| Project/Area Number |
25670064
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Environmental and hygienic pharmacy
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| Research Institution | Nihon Pharmaceutical University |
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Principal Investigator |
Negishi Kazuo 日本薬科大学, 薬学部, 教授 (70116490)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA MICHIO 日本薬科大学, 薬学部, 准教授 (50349578)
NEGISHI TOMOE 岡山大学, 医歯薬総合研究科, 准教授 (80116491)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | UV / TOF-MS / 重水素置換 / DNA / RNA / 紫外線 / 水付加 / DNA損傷 / RNA損傷 / 質量分析 / リン酸イオン / 自己修復 / 紫外線損傷 / RNA合成 / ピリミジン |
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| Outline of Final Research Achievements |
It is well known that pyrimidines are readily hydrated upon irradiation by UV. However, its significance in UV toxicity and mutagenicity is still unclear. Among DNA components, cytosines are photohydrated efficiently, while thymine is relatively resistant to the hydration. Because this reaction is reversible and hydrated cytosines are unstable, it is not easy to measure levels of hydration in DNA. We have tried to estimate the hydration by using hydrogen-exchange at 5-position of cytosine. As a model compound, oligonucleotides containing deoxycytidine was employed to study the hydrogen-deuterium exchange. The UV-irradiated sample is dissolved with H2O, and analyzed. We found that the deuterium exchange can be monitored by MALDI-TOF MS as well as NMR measurement. The MS measurement is much easier than NMR, and suitable for multiple analyses. Another aspect is an effect of cellular components on the photohydration. Phosphate can catalyze dehydration from deoxycytidine hydrate.
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