| Project/Area Number |
14370124
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hygiene
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| Research Institution | Yamaguchi University |
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Principal Investigator |
OKUDA Masayuki Yamaguchi University, School of medicine, Associate Professor, 医学部, 助教授 (50274171)
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| Co-Investigator(Kenkyū-buntansha) |
HOBARA Tatsuya Yamaguchi University, School of medicine, Professor, 医学部, 教授 (10116501)
KUNITSUGU Ichiro Yamaguchi University, School of medicine, Research Associate, 医学部, 助手 (80314798)
SUGIYAMA Shinichi Yamaguchi University, School of medicine, Research Associate, 医学部, 助手 (50335725)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | comet assay / single cell gel electrophoresis / DNA / apoptosis / bromopropane / trichloroethylene / toxicology / 8-OHdG / ラット / 精巣 / DNA損傷 / 有機溶剤 / single gel electrophoresis / 精原細胞 / 温度 / Syber Green |
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| Research Abstract |
Comet assay (single cell gel electrophoresis) is a simple method for evaluation of DNA toxicity. DNA of rat spermatogenic cells (spermatogonia and spermatocytes) are damaged with bromopropanes in dose- and time-dependent manner. Among some kinds of bromopropanes, 1,3-bromopropane has most toxicity and 2-bromopropane and 1-boromopropane follow. Trichloroethylene, which is used as the same purposes in industry, reduces the toxicity due to 1,3-bromopropane when trichloroethylene with bromopropane is exposed at the same time to primary-cultured spermatogenic cells. But this mechanism for toxicity reduction should be further investigated. When comet assay is carried in more neutral condition than usual alkaline condition, DNA damage is not detected. DNA fragmentation due to apoptosis may not be induced with bromopropane in the condition used in this study (primary culture, Wistar rats) since rat testes DNA does not show ladder in electrophoresis. Oxidative stress markers for DNA, 8-OHdG (hydroxy-deoxy-guanosine), are elevated in DNA from testes tissue or primary-cultured spermatogenic cells. This study shows that comet assay can be used as screening method foe toxicity to testes and may differentiate DNA damage from apoptosis in some specific conditions.
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