| Project/Area Number |
01560333
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Hokkaido University |
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Principal Investigator |
KUWABARA Mikinori Hokkaido University, Faculty of Veterinary Medicine, Associate Professor, 獣医学部, 助教授 (10002081)
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| Co-Investigator(Kenkyū-buntansha) |
ENDOH Daiji Hokkaido University, Faculty of Veterinary Medicine, Instructor, 獣医学部, 助手 (40168828)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Immunodeficiency Syndrome / Granulocyte Hypofunction Syndrome / neutrophil / Superoxide / Electron Spin Resonance Spectroscopy / Spin Trapping Method / NADPH Oxidase / Phorbol 12-Myristate 13-Acetate / ホルボ-ルミリステ-トアセテ-ト |
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| Research Abstract |
Hypofunction of neutrophil cells is thought to be a cause of immunodeficiency syndrome of domestic animals. Since a function of neutrophil cells has been proved to be closely related to their ability to generate reactive oxygens, a method combining electron spin resonance spectroscopy (ESR) and spin trapping method (ST) was employed to detect and identify the reactive oxygens from neutrophil cells. On the basis of the results obtained, this EST-ST method was judged to be or not to be an appropriate method as a diagnostic technique. Neutrophil cells was collected from cow, horse and human bloods. These neutrophils were stimulated by phorbol myristate acetate. The reactive oxygens which were generated by this stimulation were detected by the ESR-ST method. 2, 2, 5, 5-Dimethylpyrroline N-oxide (DMPO) was used as a spin trapping reagent. Chemiluminescence spectroscopy was also employed as another method to detect the generation of reactive oxygens.
When the generation of reactive oxygens from human neutrophil cells was examined by the ESR-ST method, it was confirmed that the reactive oxygen which was generated from this cell was only superoxide (O_2^-). When this method was applied to the neutrophils of horse and cow, it was observed that the amount of (O_2^-) generated from horse neutrophils was the same as that of human netrophils, but that of cow was a twentieth less than those of human and horse neutrophils. This means that although the ESR-ST method is an excellent method to identify species of the generated reactive oxygens and to measure the amount of them in human and horse neutrophils, but the use of an optical method to observe chemilumineescence in addition to the ESR-ST method was necessary to quantitatively measure the amount of (O_2^-)in the case of neutrophils from cow.
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