| 研究実績の概要 |
The potent fungi from the screening processes will be applied to the isolation guided by anti-Toxoplasma and antimalaria activities (in vitro). These research steps are intended to confirm and trace the active agent(s) responsible for the anti-Toxoplasma and antimalaria activities. Firstly, each moldy media from scale-up fermentation (600 grams rice) will be extracted using methanol. The dried methanol extract will be suspended in distilled water and partitioned with hexane, chloroform, and ethyl acetate. Each fraction will be evaluated for anti-Toxoplasma activity and antimalaria. The active fraction(s) will be applied to various chromatography methods for further separations. Every step of column chromatography will be guided by anti-Toxoplasma and antimalarial activities, and only the active fractions will be further separated. LC-MS/MS profiling and molecular networking analyses for active fractions will be conducted to generate the compound's map, trace, and visualize the active components' cluster. The structures of isolated active compounds will be determined by spectroscopic methods, including nuclear magnetic resonance (NMR), high-resolution mass spectroscopy (HR-MS), and chemical derivatization. Anti-Toxoplasma and antimalaria effects on growth and cytotoxicity of isolated active compounds will also be evaluated in vitro. The efficacy of the most promising bioactive compounds will be evaluated using the mouse model. Furthermore, the potent fungi will be identified based on the ITS rDNA region.
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