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PMC:3312845 / 1819-34551 JSONTXT 26 Projects

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Id Subject Object Predicate Lexical cue
T18 0-10 Sentence denotes Background
T1140 11-89 Sentence denotes Status epilepticus (SE) is a medical emergency with significant mortality [1].
T19 11-89 Sentence denotes Status epilepticus (SE) is a medical emergency with significant mortality [1].
T1141 90-222 Sentence denotes SE has been defined as continuous seizure activity, which causes neuronal cell death, epileptogenesis and learning impairment [2,3].
T20 90-222 Sentence denotes SE has been defined as continuous seizure activity, which causes neuronal cell death, epileptogenesis and learning impairment [2,3].
T1142 223-360 Sentence denotes Some brain regions vulnerable to SE play a role in the generation and propagation of paroxysmal activity in experimental epilepsy models.
T21 223-360 Sentence denotes Some brain regions vulnerable to SE play a role in the generation and propagation of paroxysmal activity in experimental epilepsy models.
T1143 361-540 Sentence denotes The piriform cortex (PC) is one of the most susceptible brain regions to seizure-induced damage in the kainate, pilocarpine and other models of temporal lobe epilepsy (TLE) [4-6].
T22 361-540 Sentence denotes The piriform cortex (PC) is one of the most susceptible brain regions to seizure-induced damage in the kainate, pilocarpine and other models of temporal lobe epilepsy (TLE) [4-6].
T1144 541-599 Sentence denotes Pilocarpine, a cholinergic agonist, induces SE in rodents.
T23 541-599 Sentence denotes Pilocarpine, a cholinergic agonist, induces SE in rodents.
T1145 600-730 Sentence denotes This pilocarpine-induced SE, similar to human TLE, shows massive neuronal loss in the hippocampus followed by glial proliferation.
T24 600-730 Sentence denotes This pilocarpine-induced SE, similar to human TLE, shows massive neuronal loss in the hippocampus followed by glial proliferation.
T1146 731-871 Sentence denotes This neuronal damage in the pilocarpine model is not restricted to the hippocampus, but often extends to extrahippocampal limbic structures.
T25 731-871 Sentence denotes This neuronal damage in the pilocarpine model is not restricted to the hippocampus, but often extends to extrahippocampal limbic structures.
T1147 872-978 Sentence denotes Indeed, pilocarpine-induced SE results in acute neuronal damages within layers II and III of the PC [5,6].
T26 872-978 Sentence denotes Indeed, pilocarpine-induced SE results in acute neuronal damages within layers II and III of the PC [5,6].
T1148 979-1081 Sentence denotes SE also induces severe vasogenic edema in the PC accompanied by neuronal and astroglial damages [5-8].
T27 979-1081 Sentence denotes SE also induces severe vasogenic edema in the PC accompanied by neuronal and astroglial damages [5-8].
T1149 1082-1177 Sentence denotes Brain edema proceeds in two phases, early cytotoxic edema phase and late vasogenic edema phase.
T28 1082-1177 Sentence denotes Brain edema proceeds in two phases, early cytotoxic edema phase and late vasogenic edema phase.
T1150 1178-1332 Sentence denotes Early cytotoxic osmotic edema is due to excess stimulation of glutamatergic pathways during SE, which increases intracellular Na+ and Ca2+ concentrations.
T29 1178-1332 Sentence denotes Early cytotoxic osmotic edema is due to excess stimulation of glutamatergic pathways during SE, which increases intracellular Na+ and Ca2+ concentrations.
T1151 1333-1433 Sentence denotes The vasogenic edema results from dysfunction of endothelial cells and the blood-brain barrier (BBB).
T30 1333-1433 Sentence denotes The vasogenic edema results from dysfunction of endothelial cells and the blood-brain barrier (BBB).
T1152 1434-1528 Sentence denotes Many studies have reported increased permeability of the BBB during epileptic activity [9-13].
T31 1434-1528 Sentence denotes Many studies have reported increased permeability of the BBB during epileptic activity [9-13].
T1153 1529-1812 Sentence denotes A fast and significant increase in systemic blood pressure, particularly shown during tonic epileptic seizures, induces a marked vasodilation of the large cerebral arteries and an increase in blood pressure in capillaries, small arteries, and veins leading to leakage of the BBB [9].
T32 1529-1812 Sentence denotes A fast and significant increase in systemic blood pressure, particularly shown during tonic epileptic seizures, induces a marked vasodilation of the large cerebral arteries and an increase in blood pressure in capillaries, small arteries, and veins leading to leakage of the BBB [9].
T1154 1813-1959 Sentence denotes Loss of BBB integrity is not only due to an abrupt increase in the intraluminal pressure but also influenced by the properties of cerebral tissue.
T33 1813-1959 Sentence denotes Loss of BBB integrity is not only due to an abrupt increase in the intraluminal pressure but also influenced by the properties of cerebral tissue.
T1155 1960-2109 Sentence denotes Indeed, an acute increase in blood pressure or epileptic activity causes an increase in pinocytosis at the level of the cerebral endothelium [11-13].
T34 1960-2109 Sentence denotes Indeed, an acute increase in blood pressure or epileptic activity causes an increase in pinocytosis at the level of the cerebral endothelium [11-13].
T1156 2110-2380 Sentence denotes Recently, reports have also emphasized that seizure or epilepsy is a prolonged inflammatory condition, and that seizure activity rapidly increases the synthesis and release of various interleukins in rodent brain areas involved in seizure onset and their generalization.
T35 2110-2380 Sentence denotes Recently, reports have also emphasized that seizure or epilepsy is a prolonged inflammatory condition, and that seizure activity rapidly increases the synthesis and release of various interleukins in rodent brain areas involved in seizure onset and their generalization.
T1157 2381-2536 Sentence denotes Cytokines act on endothelial cells and change the permeability of the BBB, which exerts significant effects on neuronal viability and excitability [14,15].
T36 2381-2536 Sentence denotes Cytokines act on endothelial cells and change the permeability of the BBB, which exerts significant effects on neuronal viability and excitability [14,15].
T1158 2537-2673 Sentence denotes Indeed, Sztriha [16] reported that dexamethasone pretreatment reduces vasogenic edema in thalamus following kainic acid-induced seizure.
T37 2537-2673 Sentence denotes Indeed, Sztriha [16] reported that dexamethasone pretreatment reduces vasogenic edema in thalamus following kainic acid-induced seizure.
T1159 2674-2825 Sentence denotes Among cytokines, tumor necrosis factor-α (TNF-α) is a 17 kDa protein that is produced mainly by activated macrophages and T cells in the immune system.
T38 2674-2825 Sentence denotes Among cytokines, tumor necrosis factor-α (TNF-α) is a 17 kDa protein that is produced mainly by activated macrophages and T cells in the immune system.
T1160 2826-3004 Sentence denotes TNF-α is expressed at low levels in normal brain and is rapidly upregulated in glia, neurons and endothelial cells in various pathophysiological conditions, including SE [17,18].
T39 2826-3004 Sentence denotes TNF-α is expressed at low levels in normal brain and is rapidly upregulated in glia, neurons and endothelial cells in various pathophysiological conditions, including SE [17,18].
T1161 3005-3263 Sentence denotes TNF-α shows various effects on brain function depending on its local tissue concentration, the type of target cells, and especially the specific receptor subtype: TNF receptor I, or p55 receptor (TNFp55R); and TNF receptor II, or p75 receptor (TNFp75R) [19].
T40 3005-3263 Sentence denotes TNF-α shows various effects on brain function depending on its local tissue concentration, the type of target cells, and especially the specific receptor subtype: TNF receptor I, or p55 receptor (TNFp55R); and TNF receptor II, or p75 receptor (TNFp75R) [19].
T1162 3264-3411 Sentence denotes Furthermore, TNF-α induces macrophage inflammatory protein-2 (MIP-2) that recruits neutrophils under pathological conditions, including SE [14,20].
T41 3264-3411 Sentence denotes Furthermore, TNF-α induces macrophage inflammatory protein-2 (MIP-2) that recruits neutrophils under pathological conditions, including SE [14,20].
T1163 3412-3576 Sentence denotes Neurons, microglia, and astrocytes produce MIP-2 when incubated with pro-inflammatory cytokines such as TNF-α and/or interleukin-1β (IL-1β) or after injury [21-23].
T42 3412-3576 Sentence denotes Neurons, microglia, and astrocytes produce MIP-2 when incubated with pro-inflammatory cytokines such as TNF-α and/or interleukin-1β (IL-1β) or after injury [21-23].
T1164 3577-3733 Sentence denotes Indeed, we have recently reported that SE-mediated MIP-2 expression is relevant to leukocyte infiltrations following SE in an IL-1β-independent manner [20].
T43 3577-3733 Sentence denotes Indeed, we have recently reported that SE-mediated MIP-2 expression is relevant to leukocyte infiltrations following SE in an IL-1β-independent manner [20].
T1165 3734-3923 Sentence denotes However, the relationship between the TNF-α system and BBB disruption/neutrophil infiltration during vasogenic edema formation induced by epileptogenic insults has not been fully clarified.
T44 3734-3923 Sentence denotes However, the relationship between the TNF-α system and BBB disruption/neutrophil infiltration during vasogenic edema formation induced by epileptogenic insults has not been fully clarified.
T1166 3924-4092 Sentence denotes Therefore, in the present study, we investigated the roles of TNF-α in vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE.
T45 3924-4092 Sentence denotes Therefore, in the present study, we investigated the roles of TNF-α in vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE.
T46 4094-4101 Sentence denotes Methods
T2958 4103-4123 Sentence denotes Experimental animals
T47 4103-4123 Sentence denotes Experimental animals
T48 4124-4288 Sentence denotes This study utilized progeny of Sprague-Dawley (SD) rats (male, 9 - 11 weeks old) obtained from Experimental Animal Center, Hallym University, Chunchon, South Korea.
T2959 4124-4483 Sentence denotes This study utilized progeny of Sprague-Dawley (SD) rats (male, 9 - 11 weeks old) obtained from Experimental Animal Center, Hallym University, Chunchon, South Korea. The animals were provided with a commercial diet and water ad libitum under controlled temperature, humidity and lighting conditions (22 ± 2°C, 55 ± 5% and a 12:12 light/dark cycle with lights).
T49 4289-4483 Sentence denotes The animals were provided with a commercial diet and water ad libitum under controlled temperature, humidity and lighting conditions (22 ± 2°C, 55 ± 5% and a 12:12 light/dark cycle with lights).
T50 4484-4587 Sentence denotes Animal protocols were approved by the Institutional Animal Care and Use Committee of Hallym University.
T2960 4484-4796 Sentence denotes Animal protocols were approved by the Institutional Animal Care and Use Committee of Hallym University. Procedures involving animals and their care were conducted in accord with our institutional guidelines that comply with NIH Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80-23, 1996).
T51 4588-4782 Sentence denotes Procedures involving animals and their care were conducted in accord with our institutional guidelines that comply with NIH Guide for the Care and Use of Laboratory Animals (NIH Publications No.
T52 4783-4796 Sentence denotes 80-23, 1996).
T2961 4797-4894 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T53 4797-4894 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T3276 4896-4933 Sentence denotes Intracerebroventricular drug infusion
T54 4896-4933 Sentence denotes Intracerebroventricular drug infusion
T3277 4934-5092 Sentence denotes Rats were divided into two groups: vehicle (saline)-treated and soluble TNFp55 receptor (sTNFp55R, 50 μg/ml; Sigma-Aldrich Co., St. Louis, MO)-treated groups.
T55 4934-5092 Sentence denotes Rats were divided into two groups: vehicle (saline)-treated and soluble TNFp55 receptor (sTNFp55R, 50 μg/ml; Sigma-Aldrich Co., St. Louis, MO)-treated groups.
T3278 5093-5252 Sentence denotes The dosage of sTNFp55R was determined as the highest dose that induced SE of comparable severity in 100% of animals with 5% mortality in the preliminary study.
T56 5093-5252 Sentence denotes The dosage of sTNFp55R was determined as the highest dose that induced SE of comparable severity in 100% of animals with 5% mortality in the preliminary study.
T3279 5253-5370 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T57 5253-5370 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T3280 5371-5681 Sentence denotes For the osmotic pump implantation, holes were drilled through the skull to introduce a brain infusion kit 1 (Alzet, Cupertino, CA) into the right lateral ventricle (1 mm posterior; 1.5 mm lateral; - 3.5 mm depth; flat skull position with bregma as reference), according to the atlas of Paxinos and Watson [24].
T58 5371-5681 Sentence denotes For the osmotic pump implantation, holes were drilled through the skull to introduce a brain infusion kit 1 (Alzet, Cupertino, CA) into the right lateral ventricle (1 mm posterior; 1.5 mm lateral; - 3.5 mm depth; flat skull position with bregma as reference), according to the atlas of Paxinos and Watson [24].
T3281 5682-5792 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T59 5682-5792 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T3282 5793-5859 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T60 5793-5859 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T3283 5860-5921 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T61 5860-5921 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T3284 5922-5985 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T62 5922-5985 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T3285 5986-6046 Sentence denotes The compounds began to be immediately infused after surgery.
T63 5986-6046 Sentence denotes The compounds began to be immediately infused after surgery.
T3286 6047-6170 Sentence denotes Since the volume of vasogenic edema peaked at 2-3 days after SE in our previous studies [5-8,20], we chose this time point.
T64 6047-6170 Sentence denotes Since the volume of vasogenic edema peaked at 2-3 days after SE in our previous studies [5-8,20], we chose this time point.
T3287 6171-6328 Sentence denotes Thus, our experimental schedules at least inhibit the function of TNF-α from 3 days prior to SE to 4 days after SE when the volume of vasogenic edema peaked.
T65 6171-6328 Sentence denotes Thus, our experimental schedules at least inhibit the function of TNF-α from 3 days prior to SE to 4 days after SE when the volume of vasogenic edema peaked.
T3941 6330-6347 Sentence denotes Seizure induction
T66 6330-6347 Sentence denotes Seizure induction
T3942 6348-6545 Sentence denotes Three days after surgery, rats were treated with pilocarpine (380 mg/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO) at 20 min after methylscopolamine (5 mg/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO).
T67 6348-6545 Sentence denotes Three days after surgery, rats were treated with pilocarpine (380 mg/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO) at 20 min after methylscopolamine (5 mg/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO).
T3943 6546-6630 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T68 6546-6630 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T3944 6631-6869 Sentence denotes Approximately 80% of pilocarpine treated rats showed acute behavioral features of SE (including akinesia, facial automatisms, limbic seizures consisting of forelimb clonus with rearing, salivation, masticatory jaw movements, and falling).
T69 6631-6869 Sentence denotes Approximately 80% of pilocarpine treated rats showed acute behavioral features of SE (including akinesia, facial automatisms, limbic seizures consisting of forelimb clonus with rearing, salivation, masticatory jaw movements, and falling).
T3945 6870-7082 Sentence denotes We applied the 2 hr-SE rat model, because > 90% of the rats that we monitored in our previous studies [25] displayed spontaneous, recurrent seizures within 1-3 months after pilocarpine-induced status epilepticus.
T70 6870-7082 Sentence denotes We applied the 2 hr-SE rat model, because > 90% of the rats that we monitored in our previous studies [25] displayed spontaneous, recurrent seizures within 1-3 months after pilocarpine-induced status epilepticus.
T3946 7083-7213 Sentence denotes Diazepam (10 mg/kg, i.p.; Hoffman Ia Roche, Neuilly sur-Seine) was administered 2 hours after onset of SE and repeated, as needed.
T71 7083-7213 Sentence denotes Diazepam (10 mg/kg, i.p.; Hoffman Ia Roche, Neuilly sur-Seine) was administered 2 hours after onset of SE and repeated, as needed.
T3947 7214-7336 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T72 7214-7336 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T3948 7337-7493 Sentence denotes Non-experienced SE (non-SE) rats (showing only acute seizure behaviors during 10 - 30 min, n = 8) and age-matched normal rats were used as controls (n = 7).
T73 7337-7493 Sentence denotes Non-experienced SE (non-SE) rats (showing only acute seizure behaviors during 10 - 30 min, n = 8) and age-matched normal rats were used as controls (n = 7).
T4413 7495-7512 Sentence denotes Tissue processing
T74 7495-7512 Sentence denotes Tissue processing
T4414 7513-7547 Sentence denotes At designated time points (non-SE:
T75 7513-7547 Sentence denotes At designated time points (non-SE:
T4415 7548-7857 Sentence denotes 12 hr, 1 day, 2 days, 3 days, 4 days and 1 week after SE; n = 5, for each time point), animals were perfused transcardially with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB, pH 7.4) under urethane anesthesia (1.5 g/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO).
T76 7548-7857 Sentence denotes 12 hr, 1 day, 2 days, 3 days, 4 days and 1 week after SE; n = 5, for each time point), animals were perfused transcardially with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB, pH 7.4) under urethane anesthesia (1.5 g/kg, i.p.; Sigma-Aldrich Co., St. Louis, MO).
T4416 7858-7927 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T77 7858-7927 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T4417 7928-8008 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T78 7928-8008 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T4418 8009-8168 Sentence denotes Thereafter, the entire hippocampus was frozen and sectioned with a cryostat at 30 μm and consecutive sections were contained in six-well plates containing PBS.
T79 8009-8168 Sentence denotes Thereafter, the entire hippocampus was frozen and sectioned with a cryostat at 30 μm and consecutive sections were contained in six-well plates containing PBS.
T4419 8169-8287 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T80 8169-8287 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T4748 8289-8309 Sentence denotes Immunohistochemistry
T81 8289-8309 Sentence denotes Immunohistochemistry
T4749 8310-8412 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T82 8310-8412 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T4750 8413-8714 Sentence denotes They were then incubated in rabbit anti-MPO IgG (1:100, Thermo fisher scientific) or rabbit anti-MIP-2 IgG (1:200, Invitrogen, Carlsbad, CA) in PBS containing 0.3% Triton X-100 (Sigma-Aldrich Co., St. Louis, MO) and 2% normal goat serum(Sigma-Aldrich Co., St. Louis, MO) overnight at room temperature.
T83 8413-8714 Sentence denotes They were then incubated in rabbit anti-MPO IgG (1:100, Thermo fisher scientific) or rabbit anti-MIP-2 IgG (1:200, Invitrogen, Carlsbad, CA) in PBS containing 0.3% Triton X-100 (Sigma-Aldrich Co., St. Louis, MO) and 2% normal goat serum(Sigma-Aldrich Co., St. Louis, MO) overnight at room temperature.
T4751 8715-8975 Sentence denotes After washing three times for 10 min with PBS, the sections were incubated sequentially, in goat anti-rabbit or horse anti-mouse IgG (Vector, Burlingame, CA) and ABC complex (Vector, Burlingame, CA), diluted 1:200 in the same solution as the primary antiserum.
T84 8715-8975 Sentence denotes After washing three times for 10 min with PBS, the sections were incubated sequentially, in goat anti-rabbit or horse anti-mouse IgG (Vector, Burlingame, CA) and ABC complex (Vector, Burlingame, CA), diluted 1:200 in the same solution as the primary antiserum.
T4752 8976-9062 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T85 8976-9062 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T4753 9063-9216 Sentence denotes To confirm vasogenic edema, some tissue sections were reacted for serum-proteins using horse anti-rat IgG (Vector, Burlingame, CA) as a primary antibody.
T86 9063-9216 Sentence denotes To confirm vasogenic edema, some tissue sections were reacted for serum-proteins using horse anti-rat IgG (Vector, Burlingame, CA) as a primary antibody.
T4754 9217-9375 Sentence denotes The sections were visualized with 3,3'-diaminobenzidine (DAB, Sigma-Aldrich Co., St. Louis, MO) in 0.1 M Tris buffer and mounted on the gelatin-coated slides.
T87 9217-9375 Sentence denotes The sections were visualized with 3,3'-diaminobenzidine (DAB, Sigma-Aldrich Co., St. Louis, MO) in 0.1 M Tris buffer and mounted on the gelatin-coated slides.
T4755 9376-9476 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T88 9376-9476 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T4756 9477-9713 Sentence denotes For negative controls, rat hippocampal tissues were incubated with 1 μg of the antibody that was preincubated with 1 μg of purified peptide for 1 hr at room temperature or incubated with pre-immune serum instead of the primary antibody.
T89 9477-9713 Sentence denotes For negative controls, rat hippocampal tissues were incubated with 1 μg of the antibody that was preincubated with 1 μg of purified peptide for 1 hr at room temperature or incubated with pre-immune serum instead of the primary antibody.
T4757 9714-9810 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T90 9714-9810 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T5369 9812-9843 Sentence denotes Double immunofluorescence study
T91 9812-9843 Sentence denotes Double immunofluorescence study
T5370 9844-9935 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T92 9844-9935 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T5371 9936-10771 Sentence denotes Sections were then incubated in a mixture of goat anti-TNF-α IgG (1:1000, R&D systems, Minneapolis, MN)/mouse anti-OX-42 IgG (1:100, Serotec, Cambridge, UK), mouse anti-GFAP IgG (1:1000, an astroglial marker, Millipore Corporation, Billerica, MA)/rabbit anti-TNFp55R IgG (1:1000, Abcam, Cambridge, UK), mouse anti-GFAP IgG/rabbit anti-TNFp75R IgG (1:1000, Abcam, Cambridge, UK), mouse anti-SMI-71 IgM (1:1000, Covance, Berkeley, CA)/rabbit anti-TNFp75R IgG, mouse anti-GFAP IgG/rabbit anti-NF-κB (p65-Ser276, p65-Ser311, p65-Ser529, and p65-Thr435) IgG (1:100, Abcam, Cambridge, UK), mouse anti-SMI-71 IgM/rabbit anti-p65-Thr435 NF-κB IgG, mouse anti-SMI-71 IgM/rabbit anti-GLUT-1 IgG (1:100, Abcam, Cambridge, UK), or mouse anti-GFAP IgG/rabbit anti-MIP-2 IgG (1:100) in PBS containing 0.3% triton X-100 overnight at room temperature.
T93 9936-10771 Sentence denotes Sections were then incubated in a mixture of goat anti-TNF-α IgG (1:1000, R&D systems, Minneapolis, MN)/mouse anti-OX-42 IgG (1:100, Serotec, Cambridge, UK), mouse anti-GFAP IgG (1:1000, an astroglial marker, Millipore Corporation, Billerica, MA)/rabbit anti-TNFp55R IgG (1:1000, Abcam, Cambridge, UK), mouse anti-GFAP IgG/rabbit anti-TNFp75R IgG (1:1000, Abcam, Cambridge, UK), mouse anti-SMI-71 IgM (1:1000, Covance, Berkeley, CA)/rabbit anti-TNFp75R IgG, mouse anti-GFAP IgG/rabbit anti-NF-κB (p65-Ser276, p65-Ser311, p65-Ser529, and p65-Thr435) IgG (1:100, Abcam, Cambridge, UK), mouse anti-SMI-71 IgM/rabbit anti-p65-Thr435 NF-κB IgG, mouse anti-SMI-71 IgM/rabbit anti-GLUT-1 IgG (1:100, Abcam, Cambridge, UK), or mouse anti-GFAP IgG/rabbit anti-MIP-2 IgG (1:100) in PBS containing 0.3% triton X-100 overnight at room temperature.
T5372 10772-10987 Sentence denotes After washing three times for 10 minutes with PBS, sections were also incubated in a mixture of FITC- and Cy3-conjugated secondary antisera (Amersham, San Francisco, CA), diluted 1:200, for 2 hr at room temperature.
T94 10772-10987 Sentence denotes After washing three times for 10 minutes with PBS, sections were also incubated in a mixture of FITC- and Cy3-conjugated secondary antisera (Amersham, San Francisco, CA), diluted 1:200, for 2 hr at room temperature.
T5373 10988-11083 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T95 10988-11083 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T5374 11084-11183 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T96 11084-11183 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T97 11184-11261 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel.
T5375 11184-11275 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel. 4.8 software.
T98 11262-11275 Sentence denotes 4.8 software.
T5848 11277-11299 Sentence denotes Fluoro-Jade B staining
T99 11277-11299 Sentence denotes Fluoro-Jade B staining
T5849 11300-11445 Sentence denotes Fluoro-Jade B (FJB) staining was used to identify degenerating neurons in tissues obtained from non-SE and 3 days post-SE animals in every group.
T100 11300-11445 Sentence denotes Fluoro-Jade B (FJB) staining was used to identify degenerating neurons in tissues obtained from non-SE and 3 days post-SE animals in every group.
T5850 11446-11548 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T101 11446-11548 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T5851 11549-11625 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T102 11549-11625 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T5852 11626-11748 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T103 11626-11748 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T5853 11749-11865 Sentence denotes The slides were immersed in 100% ethanol for 3 min, followed by 70% ethanol for 2 min and distilled water for 2 min.
T104 11749-11865 Sentence denotes The slides were immersed in 100% ethanol for 3 min, followed by 70% ethanol for 2 min and distilled water for 2 min.
T5854 11866-11962 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T105 11866-11962 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T5855 11963-12218 Sentence denotes After rinsing in distilled water for 2 min, the slides were incubated for 30 min in 0.001% FJB (Histo-Chem Inc., Jefferson, AR), freshly prepared by adding 20 ml of a 0.01% stock FJB solution to 180 ml of 0.1% acetic acid, with gentle shaking in the dark.
T106 11963-12218 Sentence denotes After rinsing in distilled water for 2 min, the slides were incubated for 30 min in 0.001% FJB (Histo-Chem Inc., Jefferson, AR), freshly prepared by adding 20 ml of a 0.01% stock FJB solution to 180 ml of 0.1% acetic acid, with gentle shaking in the dark.
T5856 12219-12393 Sentence denotes After rinsing for 1 min in each of three changes of distilled water, the slides were dried, dehydrated in xylene and coverslipped with DPX (Sigma-Aldrich Co., St. Louis, MO).
T107 12219-12393 Sentence denotes After rinsing for 1 min in each of three changes of distilled water, the slides were dried, dehydrated in xylene and coverslipped with DPX (Sigma-Aldrich Co., St. Louis, MO).
T5857 12394-12499 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T108 12394-12499 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T6363 12501-12536 Sentence denotes Volumetric analysis and cell counts
T109 12501-12536 Sentence denotes Volumetric analysis and cell counts
T110 12537-12692 Sentence denotes To measure vasogenic edema, the volume of anti-rat IgG positive region in PC was estimated according to the formula based on the modified Cavalieri method:
T6364 12537-12979 Sentence denotes To measure vasogenic edema, the volume of anti-rat IgG positive region in PC was estimated according to the formula based on the modified Cavalieri method: V = Σa × tnom × 1/ssf, where a is area of the region of the delineated subfield measured by AxioVision Rel. 4.8 software, tnom is the nominal section thickness (of 30 μm in this study), and ssf is the fraction of the sections sampled or section sampling fraction (of 1/6 in this study).
T111 12693-12800 Sentence denotes V = Σa × tnom × 1/ssf, where a is area of the region of the delineated subfield measured by AxioVision Rel.
T112 12801-12979 Sentence denotes 4.8 software, tnom is the nominal section thickness (of 30 μm in this study), and ssf is the fraction of the sections sampled or section sampling fraction (of 1/6 in this study).
T6365 12980-13057 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T113 12980-13057 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T6366 13058-13090 Sentence denotes The volumes are reported as mm3.
T114 13058-13090 Sentence denotes The volumes are reported as mm3.
T6367 13091-13149 Sentence denotes An optical fractionator was used to estimate cell numbers.
T115 13091-13149 Sentence denotes An optical fractionator was used to estimate cell numbers.
T6368 13150-13424 Sentence denotes The optical fractionator (a combination of performing counting with the optical dissector, with fractionator sampling) is a stereological method based on a properly designed systematic random sampling method that by definition yields unbiased estimates of population number.
T116 13150-13424 Sentence denotes The optical fractionator (a combination of performing counting with the optical dissector, with fractionator sampling) is a stereological method based on a properly designed systematic random sampling method that by definition yields unbiased estimates of population number.
T6369 13425-13577 Sentence denotes The sampling procedure is accomplished by focusing through the depth of the tissue (the optical dissector height, h; 15 μm in all cases for this study).
T117 13425-13577 Sentence denotes The sampling procedure is accomplished by focusing through the depth of the tissue (the optical dissector height, h; 15 μm in all cases for this study).
T118 13578-13653 Sentence denotes The number of each cell type (C) in each of the subregions is estimated as:
T6370 13578-14112 Sentence denotes The number of each cell type (C) in each of the subregions is estimated as: C = ΣQ- × t/h × 1/asf × 1/ssf, where Q- is the number of cells actually counted in the dissectors that fell within the sectional profiles of the subregion seen on the sampled sections, and Asf is the areal sampling fraction calculated by the area of the counting frame of the dissector, a(frame) (of 50 × 50 μm2 in this study) and the area associated with each x, y movement, grid (x, y step) (of 250 × 250 μm2 in this study) {asf = (a(frame)/a(x, y step))}.
T119 13654-14112 Sentence denotes C = ΣQ- × t/h × 1/asf × 1/ssf, where Q- is the number of cells actually counted in the dissectors that fell within the sectional profiles of the subregion seen on the sampled sections, and Asf is the areal sampling fraction calculated by the area of the counting frame of the dissector, a(frame) (of 50 × 50 μm2 in this study) and the area associated with each x, y movement, grid (x, y step) (of 250 × 250 μm2 in this study) {asf = (a(frame)/a(x, y step))}.
T6371 14113-14177 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T120 14113-14177 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6372 14178-14242 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T121 14178-14242 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6373 14243-14318 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T122 14243-14318 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T6374 14319-14425 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T123 14319-14425 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T6375 14426-14546 Sentence denotes SE-induced PC atrophy is evident [8], so changes in cell number may be caused by an alterations in the volume of the PC.
T124 14426-14546 Sentence denotes SE-induced PC atrophy is evident [8], so changes in cell number may be caused by an alterations in the volume of the PC.
T6376 14547-14730 Sentence denotes Therefore, the total number of cells was corrected by multiplying with appropriate correction factors (CF) representing the degree of shrinkage (or swelling) compared with the Non-SE.
T125 14547-14730 Sentence denotes Therefore, the total number of cells was corrected by multiplying with appropriate correction factors (CF) representing the degree of shrinkage (or swelling) compared with the Non-SE.
T7168 14732-14754 Sentence denotes Quantification of data
T126 14732-14754 Sentence denotes Quantification of data
T127 14755-14964 Sentence denotes The fluorescence intensities of SMI-71/p65-Thr435 phosphorylation or GFPA/p65-Thr435 phosphorylation were measured using a computer-assisted image analysis program (The University of Texas ImageTool program V.
T7169 14755-15003 Sentence denotes The fluorescence intensities of SMI-71/p65-Thr435 phosphorylation or GFPA/p65-Thr435 phosphorylation were measured using a computer-assisted image analysis program (The University of Texas ImageTool program V. 3.0 and AxioVision Rel. 4.8 software).
T128 14965-14988 Sentence denotes 3.0 and AxioVision Rel.
T129 14989-15003 Sentence denotes 4.8 software).
T7170 15004-15198 Sentence denotes After regions were outlined, 30 areas/rat (300 μm2/area) were randomly selected within the PC, and double immunofluorescent merge images were captured from the PC (15 sections from each animal).
T130 15004-15198 Sentence denotes After regions were outlined, 30 areas/rat (300 μm2/area) were randomly selected within the PC, and double immunofluorescent merge images were captured from the PC (15 sections from each animal).
T7171 15199-15381 Sentence denotes Merge images were digitally separated to red or green image, and converted to grayscale images, respectively (n = 36 per region examined, in non-SE, 12 hr post-SE and 1 day post-SE).
T131 15199-15381 Sentence denotes Merge images were digitally separated to red or green image, and converted to grayscale images, respectively (n = 36 per region examined, in non-SE, 12 hr post-SE and 1 day post-SE).
T7172 15382-15450 Sentence denotes The range of intensity values was obtained from the selected images.
T132 15382-15450 Sentence denotes The range of intensity values was obtained from the selected images.
T7173 15451-15574 Sentence denotes Based on the mean range of intensity values, each image was normalized by adjusting the black and white range of the image.
T133 15451-15574 Sentence denotes Based on the mean range of intensity values, each image was normalized by adjusting the black and white range of the image.
T7174 15575-15676 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T134 15575-15676 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T135 15677-15799 Sentence denotes Intensity measurements are represented as the mean number of a 256 gray scale (NIH Image 1.59 software and AxioVision Rel.
T7175 15677-15814 Sentence denotes Intensity measurements are represented as the mean number of a 256 gray scale (NIH Image 1.59 software and AxioVision Rel. 4.8 software).
T136 15800-15814 Sentence denotes 4.8 software).
T7176 15815-15885 Sentence denotes Values for background staining were obtained from the corpus callosum.
T137 15815-15885 Sentence denotes Values for background staining were obtained from the corpus callosum.
T7177 15886-16013 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T138 15886-16013 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T7659 16015-16028 Sentence denotes Data analysis
T139 16015-16028 Sentence denotes Data analysis
T7660 16029-16187 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T140 16029-16187 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T7661 16188-16350 Sentence denotes Linear regression analysis was also performed to determine correlations with SMI-71/p65-Thr435 phosphorylation, and the number of MPO cells/vasogenic edema areas.
T141 16188-16350 Sentence denotes Linear regression analysis was also performed to determine correlations with SMI-71/p65-Thr435 phosphorylation, and the number of MPO cells/vasogenic edema areas.
T142 16352-16359 Sentence denotes Results
T7823 16361-16395 Sentence denotes TNF-α, and TNF receptor expression
T143 16361-16395 Sentence denotes TNF-α, and TNF receptor expression
T7824 16396-16526 Sentence denotes In non-SE induced animals of the saline-infused groups, TNF-α immunoreactivity was weakly detected in PC neurons (data not shown).
T144 16396-16526 Sentence denotes In non-SE induced animals of the saline-infused groups, TNF-α immunoreactivity was weakly detected in PC neurons (data not shown).
T7825 16527-16662 Sentence denotes In 12 hr-post SE animals of the saline-infused group, most of the activated microglia showed strong TNF-α immunoreactivity (Figure 1A).
T145 16527-16662 Sentence denotes In 12 hr-post SE animals of the saline-infused group, most of the activated microglia showed strong TNF-α immunoreactivity (Figure 1A).
T7826 16663-16724 Sentence denotes This expression pattern was maintained up to 1 week after SE.
T146 16663-16724 Sentence denotes This expression pattern was maintained up to 1 week after SE.
T7827 16725-16877 Sentence denotes In non-SE-induced animals of the saline-infused groups, TNFp55R and TNFp75R immunoreactivities were also weakly observed in astrocytes (data not shown).
T147 16725-16877 Sentence denotes In non-SE-induced animals of the saline-infused groups, TNFp55R and TNFp75R immunoreactivities were also weakly observed in astrocytes (data not shown).
T7828 16878-16996 Sentence denotes In 12 hr-post SE animals of the saline-infused group, TNFp55R immunoreactivity was observed in astrocytes (Figure 1B).
T148 16878-16996 Sentence denotes In 12 hr-post SE animals of the saline-infused group, TNFp55R immunoreactivity was observed in astrocytes (Figure 1B).
T7829 16997-17109 Sentence denotes Unlike TNFp55R, TNFp75R immunoreactivity was detected in endothelial cells as well as astrocytes (Figures 1C-E).
T149 16997-17109 Sentence denotes Unlike TNFp55R, TNFp75R immunoreactivity was detected in endothelial cells as well as astrocytes (Figures 1C-E).
T7830 17110-17309 Sentence denotes One day to 1 week after SE, both TNFp55R and TNFp75R immunoreactivities were significantly reduced in astrocytes, not in endothelial cells, due to massive astroglial loss (data not shown) [5,7,8,20].
T150 17110-17309 Sentence denotes One day to 1 week after SE, both TNFp55R and TNFp75R immunoreactivities were significantly reduced in astrocytes, not in endothelial cells, due to massive astroglial loss (data not shown) [5,7,8,20].
T151 17310-17571 Sentence denotes Figure 1 Expression of TNF-α and TNF receptor in the PC 12 hr-post SE. (A) TNF-α immunoreactivity in Ox-42-positive microglia (arrows). (B) TNFp55R expression in astrocytes (arrows). (C-D) TNFp75R expression in astrocytes as well as endothelial cells (arrows).
T14488 17320-17571 Sentence denotes Expression of TNF-α and TNF receptor in the PC 12 hr-post SE. (A) TNF-α immunoreactivity in Ox-42-positive microglia (arrows). (B) TNFp55R expression in astrocytes (arrows). (C-D) TNFp75R expression in astrocytes as well as endothelial cells (arrows).
T14489 17572-17590 Sentence denotes Bar = 25 (A-D) μm.
T152 17572-17590 Sentence denotes Bar = 25 (A-D) μm.
T8344 17592-17681 Sentence denotes Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage
T153 17592-17681 Sentence denotes Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage
T8345 17682-17826 Sentence denotes In our previous [5,7,8,20] and preliminary data, vasogenic edema and neuronal damage were noticeable at 1 day and 3 days after SE, respectively.
T154 17682-17826 Sentence denotes In our previous [5,7,8,20] and preliminary data, vasogenic edema and neuronal damage were noticeable at 1 day and 3 days after SE, respectively.
T8346 17827-18000 Sentence denotes Therefore, we determined that 3 days after SE was the best time point to evaluate the effect of sTNFp55R infusion on both vasogenic edema and neuronal damages induced by SE.
T155 17827-18000 Sentence denotes Therefore, we determined that 3 days after SE was the best time point to evaluate the effect of sTNFp55R infusion on both vasogenic edema and neuronal damages induced by SE.
T8347 18001-18087 Sentence denotes In saline-treated animals, the PC was stained diffusely with anti-rat IgG (Figure 2A).
T156 18001-18087 Sentence denotes In saline-treated animals, the PC was stained diffusely with anti-rat IgG (Figure 2A).
T8348 18088-18149 Sentence denotes The volume of vasogenic edema was 17.1 ± 1.5 mm3 (Figure 2E).
T157 18088-18149 Sentence denotes The volume of vasogenic edema was 17.1 ± 1.5 mm3 (Figure 2E).
T8349 18150-18228 Sentence denotes The number of FJB-positive neurons in the PC was 236,145 ± 49,469 (Figure 2B).
T158 18150-18228 Sentence denotes The number of FJB-positive neurons in the PC was 236,145 ± 49,469 (Figure 2B).
T8350 18229-18337 Sentence denotes In sTNFp55R-treated animals, SE-induced vasogenic edema was attenuated to 9.8 ± 0.7 mm3 (Figures 2C and 2E).
T159 18229-18337 Sentence denotes In sTNFp55R-treated animals, SE-induced vasogenic edema was attenuated to 9.8 ± 0.7 mm3 (Figures 2C and 2E).
T8351 18338-18430 Sentence denotes In addition, the number of FJB-positive neurons in the PC was 89,138 ± 5,698 (Figures 2D-E).
T160 18338-18430 Sentence denotes In addition, the number of FJB-positive neurons in the PC was 89,138 ± 5,698 (Figures 2D-E).
T8352 18431-18559 Sentence denotes Thus, sTNFp55R infusion attenuated SE-induced vasogenic edema and neuronal damage compared to saline-infused animals (p < 0.05).
T161 18431-18559 Sentence denotes Thus, sTNFp55R infusion attenuated SE-induced vasogenic edema and neuronal damage compared to saline-infused animals (p < 0.05).
T162 18560-18755 Sentence denotes Figure 2 Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage. (A-D) Serum-protein extravasation and FJB-positive neuronal damages in the PC 3 days after SE.
T14706 18570-18755 Sentence denotes Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage. (A-D) Serum-protein extravasation and FJB-positive neuronal damages in the PC 3 days after SE.
T14707 18756-18905 Sentence denotes Compared to saline-infused animals, serum-protein extravasation and FJB-positive neuronal damage is markedly ameliorated in sTNFp55R-infused animals.
T163 18756-18905 Sentence denotes Compared to saline-infused animals, serum-protein extravasation and FJB-positive neuronal damage is markedly ameliorated in sTNFp55R-infused animals.
T14708 18906-19079 Sentence denotes Bars = 400 (A and C) and 50 (B and D) μm. (E) Quantitative analyses of serum-protein extravasation and FJB-positive neuronal damage in the PC 3 days after SE (mean ± S.E.M).
T164 18906-19079 Sentence denotes Bars = 400 (A and C) and 50 (B and D) μm. (E) Quantitative analyses of serum-protein extravasation and FJB-positive neuronal damage in the PC 3 days after SE (mean ± S.E.M).
T14709 19080-19143 Sentence denotes Significant differences from saline-treated animals, *p < 0.05.
T165 19080-19143 Sentence denotes Significant differences from saline-treated animals, *p < 0.05.
T8685 19145-19166 Sentence denotes NF-κB phosphorylation
T166 19145-19166 Sentence denotes NF-κB phosphorylation
T8686 19167-19261 Sentence denotes It is well established that TNF-α is one of the major stimuli toward phosphorylation of NF-κB.
T167 19167-19261 Sentence denotes It is well established that TNF-α is one of the major stimuli toward phosphorylation of NF-κB.
T8687 19262-19390 Sentence denotes To confirm TNF-α-mediated signaling following SE, we performed an immunohistochemical study using five phospho-NF-κB antibodies.
T168 19262-19390 Sentence denotes To confirm TNF-α-mediated signaling following SE, we performed an immunohistochemical study using five phospho-NF-κB antibodies.
T8688 19391-19791 Sentence denotes Compared to non-SE animals (data not shown), 12 hr-post SE animals of the saline-infused group showed p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation in astrocytes (not endothelial cells). sTNFp55R infusion effectively reduced p65-Ser276 and p65-Ser311 phosphorylation (p < 0.05, respectively), while it could not affect p65-Ser529 or p65-Ser536 phosphorylation (Figures 3 and 4A).
T169 19391-19791 Sentence denotes Compared to non-SE animals (data not shown), 12 hr-post SE animals of the saline-infused group showed p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation in astrocytes (not endothelial cells). sTNFp55R infusion effectively reduced p65-Ser276 and p65-Ser311 phosphorylation (p < 0.05, respectively), while it could not affect p65-Ser529 or p65-Ser536 phosphorylation (Figures 3 and 4A).
T8689 19792-19951 Sentence denotes In contrast, p65-Thr435 phosphorylation was increased in endothelial cells (not astrocytes) within the PC of saline-infused animals 12 hr after SE (Figure 5A).
T170 19792-19951 Sentence denotes In contrast, p65-Thr435 phosphorylation was increased in endothelial cells (not astrocytes) within the PC of saline-infused animals 12 hr after SE (Figure 5A).
T8690 19952-20116 Sentence denotes In addition, sTNFp55R infusion effectively alleviated SE-induced p65-Thr435 phosphorylation in endothelial cells, compared to saline infusion (p < 0.05, Figure 5B).
T171 19952-20116 Sentence denotes In addition, sTNFp55R infusion effectively alleviated SE-induced p65-Thr435 phosphorylation in endothelial cells, compared to saline infusion (p < 0.05, Figure 5B).
T172 20117-20209 Sentence denotes Figure 3 Effect of sTNFp55R infusion on NF-κB phosphorylation in astrocytes 12 hr after SE.
T15005 20127-20209 Sentence denotes Effect of sTNFp55R infusion on NF-κB phosphorylation in astrocytes 12 hr after SE.
T15006 20210-20578 Sentence denotes In 12 hr-post SE animals of the saline-infused group (A, C, E and G), astrocytes show p65-Ser276 (A), p65-Ser311 (C), p65-Ser529 (E), and p65-Ser536 (G) phosphorylation (arrows). sTNFp55R infusion (B, D, F and H) effectively reduces p65-Ser276 (B) and p65-Ser311 (D) phosphorylation, while it does not affect p65-Ser529 (F) and p65-Ser536 (H) phosphorylation (arrows).
T173 20210-20578 Sentence denotes In 12 hr-post SE animals of the saline-infused group (A, C, E and G), astrocytes show p65-Ser276 (A), p65-Ser311 (C), p65-Ser529 (E), and p65-Ser536 (G) phosphorylation (arrows). sTNFp55R infusion (B, D, F and H) effectively reduces p65-Ser276 (B) and p65-Ser311 (D) phosphorylation, while it does not affect p65-Ser529 (F) and p65-Ser536 (H) phosphorylation (arrows).
T15007 20579-20593 Sentence denotes Bar = 12.5 μm.
T174 20579-20593 Sentence denotes Bar = 12.5 μm.
T175 20594-20790 Sentence denotes Figure 4 Quantitative analyses of the effect of sTNFp55R infusion on NF-κB phosphorylation and SMI-71 expression. (A) Quantitative analysis of NF-κB phosphorylation 12 hr after SE (mean ± S.E.M).
T15460 20604-20790 Sentence denotes Quantitative analyses of the effect of sTNFp55R infusion on NF-κB phosphorylation and SMI-71 expression. (A) Quantitative analysis of NF-κB phosphorylation 12 hr after SE (mean ± S.E.M).
T15461 20791-20932 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (B) Quantitative analysis of SMI-71 expression 1 day after SE (mean ± S.E.M).
T176 20791-20932 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (B) Quantitative analysis of SMI-71 expression 1 day after SE (mean ± S.E.M).
T15462 20933-21097 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (C) Linear regression analysis between p65-Thr435 phosphorylation and SMI-71 in the PC following SE.
T177 20933-21097 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (C) Linear regression analysis between p65-Thr435 phosphorylation and SMI-71 in the PC following SE.
T178 21098-21841 Sentence denotes Figure 5 Effect of sTNFp55R infusion on p65-Thr435 phosphorylation in endothelial cells following SE. (A-B) Inhibition of p65-Thr435 phosphorylation by sTNFp55R infusion 12 hr after SE. p65-Thr435 phosphorylation is rarely observed in astrocytes (arrows). (C) Endothelial p65-Thr435 phosphorylation in non-SE animals. (D-E) Endothelial p65-Thr435 phosphorylation in saline-infused animals 1 day after SE. p65-Thr435 phosphorylation is enhanced, while SMI-71 expression is reduced in GLUT-1-positive endothelial cells (arrows). (F-G) Endothelial p65-Thr435 phosphorylation in sTNFp55R-infused animal 1 day after SE. sTNFp55R infusion effectively reduces p65-Thr435 and preserves SMI-71 expression in GLUT-1-positive endothelial cells (arrows).
T15783 21108-21841 Sentence denotes Effect of sTNFp55R infusion on p65-Thr435 phosphorylation in endothelial cells following SE. (A-B) Inhibition of p65-Thr435 phosphorylation by sTNFp55R infusion 12 hr after SE. p65-Thr435 phosphorylation is rarely observed in astrocytes (arrows). (C) Endothelial p65-Thr435 phosphorylation in non-SE animals. (D-E) Endothelial p65-Thr435 phosphorylation in saline-infused animals 1 day after SE. p65-Thr435 phosphorylation is enhanced, while SMI-71 expression is reduced in GLUT-1-positive endothelial cells (arrows). (F-G) Endothelial p65-Thr435 phosphorylation in sTNFp55R-infused animal 1 day after SE. sTNFp55R infusion effectively reduces p65-Thr435 and preserves SMI-71 expression in GLUT-1-positive endothelial cells (arrows).
T15784 21842-21876 Sentence denotes Bars = 12.5 (A-D) and 25 (E-G) μm.
T179 21842-21876 Sentence denotes Bars = 12.5 (A-D) and 25 (E-G) μm.
T9403 21878-21895 Sentence denotes SMI-71 expression
T180 21878-21895 Sentence denotes SMI-71 expression
T9404 21896-22021 Sentence denotes Previously, we reported that SMI-71 (an endothelial barrier antigen) immunoreactivity decreased in the PC 1 day after SE [5].
T181 21896-22021 Sentence denotes Previously, we reported that SMI-71 (an endothelial barrier antigen) immunoreactivity decreased in the PC 1 day after SE [5].
T9405 22022-22220 Sentence denotes Similarly, in 1 day-post SE animals of the saline-infused group, loss of SMI-71 immunoreactivity was detected in layer III/IV of the PC as compared to non-SE animals (Figures 4B and 5C-E, p < 0.05).
T182 22022-22220 Sentence denotes Similarly, in 1 day-post SE animals of the saline-infused group, loss of SMI-71 immunoreactivity was detected in layer III/IV of the PC as compared to non-SE animals (Figures 4B and 5C-E, p < 0.05).
T9406 22221-22314 Sentence denotes Thus, loss of SMI-71 immunoreactivity correlated with volume of vasogenic edema following SE.
T183 22221-22314 Sentence denotes Thus, loss of SMI-71 immunoreactivity correlated with volume of vasogenic edema following SE.
T9407 22315-22411 Sentence denotes This reduction in SMI-71 was accompanied by increased p65-Thr435 phosphorylation (Figures 5C-D).
T184 22315-22411 Sentence denotes This reduction in SMI-71 was accompanied by increased p65-Thr435 phosphorylation (Figures 5C-D).
T9408 22412-22590 Sentence denotes Therefore, the degree of SMI-71 immunoreactivity was inversely correlated to p65-Thr435 phosphorylation with a linear coefficient of correlation of -0.6324 (p < 0.05; Figure 4C).
T185 22412-22590 Sentence denotes Therefore, the degree of SMI-71 immunoreactivity was inversely correlated to p65-Thr435 phosphorylation with a linear coefficient of correlation of -0.6324 (p < 0.05; Figure 4C).
T9409 22591-22812 Sentence denotes In addition, sTNFp55R infusion effectively alleviated p65-Thr435 phosphorylation and preserved SMI-71 immunoreactivity in endothelial cells following SE, as compared to saline infusion (p < 0.05; Figures 4A-B, 5D and 5F).
T186 22591-22812 Sentence denotes In addition, sTNFp55R infusion effectively alleviated p65-Thr435 phosphorylation and preserved SMI-71 immunoreactivity in endothelial cells following SE, as compared to saline infusion (p < 0.05; Figures 4A-B, 5D and 5F).
T9880 22814-22837 Sentence denotes Neutrophil infiltration
T187 22814-22837 Sentence denotes Neutrophil infiltration
T9881 22838-22948 Sentence denotes Recent studies have reported that neutrophils infiltrate the brain under certain pathological conditions [26].
T188 22838-22948 Sentence denotes Recent studies have reported that neutrophils infiltrate the brain under certain pathological conditions [26].
T9882 22949-23052 Sentence denotes Indeed, we have reported massive neutrophil infiltration in layer III/IV of the PC 1 day after SE [20].
T189 22949-23052 Sentence denotes Indeed, we have reported massive neutrophil infiltration in layer III/IV of the PC 1 day after SE [20].
T9883 23053-23185 Sentence denotes In the present study, 1 day-post-SE animals of the saline-infused group showed infiltration of MPO-positive neutrophils into the PC.
T190 23053-23185 Sentence denotes In the present study, 1 day-post-SE animals of the saline-infused group showed infiltration of MPO-positive neutrophils into the PC.
T9884 23186-23319 Sentence denotes Similarly, 1 day-post-SE animals of the sTNFp55R-infused group showed neutrophil infiltration into the PC 1 day after SE (Figure 6A).
T191 23186-23319 Sentence denotes Similarly, 1 day-post-SE animals of the sTNFp55R-infused group showed neutrophil infiltration into the PC 1 day after SE (Figure 6A).
T9885 23320-23554 Sentence denotes The number of neutrophils/area in the PC region (including the vasogenic edema region and the non-vasogenic edema region) of sTNFp55R-infused animals was significantly lower than that of the saline-infused group (Figure 6D, p < 0.05).
T192 23320-23554 Sentence denotes The number of neutrophils/area in the PC region (including the vasogenic edema region and the non-vasogenic edema region) of sTNFp55R-infused animals was significantly lower than that of the saline-infused group (Figure 6D, p < 0.05).
T9886 23555-23708 Sentence denotes However, there was no difference in neutrophil infiltration per unit area of vasogenic edema between the saline- and sTNFp55R-infused groups (Figure 6D).
T193 23555-23708 Sentence denotes However, there was no difference in neutrophil infiltration per unit area of vasogenic edema between the saline- and sTNFp55R-infused groups (Figure 6D).
T9887 23709-23878 Sentence denotes Furthermore, neutrophil infiltration showed a direct proportion to the area of vasogenic edema, with a linear coefficient of correlation of 0.8631 (p < 0.05, Figure 6E).
T194 23709-23878 Sentence denotes Furthermore, neutrophil infiltration showed a direct proportion to the area of vasogenic edema, with a linear coefficient of correlation of 0.8631 (p < 0.05, Figure 6E).
T9888 23879-24026 Sentence denotes Therefore, our findings indicate that SE-induced neutrophil infiltration into the PC may be correlated to TNF-α-mediated vasogenic edema formation.
T195 23879-24026 Sentence denotes Therefore, our findings indicate that SE-induced neutrophil infiltration into the PC may be correlated to TNF-α-mediated vasogenic edema formation.
T196 24027-24288 Sentence denotes Figure 6 Effect of sTNFp55R infusion on neutrophil infiltration and MIP-2 expression following SE. (A) Neutrophil infiltration in vasogenic edema lesion 1 day after SE. (B) MIP-2 expression in the PC 1 day after SE. (C) Astroglial expression of MIP-2 (arrows).
T16281 24037-24288 Sentence denotes Effect of sTNFp55R infusion on neutrophil infiltration and MIP-2 expression following SE. (A) Neutrophil infiltration in vasogenic edema lesion 1 day after SE. (B) MIP-2 expression in the PC 1 day after SE. (C) Astroglial expression of MIP-2 (arrows).
T16282 24289-24410 Sentence denotes Bars = 12.5 (A and C) and 150 (B) μm. (D) Quantitative analysis of neutrophil infiltration 1 day after SE (mean ± S.E.M).
T197 24289-24410 Sentence denotes Bars = 12.5 (A and C) and 150 (B) μm. (D) Quantitative analysis of neutrophil infiltration 1 day after SE (mean ± S.E.M).
T16283 24411-24761 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (E) Linear regression analysis between the number of infiltrated neutrophils/area in the vasogenic edema region and the area of vasogenic edema in the PC. (F) Quantitative analysis of the number of MIP-2 positive cells per the unit area of vasogenic edema 1 day after SE (mean ± S.E.M).
T198 24411-24761 Sentence denotes Significant differences from saline-infused animals, *p < 0.05. (E) Linear regression analysis between the number of infiltrated neutrophils/area in the vasogenic edema region and the area of vasogenic edema in the PC. (F) Quantitative analysis of the number of MIP-2 positive cells per the unit area of vasogenic edema 1 day after SE (mean ± S.E.M).
T16284 24762-25073 Sentence denotes There is no difference in the number of MIP-2-positive cells per unit area of vasogenic edema between sTNFp55R-infused animals and saline-infused animals. (G) Linear regression analysis between the number of MIP-2 positive cells per unit area in vasogenic edema region and the area of vasogenic edema in the PC.
T199 24762-25073 Sentence denotes There is no difference in the number of MIP-2-positive cells per unit area of vasogenic edema between sTNFp55R-infused animals and saline-infused animals. (G) Linear regression analysis between the number of MIP-2 positive cells per unit area in vasogenic edema region and the area of vasogenic edema in the PC.
T10475 25075-25091 Sentence denotes MIP-2 expression
T200 25075-25091 Sentence denotes MIP-2 expression
T10476 25092-25174 Sentence denotes MIP-2 is a powerful chemokine that contributes to recruitment of neutrophils [27].
T201 25092-25174 Sentence denotes MIP-2 is a powerful chemokine that contributes to recruitment of neutrophils [27].
T10477 25175-25380 Sentence denotes MIP-2 is undetectable or present at low levels under physiological conditions, and shows transient increases under pathological conditions via TNF-α and/or interleukin-1β (IL-1β)-dependent mechanisms [14].
T202 25175-25380 Sentence denotes MIP-2 is undetectable or present at low levels under physiological conditions, and shows transient increases under pathological conditions via TNF-α and/or interleukin-1β (IL-1β)-dependent mechanisms [14].
T10478 25381-25494 Sentence denotes Thus, it would be plausible that TNF-α-mediated MIP-2 expression may provoke SE-induced neutrophil infiltrations.
T203 25381-25494 Sentence denotes Thus, it would be plausible that TNF-α-mediated MIP-2 expression may provoke SE-induced neutrophil infiltrations.
T10479 25495-25566 Sentence denotes To confirm this hypothesis, we investigated MIP-2 expression in the PC.
T204 25495-25566 Sentence denotes To confirm this hypothesis, we investigated MIP-2 expression in the PC.
T10480 25567-25772 Sentence denotes Consistent with our previous study [20], some MIP-2-positive astrocytes were observed in the core and periphery of the vasogenic edema lesions, but not in the non-vasogenic edema region (Figure 6B and 6C).
T205 25567-25772 Sentence denotes Consistent with our previous study [20], some MIP-2-positive astrocytes were observed in the core and periphery of the vasogenic edema lesions, but not in the non-vasogenic edema region (Figure 6B and 6C).
T10481 25773-26154 Sentence denotes Although the number of MIP-2 positive cells per unit area in the PC region of sTNFp55R-infused animals was significantly lower than that of the saline-infused group due to reduction of the area of vasogenic edema, there was no difference in the number of MIP-2 positive cells per unit area of vasogenic edema between sTNFp55R-infused animals and saline-infused animals (Figure 6F).
T206 25773-26154 Sentence denotes Although the number of MIP-2 positive cells per unit area in the PC region of sTNFp55R-infused animals was significantly lower than that of the saline-infused group due to reduction of the area of vasogenic edema, there was no difference in the number of MIP-2 positive cells per unit area of vasogenic edema between sTNFp55R-infused animals and saline-infused animals (Figure 6F).
T10482 26155-26338 Sentence denotes Furthermore, the number of MIP-2-positive cells showed a direct proportion to the unit area of vasogenic edema with a linear coefficient of correlation of 0.682 (p < 0.05, Figure 6G).
T207 26155-26338 Sentence denotes Furthermore, the number of MIP-2-positive cells showed a direct proportion to the unit area of vasogenic edema with a linear coefficient of correlation of 0.682 (p < 0.05, Figure 6G).
T10483 26339-26650 Sentence denotes Therefore, together with reduction in neutrophil infiltration in the PC region of sTNFp55R-infused animals, our findings provide evidence that TNF-α may regulate SE-induced neutrophil infiltration at least in the PC via vasogenic edema formation and not via direct TNF-α-mediated MIP-2 expression in astrocytes.
T208 26339-26650 Sentence denotes Therefore, together with reduction in neutrophil infiltration in the PC region of sTNFp55R-infused animals, our findings provide evidence that TNF-α may regulate SE-induced neutrophil infiltration at least in the PC via vasogenic edema formation and not via direct TNF-α-mediated MIP-2 expression in astrocytes.
T209 26652-26662 Sentence denotes Discussion
T11625 26663-26898 Sentence denotes The major findings in the present study are that TNF-α signaling showed cellular specific responses of NF-κB phosphorylation in the PC following SE, which may be related to vasogenic edema formation followed by neutrophil infiltration.
T210 26663-26898 Sentence denotes The major findings in the present study are that TNF-α signaling showed cellular specific responses of NF-κB phosphorylation in the PC following SE, which may be related to vasogenic edema formation followed by neutrophil infiltration.
T11626 26899-26984 Sentence denotes BBB disruption has been reported in experimental and human epilepsy [12,13,15,16,28].
T211 26899-26984 Sentence denotes BBB disruption has been reported in experimental and human epilepsy [12,13,15,16,28].
T11627 26985-27122 Sentence denotes Leakage of serum-derived components into the extracellular space is associated with hyperexcitability and seizure onset [12,13,15,16,28].
T212 26985-27122 Sentence denotes Leakage of serum-derived components into the extracellular space is associated with hyperexcitability and seizure onset [12,13,15,16,28].
T11628 27123-27244 Sentence denotes Furthermore, dysfunction of the BBB leads to epileptogenesis and contributes to progression of epilepsy [12,13,15,16,28].
T213 27123-27244 Sentence denotes Furthermore, dysfunction of the BBB leads to epileptogenesis and contributes to progression of epilepsy [12,13,15,16,28].
T11629 27245-27349 Sentence denotes In the present study, TNF-α immunoreactivity was obviously observed in microglia in the PC following SE.
T214 27245-27349 Sentence denotes In the present study, TNF-α immunoreactivity was obviously observed in microglia in the PC following SE.
T11630 27350-27466 Sentence denotes TNF receptor expressions were also up-regulated in astrocytes (TNFp55R and TNFp75R) and endothelial cells (TNFp75R).
T215 27350-27466 Sentence denotes TNF receptor expressions were also up-regulated in astrocytes (TNFp55R and TNFp75R) and endothelial cells (TNFp75R).
T11631 27467-27637 Sentence denotes Furthermore, blockade of TNF-α signaling by sTNFp55 infusion effectively (but not completely) reduced volumes of SE-induced vasogenic edema and neuronal damage in the PC.
T216 27467-27637 Sentence denotes Furthermore, blockade of TNF-α signaling by sTNFp55 infusion effectively (but not completely) reduced volumes of SE-induced vasogenic edema and neuronal damage in the PC.
T11632 27638-27801 Sentence denotes These findings indicate that TNF-α may participate in astroglial and endothelial responses to SE, which are relevant to SE-induced vasogenic edema formation [5-8].
T217 27638-27801 Sentence denotes These findings indicate that TNF-α may participate in astroglial and endothelial responses to SE, which are relevant to SE-induced vasogenic edema formation [5-8].
T11633 27802-27897 Sentence denotes Indeed, TNF-α signaling increases BBB permeability in various experimental disease models [29].
T218 27802-27897 Sentence denotes Indeed, TNF-α signaling increases BBB permeability in various experimental disease models [29].
T11634 27898-28023 Sentence denotes In the present study, sTNFp55 infusion could not completely prevent SE-induced vasogenic edema and neuronal damage in the PC.
T219 27898-28023 Sentence denotes In the present study, sTNFp55 infusion could not completely prevent SE-induced vasogenic edema and neuronal damage in the PC.
T11635 28024-28143 Sentence denotes Therefore, our findings suggest that TNF-α signaling may not be a unique upstream event in vasogenic edema development.
T220 28024-28143 Sentence denotes Therefore, our findings suggest that TNF-α signaling may not be a unique upstream event in vasogenic edema development.
T11636 28144-28288 Sentence denotes p65 phosphorylation of NF-κB enhances its transactivation potential, and p65 phosphorylation occurs in either the cytoplasm or the nucleus [30].
T221 28144-28288 Sentence denotes p65 phosphorylation of NF-κB enhances its transactivation potential, and p65 phosphorylation occurs in either the cytoplasm or the nucleus [30].
T11637 28289-28464 Sentence denotes In the present study, p65-Thr435 immunoreactivity was detected in endothelial cells, and its immunoreactivity showed an inverse correlation to the degree of SMI-71 expression.
T222 28289-28464 Sentence denotes In the present study, p65-Thr435 immunoreactivity was detected in endothelial cells, and its immunoreactivity showed an inverse correlation to the degree of SMI-71 expression.
T11638 28465-28565 Sentence denotes SMI-71, an endothelial barrier antigen, is a protein expressed by endothelial cells of rat BBB [31].
T223 28465-28565 Sentence denotes SMI-71, an endothelial barrier antigen, is a protein expressed by endothelial cells of rat BBB [31].
T11639 28566-28658 Sentence denotes Under pathological conditions, SMI-71 expression is lost in endothelial cells [5,7,8,30,32].
T224 28566-28658 Sentence denotes Under pathological conditions, SMI-71 expression is lost in endothelial cells [5,7,8,30,32].
T11640 28659-28787 Sentence denotes Acute phases of the above pathological conditions are accompanied by opening of the BBB and development of vasogenic edema [33].
T225 28659-28787 Sentence denotes Acute phases of the above pathological conditions are accompanied by opening of the BBB and development of vasogenic edema [33].
T11641 28788-29027 Sentence denotes Indeed, neutralization of SMI-71 in vivo leads to widening of intercellular junctions between endothelial cells and swelling of perivascular astrocytic processes [34], although SMI-71 is not localized at endothelial cell junctions [35-38].
T226 28788-29027 Sentence denotes Indeed, neutralization of SMI-71 in vivo leads to widening of intercellular junctions between endothelial cells and swelling of perivascular astrocytic processes [34], although SMI-71 is not localized at endothelial cell junctions [35-38].
T11642 29028-29187 Sentence denotes In the present study, SMI-71 immunoreactivity was significantly reduced in blood vessels 1 day after SE when vasogenic edema and neuronal damage were observed.
T227 29028-29187 Sentence denotes In the present study, SMI-71 immunoreactivity was significantly reduced in blood vessels 1 day after SE when vasogenic edema and neuronal damage were observed.
T11643 29188-29275 Sentence denotes Furthermore, sTNFp55R infusion effectively prevented SE-induced SMI-71 down-regulation.
T228 29188-29275 Sentence denotes Furthermore, sTNFp55R infusion effectively prevented SE-induced SMI-71 down-regulation.
T11644 29276-29576 Sentence denotes With respect to the phosphorylation of p65-Thr435 by TNF-α [39], our findings indicate that TNF-α-mediated p65-Thr435 phosphorylation in endothelial cells may play an important role in vasogenic edema induction via SMI-71 degradation or its posttranslational dysfunction influencing BBB permeability.
T229 29276-29576 Sentence denotes With respect to the phosphorylation of p65-Thr435 by TNF-α [39], our findings indicate that TNF-α-mediated p65-Thr435 phosphorylation in endothelial cells may play an important role in vasogenic edema induction via SMI-71 degradation or its posttranslational dysfunction influencing BBB permeability.
T11645 29577-29868 Sentence denotes In our previous studies [5,8], dystrophin (an actin-binding protein [40]) immunoreactivity was detected in blood vessels and in astrocytic perivascular end-feet, and was down-regulated 12 hrs after SE prior to the appearance of vasogenic edema and down-regulation of SMI-71 immunoreactivity.
T230 29577-29868 Sentence denotes In our previous studies [5,8], dystrophin (an actin-binding protein [40]) immunoreactivity was detected in blood vessels and in astrocytic perivascular end-feet, and was down-regulated 12 hrs after SE prior to the appearance of vasogenic edema and down-regulation of SMI-71 immunoreactivity.
T11646 29869-30032 Sentence denotes With respect to this previous report, changes in SMI immunoreactivity would be causes/results of interaction between endothelial cells and perivascular astrocytes.
T231 29869-30032 Sentence denotes With respect to this previous report, changes in SMI immunoreactivity would be causes/results of interaction between endothelial cells and perivascular astrocytes.
T11647 30033-30162 Sentence denotes In the present study, p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation was observed in astrocytes following SE.
T232 30033-30162 Sentence denotes In the present study, p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation was observed in astrocytes following SE.
T11648 30163-30284 Sentence denotes Furthermore, sTNFp55R infusion effectively inhibited p65-Ser276 and p65-Ser311phosphorylation in astrocytes following SE.
T233 30163-30284 Sentence denotes Furthermore, sTNFp55R infusion effectively inhibited p65-Ser276 and p65-Ser311phosphorylation in astrocytes following SE.
T11649 30285-30414 Sentence denotes Therefore, it is likely that enhanced p65-Ser276 and p65-Ser311 phosphorylation may be involved in TNF-α-mediated BBB disruption.
T234 30285-30414 Sentence denotes Therefore, it is likely that enhanced p65-Ser276 and p65-Ser311 phosphorylation may be involved in TNF-α-mediated BBB disruption.
T11650 30415-30519 Sentence denotes However, sTNFp55R infusion could not prevent p65-Ser529 and p65-Ser536 phosphorylations from SE insults.
T235 30415-30519 Sentence denotes However, sTNFp55R infusion could not prevent p65-Ser529 and p65-Ser536 phosphorylations from SE insults.
T11651 30520-30714 Sentence denotes Since p65-Ser529 and p65-Ser536 are phosphorylated by TNF-α and IL-1β [41], it is likely that IL-1β-mediated p65-Ser529/Ser536 phosphorylation may also play a role in SE-induced vasogenic edema.
T236 30520-30714 Sentence denotes Since p65-Ser529 and p65-Ser536 are phosphorylated by TNF-α and IL-1β [41], it is likely that IL-1β-mediated p65-Ser529/Ser536 phosphorylation may also play a role in SE-induced vasogenic edema.
T11652 30715-30911 Sentence denotes Therefore, our findings indicate that both TNF-α and IL-1β may be synergists to play either a direct (by endothelial cells) or indirect (by astrocytes) role in the maintenance of BBB permeability.
T237 30715-30911 Sentence denotes Therefore, our findings indicate that both TNF-α and IL-1β may be synergists to play either a direct (by endothelial cells) or indirect (by astrocytes) role in the maintenance of BBB permeability.
T11653 30912-31063 Sentence denotes Neutrophil infiltration into brain parenchyma is transiently observed during the acute phase of SE (4 - 36 hr after SE) and disappears thereafter [20].
T238 30912-31063 Sentence denotes Neutrophil infiltration into brain parenchyma is transiently observed during the acute phase of SE (4 - 36 hr after SE) and disappears thereafter [20].
T11654 31064-31163 Sentence denotes SE rapidly increases synthesis and release of chemokines in various areas of the rodent brain [42].
T239 31064-31163 Sentence denotes SE rapidly increases synthesis and release of chemokines in various areas of the rodent brain [42].
T11655 31164-31270 Sentence denotes Among them, MIP-2 is required for efficient neutrophil or lymphocyte recruitment to brain parenchyma [43].
T240 31164-31270 Sentence denotes Among them, MIP-2 is required for efficient neutrophil or lymphocyte recruitment to brain parenchyma [43].
T11656 31271-31409 Sentence denotes In our previous study [20], neutrophil infiltration in the frontoparietal cortex was regulated by P2X7 receptor-mediated MIP-2 expression.
T241 31271-31409 Sentence denotes In our previous study [20], neutrophil infiltration in the frontoparietal cortex was regulated by P2X7 receptor-mediated MIP-2 expression.
T11657 31410-31555 Sentence denotes In the PC, however, neither a P2X7 receptor agonist/antagonist nor IL-1Ra (an IL-1β antagonist) infusion could not affect leukocyte infiltration.
T242 31410-31555 Sentence denotes In the PC, however, neither a P2X7 receptor agonist/antagonist nor IL-1Ra (an IL-1β antagonist) infusion could not affect leukocyte infiltration.
T11658 31556-31720 Sentence denotes In the present study, sTNFp55R infusion effectively inhibited neutrophil infiltration in the PC by reducing vasogenic edema formation in a MIP-2-independent manner.
T243 31556-31720 Sentence denotes In the present study, sTNFp55R infusion effectively inhibited neutrophil infiltration in the PC by reducing vasogenic edema formation in a MIP-2-independent manner.
T11659 31721-31960 Sentence denotes With respect to the present and our previous reports, it is therefore likely that vasogenic edema induced by TNF-α can induce neutrophil infiltration and press injury to evoke neuronal-astroglial loss in the PC, unlike other brain regions.
T244 31721-31960 Sentence denotes With respect to the present and our previous reports, it is therefore likely that vasogenic edema induced by TNF-α can induce neutrophil infiltration and press injury to evoke neuronal-astroglial loss in the PC, unlike other brain regions.
T11660 31961-32216 Sentence denotes In conclusion, our findings reveal that impairments of endothelial cell function via TNF-α mediated p65-Thr 435 NF-κB phosphorylation may be involved in SE-induced vasogenic edema, which is relevant to neutrophil infiltration and neuronal-astroglial loss.
T245 31961-32216 Sentence denotes In conclusion, our findings reveal that impairments of endothelial cell function via TNF-α mediated p65-Thr 435 NF-κB phosphorylation may be involved in SE-induced vasogenic edema, which is relevant to neutrophil infiltration and neuronal-astroglial loss.
T246 32218-32237 Sentence denotes Competing interests
T247 32238-32296 Sentence denotes The authors declare that they have no competing interests.
T248 32298-32320 Sentence denotes Authors' contributions
T249 32321-32391 Sentence denotes JEK and HJR were involved in designing and performing all experiments.
T250 32392-32438 Sentence denotes SYC and TCK helped in drafting the manuscript.
T251 32439-32585 Sentence denotes JEK and HJR did the immunohistochemistry, the intracerebroventricular drug infusion, the seizure studies and the acquisition of data and analyses.
T252 32586-32680 Sentence denotes SYC and TCK provided continuous intellectual input, and evaluation and interpretation of data.
T253 32681-32732 Sentence denotes All authors read and approved the final manuscript.