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T46 0-7 Sentence denotes Methods
T2958 9-29 Sentence denotes Experimental animals
T47 9-29 Sentence denotes Experimental animals
T48 30-194 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 30-389 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 195-389 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 390-493 Sentence denotes Animal protocols were approved by the Institutional Animal Care and Use Committee of Hallym University.
T2960 390-702 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 494-688 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 689-702 Sentence denotes 80-23, 1996).
T2961 703-800 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T53 703-800 Sentence denotes In addition, we have made all efforts to minimize the number of animals used and their suffering.
T3276 802-839 Sentence denotes Intracerebroventricular drug infusion
T54 802-839 Sentence denotes Intracerebroventricular drug infusion
T3277 840-998 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 840-998 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 999-1158 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 999-1158 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 1159-1276 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T57 1159-1276 Sentence denotes Animals were anesthetized (Zolretil, 50 mg/kg, i.m.; Virbac Laboratories, France) and placed in a stereotaxic frames.
T3280 1277-1587 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 1277-1587 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 1588-1698 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T59 1588-1698 Sentence denotes The infusion kit was sealed with dental cement and connected to an osmotic pump (1007D, Alzet, Cupertino, CA).
T3282 1699-1765 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T60 1699-1765 Sentence denotes The pump was placed in a subcutaneous pocket in the dorsal region.
T3283 1766-1827 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T61 1766-1827 Sentence denotes Animals received 0.5 μl/hr of vehicle or compound for 1 week.
T3284 1828-1891 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T62 1828-1891 Sentence denotes Therefore, the dose of sTNFp55R was 0.6 μg/day per each animal.
T3285 1892-1952 Sentence denotes The compounds began to be immediately infused after surgery.
T63 1892-1952 Sentence denotes The compounds began to be immediately infused after surgery.
T3286 1953-2076 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 1953-2076 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 2077-2234 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 2077-2234 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 2236-2253 Sentence denotes Seizure induction
T66 2236-2253 Sentence denotes Seizure induction
T3942 2254-2451 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 2254-2451 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 2452-2536 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T68 2452-2536 Sentence denotes Using this treatment paradigm, behavioral seizures typically began within 20-40 min.
T3944 2537-2775 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 2537-2775 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 2776-2988 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 2776-2988 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 2989-3119 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 2989-3119 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 3120-3242 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T72 3120-3242 Sentence denotes The rats were then observed 3 - 4 hours a day in the vivarium for general behavior and occurrence of spontaneous seizures.
T3948 3243-3399 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 3243-3399 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 3401-3418 Sentence denotes Tissue processing
T74 3401-3418 Sentence denotes Tissue processing
T4414 3419-3453 Sentence denotes At designated time points (non-SE:
T75 3419-3453 Sentence denotes At designated time points (non-SE:
T4415 3454-3763 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 3454-3763 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 3764-3833 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T77 3764-3833 Sentence denotes The brains were removed, and postfixed in the same fixative for 4 hr.
T4417 3834-3914 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T78 3834-3914 Sentence denotes The brain tissues were cryoprotected by infiltration with 30% sucrose overnight.
T4418 3915-4074 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 3915-4074 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 4075-4193 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T80 4075-4193 Sentence denotes For stereological study, every sixth section in the series throughout the entire hippocampus was used in some animals.
T4748 4195-4215 Sentence denotes Immunohistochemistry
T81 4195-4215 Sentence denotes Immunohistochemistry
T4749 4216-4318 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T82 4216-4318 Sentence denotes Free-floating sections were first incubated with 10% normal goat serum for 30 min at room temperature.
T4750 4319-4620 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 4319-4620 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 4621-4881 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 4621-4881 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 4882-4968 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T85 4882-4968 Sentence denotes Between the incubations, the tissues were washed with PBS three times for 10 min each.
T4753 4969-5122 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 4969-5122 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 5123-5281 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 5123-5281 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 5282-5382 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T88 5282-5382 Sentence denotes The immunoreactions were observed under the Axioscope microscope (Carl Zeiss, Munchen-Hallbergmoos).
T4756 5383-5619 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 5383-5619 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 5620-5716 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T90 5620-5716 Sentence denotes For negative controls, tissues were incubated with pre-immune serum instead of primary antibody.
T5369 5718-5749 Sentence denotes Double immunofluorescence study
T91 5718-5749 Sentence denotes Double immunofluorescence study
T5370 5750-5841 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T92 5750-5841 Sentence denotes Sections were incubated with 3% bovine serum albumin in PBS for 30 min at room temperature.
T5371 5842-6677 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 5842-6677 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 6678-6893 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 6678-6893 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 6894-6989 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T95 6894-6989 Sentence denotes The sections were washed three times for 10 min with PBS, and mounted on gelatin-coated slides.
T5374 6990-7089 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T96 6990-7089 Sentence denotes For nuclei counterstaining, we used Vectashield mounting medium with DAPI (Vector, Burlingame, CA).
T97 7090-7167 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel.
T5375 7090-7181 Sentence denotes All images were captured using an AxioImage M2 microscope and AxioVision Rel. 4.8 software.
T98 7168-7181 Sentence denotes 4.8 software.
T5848 7183-7205 Sentence denotes Fluoro-Jade B staining
T99 7183-7205 Sentence denotes Fluoro-Jade B staining
T5849 7206-7351 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 7206-7351 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 7352-7454 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T101 7352-7454 Sentence denotes In our previous [18,25] and preliminary data, neuronal damage was first detectable at 3 days after SE.
T5851 7455-7531 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T102 7455-7531 Sentence denotes Therefore, we determined 3 days after SE as the best time point to look FJB.
T5852 7532-7654 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T103 7532-7654 Sentence denotes Briefly, sections were rinsed in distilled water, and mounted onto gelatin-coated slides and then dried on a slide warmer.
T5853 7655-7771 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 7655-7771 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 7772-7868 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T105 7772-7868 Sentence denotes The slides were then transferred to 0.06% potassium permanganate for 15 min and gently agitated.
T5855 7869-8124 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 7869-8124 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 8125-8299 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 8125-8299 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 8300-8405 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T108 8300-8405 Sentence denotes For stereological study, every sixth section in the series throughout the entire PC was used (see below).
T6363 8407-8442 Sentence denotes Volumetric analysis and cell counts
T109 8407-8442 Sentence denotes Volumetric analysis and cell counts
T110 8443-8598 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 8443-8885 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 8599-8706 Sentence denotes V = Σa × tnom × 1/ssf, where a is area of the region of the delineated subfield measured by AxioVision Rel.
T112 8707-8885 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 8886-8963 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T113 8886-8963 Sentence denotes The subfield areas were delineated with a 2.5 × objective lens [5,7,8,18,25].
T6366 8964-8996 Sentence denotes The volumes are reported as mm3.
T114 8964-8996 Sentence denotes The volumes are reported as mm3.
T6367 8997-9055 Sentence denotes An optical fractionator was used to estimate cell numbers.
T115 8997-9055 Sentence denotes An optical fractionator was used to estimate cell numbers.
T6368 9056-9330 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 9056-9330 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 9331-9483 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 9331-9483 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 9484-9559 Sentence denotes The number of each cell type (C) in each of the subregions is estimated as:
T6370 9484-10018 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 9560-10018 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 10019-10083 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T120 10019-10083 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6372 10084-10148 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T121 10084-10148 Sentence denotes The immunoreactive cells were counted with a 40× objective lens.
T6373 10149-10224 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T122 10149-10224 Sentence denotes All immunoreactive cells were counted regardless the intensity of labeling.
T6374 10225-10331 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T123 10225-10331 Sentence denotes Cell counts were performed by two different investigators who were blind to the classification of tissues.
T6375 10332-10452 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 10332-10452 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 10453-10636 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 10453-10636 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 10638-10660 Sentence denotes Quantification of data
T126 10638-10660 Sentence denotes Quantification of data
T127 10661-10870 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 10661-10909 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 10871-10894 Sentence denotes 3.0 and AxioVision Rel.
T129 10895-10909 Sentence denotes 4.8 software).
T7170 10910-11104 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 10910-11104 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 11105-11287 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 11105-11287 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 11288-11356 Sentence denotes The range of intensity values was obtained from the selected images.
T132 11288-11356 Sentence denotes The range of intensity values was obtained from the selected images.
T7173 11357-11480 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 11357-11480 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 11481-11582 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T134 11481-11582 Sentence denotes Manipulation of the images was restricted to threshold and brightness adjustments to the whole image.
T135 11583-11705 Sentence denotes Intensity measurements are represented as the mean number of a 256 gray scale (NIH Image 1.59 software and AxioVision Rel.
T7175 11583-11720 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 11706-11720 Sentence denotes 4.8 software).
T7176 11721-11791 Sentence denotes Values for background staining were obtained from the corpus callosum.
T137 11721-11791 Sentence denotes Values for background staining were obtained from the corpus callosum.
T7177 11792-11919 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T138 11792-11919 Sentence denotes Optical density values were then corrected by subtracting the average values of background noise obtained from 15 image inputs.
T7659 11921-11934 Sentence denotes Data analysis
T139 11921-11934 Sentence denotes Data analysis
T7660 11935-12093 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T140 11935-12093 Sentence denotes Data obtained from volumetric analysis, cell counts, and quantitative measurements were analyzed using Student's t-test to determine statistical significance.
T7661 12094-12256 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 12094-12256 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.