Development of non-invasive evaluation of oxidative stress against living organisms by in vivo ESR

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1)New trapping agents for nitric oxide(NO) were developed and their function was elucidated. Further, using these trapping agents NO was detected directly in living animals exposed by whole-body X-irradiation.2)It was shown that oxidative stress due to X-irradiation could be non-invasively evaluated in living animals by L-band ESR.3)As a novel nitroxyl probe, acetoxymethyl derivative of Carboxy-PROXYL which can be remained in the brain was developed. This new probe could be well distributed into the mouse brain than other probes. Further, imaging analyzer using ESR-CT method showed that this new probe could be gathered in the brain.4)A method of separable ESR-CT (electron spin resonance-computed tomography) imaging for multiple radical species was developed and applied to imaging of hydroxyl radical (OH) and nitric oxide (NO). As the result, the position and size of the individual images of OH and NO were in very good agreement with the findings for the sample.5)A simple and sensitive assay and a cellular bioimaging of a novel diaminofluorescein DAF-FM (3-amino-4-(N-methylamino)-2',7' -difluororescein) and its diacetate. It was found that this novel diaminofluorescein fluorometry should be useful not only for sensitive NO assay, but also for NO imaging in a variety of biological speciemens.6)In vivo electron spin resonance(ESR) imaging was applied to living mice after peroral administration of a nitroxide radical spin probe. A 3D ESR imaging procedure was applied in vivo in order to obtain the exact distribution of the spin probe in a living animal. The imaging pictures demonstrated that the administered spin probe was firstly located in the stomach, then delivered to the lliver, kidney and heart of the animal.