| Project/Area Number |
02670176
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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 |
細菌学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
UENO Ikuko Institute of Medical Science, Lecturer Univ. of Tokyo,, 医科学研究所, 講師 (60012738)
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| Co-Investigator(Kenkyū-buntansha) |
KANEGASAKI Shiro Institute of Medical Science, Univ. of Tokyo, Professor, 医科学研究所, 教授 (10012767)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1990: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Hydroxyl radical / Phagocytosis / Neutrophils / Macrophages / ESR spin-trapping / Superoxide / PMA / Zymosan / DMPO / zymosan. / マクロファ-ジ / ス-パ-オキシド / ビドロキシラジカル / ESRスピントラップ法 / チモ-ザン / DMPO / ヒドロキシラジカル / スピントラップ / 8ーハイドロキシグアニン |
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| Research Abstract |
Upon phagocytosis, phagocytes such as neutrophils and macrophages produce superoxide anion, from which other reactive oxygen species such as H_2O_2 and HOCl are generated. Although it was assumed that OH is also generated, no proof has been provided probably because of its high sensitivity. We investigated whether OH is generated by porcine neutrophils, mouse and rat peritoneal macrophages and lung macrophages. Following four sensitive analytical methods were employed to detect OH. (1) Detection of DMPO-CH_3 derived from dimethylsulfoxide and DMPO in the reaction mixture and OH by ESR spin-trapping technique. (2) Colorimetric detection of thiobarbituric acid-reactive material formed in added deoxyribose with OH. (3) Quantitation of 8-hydroxydeoxyguanosine formed in cell DNA and added DNA. (4) Quantitation of hydroxy derivatives of phenylalanine and tyrosine in phagocytes. However, no indication of OH generation was found when the cells were stimulated with PMA or zymosan. On the other hand, when membrane fraction obtained from the stimulated neutrophils, was used as O_2 generating system OH adduct was detected by ESR spin-trapping technique. The result suggest formation of OH from O_2 with the aid of catalytic amount of iron-complexes released from the damaged cells.
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