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T102 0-224 Sentence denotes Tumor necrosis factor-α-mediated threonine 435 phosphorylation of p65 nuclear factor-κB subunit in endothelial cells induces vasogenic edema and neutrophil infiltration in the rat piriform cortex following status epilepticus
T1 0-224 Sentence denotes Tumor necrosis factor-α-mediated threonine 435 phosphorylation of p65 nuclear factor-κB subunit in endothelial cells induces vasogenic edema and neutrophil infiltration in the rat piriform cortex following status epilepticus
T2 226-234 Sentence denotes Abstract
T103 235-245 Sentence denotes Background
T3 235-245 Sentence denotes Background
T104 246-376 Sentence denotes Status epilepticus (SE) induces severe vasogenic edema in the piriform cortex (PC) accompanied by neuronal and astroglial damages.
T4 246-376 Sentence denotes Status epilepticus (SE) induces severe vasogenic edema in the piriform cortex (PC) accompanied by neuronal and astroglial damages.
T105 377-638 Sentence denotes To elucidate the mechanism of SE-induced vasogenic edema, we investigated the roles of tumor necrosis factor (TNF)-α in blood-brain barrier (BBB) disruption during vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE.
T5 377-638 Sentence denotes To elucidate the mechanism of SE-induced vasogenic edema, we investigated the roles of tumor necrosis factor (TNF)-α in blood-brain barrier (BBB) disruption during vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE.
T106 640-647 Sentence denotes Methods
T6 640-647 Sentence denotes Methods
T107 648-806 Sentence denotes SE was induced by pilocarpine in rats that were intracerebroventricularly infused with saline-, and soluble TNF p55 receptor (sTNFp55R) prior to SE induction.
T7 648-806 Sentence denotes SE was induced by pilocarpine in rats that were intracerebroventricularly infused with saline-, and soluble TNF p55 receptor (sTNFp55R) prior to SE induction.
T108 807-916 Sentence denotes Thereafter, we performed Fluoro-Jade B staining and immunohistochemical studies for TNF-α and NF-κB subunits.
T8 807-916 Sentence denotes Thereafter, we performed Fluoro-Jade B staining and immunohistochemical studies for TNF-α and NF-κB subunits.
T109 918-925 Sentence denotes Results
T9 918-925 Sentence denotes Results
T110 926-1002 Sentence denotes Following SE, most activated microglia showed strong TNF-α immunoreactivity.
T10 926-1002 Sentence denotes Following SE, most activated microglia showed strong TNF-α immunoreactivity.
T111 1003-1100 Sentence denotes In addition, TNF p75 receptor expression was detected in endothelial cells as well as astrocytes.
T11 1003-1100 Sentence denotes In addition, TNF p75 receptor expression was detected in endothelial cells as well as astrocytes.
T112 1101-1247 Sentence denotes In addition, only p65-Thr435 phosphorylation was increased in endothelial cells accompanied by SMI-71 expression (an endothelial barrier antigen).
T12 1101-1247 Sentence denotes In addition, only p65-Thr435 phosphorylation was increased in endothelial cells accompanied by SMI-71 expression (an endothelial barrier antigen).
T113 1248-1445 Sentence denotes Neutralization of TNF-α by soluble TNF p55 receptor (sTNFp55R) infusion attenuated SE-induced vasogenic edema and neuronal damages via inhibition of p65-Thr435 phosphorylation in endothelial cells.
T13 1248-1445 Sentence denotes Neutralization of TNF-α by soluble TNF p55 receptor (sTNFp55R) infusion attenuated SE-induced vasogenic edema and neuronal damages via inhibition of p65-Thr435 phosphorylation in endothelial cells.
T114 1446-1530 Sentence denotes Furthermore, sTNFp55R infusion reduced SE-induced neutrophil infiltration in the PC.
T14 1446-1530 Sentence denotes Furthermore, sTNFp55R infusion reduced SE-induced neutrophil infiltration in the PC.
T115 1532-1542 Sentence denotes Conclusion
T15 1532-1542 Sentence denotes Conclusion
T116 1543-1712 Sentence denotes These findings suggest that impairments of endothelial cell functions via TNF-α-mediated p65-Thr 485 NF-κB phosphorylation may be involved in SE-induced vasogenic edema.
T16 1543-1712 Sentence denotes These findings suggest that impairments of endothelial cell functions via TNF-α-mediated p65-Thr 485 NF-κB phosphorylation may be involved in SE-induced vasogenic edema.
T117 1713-1817 Sentence denotes Subsequently, vasogenic edema results in extensive neutrophil infiltration and neuronal-astroglial loss.
T17 1713-1817 Sentence denotes Subsequently, vasogenic edema results in extensive neutrophil infiltration and neuronal-astroglial loss.
T18 1819-1829 Sentence denotes Background
T1140 1830-1908 Sentence denotes Status epilepticus (SE) is a medical emergency with significant mortality [1].
T19 1830-1908 Sentence denotes Status epilepticus (SE) is a medical emergency with significant mortality [1].
T1141 1909-2041 Sentence denotes SE has been defined as continuous seizure activity, which causes neuronal cell death, epileptogenesis and learning impairment [2,3].
T20 1909-2041 Sentence denotes SE has been defined as continuous seizure activity, which causes neuronal cell death, epileptogenesis and learning impairment [2,3].
T1142 2042-2179 Sentence denotes Some brain regions vulnerable to SE play a role in the generation and propagation of paroxysmal activity in experimental epilepsy models.
T21 2042-2179 Sentence denotes Some brain regions vulnerable to SE play a role in the generation and propagation of paroxysmal activity in experimental epilepsy models.
T1143 2180-2359 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 2180-2359 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 2360-2418 Sentence denotes Pilocarpine, a cholinergic agonist, induces SE in rodents.
T23 2360-2418 Sentence denotes Pilocarpine, a cholinergic agonist, induces SE in rodents.
T1145 2419-2549 Sentence denotes This pilocarpine-induced SE, similar to human TLE, shows massive neuronal loss in the hippocampus followed by glial proliferation.
T24 2419-2549 Sentence denotes This pilocarpine-induced SE, similar to human TLE, shows massive neuronal loss in the hippocampus followed by glial proliferation.
T1146 2550-2690 Sentence denotes This neuronal damage in the pilocarpine model is not restricted to the hippocampus, but often extends to extrahippocampal limbic structures.
T25 2550-2690 Sentence denotes This neuronal damage in the pilocarpine model is not restricted to the hippocampus, but often extends to extrahippocampal limbic structures.
T1147 2691-2797 Sentence denotes Indeed, pilocarpine-induced SE results in acute neuronal damages within layers II and III of the PC [5,6].
T26 2691-2797 Sentence denotes Indeed, pilocarpine-induced SE results in acute neuronal damages within layers II and III of the PC [5,6].
T1148 2798-2900 Sentence denotes SE also induces severe vasogenic edema in the PC accompanied by neuronal and astroglial damages [5-8].
T27 2798-2900 Sentence denotes SE also induces severe vasogenic edema in the PC accompanied by neuronal and astroglial damages [5-8].
T1149 2901-2996 Sentence denotes Brain edema proceeds in two phases, early cytotoxic edema phase and late vasogenic edema phase.
T28 2901-2996 Sentence denotes Brain edema proceeds in two phases, early cytotoxic edema phase and late vasogenic edema phase.
T1150 2997-3151 Sentence denotes Early cytotoxic osmotic edema is due to excess stimulation of glutamatergic pathways during SE, which increases intracellular Na+ and Ca2+ concentrations.
T29 2997-3151 Sentence denotes Early cytotoxic osmotic edema is due to excess stimulation of glutamatergic pathways during SE, which increases intracellular Na+ and Ca2+ concentrations.
T1151 3152-3252 Sentence denotes The vasogenic edema results from dysfunction of endothelial cells and the blood-brain barrier (BBB).
T30 3152-3252 Sentence denotes The vasogenic edema results from dysfunction of endothelial cells and the blood-brain barrier (BBB).
T1152 3253-3347 Sentence denotes Many studies have reported increased permeability of the BBB during epileptic activity [9-13].
T31 3253-3347 Sentence denotes Many studies have reported increased permeability of the BBB during epileptic activity [9-13].
T1153 3348-3631 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 3348-3631 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 3632-3778 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 3632-3778 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 3779-3928 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 3779-3928 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 3929-4199 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 3929-4199 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 4200-4355 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 4200-4355 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 4356-4492 Sentence denotes Indeed, Sztriha [16] reported that dexamethasone pretreatment reduces vasogenic edema in thalamus following kainic acid-induced seizure.
T37 4356-4492 Sentence denotes Indeed, Sztriha [16] reported that dexamethasone pretreatment reduces vasogenic edema in thalamus following kainic acid-induced seizure.
T1159 4493-4644 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 4493-4644 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 4645-4823 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 4645-4823 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 4824-5082 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 4824-5082 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 5083-5230 Sentence denotes Furthermore, TNF-α induces macrophage inflammatory protein-2 (MIP-2) that recruits neutrophils under pathological conditions, including SE [14,20].
T41 5083-5230 Sentence denotes Furthermore, TNF-α induces macrophage inflammatory protein-2 (MIP-2) that recruits neutrophils under pathological conditions, including SE [14,20].
T1163 5231-5395 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 5231-5395 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 5396-5552 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 5396-5552 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 5553-5742 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 5553-5742 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 5743-5911 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 5743-5911 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 5913-5920 Sentence denotes Methods
T2958 5922-5942 Sentence denotes Experimental animals
T47 5922-5942 Sentence denotes Experimental animals
T48 5943-6107 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 5943-6302 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 6108-6302 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 6303-6406 Sentence denotes Animal protocols were approved by the Institutional Animal Care and Use Committee of Hallym University.
T2960 6303-6615 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 6407-6601 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 6602-6615 Sentence denotes 80-23, 1996).
T2961 6616-6713 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T53 6616-6713 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T3276 6715-6752 Sentence denotes Intracerebroventricular drug infusion
T54 6715-6752 Sentence denotes Intracerebroventricular drug infusion
T3277 6753-6911 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 6753-6911 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 6912-7071 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 6912-7071 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 7072-7189 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T57 7072-7189 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T3280 7190-7500 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 7190-7500 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 7501-7611 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T59 7501-7611 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T3282 7612-7678 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T60 7612-7678 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T3283 7679-7740 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T61 7679-7740 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T3284 7741-7804 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T62 7741-7804 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T3285 7805-7865 Sentence denotes The compounds began to be immediately infused after surgery.
T63 7805-7865 Sentence denotes The compounds began to be immediately infused after surgery.
T3286 7866-7989 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 7866-7989 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 7990-8147 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 7990-8147 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 8149-8166 Sentence denotes Seizure induction
T66 8149-8166 Sentence denotes Seizure induction
T3942 8167-8364 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 8167-8364 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 8365-8449 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T68 8365-8449 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T3944 8450-8688 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 8450-8688 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 8689-8901 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 8689-8901 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 8902-9032 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 8902-9032 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 9033-9155 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T72 9033-9155 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T3948 9156-9312 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 9156-9312 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 9314-9331 Sentence denotes Tissue processing
T74 9314-9331 Sentence denotes Tissue processing
T4414 9332-9366 Sentence denotes At designated time points (non-SE:
T75 9332-9366 Sentence denotes At designated time points (non-SE:
T4415 9367-9676 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 9367-9676 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 9677-9746 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T77 9677-9746 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T4417 9747-9827 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T78 9747-9827 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T4418 9828-9987 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 9828-9987 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 9988-10106 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T80 9988-10106 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T4748 10108-10128 Sentence denotes Immunohistochemistry
T81 10108-10128 Sentence denotes Immunohistochemistry
T4749 10129-10231 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T82 10129-10231 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T4750 10232-10533 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 10232-10533 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 10534-10794 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 10534-10794 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 10795-10881 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T85 10795-10881 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T4753 10882-11035 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 10882-11035 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 11036-11194 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 11036-11194 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 11195-11295 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T88 11195-11295 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T4756 11296-11532 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 11296-11532 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 11533-11629 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T90 11533-11629 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T5369 11631-11662 Sentence denotes Double immunofluorescence study
T91 11631-11662 Sentence denotes Double immunofluorescence study
T5370 11663-11754 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T92 11663-11754 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T5371 11755-12590 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 11755-12590 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 12591-12806 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 12591-12806 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 12807-12902 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T95 12807-12902 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T5374 12903-13002 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T96 12903-13002 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T97 13003-13080 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel.
T5375 13003-13094 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel. 4.8 software.
T98 13081-13094 Sentence denotes 4.8 software.
T5848 13096-13118 Sentence denotes Fluoro-Jade B staining
T99 13096-13118 Sentence denotes Fluoro-Jade B staining
T5849 13119-13264 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 13119-13264 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 13265-13367 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T101 13265-13367 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T5851 13368-13444 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T102 13368-13444 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T5852 13445-13567 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T103 13445-13567 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T5853 13568-13684 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 13568-13684 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 13685-13781 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T105 13685-13781 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T5855 13782-14037 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 13782-14037 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 14038-14212 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 14038-14212 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 14213-14318 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T108 14213-14318 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T6363 14320-14355 Sentence denotes Volumetric analysis and cell counts
T109 14320-14355 Sentence denotes Volumetric analysis and cell counts
T110 14356-14511 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 14356-14798 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 14512-14619 Sentence denotes V = Σa × tnom × 1/ssf, where a is area of the region of the delineated subfield measured by AxioVision Rel.
T112 14620-14798 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 14799-14876 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T113 14799-14876 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T6366 14877-14909 Sentence denotes The volumes are reported as mm3.
T114 14877-14909 Sentence denotes The volumes are reported as mm3.
T6367 14910-14968 Sentence denotes An optical fractionator was used to estimate cell numbers.
T115 14910-14968 Sentence denotes An optical fractionator was used to estimate cell numbers.
T6368 14969-15243 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 14969-15243 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 15244-15396 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 15244-15396 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 15397-15472 Sentence denotes The number of each cell type (C) in each of the subregions is estimated as:
T6370 15397-15931 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 15473-15931 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 15932-15996 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T120 15932-15996 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6372 15997-16061 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T121 15997-16061 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6373 16062-16137 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T122 16062-16137 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T6374 16138-16244 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T123 16138-16244 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T6375 16245-16365 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 16245-16365 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 16366-16549 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 16366-16549 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 16551-16573 Sentence denotes Quantification of data
T126 16551-16573 Sentence denotes Quantification of data
T127 16574-16783 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 16574-16822 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 16784-16807 Sentence denotes 3.0 and AxioVision Rel.
T129 16808-16822 Sentence denotes 4.8 software).
T7170 16823-17017 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 16823-17017 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 17018-17200 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 17018-17200 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 17201-17269 Sentence denotes The range of intensity values was obtained from the selected images.
T132 17201-17269 Sentence denotes The range of intensity values was obtained from the selected images.
T7173 17270-17393 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 17270-17393 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 17394-17495 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T134 17394-17495 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T135 17496-17618 Sentence denotes Intensity measurements are represented as the mean number of a 256 gray scale (NIH Image 1.59 software and AxioVision Rel.
T7175 17496-17633 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 17619-17633 Sentence denotes 4.8 software).
T7176 17634-17704 Sentence denotes Values for background staining were obtained from the corpus callosum.
T137 17634-17704 Sentence denotes Values for background staining were obtained from the corpus callosum.
T7177 17705-17832 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T138 17705-17832 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T7659 17834-17847 Sentence denotes Data analysis
T139 17834-17847 Sentence denotes Data analysis
T7660 17848-18006 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T140 17848-18006 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T7661 18007-18169 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 18007-18169 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 18171-18178 Sentence denotes Results
T7823 18180-18214 Sentence denotes TNF-α, and TNF receptor expression
T143 18180-18214 Sentence denotes TNF-α, and TNF receptor expression
T7824 18215-18345 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 18215-18345 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 18346-18481 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 18346-18481 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 18482-18543 Sentence denotes This expression pattern was maintained up to 1 week after SE.
T146 18482-18543 Sentence denotes This expression pattern was maintained up to 1 week after SE.
T7827 18544-18696 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 18544-18696 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 18697-18815 Sentence denotes In 12 hr-post SE animals of the saline-infused group, TNFp55R immunoreactivity was observed in astrocytes (Figure 1B).
T148 18697-18815 Sentence denotes In 12 hr-post SE animals of the saline-infused group, TNFp55R immunoreactivity was observed in astrocytes (Figure 1B).
T7829 18816-18928 Sentence denotes Unlike TNFp55R, TNFp75R immunoreactivity was detected in endothelial cells as well as astrocytes (Figures 1C-E).
T149 18816-18928 Sentence denotes Unlike TNFp55R, TNFp75R immunoreactivity was detected in endothelial cells as well as astrocytes (Figures 1C-E).
T7830 18929-19128 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 18929-19128 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 19129-19390 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 19139-19390 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 19391-19409 Sentence denotes Bar = 25 (A-D) μm.
T152 19391-19409 Sentence denotes Bar = 25 (A-D) μm.
T8344 19411-19500 Sentence denotes Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage
T153 19411-19500 Sentence denotes Effect of sTNFp55R infusion on SE-induced serum-protein extravasation and neuronal damage
T8345 19501-19645 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 19501-19645 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 19646-19819 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 19646-19819 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 19820-19906 Sentence denotes In saline-treated animals, the PC was stained diffusely with anti-rat IgG (Figure 2A).
T156 19820-19906 Sentence denotes In saline-treated animals, the PC was stained diffusely with anti-rat IgG (Figure 2A).
T8348 19907-19968 Sentence denotes The volume of vasogenic edema was 17.1 ± 1.5 mm3 (Figure 2E).
T157 19907-19968 Sentence denotes The volume of vasogenic edema was 17.1 ± 1.5 mm3 (Figure 2E).
T8349 19969-20047 Sentence denotes The number of FJB-positive neurons in the PC was 236,145 ± 49,469 (Figure 2B).
T158 19969-20047 Sentence denotes The number of FJB-positive neurons in the PC was 236,145 ± 49,469 (Figure 2B).
T8350 20048-20156 Sentence denotes In sTNFp55R-treated animals, SE-induced vasogenic edema was attenuated to 9.8 ± 0.7 mm3 (Figures 2C and 2E).
T159 20048-20156 Sentence denotes In sTNFp55R-treated animals, SE-induced vasogenic edema was attenuated to 9.8 ± 0.7 mm3 (Figures 2C and 2E).
T8351 20157-20249 Sentence denotes In addition, the number of FJB-positive neurons in the PC was 89,138 ± 5,698 (Figures 2D-E).
T160 20157-20249 Sentence denotes In addition, the number of FJB-positive neurons in the PC was 89,138 ± 5,698 (Figures 2D-E).
T8352 20250-20378 Sentence denotes Thus, sTNFp55R infusion attenuated SE-induced vasogenic edema and neuronal damage compared to saline-infused animals (p < 0.05).
T161 20250-20378 Sentence denotes Thus, sTNFp55R infusion attenuated SE-induced vasogenic edema and neuronal damage compared to saline-infused animals (p < 0.05).
T162 20379-20574 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 20389-20574 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 20575-20724 Sentence denotes Compared to saline-infused animals, serum-protein extravasation and FJB-positive neuronal damage is markedly ameliorated in sTNFp55R-infused animals.
T163 20575-20724 Sentence denotes Compared to saline-infused animals, serum-protein extravasation and FJB-positive neuronal damage is markedly ameliorated in sTNFp55R-infused animals.
T14708 20725-20898 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 20725-20898 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 20899-20962 Sentence denotes Significant differences from saline-treated animals, *p < 0.05.
T165 20899-20962 Sentence denotes Significant differences from saline-treated animals, *p < 0.05.
T8685 20964-20985 Sentence denotes NF-κB phosphorylation
T166 20964-20985 Sentence denotes NF-κB phosphorylation
T8686 20986-21080 Sentence denotes It is well established that TNF-α is one of the major stimuli toward phosphorylation of NF-κB.
T167 20986-21080 Sentence denotes It is well established that TNF-α is one of the major stimuli toward phosphorylation of NF-κB.
T8687 21081-21209 Sentence denotes To confirm TNF-α-mediated signaling following SE, we performed an immunohistochemical study using five phospho-NF-κB antibodies.
T168 21081-21209 Sentence denotes To confirm TNF-α-mediated signaling following SE, we performed an immunohistochemical study using five phospho-NF-κB antibodies.
T8688 21210-21610 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 21210-21610 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 21611-21770 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 21611-21770 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 21771-21935 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 21771-21935 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 21936-22028 Sentence denotes Figure 3 Effect of sTNFp55R infusion on NF-κB phosphorylation in astrocytes 12 hr after SE.
T15005 21946-22028 Sentence denotes Effect of sTNFp55R infusion on NF-κB phosphorylation in astrocytes 12 hr after SE.
T15006 22029-22397 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 22029-22397 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 22398-22412 Sentence denotes Bar = 12.5 μm.
T174 22398-22412 Sentence denotes Bar = 12.5 μm.
T175 22413-22609 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 22423-22609 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 22610-22751 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 22610-22751 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 22752-22916 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 22752-22916 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 22917-23660 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 22927-23660 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 23661-23695 Sentence denotes Bars = 12.5 (A-D) and 25 (E-G) μm.
T179 23661-23695 Sentence denotes Bars = 12.5 (A-D) and 25 (E-G) μm.
T9403 23697-23714 Sentence denotes SMI-71 expression
T180 23697-23714 Sentence denotes SMI-71 expression
T9404 23715-23840 Sentence denotes Previously, we reported that SMI-71 (an endothelial barrier antigen) immunoreactivity decreased in the PC 1 day after SE [5].
T181 23715-23840 Sentence denotes Previously, we reported that SMI-71 (an endothelial barrier antigen) immunoreactivity decreased in the PC 1 day after SE [5].
T9405 23841-24039 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 23841-24039 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 24040-24133 Sentence denotes Thus, loss of SMI-71 immunoreactivity correlated with volume of vasogenic edema following SE.
T183 24040-24133 Sentence denotes Thus, loss of SMI-71 immunoreactivity correlated with volume of vasogenic edema following SE.
T9407 24134-24230 Sentence denotes This reduction in SMI-71 was accompanied by increased p65-Thr435 phosphorylation (Figures 5C-D).
T184 24134-24230 Sentence denotes This reduction in SMI-71 was accompanied by increased p65-Thr435 phosphorylation (Figures 5C-D).
T9408 24231-24409 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 24231-24409 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 24410-24631 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 24410-24631 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 24633-24656 Sentence denotes Neutrophil infiltration
T187 24633-24656 Sentence denotes Neutrophil infiltration
T9881 24657-24767 Sentence denotes Recent studies have reported that neutrophils infiltrate the brain under certain pathological conditions [26].
T188 24657-24767 Sentence denotes Recent studies have reported that neutrophils infiltrate the brain under certain pathological conditions [26].
T9882 24768-24871 Sentence denotes Indeed, we have reported massive neutrophil infiltration in layer III/IV of the PC 1 day after SE [20].
T189 24768-24871 Sentence denotes Indeed, we have reported massive neutrophil infiltration in layer III/IV of the PC 1 day after SE [20].
T9883 24872-25004 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 24872-25004 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 25005-25138 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 25005-25138 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 25139-25373 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 25139-25373 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 25374-25527 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 25374-25527 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 25528-25697 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 25528-25697 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 25698-25845 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 25698-25845 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 25846-26107 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 25856-26107 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 26108-26229 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 26108-26229 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 26230-26580 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 26230-26580 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 26581-26892 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 26581-26892 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 26894-26910 Sentence denotes MIP-2 expression
T200 26894-26910 Sentence denotes MIP-2 expression
T10476 26911-26993 Sentence denotes MIP-2 is a powerful chemokine that contributes to recruitment of neutrophils [27].
T201 26911-26993 Sentence denotes MIP-2 is a powerful chemokine that contributes to recruitment of neutrophils [27].
T10477 26994-27199 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 26994-27199 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 27200-27313 Sentence denotes Thus, it would be plausible that TNF-α-mediated MIP-2 expression may provoke SE-induced neutrophil infiltrations.
T203 27200-27313 Sentence denotes Thus, it would be plausible that TNF-α-mediated MIP-2 expression may provoke SE-induced neutrophil infiltrations.
T10479 27314-27385 Sentence denotes To confirm this hypothesis, we investigated MIP-2 expression in the PC.
T204 27314-27385 Sentence denotes To confirm this hypothesis, we investigated MIP-2 expression in the PC.
T10480 27386-27591 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 27386-27591 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 27592-27973 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 27592-27973 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 27974-28157 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 27974-28157 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 28158-28469 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 28158-28469 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 28471-28481 Sentence denotes Discussion
T11625 28482-28717 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 28482-28717 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 28718-28803 Sentence denotes BBB disruption has been reported in experimental and human epilepsy [12,13,15,16,28].
T211 28718-28803 Sentence denotes BBB disruption has been reported in experimental and human epilepsy [12,13,15,16,28].
T11627 28804-28941 Sentence denotes Leakage of serum-derived components into the extracellular space is associated with hyperexcitability and seizure onset [12,13,15,16,28].
T212 28804-28941 Sentence denotes Leakage of serum-derived components into the extracellular space is associated with hyperexcitability and seizure onset [12,13,15,16,28].
T11628 28942-29063 Sentence denotes Furthermore, dysfunction of the BBB leads to epileptogenesis and contributes to progression of epilepsy [12,13,15,16,28].
T213 28942-29063 Sentence denotes Furthermore, dysfunction of the BBB leads to epileptogenesis and contributes to progression of epilepsy [12,13,15,16,28].
T11629 29064-29168 Sentence denotes In the present study, TNF-α immunoreactivity was obviously observed in microglia in the PC following SE.
T214 29064-29168 Sentence denotes In the present study, TNF-α immunoreactivity was obviously observed in microglia in the PC following SE.
T11630 29169-29285 Sentence denotes TNF receptor expressions were also up-regulated in astrocytes (TNFp55R and TNFp75R) and endothelial cells (TNFp75R).
T215 29169-29285 Sentence denotes TNF receptor expressions were also up-regulated in astrocytes (TNFp55R and TNFp75R) and endothelial cells (TNFp75R).
T11631 29286-29456 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 29286-29456 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 29457-29620 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 29457-29620 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 29621-29716 Sentence denotes Indeed, TNF-α signaling increases BBB permeability in various experimental disease models [29].
T218 29621-29716 Sentence denotes Indeed, TNF-α signaling increases BBB permeability in various experimental disease models [29].
T11634 29717-29842 Sentence denotes In the present study, sTNFp55 infusion could not completely prevent SE-induced vasogenic edema and neuronal damage in the PC.
T219 29717-29842 Sentence denotes In the present study, sTNFp55 infusion could not completely prevent SE-induced vasogenic edema and neuronal damage in the PC.
T11635 29843-29962 Sentence denotes Therefore, our findings suggest that TNF-α signaling may not be a unique upstream event in vasogenic edema development.
T220 29843-29962 Sentence denotes Therefore, our findings suggest that TNF-α signaling may not be a unique upstream event in vasogenic edema development.
T11636 29963-30107 Sentence denotes p65 phosphorylation of NF-κB enhances its transactivation potential, and p65 phosphorylation occurs in either the cytoplasm or the nucleus [30].
T221 29963-30107 Sentence denotes p65 phosphorylation of NF-κB enhances its transactivation potential, and p65 phosphorylation occurs in either the cytoplasm or the nucleus [30].
T11637 30108-30283 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 30108-30283 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 30284-30384 Sentence denotes SMI-71, an endothelial barrier antigen, is a protein expressed by endothelial cells of rat BBB [31].
T223 30284-30384 Sentence denotes SMI-71, an endothelial barrier antigen, is a protein expressed by endothelial cells of rat BBB [31].
T11639 30385-30477 Sentence denotes Under pathological conditions, SMI-71 expression is lost in endothelial cells [5,7,8,30,32].
T224 30385-30477 Sentence denotes Under pathological conditions, SMI-71 expression is lost in endothelial cells [5,7,8,30,32].
T11640 30478-30606 Sentence denotes Acute phases of the above pathological conditions are accompanied by opening of the BBB and development of vasogenic edema [33].
T225 30478-30606 Sentence denotes Acute phases of the above pathological conditions are accompanied by opening of the BBB and development of vasogenic edema [33].
T11641 30607-30846 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 30607-30846 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 30847-31006 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 30847-31006 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 31007-31094 Sentence denotes Furthermore, sTNFp55R infusion effectively prevented SE-induced SMI-71 down-regulation.
T228 31007-31094 Sentence denotes Furthermore, sTNFp55R infusion effectively prevented SE-induced SMI-71 down-regulation.
T11644 31095-31395 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 31095-31395 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 31396-31687 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 31396-31687 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 31688-31851 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 31688-31851 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 31852-31981 Sentence denotes In the present study, p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation was observed in astrocytes following SE.
T232 31852-31981 Sentence denotes In the present study, p65-Ser276, p65-Ser311, p65-Ser529, and p65-Ser536 phosphorylation was observed in astrocytes following SE.
T11648 31982-32103 Sentence denotes Furthermore, sTNFp55R infusion effectively inhibited p65-Ser276 and p65-Ser311phosphorylation in astrocytes following SE.
T233 31982-32103 Sentence denotes Furthermore, sTNFp55R infusion effectively inhibited p65-Ser276 and p65-Ser311phosphorylation in astrocytes following SE.
T11649 32104-32233 Sentence denotes Therefore, it is likely that enhanced p65-Ser276 and p65-Ser311 phosphorylation may be involved in TNF-α-mediated BBB disruption.
T234 32104-32233 Sentence denotes Therefore, it is likely that enhanced p65-Ser276 and p65-Ser311 phosphorylation may be involved in TNF-α-mediated BBB disruption.
T11650 32234-32338 Sentence denotes However, sTNFp55R infusion could not prevent p65-Ser529 and p65-Ser536 phosphorylations from SE insults.
T235 32234-32338 Sentence denotes However, sTNFp55R infusion could not prevent p65-Ser529 and p65-Ser536 phosphorylations from SE insults.
T11651 32339-32533 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 32339-32533 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 32534-32730 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 32534-32730 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 32731-32882 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 32731-32882 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 32883-32982 Sentence denotes SE rapidly increases synthesis and release of chemokines in various areas of the rodent brain [42].
T239 32883-32982 Sentence denotes SE rapidly increases synthesis and release of chemokines in various areas of the rodent brain [42].
T11655 32983-33089 Sentence denotes Among them, MIP-2 is required for efficient neutrophil or lymphocyte recruitment to brain parenchyma [43].
T240 32983-33089 Sentence denotes Among them, MIP-2 is required for efficient neutrophil or lymphocyte recruitment to brain parenchyma [43].
T11656 33090-33228 Sentence denotes In our previous study [20], neutrophil infiltration in the frontoparietal cortex was regulated by P2X7 receptor-mediated MIP-2 expression.
T241 33090-33228 Sentence denotes In our previous study [20], neutrophil infiltration in the frontoparietal cortex was regulated by P2X7 receptor-mediated MIP-2 expression.
T11657 33229-33374 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 33229-33374 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 33375-33539 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 33375-33539 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 33540-33779 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 33540-33779 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 33780-34035 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 33780-34035 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 34037-34056 Sentence denotes Competing interests
T247 34057-34115 Sentence denotes The authors declare that they have no competing interests.
T248 34117-34139 Sentence denotes Authors' contributions
T249 34140-34210 Sentence denotes JEK and HJR were involved in designing and performing all experiments.
T250 34211-34257 Sentence denotes SYC and TCK helped in drafting the manuscript.
T251 34258-34404 Sentence denotes JEK and HJR did the immunohistochemistry, the intracerebroventricular drug infusion, the seizure studies and the acquisition of data and analyses.
T252 34405-34499 Sentence denotes SYC and TCK provided continuous intellectual input, and evaluation and interpretation of data.
T253 34500-34551 Sentence denotes All authors read and approved the final manuscript.