| Project/Area Number |
08672142
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional basic dentistry
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| Research Institution | Kanagawa Dental College |
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Principal Investigator |
TODOKI Kazuo Assistant Professor, Kanagawa Dental College, Pharmacology, 歯学部, 講師 (90139577)
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| Co-Investigator(Kenkyū-buntansha) |
SHOJI Hirofumi Instructor, Kanagawa Dental College, 歯学部, 助手 (90277913)
LEE Chang-il Instructor, Kanagawa Dental College, 歯学部, 助手 (60220795)
TAKAHASHI Syun-suke Instructor, Kanagawa Dental College, 歯学部, 助手 (60206810)
OKABE Eiichiro Professor, Kanagawa Dental College, Pharmacology, 歯学部, 教授 (50097276)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Optical biosensor / Hypoxanthine / Ischemia / Chemiluminescence / Hydrogen peroxide / Luminol / Oxygen free radicals / Plasuma / フリーラジカル |
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| Research Abstract |
The purpose of this study is twofold : 1) to design and develop an optical biosensor that can be used without renewing reagents in the probe for detection of biologically important analytes, and 2) to apply the hypoxanthine affinity optical biosensor to measuring the purine metabolism in an ischemia-reperfusion models. We developed the chemiluminometric bienzyme sensors on the basis of the horseradish peroxidase catalyzed H_2O_2/luminol reaction ; horseradish peroxidase was coimmobilized with xanthine oxidase. Optical biosensor was developed for the batch analysis system of hypoxanthine in the concentration range between 10^<-8> and 10^<-4> M.Optical biosensor probe with photomultiplier tube (PMP) with an integrated preamplifier was investigated to detect the chemiluminescence (which is enhanced by horseradish peroxidase). Horseradish peroxidase and xanthine oxidase were immobilized entrapping sodium alginate gel. The probe based superoxide anion radicals or H_2O_2 sensor achieve detection limits 10^<-8> M.The concentration of hypoxanthine in rabbit serum was also determined in peripheral blood samples (obtained from auricular vein). Increase in hypoxanthine levels was observed in serum obtained immediately after 90 min of auricular artery occlusion. Although there is conflicting evidence as to whether oxygen-derived free radicals play a major role in reperfusion injury, the relatively sustained nature of enhanced oxygen-derived free radical generation during reperfusion has potential clinical implications. With this optical biosensor, it should be possible to measure oxygen-derived free radical generation in a variety of conditions and with multiple interventions. Such studies may further characterize the role of oxygen free radicals in ischemiaミreperfusion injury.
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