著者

Yuki HIRAKAWA Tomomi YAMASAKI Ayako HARADA Seiji IWASA Hiroshi NARITA Shiro MIYAKE

出版者

The Japan Society for Analytical Chemistry

雑誌

Analytical Sciences (ISSN:09106340)

巻号頁・発行日

vol.34, no.5, pp.533-539, 2018-05-10 (Released:2018-05-10)

参考文献数

30

被引用文献数

18

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A simultaneous immunosensor based on surface plasmon resonance (SPR) was developed for determination of 3 pesticides —boscalid, clothianidin and nitenpyram— instead of the direct competitive enzyme-linked immunosorbent assays (dcELISAs) widely used as individual determination methods. Carboxy groups that introduced compounds to their pesticides were designed, and conjugates of them and bovine serum albumin were immobilized onto separate channels of the same sensor chip. When a mixture of 3 monoclonal antibodies reacted to each pesticide, and 3 pesticides were injected into the SPR immunosensor, each channel showed specific reactivity at 15 – 93 ng mL−1 for boscalid, 6.7 – 27 ng mL−1 for clothianidin, and 7.3 – 62 ng mL−1 for nitenpyram. Recovery tests using vegetables spiked with a mixture of 3 pesticides showed good results: 75 – 90%, 88 – 104%, and 72 – 105%, respectively, with a high correlation to results of the dcELISAs. The SPR immunosensor would be useful for the determination of pesticide residues in vegetables.

著者

Asuka Kato Yuki Hirakawa Wakako Hiraoka

出版者

Fujita Medical Society

雑誌

Fujita Medical Journal (ISSN:21897247)

巻号頁・発行日

vol.5, no.4, pp.98-103, 2019 (Released:2019-11-01)

参考文献数

36

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Objectives: The objective of this study was to identify the role of reactive oxygen species (ROS) generated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, in inositol hexaphosphate (IP6)-induced metabolic disruption in human leukemia PLB-985 cells.Methods: PLB-985 and X chromosome linked gp91-phox gene knockout (X-CGD) cells were treated with 5, 10, or 20 mM IP6 for 24 to 72 h. Cell growth was assayed using a highly water-soluble tetrazolium salt. The rate of apoptotic and necrotic cell death was determined with an Annexin V-fluorescein isothiocyanate/propidium iodide kit. The expression of CD11b as a marker of monocytic property and of LC3 as an autophagy marker was tested, using flow cytometry combined with fluorescent antibodies.Results: Treatment with 5 and 10 mM IP6 for 24 h was found to suppress the growth of both cell lines, though the effect was more dramatic in PLB-985 cells. After 6-h treatment with 20 mM IP6, the necrosis rate of PLB-985 cells was significantly greater than that of X-CGD cells. Further, after 72-h treatment with 10 mM IP6, CD11b expression was observed in PLB-985 cells but inhibited in X-CGD cells. Autophagy monitoring after 6-h treatment with 10 mM IP6 revealed that LC3 expression was suppressed in PLB-985 cells, whereas it was somewhat increased in X-CGD cells.Conclusions: Our results suggest that NADPH oxidase activation mediates IP6-induced metabolic disruption associated with necrosis, differentiation, cell growth, and autophagy in PLB-985 cells.