilis diagnostics.
T249 753509-753703 Sentence denotes External quality control of immunodiagnostics in rare and fastidious pathogens (bordetella, mycoplasma, coxiella, campylobacter, and yersinia): a metaanalysis of proficiency testing trials K.-P.
T250 753704-753719 Sentence denotes Hunfeld* 1 , I.
T251 753720-753730 Sentence denotes Müller 1 1
T252 753732-753745 Sentence denotes Introduction:
T253 753746-753926 Sentence denotes There is a long tradition in the participation of medical laboratories in external quality control of well established parameters and diagnostic procedures in medical microbiology.
T254 753927-754029 Sentence denotes This tradition is now additionally enforced by the new guidelines of the German chamber of physicians.
T255 754030-754155 Sentence denotes Evaluations of proficiency testing trials of indirect detection methods for rare and fastidious pathogens however are sparse.
T256 754156-754346 Sentence denotes Most importantly the participation in such surveys is not mandatory in Germany although there is a lack of test standardization and very few data are available on the quality of such assays.
T257 754347-754547 Sentence denotes Here, we present a summary report on the results of the INSTAND proficiency testing trials in the serodiagnostics of bordetella, mycoplasma, coxiella, campylobacter and yersinia Materials and methods:
T258 754548-755101 Sentence denotes From 2000 to 2010 twenty-five proficiency testing surveys addressing the serodiagnostics of the above mentioned pathogens were conducted in Germany by the central reference laboratory for bacteriological serodiagnostics now situated at the Institute for Laboratory Medicine at the Northwest Medical Centre in Frankfurt/Main, Germany, in cooperation with the DGHM, the Institute of Standardization in the Medical Laboratory (INSTAND e.V.), Düsseldorf and the reference laboratories of the Bacteriologic Infection Serology Study Group of Germany (BISSGG).
T259 755102-755263 Sentence denotes In each survey, two samples obtained from infected patients or healthy donors were sent out to the participants for analysis under routine laboratory conditions.
T260 755264-755450 Sentence denotes Assessment of reference test results was performed according to standard operating procedures of INSTAND and current guidelines of the German chamber of physicians as published recently.
T261 755451-755459 Sentence denotes Results:
T262 755460-755617 Sentence denotes During the study period on average the following numbers of laboratories participated in the different serodiagnostic trials: yersinia: N=197; campylobacter:
T263 755618-755652 Sentence denotes 73; bordetella: N=117; mycoplasma:
T264 755653-755667 Sentence denotes 118; coxiella:
T265 755668-755671 Sentence denotes 47.
T266 755672-755925 Sentence denotes For the detection of specific antibodies in yersinia-, campylobacter-, and bordetella-serology mainly ELISA and immunoblots were used by the participants whereas in coxiella-serology more traditional tests such as CFT and IFA were predominantly applied.
T267 755926-756028 Sentence denotes Ranges of average pass rates for diagnostic testing and diagnostic comments were as follows: yersinia:
T268 756029-756048 Sentence denotes 58-85%; bordetella:
T269 756049-756068 Sentence denotes 70-95%; mycoplasma:
T270 756069-756086 Sentence denotes 56-90%; coxiella:
T271 756087-756110 Sentence denotes 60-100%; campylobacter:
T272 756111-756118 Sentence denotes 43-93%.
T273 756119-756130 Sentence denotes Discussion:
T274 756131-756410 Sentence denotes Although, proficiency testing trials cannot fully substitute clinical evaluations of immunodiagnostic tests, the results of our metaanalysis demonstrate a very variable degree of standardization and assay quality in the serodiagnostics of rare and fastidious bacterial pathogens.
T275 756411-756771 Sentence denotes Most importantly, our observations provide interesting insights into the comparability of different assay systems the interpretation of quantitative and semi quantitative test results and the correct clinical interpretation of the serological findings pointing to limitations of these diagnostic procedures in the routine diagnostic microbiological laboratory.
T276 756772-756882 Sentence denotes Staphylococcus aureus is a leading cause of human disease in the hospital setting as well as in the community.
T277 756883-756991 Sentence denotes It has been shown an association between nasal colonization of S. aureus and infection with the same strain.
T278 756992-757114 Sentence denotes Of particular concern is a worldwide increase in the number of infections caused by methicllin-resistant S. aureus (MRSA).
T279 757115-757261 Sentence denotes In addition, to the longknown healthcare-associated clones, there is observed the emergence and spread of community-and livestock-associated MRSA.
T280 757262-757495 Sentence denotes The ability of the clinically important strains to persist within hosts is largely due to the expression of a battery of virulence factors which promote adhesion, acquisition of nutrients, and evasion of host immunological responses.
T281 757496-757793 Sentence denotes The objective of this study was to determine prevalence of the virulence genes among the most successful and sporadic MRSA clones in the Dutch-German Euregio A total of 52 non-repetitive MRSA isolates recovered from nasal carriage from healthy persons in Euregio between 2003 and 2009 were tested.
T282 757794-758034 Sentence denotes Isolates selection was based on the spa typing results and was intended to include representatives of the most predominant spa types recovered in Euregio (t003, t032, t011, t034, t004, t008) as well as representatives of sporadic spa types.
T283 758035-758357 Sentence denotes The virulence genes (tst1, sea, seb, sec, sed, sef, seg, seh, sei, sej, sek, sel, selm, seln, selo, seq, ser, selu, PVL, lukM, lukD, lukE, sak, chp, scn, etA, etB, etD, edinA, edinB, edinC, arcA-ACME, arcB-ACME, arcC-ACME, arcD-ACME) were detected by the microarrays (StaphyType by Alere Technologies GmbH, Jena, Germany).
T284 758358-758436 Sentence denotes Different combinations of virulence genes were identified among MRSA isolates.
T285 758437-758583 Sentence denotes All isolates carried at least 5 virulence genes with the exception of MRSA isolates of spa-Clonal Complex (CC) 011 (with spa types t011 and t034).
T286 758584-758744 Sentence denotes Isolates of spa-CC 011 did not carry any virulence gene with the exception of a single isolate (with spa type t011) which harboured the sed and arcB-ACME genes.
T287 758745-759090 Sentence denotes We observed association the more predominant MRSA clone the more virulence genes were detected: t003 -15 genes (sea, sed, seg, sei, sej, selm, seln, selo, ser, selu, lukD, lukE, sak, chp, scn); t032 -12 genes (sec, sed, seg, sei, sel, selm, seln, selo, selu, sak, chp, scn); t004 -10 genes (sed, seg, sei, selm, seln, selo, selu, sak, chp, scn).
T288 759091-759305 Sentence denotes Although, most of the la-MRSA carry no tested virulence factors, some la-MRSA harbour important virulence factors which shows the genomic plasticity of this MRSA subtype and possible adaptive changes in the future.
T289 759306-759437 Sentence denotes Diphtheria and diphtheria-like illness is caused by Corynebacterium species harbouring the diphtheria toxin (DT) encoding tox gene.
T290 759438-759613 Sentence denotes In recent years, diphtheria-like infections with toxigenic Corynebacterium ulcerans have outnumbered those caused by toxigenic C. diphtheriae in many industrialized countries.
T291 759614-759797 Sentence denotes Originally associated with consumption of raw milk and dairy products or contact to cattle, C. ulcerans has increasingly been isolated from domestic animals such as pet dogs and cats.
T292 759798-759928 Sentence denotes So far, isolation of an identical toxigenic C. ulcerans strain from an animal and its owner is documented only for dogs and a pig.
T293 759929-760089 Sentence denotes Here we report the first case of proven transmission of a toxigenic C. ulcerans strain between a pet cat and a human causing pharyngeal diphtheria-like illness.
T294 760090-760230 Sentence denotes Antimicrobial resistance patterns of Corynebacterium diphtherieae, C. ulcerans and C. pseudotuberculosis strains of human and animal origin.
T295 760231-760242 Sentence denotes Objectives:
T296 760243-760514 Sentence denotes Human infections due to Corynebacterium diphtheriae and the zoonotic germs C. ulcerans and C. pseudotuberculosis are rare but potentially life-threatening events requiring specific antibiotic treatment, and in cases suspicious for diphtheria the prompt antitoxin therapy.
T297 760515-760730 Sentence denotes In the last years, we have observed an increased number of C. ulcerans strains isolated from patients with wound infections and diphtheria-like illness, mainly harbouring the diphtheria toxin (DT) encoding tox gene.
T298 760731-760973 Sentence denotes Benzyl penicillin and erythromycin are still recommended for specific antibiotic treatment, to eradicate the pathogen in patients and to prevent infection among contacts, although only very few antimicrobial susceptibility data are available.
T299 760974-760982 Sentence denotes Methods:
T300 760983-761256 Sentence denotes In this study 106 isolates of toxigenic and non-toxigenic clinical isolates of C. diphtheriae (n=35), C. pseudotuberculosis (n=47) and C. ulcerans (n=24) from humans and animals collected in the National Consiliary Laboratory for Diphtheria from 2001 to 2010 were examined.
T301 761257-761429 Sentence denotes Minimal inhibitory concentration (MIC) of benzyl penicillin, ceftriaxone, erythromycin, tetracycline, clindamycin, and ciprofloxacin were determined using the Etest system.
T302 761430-761626 Sentence denotes In the absence of standardized breakpoints, antibiotic susceptibility was determined by using the CLSI criteria for broth microbouillon dilution susceptibility testing for Corynebacterium species.
T303 761627-761635 Sentence denotes Results:
T304 761636-761819 Sentence denotes Benzyl penicillin, erythromycin and ciprofloxacin seem to be highly active substances in all three Corynebacterium species (susceptibility rates of 98.1, 98.1 and 96.2% respectively).
T305 761820-762029 Sentence denotes For C. diphtheriae tetracycline and ceftriaxone showed lowest activity in vitro (susceptibility rates of 71.4% and 42.9%), while clindamycin was least active against C. ulcerans (susceptibility rate of 16.7%).
T306 762030-762129 Sentence denotes In contrast, C. pseudotuberculosis strains were completely susceptible to all examined antibiotics.
T307 762130-762141 Sentence denotes Conclusion:
T308 762142-762300 Sentence denotes Our data emphasize the need for an accurate species identification of potentially toxigenic corynebacteria and underscore the necessity of antibiotic testing.
T309 762301-762499 Sentence denotes Although clindamycin is not a first-line drug in diphtheria therapy, it is favoured for the treatment of wound infections, but due to our results C. ulcerans is not surely covered by this substance.
T310 762500-762642 Sentence denotes Benzyl penicillin and erythromycin can be still recommended for specific antibiotic treatment in Germany, but antibiotic resistance may occur.
T311 762643-762655 Sentence denotes Case report:
T312 762656-762754 Sentence denotes A 14 year old girl presented with persistent epigastric pain since several months in our hospital.
T313 762755-762826 Sentence denotes A treatment with omeprazole for 14 days showed no clinical improvement.
T314 762827-762900 Sentence denotes An esophagogastroscopy was performed, revealing a mild erosive gastritis.
T315 762901-763016 Sentence denotes Biopsy specimens from the stomach were sent for histopathology and helicobacter culture and susceptibility testing.
T316 763017-763156 Sentence denotes In a direct gram stain of the homogenized gastric biopsy specimen long gram negative corkscrew-like rods atypical for H. pylori were found.
T317 763157-763280 Sentence denotes After 7 days of incubation on agar media containing human erythrocytes and inactivated horse serum first colonies appeared.
T318 763281-763375 Sentence denotes Cytochrome oxidase, catalase and urease tests performed from the grown bacteria were positive.
T319 763376-763507 Sentence denotes The strain was susceptible to clarithromycin, metronidazole, amoxicillin, levofloxacin, tetracycline and rifampicin (Etest method).
T320 763508-763643 Sentence denotes For species identification partial ureA and ureB gene PCR followed by sequencing was performed as described by O`Rourke et al. (2004) .
T321 763644-763734 Sentence denotes Following our phylogenetic analyses, this isolate can be identified as Helicobacter felis.
T322 763735-763877 Sentence denotes Helicobacter felis belongs to the "gastric non-H. pylori helicobacters", formerly also classified as H. heilmannii type II (Haesebrouck et al.
T323 763878-763885 Sentence denotes 2009 ).
T324 763886-763947 Sentence denotes H. felis is frequently found in the stomach of cats and dogs.
T325 763948-764060 Sentence denotes Infections in humans have been reported, but culture is difficult due to the fastidious nature of this organism.
T326 764061-764165 Sentence denotes To our knowledge only two non-H. pylori strains were cultured from human gastric biopsy specimen so far.
T327 764166-764228 Sentence denotes Both strains were classified as H. bizzozeronii (Jalava et al.
T328 764229-764249 Sentence denotes 2001; Kivistö et al.
T329 764250-764257 Sentence denotes 2008) .
T330 764258-764399 Sentence denotes This case report demonstrates that direct gram stain of gastric biopsy specimen is important to identify gastric non-H. pylori helicobacters.
T331 764400-764503 Sentence denotes Furthermore, culture of fastidious non-H. pylori helicobacters using our standard protocol is possible.
T332 764504-764603 Sentence denotes Which factors interfere with metronidazole susceptibility testing of H. pylori by the Etest method?
T333 764604-764606 Sentence denotes N.
T334 764607-764626 Sentence denotes Wüppenhorst* 1 , B.
T335 764627-764644 Sentence denotes Hobmaier 1 , H.P.
T336 764645-764658 Sentence denotes Stüger 2 , E.
T337 764659-764673 Sentence denotes Glocker 1 , M.
T338 764674-764880 Sentence denotes Kist 1 1 Institute of Medical Microbiology and Hygiene, National Reference Centre for Helicobacter pylori, Freiburg, Deutschland 2 Austrian Agency for Health and Food Safety, Graz, Oesterreich Introduction:
T339 764881-765001 Sentence denotes Metronidazole is part of the Italian triple therapy, which is a recommended first line scheme for H. pylori eradication.
T340 765002-765157 Sentence denotes In Germany, the primary resistance rate for metronidazole is 29% (data from the 3rd European multi-centre study on antibiotic susceptibility of H. pylori).
T341 765158-765320 Sentence denotes However, phenotypic susceptibility testing is currently not approved by the FDA due to "variance in test results, lack of reproducibility and clinical relevance".
T342 765321-765485 Sentence denotes But phenotypic susceptibility testing is the only option for detection of resistance, because the molecular mechanisms of resistance are not conclusively explained.
T343 765486-765653 Sentence denotes Therefore, the aim of our study is to evaluate factors that influence the precision and reproducibility of metronidazole susceptibility testing using the Etest method.
T344 765654-765675 Sentence denotes Material and Methods:
T345 765676-765869 Sentence denotes 20 randomly selected H. pylori isolates cultured from routine gastric biopsy specimens (no selection concerning resistance patterns, pre-treatments, patient data etc.) are actually under study.
T346 765870-766000 Sentence denotes Etests of each isolate are performed repeatedly at the same time point (precision) and at different time points (reproducibility).
T347 766001-766170 Sentence denotes Further variables included are the culture media (Mueller-Hinton + 10% horse blood vs. Isosensitest + 10% horse blood) and the incubation period (48 hours vs. 72 hours).
T348 766171-766179 Sentence denotes Results:
T349 766180-766331 Sentence denotes The minimal inhibitory concentrations (MIC) detected on Mueller-Hinton and Isosensitest agar after 48h and 72h of incubation showed a high correlation.
T350 766332-766417 Sentence denotes A small but systematic difference between the media and incubation periods was found.
T351 766418-766579 Sentence denotes MICs on Isosensitest agar were higher compared to Mueller-Hinton agar (factor 1.1), and 48h of incubation led to lower MICs than 72h of incubation (factor 0.86).
T352 766580-766687 Sentence denotes The range of MICs was significantly broader between series at different time points than within one series.
T353 766688-766699 Sentence denotes Discussion:
T354 766700-766862 Sentence denotes There is a small but systematic influence of culture media and incubation period on metronidazole testing using the Etest method, but correlation of MICs is high.
T355 766863-766919 Sentence denotes To evaluate the reproducibility further data are needed.
T1 766920-766922 Sentence denotes J.
T2 766923-766935 Sentence denotes Kolb* 1 , D.
T3 766936-766952 Sentence denotes Hillemann 1 , J.
T4 766953-766965 Sentence denotes Reetz 2 , S.
T5 766966-766985 Sentence denotes Rüsch-Gerdes 1 , E.
T6 766986-767163 Sentence denotes Richter 1 1 Forschungszentrum Borstel, Nationales Referenzzentrum für Mykobakterien, Borstel, Deutschland 2 Bundesinstitut für Risikobewertung, Berlin, Deutschland Introduction:
T7 767164-767464 Sentence denotes One of the most frequent isolated species within nontuberculous mycobacteria is M. avium, which is ubiquitously found in the environment and has been shown to be responsible for a variety of human diseases, mainly in immunocomprised patients, but also in patients with no obvious immune deficiencies.
T8 767465-767616 Sentence denotes M. avium comprises the subspecies M. avium subsp. avium, M. avium subsp. silvaticum, M. avium subsp. paratuberculosis, and M. avium subsp. hominissuis.
T9 767617-767654 Sentence denotes Routes of infection are rarely known.
T10 767655-767816 Sentence denotes For understanding of transmission mechanisms and for deeper insights into the pathogenicity genetic characterization of the infectious strains is a prerequisite.
T11 767817-767939 Sentence denotes Therefore, the aim of our study was to assess the applicability of MLST and MIRU-VNTR for distinguishing M. avium strains.
T12 767940-767961 Sentence denotes Material and Methods:
T13 767962-767995 Sentence denotes Genotyping was performed by MLST.
T14 767996-768081 Sentence denotes For this, fragments of several housekeeping genes were analyzed by sequence analysis.
T15 768082-768223 Sentence denotes Comparing the DNA sequences of each locus we estimated the degree of variability of these genes in isolates of different M. avium subspecies.
T16 768224-768291 Sentence denotes By MIRU variable numbers of tandem repeats of 6 loci were analyzed.
T17 768292-768438 Sentence denotes Into the study we included M. avium isolates of children suffering from M. avium lymphadenitis sent to the NRZ in the years 2003, 2004, and 2009 .
T18 768439-768518 Sentence denotes Furthermore, strains isolated from slaughter pigs and from birds were analyzed.
T19 768519-768584 Sentence denotes Subspecies analysis was done by PCR targeting IS900/901 elements.
T20 768585-768593 Sentence denotes Results:
T21 768594-768673 Sentence denotes Firstly bird isolates were proven to be M. avium subsp. avium by IS900/901 PCR.
T22 768674-768775 Sentence denotes Various single nucleotide polymorphisms could be detected by MLST analysis in the genes investigated.
T23 768776-768894 Sentence denotes Differences in the genes concerning their variability were used to decide the applicability for future investigations.
T24 768895-768971 Sentence denotes The MIRU-VNTR analysis showed no great impact due to a moderate variability.
T25 768972-768983 Sentence denotes Discussion:
T26 768984-769046 Sentence denotes By molecular methods subspecies of M. avium can be identified.
T27 769047-769180 Sentence denotes MLST enables typing of strains that in future may be used for example for analysis of transmission or reactivation vs. new infection.
T28 769181-769368 Sentence denotes Reduction of antibiotic resistance among S. pneumoniae isolated from IPD in children and adults as a result of the German national immunization program for pneumococcal conjugate vaccine.
T29 769369-769394 Sentence denotes M. van der Linden* 1 , M.
T30 769395-769500 Sentence denotes Imöhl 1 1 Universitätsklinikum RWTH Aachen, Medizinische Mikrobiologie, Aachen, Deutschland Introduction:
T31 769501-769767 Sentence denotes We evaluated the impact of the immunization of children <2 with pneumococcal conjugate vaccine (3+1 doses) started in July 2006 on antibiotic resistance levels among S. pneumoniae isolated from children and adults with invasive pneumococcal disease (IPD) in Germany.
T32 769768-769790 Sentence denotes Materials and methods:
T33 769791-769976 Sentence denotes Isolates from IPD cases in children <16 years and adults >= 16 years were sent to the German National Reference Center for Streptococci by microbiological laboratories all over Germany.
T34 769977-770094 Sentence denotes Serotyping was performed using the Neufeld Quellung reaction and MICs were determined using the microdilution method.
T35 770095-770103 Sentence denotes Results:
T36 770104-770294 Sentence denotes Numbers of IPD cases with serotypes included in the 7-valent pneumococcal conjugate vaccine (PCV7) were strongly reduced in children under two years after the start of childhood vaccination.
T37 770295-770458 Sentence denotes In the years before vaccination (1997) (1998) (1999) (2000) (2001) (2002) (2003) (2004) (2005) (2006) an average 66.4% were PCV7 serotypes, in 2010-2011 only 7.5%.
T38 770459-770585 Sentence denotes In children 2-4 years these percentages were 72.1% before and 4.6% after, in children 5-15 years, 36.0% before and 9.8% after.
T39 770586-770681 Sentence denotes In adults (16 years and older) an average 46.0% of cases had PCV7 serotypes before vaccination.
T40 770682-770726 Sentence denotes After vaccination this was reduced to 12.6%.
T41 770727-770844 Sentence denotes Among isolates from children macrolide resistance was reduced from an average 26,1% in 1997-2006 to 10,5% in 2010-11.
T42 770845-770935 Sentence denotes Among adults macrolide resistance was reduced from 17.4% before to 9.0% after vaccination.
T43 770936-771034 Sentence denotes Penicillin resistance levels have not changed much after vaccination in either children or adults.
T44 771035-771124 Sentence denotes However, the serotype distribution among these isolates has changed considerably as well.
T45 771125-771136 Sentence denotes Discussion:
T46 771137-771294 Sentence denotes Five years after the introduction of PCV7, the reduction in numbers of reported cases of IPD with vaccine serotypes in children under 2 years was almost 90%.
T47 771295-771418 Sentence denotes The observed reduction of IPD cases with vaccine serotypes in older children and adults indicates a herd protection effect.
T48 771419-771524 Sentence denotes Due to the disappearance of vaccine serotypes macrolide resistance levels have been reduced considerably.
T49 771525-771709 Sentence denotes Penicillin resistance levels have not changed, since the disappearance of penicillin resistant vaccine serotypes was compensated by the appearance of penicillin resistant serotype 19A.
T50 771710-771826 Sentence denotes Establishment of a red deer DNA banc as source for future studies on emerging zoonotic and vector borne pathogens C.
T51 771827-771840 Sentence denotes Klier* 1 , J.
T52 771841-771856 Sentence denotes Brenauer 1 , B.
T53 771857-771873 Sentence denotes Fleischer 2 , E.
T54 771874-771890 Sentence denotes Gerstmair 1 , M.
T55 771891-771905 Sentence denotes Niedrig 3 , Y.
T56 771906-771920 Sentence denotes Schulze 3 , A.
T57 771921-771932 Sentence denotes Sing 1 , V.
T58 771933-772193 Sentence denotes Fingerle 1 1 LGL Bayern, NRZ Borrelien,, Oberschleißheim, Deutschland 2 Bernhard-Nocht-Institut , Nationales Referenzzentrum für tropische Infektionserreger, Hamburg, Deutschland 3 Robert Koch Institut, Konsiliarlabor für FSME , Berlin, Deutschland Background:
T59 772194-772398 Sentence denotes Ongoing global warming is thought to cause environmental changes which could foster the emergence and re-emergence of zoonotic as well as vectorborne infectious diseases affecting human and animal health.
T60 772399-772514 Sentence denotes Besides domestic livestock, wild animals such as deer or wild boar serve as reservoir for such infectious diseases.
T61 772515-772712 Sentence denotes To date there is little data available on the prevalence of zoonotic and vector borne pathogens such as Anaplasma phagocytophilum, Francisella tularensis, Babesia spp, Rickettsia spp, or TBE-Virus.
T62 772713-772928 Sentence denotes Data obtained in seroprevalence studies indicate that wild animals are susceptible to several zoonotic and vector borne pathogens, however do not provide informations to which extend wild animals serve as reservoir.
T63 772929-773136 Sentence denotes In this study we aim at 1) establishing a DNA banc from well defined material from red deer and 2) to perform first investigations on the prevalence of zoonotic and vector borne pathogens in these materials.
T64 773137-773303 Sentence denotes Standardized collected and stored DNA-samples may serve as a material collection for longitudinal studies on endemic and emerging zoonotic and vector borne pathogens.
T65 773304-773312 Sentence denotes Methods:
T66 773313-773432 Sentence denotes Lymph nodes from red deer (Cervus elapus) were collected directly after shooting and stored at -20°C until further use.
T67 773433-773662 Sentence denotes DNA (600μl) was extracted (Biosprint, Quiagen) from each single lymph node and stored in 20 vials as a DNA-banc, which is connected to a data-banc containing data like sex, age, health status, or place of shot regarding the deer.
T68 773663-773787 Sentence denotes These samples are used to screen for zoonotic and vector borne pathogens like Anaplasma, Francisella, or DNA-viruses by PCR.
T69 773788-773796 Sentence denotes Results:
T70 773797-773901 Sentence denotes So far, around 1500 lymph nodes from 323 red deer were collected and the deer data banc was established.
T71 773902-773979 Sentence denotes The DNA banc actually consists of 323 samples, one lymph node from each deer.
T72 773980-774172 Sentence denotes First screening revealed that none of the 323 tested samples contained measurable F. tularensis DNA, but 39 (12.1%) samples were positive for A. phagocytophilum DNA as confirmed by sequencing.
T73 774173-774248 Sentence denotes Further pathogenscreening actually is on the way and data will be provided.
T74 774249-774372 Sentence denotes Moreover, 200 tissue samples were subjected to RNA-extraction and are currently screened for tick borne encephalitis virus.
T75 774373-774384 Sentence denotes Conclusion:
T76 774385-774560 Sentence denotes The detection of A. phagocytophilum DNA in the screened samples indicates that lymph nodes from red deer is suitable for screening purposes with respect to A. phagocytophilum.
T77 774561-774670 Sentence denotes The absence of F. tularensis DNA in lymph nodes however does not conclusively rule out red deer as reservoir.
T78 774671-774809 Sentence denotes Due to the high mobility and broad distribution of red deer, it might serve as a sentinel for emerging zoonotic and vectorborne pathogens.
T79 774810-774923 Sentence denotes The presented DNA banc is an important prerequisite for future investigations on emerging pathogens. bavariensis.
T80 774924-774993 Sentence denotes In Germany Borrelia are transmitted by the hard tick Ixodes ricinus .
T81 774994-775232 Sentence denotes Infection with Borrelia can result in clinical manifestations of the skin (Erythema migrans, Borrelia lymphocytoma, Acrodermatitis chronica atrophicans), nervous system (neuroborreliosis), joints (Lyme Arthritis) or heart (Lyme Carditis).
T82 775233-775310 Sentence denotes Currently concise data on the incidences of LB in European countries is rare.
T83 775311-775440 Sentence denotes In this review we aimed at collecting data on the incidence of LB in all European countries emphasising data on neuroborreliosis.
T84 775441-775449 Sentence denotes Methods:
T85 775450-775596 Sentence denotes Data were collected by structured literature search in PubMed, national health registries, public health institutions and personal expertcontacts.
T86 775597-775605 Sentence denotes Results:
T87 775606-775673 Sentence denotes Data on the incidence of LB was obtained for 29 European countries.
T88 775674-775849 Sentence denotes Reporting regimens are very heterogenous among European countries and there is little information on the reliability of the employed reporting systems (e.g. case definitions).
T89 775850-775988 Sentence denotes Geographic distribution shows lowest incidences of LB in southwest Europe (Portugal < 0.7 x 10 5 , 2007) and along the Mediteranian coast.
T90 775989-776167 Sentence denotes Incidences increase northward (Sweden 69 x 10 5 , 1995) as well as from Western Europe (England & Wales,1.72 x10 5 , 2009) towards Central Europe (Switzerland, 115x 10 5 , 2008).
T91 776168-776231 Sentence denotes The highest incidence was reported from Slovenia (312 x 10 5 ).
T92 776232-776310 Sentence denotes With respect to Neuroborreliosis (NB) there is data from 9 European countries.
T93 776311-776456 Sentence denotes Data collected in studies and national registries show that reported incidence of NB varies between 0.6 x 10 5 in France and 11 x 10 5 in Sweden.
T94 776457-776530 Sentence denotes In 2009 incidence of NB was 1.1x10 5 in Norway and 3.8 x 10 5 in Denmark.
T95 776531-776670 Sentence denotes Incidence of NB in Germany was 3.3 x 10 5 in 1998; more recent data from Eastern Germany show an incidence of early NB of 0.3 x10 5 (2009).
T96 776671-776885 Sentence denotes Longitudinal data on the incidence of LB in Europe indicate that some countries observe a clear increase, e.g. Slovenia, whereas incidence data from other countries remained fairly constant over the past ten years.
T97 776886-777044 Sentence denotes Some countries reported a rise in LB incidence until 2006, since then reported data either remained within the same range (Norway) or even declined (Germany).
T98 777045-777057 Sentence denotes Conclusions:
T99 777058-777283 Sentence denotes Although data on the incidence of LB are available from many European countries, currently it is hard to provide a reliable picture mainly due to the various methodologies of data acquisition preventing conclusive comparison.
T100 777284-777387 Sentence denotes For the purpose of comparable data, well designed sentinels or prospective studies are urgently needed.
T101 777388-777555 Sentence denotes Climate change -What about borreliae and ticks? stricto(6.9%; for the first time identified in Romania), B. spielmanii/B. valaisiana (11,6%) and B. lusitaniae (2,3%) .
T102 777556-777726 Sentence denotes In case of infection with B. spielmanii or B. valaisiana the melting curve analysis could not differentiate between the two species because of overlapping melting curves.
T103 777727-777759 Sentence denotes No mixed infection was detected.
T104 777760-777919 Sentence denotes Results on the clinical outcome and Borrelia serology of the individuals bitten by Borrelia-positive ticks will be presented and compared to the control group.
T105 777920-778048 Sentence denotes Preliminary results suggest that there is a low risk to develop Lyme borreliosis after a Borrelia-infected tick bite in Romania.
T106 778049-778059 Sentence denotes Objective:
T107 778060-778188 Sentence denotes Testing pertussis serology performance in routine laboratories after the availability of an International Reference Preparation.
T108 778189-778197 Sentence denotes Methods:
T109 778198-778310 Sentence denotes In cooperation with INSTAND, reference preparations were used in an external quality assessment programme (EQA).
T110 778311-778524 Sentence denotes Two reference preparations, the WHO Reference Reagent (06/142) and a preparation from NIBSC with very low level of antibodies were reconstituted centrally and sent as liquid samples to a total of 200 participants.
T111 778525-778580 Sentence denotes Results were reported qualitatively and quantitatively.
T112 778581-778706 Sentence denotes Quantitative results were converted into IU/ml wherever possible using the suggestions from the producers of diagnostic kits.
T113 778707-778715 Sentence denotes Results:
T114 778716-778771 Sentence denotes Results were reported from a total of 183 participants.
T115 778772-778858 Sentence denotes ELISAs with mixed antigens were used for measuring IgG-antibodies by 111 participants.
T116 778859-779004 Sentence denotes IgA-antibodies were measured by 110 participants using mixed antigen ELISAs, and 113 participants used these ELISAs for measuring IgM-antibodies.
T117 779005-779117 Sentence denotes A total of 47 or 44 participants used ELISAs with purified PT for measuring IgG-or IgA-antibodies, respectively.
T118 779118-779231 Sentence denotes Immunoblot assays were used to estimate IgG, IgA and IgM antibodies by 62, 63, and 11 participants, respectively.
T119 779232-779366 Sentence denotes Five participants used indirect immunofluorescence measuring IgG, IgA and IgM antibodies, and one laboratory used complement fixation.
T120 779367-779526 Sentence denotes Quantitative results were reported in different units, and only the ELISAs using purified PT gave results that were comparable to the expected values in IU/ml.
T121 779527-779539 Sentence denotes Conclusions:
T122 779540-779704 Sentence denotes Many laboratories used methods that are not recommended by the EU reference laboratories and report results that cannot be related to the WHO reference preparation.
T123 779705-779819 Sentence denotes Retrospective performance of HRM-PCR to discover two outbreaks of listeriosis in Austria, 2008 -2010 Introduction:
T124 779820-780079 Sentence denotes For public health laboratories in charge of listeriosis surveillance, high-resolution melting PCR (HRM-PCR) analysis is a promising new technique to get by with low specificity of serotyping and various disadvantages of pulsed field gelelectrophoresis (PFGE).
T125 780080-780101 Sentence denotes Material and Methods:
T126 780102-780192 Sentence denotes From 2008 to 2010, two different outbreaks of listeriosis have been reported from Austria:
T127 780193-780195 Sentence denotes 1.
T128 780196-780394 Sentence denotes An outbreak of febrile gastroenteritis in guests of a local restaurant after consumption of contaminated home-made jellied pork, including two cases of bacteriologically proven invasive listeriosis.
T129 780395-780397 Sentence denotes 2.
T130 780398-780506 Sentence denotes A multi-national outbreak of invasive listeriosis due to contaminated acid curd cheese sold by supermarkets.
T131 780507-780707 Sentence denotes The product carried two genetically different outbreak strains that were responsible for 12 cases (clone 1) and 13 cases (clone 2) of bacteriologically proven invasive listeriosis from Austria (total:
T132 780708-780718 Sentence denotes 25 cases).
T133 780719-780833 Sentence denotes For all samples belonging to the outbreaks the inlB HRM-PCR products were subsequently sequenced for verification.
T134 780834-780994 Sentence denotes Retrospectively, we studied performance of serotyping and HRM-PCR inlB sequence-type (inlB) analysis to detect these outbreaks that have been confirmed by PFGE.
T135 780995-781003 Sentence denotes Results:
T136 781004-781133 Sentence denotes 112 isolates of Listeria monocytogenes from patients with invasive disease from Austria have been tested during the study period.
T137 781134-781186 Sentence denotes Serotype distribution in the total study population:
T138 781187-781239 Sentence denotes 1/2a -55.3 %, 4b -28.6 %, 1/2b -15.2 %, 1/2c -0.9 %.
T139 781240-781258 Sentence denotes InlB distribution:
T140 781259-781380 Sentence denotes 15 -18.8 %, 9 -17.9 %, 1 -13.9 %, 5 -9.8 %, non-typable -8.0 %, 2, 3 and 7 -7.1 % each, and else types 10.3 % altogether.
T141 781381-781419 Sentence denotes Both invasive isolates of outbreak no.
T142 781420-781450 Sentence denotes 1 were serotype 4b and inlB 2.
T143 781451-781493 Sentence denotes All 12 isolates of clone 1 of outbreak no.
T144 781494-781589 Sentence denotes 2 were serotype 1/2a and inlB 15, and all 13 isolates of clone 2 were serotype 1/2a and inlB 9.
T145 781590-781727 Sentence denotes In addition, we found 30 isolates of serotype 4b and 37 isolates of serotype 1/2a that were not related to the mentioned outbreak clones.
T146 781728-781871 Sentence denotes With regard to HRM-PCR analysis, 6 of 8 inlB 2 isolates, 9 of 21 inlB 15 isolates, and 7 of 20 inlB 9 isolates did not belong to the outbreaks.
T147 781872-781998 Sentence denotes Conclusions: HRM-PCR analysis appears to be a sensitive, fast and inexpensive screening method to discover possible outbreaks.
T148 781999-782169 Sentence denotes In our study, it better distinguished outbreak clones from unrelated strains than serotyping, but it still requires confirmation by more discriminatory methods like PFGE.
T149 782170-782228 Sentence denotes Population analysis of community-acquired MRSA in Germany:
T150 782229-782289 Sentence denotes Data from the National Reference Centre for Staphylococci F.
T151 782290-782303 Sentence denotes Layer* 1 , C.
T152 782304-782315 Sentence denotes Cuny 1 , B.
T153 782316-782334 Sentence denotes Strommenger 1 , W.
T154 782335-782450 Sentence denotes Witte 1 1 Robert Koch Institut, National Reference Centre for Staphylococci, Wernigerode, Deutschland Introduction:
T155 782451-782539 Sentence denotes Community associated MRSA (CA-MRSA) have emerged worldwide from the end of the 1990ties.
T156 782540-782773 Sentence denotes Although particular clonal lineages were associated with particular geographic areas (e.g. ST30: "Oceanic", ST8: "USA300", ST80: "European", ST59 "Asiatic") further spread leading to dynamics of clonal lineages over time is observed.
T157 782774-782862 Sentence denotes Here we report on CA-MRSA clonal lineages recorded in Germany over the past three years.
T158 782863-782871 Sentence denotes Methods:
T159 782872-783003 Sentence denotes The respective isolates were sent to the Reference Centre for further characterisation between January 2009 and May 2011 (n = 320).
T160 783004-783173 Sentence denotes The antimicrobial susceptibility, the presence of several resistance and pathogenicity associated genes, the SCCmec-type, spa-and multilocus sequence-type were analyzed.
T161 783174-783259 Sentence denotes For selected isolates lukSF-PV was sequenced to determine the PVL sequence variation.
T162 783260-783374 Sentence denotes PFGE after SmaIdigestion of the chromosomal DNA was also performed for the comparison of selected clonal lineages.
T163 783375-783383 Sentence denotes Results:
T164 783384-783475 Sentence denotes The dominant CA-MRSA were the European clone ST80, isolates attributed to ST8, and to ST30.
T165 783476-783577 Sentence denotes Compared to data of the last years, we observed an increasing frequency of ST8 and surprisingly ST30.
T166 783578-783647 Sentence denotes At the first look CA-MRSA ST8 were suspicious for the "USA300" clone.
T167 783648-783714 Sentence denotes However, a considerable portion of them was lacking arcA and mphB.
T168 783715-783862 Sentence denotes These isolates do not represent a subpopulation of "degenerated" ST8 (loss of the ACME cluster and of the resistance plasmid) as initially assumed.
T169 783863-783980 Sentence denotes Although finding of the same rudimentary inverted repeat in the fosB vicinity and of sek and set suggest relatedness.
T170 783981-784103 Sentence denotes SCCmecIVc in the arcA-negative isolates instead of SCCmecIVa, which is typical for USA300, indicates convergent evolution.
T171 784104-784308 Sentence denotes Furthermore CA-MRSA clones which belonged to ST1, ST5, ST22, ST152 The capsule-specific synthesis (cap) regions II of Haemophilus influenzae serotypes c and d (Hic and Hid) were unknown before this study.
T172 784309-784558 Sentence denotes The Hic capsule consists of dimeric repeating units of O-acetylated N-acetyl--Dglucosamine ( -D-GlcNAc) and -D-Galactose ( -D-Gal), whereas the Hid capsule is essentially a polymer of -D-GlcNAc and N-acetyl-Dmannosaminuronic acid (D-ManANAc) dimers.
T173 784559-784745 Sentence denotes The polysaccharide of Hid shares common monomers with Hie capsule, and high similarities between the capsule loci of Hie and primarily of P. multocida B and E have been found previously.
T174 784746-784905 Sentence denotes We here present an analysis of the cap regions II of Hic and Hid and, based on similarity searches, discuss the phylogenetic development of bacterial capsules.
T175 784906-785015 Sentence denotes Sequencing of the cap region II of Hic ATCC 9007 and Hid ATCC 9008 was carried out by PCR and primer walking.
T176 785016-785091 Sentence denotes The sequence of Hic revealed four open reading frames (ORFs), named ccs1-4.
T177 785092-785158 Sentence denotes Similarity searches showed a phosphotransferase function for Ccs1.
T178 785159-785217 Sentence denotes Ccs2 seems to be a member of glycosyltransferase family 2.
T179 785218-785388 Sentence denotes These findings point at a role of both proteins in the synthesis of the capsule polysaccharaide, where a phosphodiester linkage connects O-acetylated -D-GlcNAc to -D-Gal.
T180 785389-785610 Sentence denotes Ccs4 was found to be subject to phase variation and showed similarities to O-acetyl transferases of the CysE/LacA/LpxA/NodL family, which includes the capsule O-acetyltransferase of Neisseria meningitidis serogroup W-135.
T181 785611-785810 Sentence denotes Of the five ORFs dcs1-5 found in Hid, similarity searches revealed an UDP-N-acetyl-D-glucosamine 2-epimerase function for Dcs1 and UDP-Nacetyl-D-mannosaminuronic acid dehydrogenase function for Dcs2.
T182 785811-785980 Sentence denotes Their putative role in the conversion of -D-GlcNAc to -D-ManANAc corresponds to the repetitive disaccharide units -D-GlcNAc and -D-ManANAc of the capsule polysaccharide.
T183 785981-786251 Sentence denotes The similarity of genes and gene organization found in Hic and Hid to capsulation genes and operons in Actinobacillus spp. as well as to more distant genera including N. meningitidis suggest horizontal gene transfer during capsule evolution across the bacterial classes.
T184 786252-786254 Sentence denotes A.
T185 786255-786270 Sentence denotes Flieger* 1 , A.
T186 786271-786283 Sentence denotes Fruth 1 , R.
T187 786284-786297 Sentence denotes Prager 1 , W.
T188 786298-786311 Sentence denotes Rabsch 1 , E.
T189 786312-786655 Sentence denotes Tietze 1 1 RKI, FG Bakterielle Infektionen, Wernigerode, Deutschland Within the National Reference Centre (NRC) for Salmonella and other Enteric Bacterial Pathogens located at the Robert Koch-Institut, a broad spectrum of human pathogenic bacteria is investigated, such as Salmonella, intestinal E. coli, Shigella, Yersinia, and Campylobacter.
T190 786656-786846 Sentence denotes Additionally since 2009, the NRC has been the German partner of the Binational Consilary Laboratory for Listeria located at The Austrian Agency for Health and Food Safety in Vienna, Austria.
T191 786847-787082 Sentence denotes Methods for lab-based surveillance of the above mentioned pathogens are developed, validated, and used to monitor changes of pathogen occurrence, alterations within the pathogen (e.g. virulence gene profile), and antibiotic resistance.
T192 787083-787202 Sentence denotes Further, clusters or increased abundance of disease cases are investigated to examine their epidemiologic relationship.
T193 787203-787397 Sentence denotes Within the year 2010, about 6000 bacterial samples from human disease cases were analyzed and among others the following interesting observations with respect to typing of the strains were made:
T194 787398-788067 Sentence denotes 1) Reduction in occurrence of the formerly leading Salmonella enterica serovar Enteritidis 2) Increase in occurrence of monophasic Salmonella enterica serovar Typhimurium strains 3) Increased observation of Salmonella infections in children caused by rare serotypes and the link to reptile presence in households 4) Virulence gene fingerprinting of disease-associated intestinal E. coli and characterization of intermediate E. coli strains which contain virulence genes or plasmids from different pathotypes 5) Work on outbreaks or family-associated cases due to Salmonella (75), E. coli (31, EHEC and EPEC), Campylobacter (10), Shigella sonnei (2) , and Listeria (2) .
T195 788068-788144 Sentence denotes Tasks of the National Reference Centre for Helicobacter pylorian overview E.
T196 788145-788160 Sentence denotes Glocker* 1 , M.
T197 788161-788172 Sentence denotes Kist 1 , N.
T198 788173-788192 Sentence denotes Wueppenhorst 1 , B.
T199 788193-788502 Sentence denotes Hobmaier 1 1 Universitätsklinikum Freiburg, Institut für Medizinische Mikrobiologie und Hygiene, Freiburg, Deutschland In 2001 the German National Reference Centre (NRC) for Helicobacter pylori launched ResiNet, a German multicentre study aiming to keep resistance development in H. pylori under surveillance.
T200 788503-788683 Sentence denotes In collaboration with 14 microbiological centres all across Germany, ResiNet monitors antimicrobial resistance rates in H. pylori and identifies risk factors for their development.
T201 788684-788785 Sentence denotes This is of paramount importance, since resistances have a massive negative impact on therapy success.
T202 788786-789139 Sentence denotes The NRC is dedicated to constrain the increase of resistances by recommending susceptibility testing in patients after the first treatment failure and in patients who have undergone antibiotic therapies at multiple occasions due to unrelated bacterial infections, as noted down in the S3-guideline "Helicobacter pylori and gastroduodenal ulcer disease".
T203 789140-789374 Sentence denotes Even though phenotypic susceptibility testing, as standardised by the NRC, has been shown to be reliable and reproducible, it may fail due to extra-long transport times, incorrect handling of gastric tissue samples and contaminations.
T204 789375-789562 Sentence denotes To overcome these shortcomings, the NRC has set up molecular genetic methods to examine antimicrobial resistance in H. pylori genotypically by identifying resistance associated mutations.
T205 789563-789753 Sentence denotes The request for genotypic resistance testing in some 5000 patients' samples underlines the increasing demand and need for these rescue diagnostic procedures, which are continuously enhanced.
T206 789754-789978 Sentence denotes Since 2006, the NRC has been offering gastroenterologists a unique service: on request, they are sent individual treatment recommendations based on the antimicrobial resistance patterns of their patients' H. pylori isolates.
T207 789979-790180 Sentence denotes Data on the outcome of the so far more than 1500 therapy recommendations will be analysed and will provide precious data to work out optimised treatment recommendations for difficult-to-treat patients.
T208 790181-790582 Sentence denotes 10 years after being founded, the NRC for H. pylori established itself as an indispensable institution needed to (i) improve the diagnosis of H. pylori related infections; (ii) to control the development of antimicrobial resistance in H. pylori; (iii) to standardise antimicrobial susceptibility testing procedures; and (iv) to give treatment recommendations in difficultto-treat H. pylori infections.
T209 790583-790748 Sentence denotes Characterization of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) using multiple-locus variable number of tandem repeats analysis (MLVA) K.
T210 790749-790763 Sentence denotes Brandt* 1 , A.
T211 790764-790779 Sentence denotes Mellmann 1 , C.
T212 790780-790792 Sentence denotes Jenke 1 , R.
T213 790793-791049 Sentence denotes Köck 1 1 University Hospital Münster, Institute for Hygiene, Münster, Deutschland Staphylococcus aureus is an important human pathogen and the emergence of methicillin-resistant S. aureus (MRSA) has become a major concern especially in healthcare settings.
T214 791050-791170 Sentence denotes Recently, MRSA have also emerged in livestock (e. g. swine and poultry) and are spreading to humans from this reservoir.
T215 791171-791296 Sentence denotes Therefore, precise typing methods to unravel the transmission dynamics of zoonotic S. aureus strains are of great importance.
T216 791297-791451 Sentence denotes Widely used and portable tools to track epidemic isolates are multilocus sequence typing (MLST) and sequence typing of the S. aureus protein A (spa) gene.
T217 791452-791670 Sentence denotes However, studies have shown that zoonotic MRSA-isolates from livestock mainly belong to the MLST clonal complex 398 and are mostly associated with a limited subset of spa types (predominantly with t011, t108 and t034).
T218 791671-791876 Sentence denotes To assess the range of genetic diversity among these MRSA isolates, we investigate in whether multiple-locus variable number of tandem repeats analysis (MLVA) provides a greater resolution than spa-typing.
T219 791877-792060 Sentence denotes In this study, we apply the S. aureus-MLVA scheme from Schouls et al. [1] that includes eight different gene loci and is based on automated fragment analysis on a capillary sequencer.
T220 792061-792253 Sentence denotes We aim to investigate the diversity of livestock-associated MRSA isolates derived between 2005 and 2010 from more than 50 pig stables comprising locations in various federal states of Germany.
T221 792254-792414 Sentence denotes First results of MLVA show that by spa-typing indistinguishable strains of different geographic regions can be clustered into at least two different MLVA-types.
T222 792415-792544 Sentence denotes MLVA discriminated isolates associated with spa-type t011 from two different pig farms assigning them to MLVA-types 398 and 2215.
T223 792545-792685 Sentence denotes Yet, the MLVA profiles identified so far differ in only one locus which supports the theory of a clonal origin of livestock-associated MRSA.
T224 792686-792840 Sentence denotes MLVA typing of zoonotic MRSA might therefore contribute to a better understanding of its genetic diversity against a geographical and temporal background.
T225 792841-793006 Sentence denotes Our findings will finally add valuable information to investigate the transmission dynamics of a zoonotic pathogen that has only recently overcome a species barrier.
T226 793007-793109 Sentence denotes Similarities of the dense granule protein GRA9 between the close relatives T. gondii and N. caninum M.
T227 793110-793128 Sentence denotes Leineweber* 1 , K.
T228 793129-793143 Sentence denotes Spekker 1 , V.
T229 793144-793157 Sentence denotes Ince 1 , S.K.
T230 793158-793172 Sentence denotes Schmidt 1 , G.
T231 793173-793187 Sentence denotes Schares 2 , P.
T232 793188-793203 Sentence denotes Maksimov 2 , W.
T233 793204-793531 Sentence denotes Däubener 1 1 Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Düsseldorf, Deutschland 2 Institut für Epidemiologie, Friedrich Löffler Institut, Wusterhausen, Deutschland Neospora caninum (N. caninum) is an obligate intracellular apicomplexan parasite which is very close related to Toxoplasma gondii (T. gondii).
T234 793532-793623 Sentence denotes The definitive hosts of N. caninum are dogs and cattle display the main intermediate hosts.
T235 793624-793738 Sentence denotes This work deals with dense granule proteins which are known to have important roles in host-parasite-relationship.
T236 793739-793808 Sentence denotes Previously, we described the dense granule protein GRA9 in T. gondii.
T237 793809-793874 Sentence denotes In this study we identified the putative homologue in N. caninum.
T238 793875-793961 Sentence denotes The genomic sequence of NcGRA9 is located on chromosome XII and consists of two exons.
T239 793962-794034 Sentence denotes By Southern Blot it was proven that it is present as a single copy gene.
T240 794035-794211 Sentence denotes After production of a N. caninum specific anti-GRA9 antiserum against the recombinant protein it could be shown by Western Blot that the NcGRA9 gene encodes a 40,2 kDa protein.
T241 794212-794308 Sentence denotes Furthermore, we defined NcGRA9 as a member of the excreted-secreted antigens (ESA) of N.caninum.
T242 794309-794499 Sentence denotes By confocal microscopy it was observed that in extracellular parasites the protein is accumulated in a dotted pattern which points out that NcGRA9 is located in the dense granule organelles.
T243 794500-794640 Sentence denotes Furthermore, we could show that after invasion of the parasites into the host cell NcGRA9 is secreted into the parasitophorous vacuole (PV).
T244 794641-794702 Sentence denotes Thus NcGRA9 fulfils all main characteristics of GRA proteins.
T245 794703-794944 Sentence denotes In sum, we could verify in this study that NcGRA9 and TgGRA9 are structurally similar, show immunological cross reactivity and are similar distributed within the respective parasites which indicate a close relationship between both proteins.
T246 794945-795206 Sentence denotes Interaction and localization of the Toxoplasma gondii protein GRA9 within the parasitophorous vacuole The apicomplexan parasite Toxoplasma gondii (T. gondii) is one of the most successful protozoan parasites infecting all warm-blooded animals, including humans.
T247 795207-795324 Sentence denotes In host cells the parasite develops within a parasitophorous vacuole (PV), which causes handicaps in nutrient supply.
T248 795325-795496 Sentence denotes GRA-and ROP-proteins, the main components of the PV are delivered from secretory organelles and are important in the connection between parasites and host cell organelles.
T249 795497-795647 Sentence denotes We have described a new 41 kDa T. gondii protein called GRA9 which stably associates with the network of membranous tubules and the PV membrane (PVM).
T250 795648-795748 Sentence denotes We show here that GRA9 co-localize with the dense granules proteins GRA1, GRA2, GRA3, GRA5 and GRA7.
T251 795749-795949 Sentence denotes Recent coimmunoprecipitation data indicate that GRA9 is a major interaction partner of GRA7 and GRA3 which both were found to be important in nutrient transport between the host cell and the parasite.
T252 795950-796166 Sentence denotes To explore GRA9 secretion into the PV and the interaction with network structures and the PVM in more detail, we exogenously expressed HAtagged GRA9 as well as C-terminal truncated versions of GRA9 in RH tachyzoites.
T253 796167-796283 Sentence denotes Results indicate that the C-terminus of GRA9 is essential and sufficient for the correct GRA9 secretion into the PV.
T254 796284-796416 Sentence denotes GRA9-proteins lacking the last one hundred amino acids, including two hydrophobic domains show an alteration in secretion behaviour.
T255 796417-796555 Sentence denotes The C-terminal truncated GRA9-protein was found exclusively within the cytoplasm of the parasites and could not enter the lumen of the PV.
T256 796556-796866 Sentence denotes Advances in the biomolecular interaction analysis of immunoglobulin-binding protein EibG from Shiga toxinproducing Escherichia coli Shiga toxin-producing Escherichia coli (STEC) can cause diarrhea and the hemolytic uremic syndrome (HUS) in humans and belong therefore to a group of pathogens of major interest.
T257 796867-796947 Sentence denotes Several STEC strains express seven related immunoglobulin (Ig)-binding proteins:
T258 796948-796974 Sentence denotes EibA, C, D, E, F, G and H.
T259 796975-797096 Sentence denotes Sharing the facility to bind IgG by nonimmune manner, these proteins occur separately or combined within a single strain.
T260 797097-797236 Sentence denotes Additionally to the potential evasion of host immune response, EibG is involved in bacterial adherence to host intestinal epithelial cells.
T261 797237-797347 Sentence denotes Furthermore eibG-positive strains show chain-like adherence pattern (CLAP) depending on different eibG-allels.
T262 797348-797456 Sentence denotes Sequencing of entire eibG resulted in 21 different eibG-allels allocated in three subtypes (eibG-, -and -) .
T263 797457-797572 Sentence denotes Adhesion assays with human and bovine epithelial cells showed different CLAP-phenotypes depending on eibG-subtypes.
T264 797573-797669 Sentence denotes These results were confirmed by inserting eibG into E. coli with alternative genetic background.
T265 797670-797865 Sentence denotes The possible functional role of glycosylation of IgG, harboring N-glycans in the CH2 domain of the Fc fragment, in EibG affinity was investigated with native and enzymatically deglycosylated IgG.
T266 797866-798058 Sentence denotes Peptide N-glycosidase (PNGase) treatment of IgG did not influence EibG-IgG interaction, indicating an EibG binding domain different from the site of glycosylation within the CH2 domain of IgG.
T267 798059-798325 Sentence denotes Next EibG affinity to IgG subtypes of different vertebrate species was determined by ELISA under variable conditions aimed at identification of Igs with high binding affinity to EibG that may facilitate efficient isolation and purification of EibG from cell lysates.
T268 798326-798506 Sentence denotes Pending determination of the amino acid sequences of EibG variants will be performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).
T269 798507-798665 Sentence denotes Thus, peptide mapping and comparison to sequences of other Eib-proteins should help to unravel the molecular mechanisms of IgG-binding and cellular adherence.
T270 798666-798897 Sentence denotes Furthermore, in search of the cellular receptors of epithelial cells, the potential affinity of EibG to glycosphingolipids of different biological origin will be explored by thin-layer chromatography (TLC) overlay assay techniques.
T271 798898-798995 Sentence denotes Do complement factor H 402Y and C7 M allotypes predispose to (typical) hemolytic uremic syndrome?
T272 798996-799190 Sentence denotes Typical hemolytic uremic syndrome (HUS) is mainly caused by infections with enterohemorrhagic Escherichia coli, whereas in atypical, non-bacteriaassociated HUS, complement plays a dominant role.
T273 799191-799261 Sentence denotes Recently complement has also been shown to be involved in typical HUS.
T274 799262-799499 Sentence denotes In this study, mostly weakly significant associations with homozygosities of complement allotype C7 M and inversely with factor H 402H were found, suggesting that 402Y and C7 M allotypes predispose to (typical) hemolytic uremic syndrome.
T275 799500-799600 Sentence denotes Establishment of a National Giardia intestinalis Database and Biobank for functional epidemiology C.
T276 799601-799614 Sentence denotes Klotz* 1 , S.
T277 799615-799627 Sentence denotes Banik 1 , P.
T278 799628-799650 Sentence denotes Gosten-Heinrich 1 , T.
T279 799651-799863 Sentence denotes Aebischer FG16 Parasitologie, Berlin, Deutschland Giardiasis is a common and worldwide distributed protozoan infection of the gastrointestinal tract that is caused by the flagellate parasite Giardia intestinalis.
T280 799864-800079 Sentence denotes Clinical signs are extremely variable, ranging from asymptomatic to chronic symptoms including severe diarrhoea and abdominal pain, and the contributing factors and mechanisms of pathophysiology are largely unknown.
T281 800080-800317 Sentence denotes The zoonosis potential of distinct Giardia intestinalis genotypes has been highlighted, however, definition of specific parasite virulence factors and host risk factors has been hampered by the lack of functional epidemiological studies.
T282 800318-800652 Sentence denotes The aim of this project is to built-up a unique database-linked biobank collection of Giardia intestinalis from human clinical cases and isolates from the field or from domestic animals and thus connecting epidemiological data and molecular characteristics with in-vitro functional test using the respective parasite isolate cultures.
T283 800653-800857 Sentence denotes Subsequently, we will exploit the data and clonal parasite collection for functional studies, e.g. to correlate genotype-specific variations in potential virulence factors with functional characteristics.
T284 800858-801057 Sentence denotes Currently in the first phase of the project, we are collecting, characterizing and cultivating Giardia isolates from humans and animals contributed by cooperation-partners in Germany and Switzerland.
T285 801058-801200 Sentence denotes We successfully established an in-vitro culture protocol for the expansion of clonal Giardia trophozoites derived from human faecal specimens.
T286 801201-801448 Sentence denotes First genotyping results of cloned parasites at the glutamate dehydrogenase gene locus indicate a successful establishment of assemblage B sub-cultures, which are more difficult to isolate than assemblage A due to slower growth behaviour in vitro.
T287 801449-801572 Sentence denotes Eventually, this biobank collection will provide a unique tool for functional epidemiological studies of virulence factors.
T288 801573-801619 Sentence denotes Knowledge is there but where is the knowledge?
T289 801620-801659 Sentence denotes Knowledge management in microbiology M.
T290 801660-801674 Sentence denotes Walter* 1 , D.
T291 801675-801869 Sentence denotes Frangoulidis 2 1 Helmholtz-Zentrum München, Institut für Bioinformatik und Systembiologie, Neuherberg, Deutschland 2 Institut für Mikrobiologie der Bundeswehr, München, Deutschland Introduction:
T292 801870-802027 Sentence denotes The rapid expansion of biological data resulting from thousands of genome sequencing projects and recently developed highthroughput techniques is prodigious.
T293 802028-802141 Sentence denotes Furthermore, the numbers of article citations in public literature databases like PubMed increases exponentially.
T294 802142-802279 Sentence denotes Hence, the risk for the individual specialist to lose control of the available information in his field of interest is rising constantly.
T295 802280-802390 Sentence denotes To address these problems databases and software tools were constructed to structure and consolidate the data.
T296 802391-802492 Sentence denotes But the numbers of these databases is growing as well and their quality is by far not self-evidently.
T297 802493-802599 Sentence denotes Here we show our recent developments using text and data mining technologies to make knowledge accessible.
T298 802600-802608 Sentence denotes Methods:
T299 802609-802665 Sentence denotes Detailed knowledge is stored in full-text articles only.
T300 802666-802805 Sentence denotes Thus we enhanced our in-house text mining system to work on all available and licensed full-text articles in our special field of interest.
T301 802806-802893 Sentence denotes Then we extracted a large set of entity relationships and created a relational network.
T302 802894-802991 Sentence denotes Additionally, we extended a data mining platform to perform multiple genome analyses in parallel.
T303 802992-803114 Sentence denotes Finally, we integrated the text mining into the data mining platform to enhance the analytic power of the complete system.
T304 803115-803123 Sentence denotes Results:
T305 803124-803253 Sentence denotes We obtained all available full-text articles (about 1,800) of our main research field regarding to Coxiella burnetii and Q fever.
T306 803254-803421 Sentence denotes Based on selected review articles, we evaluate and compare the automatically extracted relation network with an interaction network created by human curators in-house.
T307 803422-803494 Sentence denotes This allows us to determine in-depth the bias of the automated approach.
T308 803495-803507 Sentence denotes Conclusions:
T309 803508-803608 Sentence denotes The data mining techniques used here allows scientists to deal with many species or strains at once.
T310 803609-803788 Sentence denotes With the incorporation of text mining they are further able to quickly highlight important functions, causalities and differences as well as to reveal previously hidden knowledge.
T311 803789-803999 Sentence denotes Hence the here presented tool assist users strongly during their studies on novel aspects and hypotheses in their research field when providing the complete accessible knowledge in a fast and structured manner.
T312 804000-804050 Sentence denotes Now, knowledge is there where knowledge should be.
T313 804051-804167 Sentence denotes Core genome determination of zoonotic pathogens using whole genome data in combination with a new web application S.
T314 804168-804170 Sentence denotes R.
T315 804171-804188 Sentence denotes Leopold* 1 , D.J.
T316 804189-804210 Sentence denotes Schmitz-Hübsch 2 , D.
T317 804211-804225 Sentence denotes Harmsen 2 , A.
T318 804226-804393 Sentence denotes Mellmann 1 1 Universitätsklinikum Münster, Institut für Hygiene, Münster, Deutschland 2 Universitätsklinikum Münster, Periodontologie, Münster, Deutschland Background:
T319 804394-804570 Sentence denotes The increasing accessibility of next generation sequencing tools has led to an abundance of genome data, including both fully finished genomes and unfinished assembled contigs.
T320 804571-804746 Sentence denotes Definition of a core genome, pan genome and/or singleton genes are often necessary to perform comparative or phylogenetic analyses of multiple strains within the same species.
T321 804747-804976 Sentence denotes We developed a program that facilitates comparative genome analysis utilizing both annotated as well as un-annotated sequence data, while also providing maximal options and parameters for the user for further downstream analysis.
T322 804977-804985 Sentence denotes Methods:
T323 804986-805176 Sentence denotes This program utilizes NCBI's BLAST algorithm to query the genome(s) of interest using a list of ORFs or sequences (defined by the user, or based upon derived from annotated genomes queried).
T324 805177-805302 Sentence denotes The user controls the parameters for the 'core' definition by setting a threshold for percent identity, percent overlap, etc.
T325 805303-805428 Sentence denotes Perl programs pair BLAST hit data with corresponding subject ORF data, then sort the results into separate categorized files.
T326 805429-805649 Sentence denotes If querying annotated genomes, the user may choose various output file formats such as a list of orthologous core ORFs, the sequence of the core ORFs for a reference strain, and/or these core ORFs sequences concatenated.
T327 805650-805658 Sentence denotes Results:
T328 805659-805778 Sentence denotes The Perl programs were implemented in a web interface based on Java, creating a user-friendly graphical-user interface.
T329 805779-806138 Sentence denotes An evaluation with 15 Staphylococcus aureus and 4 Escherichia coli O157:H7 genomes with default parameters (95% minimal sequence similarity, not >95% similar to another ORF within the same chromosome (percent identity and overlap), 100% presence in the investigated genomes/sequences) yielded 1,131 and 3,200 core ORFs for S. aureus and E. coli, respectively.
T330 806139-806311 Sentence denotes All non-core ORFs are reported to the User in categorized files (e.g., multi-copy ORFs, alternative start and/or stop codons, gaps in alignments, <95% sequence similarity).
T331 806312-806324 Sentence denotes Conclusions:
T332 806325-806481 Sentence denotes The Microbial Core Genome analysis tool, MCGenome, can be used to comparatively analyze multiple annotated genomes or interrogate a single un-annotated one.
T333 806482-806648 Sentence denotes The process was created to be completely transparent by giving the user control over their parameters and their data, while maintaining a user-friendly web interface.
T334 806649-806695 Sentence denotes The source code is available from the authors.
T335 806696-806709 Sentence denotes Introduction:
T336 806710-806847 Sentence denotes Sparganosis is a neglected parasitic disease caused by the plerocercoid stage (sparganum) of Spirometra sp., a pseudophyllidean tapeworm.
T337 806848-807020 Sentence denotes The disease is most often found in East Asia, were humans become infected by eating raw or undercooked meat of fish, amphibians, or reptiles which contain the larval stage.
T338 807021-807114 Sentence denotes The spargana invade the brain, eye, viscera, and the subcutis, and can cause serious illness.
T339 807115-807283 Sentence denotes A rare and complicated form, disseminated proliferative sparganosis, is caused by a branching, maldifferentiated plerocercoid provisionally termed Sparganum proliferum.
T340 807284-807292 Sentence denotes Methods:
T341 807293-807463 Sentence denotes Several 4-5 cm long, flattened pseudosegmented helminths were extracted from a subcutaneous nodule of the infraumbilical region of a 60year-old female Japanese immigrant.
T342 807464-807558 Sentence denotes The parasite was formalin-fixed and embedded in paraffin for further histological examination.
T343 807559-807727 Sentence denotes In addition, DNA was extracted and subjected to a cestode-specific mitochondrial 12S rRNA gene-and cytochrome c oxidase subunit 1 (cox1)-PCR for species identification.
T344 807728-807736 Sentence denotes Results:
T345 807737-807911 Sentence denotes Morphological and histological analysis of the helminth showed typical aspects of a cestode, i.e. calcareous corpuscles in a spongy stroma surrounded by an aspinous tegument.
T346 807912-808088 Sentence denotes As a characteristic feature of a pseudophyllidean cestode larva (plerocercoid or sparganum), the anterior region showed an invagination and no proper scolex had been developed.
T347 808089-808155 Sentence denotes The organism was pseudosegmented without any strobilar structures.
T348 808156-808341 Sentence denotes Sequence analysis of the 440 and 425 bp amplicons of the 12S rRNA geneand cox1-PCR showed 100% and 99% identity with Spirometra erinaceieuropaei (AB374543.1 and AF096237.2, AF096238.2).
T349 808342-808432 Sentence denotes Histological sections of the parasite were also used to set up an immunofluorescence test.
T350 808433-808533 Sentence denotes When incubated with the patient's serum a tegumental signal was detected at concentrations of 1:100.
T351 808534-808545 Sentence denotes Conclusion:
T352 808546-808678 Sentence denotes Spirometra erinaceieuropaei has a cosmopolitan distribution, but most human cases occur in Asia possibly due to local eating habits.
T353 808679-808891 Sentence denotes PCR analysis unambiguously identified the species responsible for the patient's disease and thus excluded the more pathogenic Sparganum proliferum, a larval cestode for which the adult strobilar stage is unknown.
T354 808892-808894 Sentence denotes I.
T355 808895-808909 Sentence denotes Popescu 1 , I.
T356 808910-808923 Sentence denotes Tudose 1 , B.
T357 808924-808937 Sentence denotes Muntau 2 , C.
T358 808938-808956 Sentence denotes Giurcaneanu 1 , S.
T359 808957-809106 Sentence denotes Poppert* 2 1 ELIAS Emergency University Hospital, Bukarest, Rumaenien 2 Bernhard-Nocht-Institut für Tropenmedizin, Hamburg, Deutschland Introduction:
T360 809107-809245 Sentence denotes Human dirofilariasis is a zoonotic infectious disease caused by the filarial nematodes of dogs Dirofilaria repens and Dirofilaria immitis.
T361 809246-809417 Sentence denotes Depending on the species involved human infections usually manifest as one cutaneous or visceral larva migrans that forms a painless nodule in the later course of disease.
T362 809418-809487 Sentence denotes Dirofilariae are endemic in the Mediterranean, particularly in Italy.
T363 809488-809571 Sentence denotes They are considered as emerging pathogens just increasing their geographical range.
T364 809572-809581 Sentence denotes The case:
T365 809582-809675 Sentence denotes We present one of the few known cases of human dirofilariasis caused by D. repens in Romania.
T366 809676-809821 Sentence denotes The patient developed unusual and severe clinical manifestations that mimicked pathological conditions like cellulitis or deep venous thrombosis.
T367 809822-809833 Sentence denotes Conclusion:
T368 809834-810009 Sentence denotes Human doctors as well as veterinarians should be prepared to come across cases of Dirofilariasis in Romania, as well as in other European countries regarded nonendemic so far.
T369 810010-810205 Sentence denotes As our case illustrates, infection should even be considered in patients with signs of parasitic infection, which do not exactly correspond to the description of Dirofilariasis in the text books.
T370 810206-810333 Sentence denotes Characterization of sorbitol-fermenting Shiga toxin-negative eae-positive Escherichia coli O157 strains isolated from humans W.
T371 810334-810347 Sentence denotes Zhang* 1 , K.
T372 810348-810361 Sentence denotes Hansen 1 , C.
T373 810362-810374 Sentence denotes Rüter 2 , M.
T374 810375-810631 Sentence denotes Bielaszewska We characterized 31 Shiga toxin (Stx)-negative sorbitol-fermenting (SF) E. coli O157 strains isolated during 1987-2007 from patients with hemolytic uremic syndrome (HUS) (n = 16), diarrhea (n = 10) and asymptomatic carriers (n = 5) in Germany.
T375 810632-810782 Sentence denotes H serotyping and fliC genotyping identified five different H types including H7, H16, H34, H39, and H45; all strains with fliCH7 were H -(non-motile).
T376 810783-811018 Sentence denotes The strains carried four different eae alleles ( , , and a) which were associated with particular serotypes/fliC types: strains O157:H -[fliCH7] carried eae-g, O157:H16/H -[fliCH16] and O157:H34 eae-, O157:H39 eae-, and O157:H45 eae-a.
T377 811019-811231 Sentence denotes All O157:H -[fliCH7] strains possessed sfpA, a marker for the gene cluster encoding Sfp fimbriae, and most of them carried EHEC-hlyA and cdtV encoding EHEC hemolysin and cytolethal distending toxin, respectively.
T378 811232-811298 Sentence denotes None of these virulence genes was present in O157:non-H7 isolates.
T379 811299-811426 Sentence denotes Moreover, strains of the different serotypes differed by their ability to adhere to human intestinal epithelial cells in vitro.
T380 811427-811631 Sentence denotes Phylogenetic analysis of the 31 Stx-negative SF O157 strains using multilocus sequence typing demonstrated five different sequence types (STs 11, 10, 752, 1041, 593) and O157:H16 were associated with HUS.
T381 811632-811838 Sentence denotes Our data demonstrate that the group of E. coli O157 pathogenic for humans is broader than previously thought and includes, besides classical nSF STEC O157:H7 also SF E. coli O157 of several other serotypes.
T382 811839-812105 Sentence denotes Stx-independent diagnostic strategies involving PCRs targeting rfbO157 and eae are method of choice to identify these pathogens Enterohemorrhagic Escherichia coli (EHEC) is a highly pathogenic and Shiga toxin (Stx) positive subgroup of intestinal pathogenic E. coli.
T383 812106-812215 Sentence denotes EHEC infections often lead to diarrhea, casually hemorrhagic colitis and the hemolytic uremic syndrome (HUS).
T384 812216-812426 Sentence denotes Whereas EHEC O157:H7 is the most common serotype in HUS patients, EHEC isolated from patients with diarrhea exhibit more frequently non-O157 serotypes; among them O26:H11 is the most common serotype in Germany.
T385 812427-812598 Sentence denotes In contrast to the wellstudied EHEC O157:H7, only little is known about the genome content and genome plasticity (e. g. loss of Stx-encoding genes [stx]) of this serotype.
T386 812599-812743 Sentence denotes We therefore investigated the genomes of two O26 strains isolated at an interval of 8 days from the same HUS-patient by whole genome sequencing.
T387 812744-812875 Sentence denotes Using 454 next generation and classical Sanger sequencing, we determined the complete genome sequences of the two O26:H11 isolates.
T388 812876-813042 Sentence denotes Whole genome comparisons using Mauve 2.3.0 software were performed to investigate differences in the gene content and to detect single nucleotide polymorphisms (SNP).
T389 813043-813127 Sentence denotes SNPs were additionally confirmed by PCR product sequencing of the particular region.
T390 813128-813237 Sentence denotes For confirmed SNPs in coding regions, phenotypic tests were applied to confirm their impact on the phenotype.
T391 813238-813356 Sentence denotes Whole genome comparisons revealed the loss of a total of 71 genes in the subsequent isolate after 8 days of infection.
T392 813357-813447 Sentence denotes The majority of them were located on a genomic island, which also included the stx operon.
T393 813448-813535 Sentence denotes Moreover, we determined three non-synonymous SNPs and three SNPs in non-coding regions.
T394 813536-813687 Sentence denotes The non-synonymous SNPs were located in rpoS (RNA polymerase sigma factor), mreB (rod shape-determining protein), and malT (transcriptional regulator).
T395 813688-813785 Sentence denotes Interestingly, the SNP in rpoS (position 443, TAG to TGG) resolved a stopcodon to an intact gene.
T396 813786-813929 Sentence denotes RpoS plays a crucial role in survival against stress and controls a large regulon, among them KatE (catalase HPII), which is dependent on RpoS.
T397 813930-814053 Sentence denotes Because of the reconstitution of the rpoS gene function, a positive catalase test was detectable in the subsequent isolate.
T398 814054-814168 Sentence denotes MalT, the transcriptional activator of maltose-regulon genes, may be also involved in protection against stresses.
T399 814169-814339 Sentence denotes In contrast, the change in MreB, which is involved in cell division, chromosome segregation, and is responsible for the typical rod shape of cells, is not understood yet.
T400 814340-814561 Sentence denotes Interestingly, synonymous SNPs, which usually represent the molecular clock and the normal mutation rate within a chromosome, were not detected indicating that external factors were responsible for the pathogens´ changes.
T401 814562-814737 Sentence denotes In conclusion, our analysis showed the enormous plasticity of genomes during infection because O26:H11 was able to undergo genomic changes with phenotypic effects within days.
T402 814738-814838 Sentence denotes Further studies will focus on the impact of the phenotypic changes to the host-pathogen interaction.
T403 814839-814841 Sentence denotes S.
T404 814842-814857 Sentence denotes Poppert* 1 , J.
T405 814858-814881 Sentence denotes Schmidt-Chanasit 1 , A.
T406 814882-814895 Sentence denotes Krüger 2 , E.
T407 814896-814917 Sentence denotes Tannich Introduction:
T408 814918-815028 Sentence denotes Dirofilaria (D.) repens is a filaria of dogs and other carnivores, which is transmitted by various mosquitoes.
T409 815029-815157 Sentence denotes D. repens can accidentally infect humans, which usually results in a subcutaneous nodule containing a single infertile parasite.
T410 815158-815254 Sentence denotes D. repens is endemic in many tropical and subtropical countries as well as in the Mediterranean.
T411 815255-815328 Sentence denotes Recently single cases of infections were reported from north of the Alps.
T412 815329-815349 Sentence denotes Methods and Results:
T413 815350-815452 Sentence denotes We recently received multiple samples of D. repens worms, which had been removed from infected humans.
T414 815453-815656 Sentence denotes The patients showed various clinical symptoms, such as dermal and subconjunctival larva migrans, formation of subcutaneous nodules at diverse locations as well as formation of nodules in lung and testis.
T415 815657-815766 Sentence denotes In most cases our phenotypic diagnosis was confirmed by PCR and sequencing of the mitochondrial 12SrRNA-gene.
T416 815767-815903 Sentence denotes In the recent years we diagnosed more cases of human D. repens infections than ever before, suggesting an increasing rate of infections.
T417 815904-816026 Sentence denotes It has been postulated that climate warming will lead to spread of D. repens infections even to regions north of the Alps.
T418 816027-816173 Sentence denotes We therefore analyzed more than 40 000 mosquitoes captured in Southwest Germany in the years 2009 and 2010 for the presence of filariae using PCR.
T419 816174-816272 Sentence denotes We did not find evidence for the presence of significant numbers of D. repens-infected mosquitoes.
T420 816273-816284 Sentence denotes Conclusion:
T421 816285-816434 Sentence denotes The number of D. repens infections seems to be increasing and doctors should be prepared to diagnose and treat corresponding infections in travelers.
T422 816435-816522 Sentence denotes In Germany mosquitoes do not (yet?) pose a significant risk for transmitting D. repens.
T1 816523-816710 Sentence denotes Protein complexes in brains are composed of heterogeneous cellular prion protein phenotypes The main characteristic of prion diseases is the accumulation of pathological prions (PrP Sc ).
T2 816711-816842 Sentence denotes The molecular mechanism leading to PrP Sc is considered to be caused by conversion of a normal host-encoded prion protein (PrP C ).
T3 816843-816954 Sentence denotes PrP C acts as substrate in this protein transformation with the result of generating few distinct PrP Sc types.
T4 816955-817096 Sentence denotes However, PrP C is composed of several overlapping types in brain protein complexes but little is known about the composition of the subtypes.
T5 817097-817231 Sentence denotes In this study we separated PrP C proteins obtained from human, sheep and cattle on the basis of differential solubility in detergents.
T6 817232-817447 Sentence denotes The subtypes were characterized by immunoblotting using various antibodies recognizing different PrP epitopes followed by densitometrical signal quantification and determination of the glycoprotein banding patterns.
T7 817448-817694 Sentence denotes Single and serial applications of the detergents N-octyl--Dglucopyranoside and CHAPS facilitated high solubility of glycosylated PrP C isoforms whereas high contingents of nonglycosylated human and half of total bovine PrP C remained non-soluble.
T8 817695-817798 Sentence denotes Most proteins of all species became highly soluble with N-lauroylsarcosine and sodium dodecyl sulphate.
T9 817799-817912 Sentence denotes Furthermore we identified an aggregated, highly soluble PrP C isoform in the supenatants of all analysed species.
T10 817913-818095 Sentence denotes Our findings demonstrate that heterogenous PrP C phenotypes overlapping in brains may be separated regarding their solubility characteristics and convey distinct solubility subtypes.
T11 818096-818347 Sentence denotes Protein differentiation by solubility experiments can therefore facilitate the separation of PrP C types, can provide detailed information on protein composition and can offer new prospects for transformation efficiency to PrP Sc by conversion assays.
T12 818348-818372 Sentence denotes Conclusion/Significance:
T13 818373-818646 Sentence denotes We conclude that gnotobiotic and "humanized" mice represent excellent novel C. jejuni-infection and -inflammation models and provide deep insights into the immunological and molecular interplays between C. jejuni, microbiota and innate immunity in human campylobacteriosis.
T14 818647-818731 Sentence denotes Helicobacter do transcribe small RNAs, they do not have a recognisable Hfq ortholog.
T15 818732-819019 Sentence denotes Using bioinformatic approaches, we have identified 3 genes (Cj0138, Cj0667 and Cj1103) in the C. jejuni genome which encode proteins with a predicted RNA-binding motif, and have investigated their role in gene expression and colonization/virulence using gnotobiotic IL-10 deficient mice.
T16 819020-819051 Sentence denotes Methodology/Principal findings:
T17 819052-819198 Sentence denotes Mutant strains were generated with an inactivated gene, with two copies of the gene, and one where the inactivated gene was complemented in trans.
T18 819199-819432 Sentence denotes Growth of the mutants was similar to that of the wild-type strain, but while inactivation of Cj1103 did not affect motility or growth phenotypes, overexpression of Cj1103 resulted in a significant decrease in autoagglutination (AAG).
T19 819433-819584 Sentence denotes Transcriptomic and proteomic characterisation of these strains suggested changes in expression of flagellar genes, which may explain the AAG phenotype.
T20 819585-819751 Sentence denotes Infection of gnotobiotic IL-10 deficient mice with the C. jejuni wildtype strain caused acute bloody diarrhea, histopathology and inflammatory responses in the colon.
T21 819752-819927 Sentence denotes All mutants colonized the mouse intestine to similar levels as the wildtype strain, but differed in their potential to induce diarrhea and inflammatory responses in the colon.
T22 819928-820093 Sentence denotes Clinical signs of disease and levels of pro-inflammatory cytokines in the colon were significantly reduced in the Cj0138 mutant when compared to the wildtype strain.
T23 820094-820118 Sentence denotes Conclusion/Significance:
T24 820119-820351 Sentence denotes Although there are subtle phenotypes associated with these putative RNA-binding proteins, there is currently no proof that these proteins function as general RNA-chaperones, as the predicted pleiotropic phenotypes were not observed.
T25 820352-820499 Sentence denotes The results obtained in the infected gnotobiotic IL-10-deficient mice indicate a role of the genes Cj0138, Cj0667 and Cj1103 in C.jejuni virulence.
T26 820500-820738 Sentence denotes Adherent-Invasive Escherichia coli (AIEC), which have been associated with Crohn's disease, show traits similar to human and animal extraintestinal pathogenic E. coli (ExPEC) regarding their phylogenetic origin and virulence gene profile.
T27 820739-820988 Sentence denotes However, previous studies have shown that human ExPEC strains rarely share the AIEC phenotype, whereas only limited data are available on the AIEC-like phenotype properties of animal E. coli strains, both from extraintestinal and intestinal sources.
T28 820989-821271 Sentence denotes We therefore analyzed the AIEC phenotype of 79 ExPEC strains from birds, cats, dogs, and swine, which were selected with respect to multi locus sequence types (STs) that had previously been linked with AIEC strains from patients with Crohn's disease, including ST131, ST73 and ST10.
T29 821272-821500 Sentence denotes In addition, 45 E. coli enteritis isolates from cats, dogs -in case of ST complex (STC) 73 (n=26) representing a phylotype typically observed among strains causing cystitis and pyelonephritis in humans -and swine, were included.
T30 821501-821825 Sentence denotes While only 1.3% of ExPEC strains showed an AIEC phenotype, as shown by the ability to adhere to and to invade intestinal epithelial cells, as well as to survive and replicate within murine macrophages, up to 57.8% of animal enteritis strains (33.3% swine; 35.3% dogs; 81.8% cats) phenotypically resembled the AIEC pathotype.
T31 821826-821978 Sentence denotes Interestingly, none of the STC73 ExPEC strains showed this phenotype, whereas 72.4% of STC73 strains from enteritis were confirmed as AIEC-like strains.
T32 821979-822234 Sentence denotes Due to the non-arbitrary strain selection no conclusion can be drawn considering the real prevalence of AIEC-like strains in the intestinal tract of animals, but their frequent detection provides support for a lack of host-specificity and a zoonotic risk.
T33 822235-822550 Sentence denotes Further studies including well characterized diseased animals are needed in order to detect putative reservoirs of AIEC strains, even in the intestine of asymptomatic carriers, and to evaluate the dimension of the risk with respect to the implementation of prevention and control measures and thus to public health.
T34 822551-822694 Sentence denotes A Enterohemorrhagic Escherichia coli (EHEC) cause hemolytic uremic syndrome (HUS), the most common cause of acute renal failure in children 1 .
T35 822695-822802 Sentence denotes Shiga toxins (Stxs) are key virulence factors of EHEC and endothelial cells are the major toxin targets 2 .
T36 822803-823022 Sentence denotes We performed a direct and comprehensive comparison of cellular injury induced by two major Stx types, Stx1 and Stx2, in human brain microvascular endothelial cells (HBMECs) and EA.hy 926 macrovascular endothelial cells.
T37 823023-823290 Sentence denotes Scanning electron microscopy of microcarrier-based cultures, digital holographic microscopy of single cells, and quantitative apoptosis/necrosis assays demonstrate that Stx1 causes both necrosis and apoptosis, whereas Stx2 induces mainly apoptosis in both cell lines.
T38 823291-823491 Sentence denotes Moreover, microvascular and macrovascular endothelial cells have different susceptibilities to the toxins: HBMECs are more susceptible to Stx2, whereas EA.hy 926 cells are more susceptible to Stx1 3 .
T39 823492-823696 Sentence denotes These findings have implications in the pathogenesis of HUS, and suggest the existence of yet to be delineated Stx type-specific mechanisms of endothelial injury beyond inhibition of protein biosynthesis.
T40 823697-823947 Sentence denotes Simultaneous detection of eight potential bioterrorism agents by real time PCR The Bacillus anthracis attacks in the United States in 2001 demonstrated the need of fast and reliable diagnostic methods for the detection of potential biological agents.
T41 823948-824263 Sentence denotes Not only Bacillus anthracis but also Burkholderia (pseudo-) mallei, Brucella ssp., Francisella tularensis, Yersinia pestis, Coxiella burnetii and the Orthopoxvirus Variola major are listed as biological agents with high risk of misuse in bioterrorism attacks by the Centers for Disease Control and Prevention (CDC).
T42 824264-824427 Sentence denotes Since then a number of PCR-based detection assays for these agents were published, some of them are designed as multiplex assay for two or three biological agents.
T43 824428-824533 Sentence denotes However, an assay for the simultaneous detection of eight different agents has not been developed so far.
T44 824534-824811 Sentence denotes We designed and optimised real-time PCR TaqMan-Assays for the detection of Bacillus anthracis, Burkholderia (pseudo-) mallei, Brucella ssp., Francisella tularensis, Yersinia pestis, Coxiella burnetii and Orthopoxviruses based on different chromosomal targets for each pathogen.
T45 824812-824907 Sentence denotes Primers and probes were lyophilizied on 96well plates for use with the LightCycler® 480-System.
T46 824908-825036 Sentence denotes In total we tested over 400 DNA samples including at minimum 20 DNA samples of each of the eight biological agents respectively.
T47 825037-825105 Sentence denotes All assays were tested for specificity and limit of detection (LOD).
T48 825106-825285 Sentence denotes Our findings show only positive PCR results for the respective target bacteria and no cross reaction with closely related species or other clinically relevant bacterial pathogens.
T49 825286-825388 Sentence denotes Sensitivity experiments showed detection limits down to 10 copies per reaction for most of the assays.
T50 825389-825545 Sentence denotes In total we designed a real-time PCR TaqMan assay combination for the detection of eight different potential bioterrorism specimen in one real-time PCR run.
T51 825546-825754 Sentence denotes This assay is easily to handle because of lyophilizied primers and probes and for the first time, it is possible to detect all these highly virulent pathogens at the same time with very good detection limits.
T52 825755-825867 Sentence denotes Analysis of antibiotic tolerance in Staphylococcus -towards the characterization of S. aureus persister cells S.
T53 825868-825883 Sentence denotes Lechner* 1 , M.
T54 825884-825896 Sentence denotes Vulic 2 , K.
T55 825897-825909 Sentence denotes Lewis 2 , R.
T56 825910-826193 Sentence denotes Bertram 1 1 IMIT, Lehrbereich mikrobielle Genetik, Eberhard Karls Universität Tübingen, Tübingen 2 Antimicrobial Discovery Center, Department of Biology, Northeastern University, Boston, MA 02115, USA Bacterial cultures contain a subpopulation of dormant cells, so called persisters.
T57 826194-826316 Sentence denotes These non-growing cells are phenotypic variants of the wild type capable of surviving killing by bactericidal antibiotics.
T58 826317-826471 Sentence denotes The persister state is reversible and dormant cells might be expected to form stochastically during growth of a culture, prior to the antibiotic exposure.
T59 826472-826612 Sentence denotes We aimed to study mechanisms governing persister formation and their cellular, physiologic, and genetic properties in Staphylococcus aureus.
T60 826613-826835 Sentence denotes Tolerance of different planktonically grown S. aureus strains (SA113, two small colony variants (SCVs) hemB and menD, as well as HG001, HG002, and HG003) to various antibiotics was tested at exponential-or stationaryphase.
T61 826836-826959 Sentence denotes Antibiotics applied were daptomycin, ciprofloxacin, tobramycin, rifampicin, and penicillin in a range of 1-to 100-fold MIC.
T62 826960-827168 Sentence denotes The biphasic killing kinetics, highly indicative of persister cells, were observed in stationary-phase upon treatment of SA113 with 100-fold MIC daptomycin and of menD challenged with 100-fold MIC tobramycin.
T63 827169-827322 Sentence denotes Time-dependent CFU analyses revealed a strong killing effect of daptomycin on exponentially growing SA113 and HG001-003 while SCVs' killing was retarded.
T64 827323-827404 Sentence denotes Tobramycin treatment appeared to eradicate SA113 less efficiently than SCV cells.
T65 827405-827513 Sentence denotes Challenging of HG001-003 with 10-fold tobramycin resulted in SCV-like cells upon cultivation on solid media.
T66 827514-827664 Sentence denotes These results indicate that stationary-phase cells are more antibiotic tolerant than exponential-phase cells due to the elevated levels of persisters.
T67 827665-827803 Sentence denotes Thus, growth phase as well as strain background and genotype appear to be important factors in the formation of S. aureus persister cells.
T68 827804-827939 Sentence denotes Crohn's disease relevant Enterococcus faecalis trigger differential host response in germfree and conventional NOD2 deficient mice I.G.
T69 827940-827952 Sentence denotes Sava* 1 , T.
T70 827953-827966 Sentence denotes Hrncir 2 , S.
T71 827967-827980 Sentence denotes Ocvirk 1 , D.
T72 827981-828329 Sentence denotes Haller 1 1 Technische Universität München, Chair for Biofunctionality, CDD -Center for Diet and Disease, ZIEL -Research Centre for Nutrition and Food Science, Freising-Weihenstephan, Germany 2 Institute of Microbiology, v.v.i., Dept. of Immunology and Gnotobiology, Academy of Sciences of the Czech Republic, Novy Hradek, Czech Republic Background:
T73 828330-828542 Sentence denotes The pattern recognition molecule NOD2 plays an important role in maintaining of intestinal homeostasis and mutation of the NOD2 gene has been associated in humans with increased susceptibility to Crohn's disease.
T74 828543-828693 Sentence denotes Mice deficient for NOD2 molecule show no symptoms of spontaneous intestinal inflammation, underscoring the multifactorial etiology of Crohn's disease.
T75 828694-829029 Sentence denotes However, several studies in mice have shown that NOD2 plays an important role in controlling the composition of the intestinal microbiota by regulating expression of antimicrobial peptides and in the host defense against several intestinal pathogens such as Yersinia pseudotuberculosis, Citrobacter rodentium or Helicobacter hepaticus.
T76 829030-829233 Sentence denotes Involvement of enterococci as common members of the intestinal microbiota in the development of intestinal inflammation has been already proved using IL10-/-mice monocolonized with Enterococcus faecalis.
T77 829234-829242 Sentence denotes Methods:
T78 829243-829391 Sentence denotes One new Enterococcus faecalis strain (CD11) was identified by screening fecal samples from patients with Crohn's disease for gelatinolytic activity.
T79 829392-829578 Sentence denotes Specific pathogen free and germ free NOD2-/-mice were orally infected either with Enterococcus faecalis CD11 or OG1RF (a wellcharacterized, fully sequenced strain) at the age of 4 weeks.
T80 829579-829587 Sentence denotes Results:
T81 829588-829854 Sentence denotes Specific pathogen free NOD2-/-mice are highly susceptible to oral inoculation with CD11 strain and die all within 3 weeks post infection, while mice infected with OG1RF strain as well as monocolonized mice (both with CD11 or with OG1RF) are protected from mortality.
T82 829855-830069 Sentence denotes In a shortterm experiment (1 week), mice infection with CD11 was followed by significant weight loss, elevated levels of serum-amyloid A and IL6 in the blood and high regulation of TNF, IL6 and IL1-beta in the gut.
T83 830070-830082 Sentence denotes Conclusions:
T84 830083-830249 Sentence denotes These results clearly demonstrate a link between E. faecalis stain specificity and the ability of this commensal to trigger disease in a genetically susceptible host.