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Injury of Neurons by Oxygen-Glucose Deprivation in Organotypic Hippocampal Slice Culture (뇌 해마조직 절편 배양에서 산소와 당 박탈에 의한 뇌신경세포 손상)

  • Chung, David Chanwook;Hong, Kyung Sik;Kang, Jihui;Chang, Young Pyo
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1112-1117
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    • 2008
  • Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

A Dynamic Behavior Evaluation of the Curved Rail according to Lateral Spring Stiffness of Track System (궤도시스템의 횡탄성에 따른 곡선부 레일의 동적거동평가)

  • Kim, Bag-Jin;Choi, Jung-Youl;Chun, Dae-Sung;Eom, Mac;Kang, Yun-Suk;Park, Yong-Gul
    • Proceedings of the KSR Conference
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    • 2007.11a
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    • pp.517-528
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    • 2007
  • Domestic or international existing researches regarding rail damage factors are focused on laying, vehicle conditions, driving speed and driving habits and overlook characteristics of track structure (elasticity, maintenance etc). Also in ballast track, as there is no special lateral spring stiffness of track also called as ballast lateral resistance in concrete track, generally, existing study shows concrete track has 2 time shorter life cycle for rail replacement than ballast track due to abrasion. As a result of domestic concrete track design and operation performance review, concrete track elasticity is lower than track elasticity of ballast track resulting higher damage on rail and tracks. Generally, concrete track has advantage in track elasticity adjustment than ballast track and in case of Europe, in concrete track design, it is recommended to have same or higher performance range of vertical elastic stiffness of ballast track but domestically or internationally review on lateral spring stiffness of track is very minimal. Therefore, through analysis of service line track on site measurement and analysis on performance of maintenance, in this research, dynamic characteristic behaviors of commonly used ballast and concrete track are studied to infer elasticity of service line track and experimentally prove effects of track lateral spring stiffness that influence curved rail damage as well as correlation between track elasticity by track system and rail damage to propose importance of appropriate elastic stiffness level for concrete and ballast track.

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Effect of Microbial Additives on Metabolic Characteristics in Sheep and Milking Performance of Lactating Dairy Cows (미생물제제의 첨가가 면양의 반추대사 및 젖소의 유생산성에 미치는 영향)

  • Kim, G.L.;Choi, S.K.;Choi, S.H.;Song, M.K.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.819-828
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    • 2007
  • Two experiments were conducted to observe the effects of direct fed microbials on metabolic characteristics in sheep and milking performance in dairy cows. A metabolic trial with four ruminally cannulated sheep(60±6kg) was conducted in a 4×4 Latin square design to investigate the supplementation effects of Saccharomyces cerevisiae, Clostridium butyricum or mixed microbes of S. cerevisiae and C. butyricum on ruminal fermentation characteristics and whole tract digestibility. Sheep were fed 1.25 kg of total mixed ration(TMR, DM basis) supplemented with S. cerevisiae (2.5g/day), C. butyricum (1.0g/day) or its mixture(S. cerevisiae 1.25g/day+C. butyricum 1g/day), twice daily in an equal volume. But control sheep were fed only TMR. A feeding trial with 28 lactating Holstein cattle was also conducted for 12 weeks to investigate the effects of the same microbial supplements as for the metabolic trial on milking performance. The cows were fed the TMR(control), and fed S. cerevisiae(50g/day), C. butyricum(15g/day) or its mixture (S. cerevisiae 25g/day + C. butyricum 7.5g/day) with upper layer dressing method. Total VFA concentration and the digestibility of whole digestive tract in the sheep increased by supplementation of S. cerevisiae, C. butyricum or their combined microbials compare to control group. The proportion of propionic acid at 1h(P<0.039) and 3h(P<0.022) decreased by supplementation of S. cerevisiae while tended to increase acetic acid proportion at the same times. Daily dry matter intake(DMI) was not influenced by the microbial treatments, but milk yield(P<0.031) and feed efficiency(milk yield/DMI, P<0.043) were higher for the cow received C. butyricum than those for other treatments. The milk fat content was higher (P<0.085) when cows fed S. cerevisiae(4.11%) than that fed the control (4.08%), the diets with C. butyricum (3.85%) and the microbial mixture. Based on the results obtained from the current experiments, supplementation of C. butyricum or mixture with S. cerevisiae might be increased milk fat content and milk productivity of lactating daily cows. (Key words:Saccharomyces cerevisiae, Clostridium butyricum, Fermentation characteristics,