Expectations for extension of cell life and next generation anticancer drugs by using secondary metabolites of actinomycetes
Inhibitory effects of the secondary metabolite of actinomycete were examined on cell cycle of the yeasts of S. pombe and S. cerevisiae. The secondary metabolite was obtained from cultivation of the actinomycete isolated from the soil of Owakudani in Hakone, Japan. The fifth fraction of the secondary metabolite by ODS column separation (HK-T5), which was soluble to pure methanol, was used in the present experiments. The HK-T5 brought about the delay of forming colonies of S. pombe for about 11 days compared to that cultivated without the HK-T5. The delay of the colony formation was longer for the S. pombe cultivated with more amount of the HK-T5. The cultivation with HK-T5 also brought about the extension of the lifespan of the S. pombe for more than 10 weeks in a liquidus medium. The cell life recovered the ordinary manner by removal of the HK-T5, meaning that the activities of the HK-T5 is reversible. These facts confirm the suppression of cell cycle, and the delay of cell growth by the HK-T5. These phenomena were similarly observed for S. cerevisiae. Comparison of the action of HK-T5 with hydroxyurea, which is an anticancer drug inhibiting the cell cycle at S phase, clarified that the inhibitory action of HK-T5 worked at the phase earlier than S phase. The combined effects of HK-T5 on the cell cycle were evaluated with triamcinolone acetonide (TA), or aspirin, the former of which is a drug synchronizing cancer cells in S phase, and the latter keeping human cells in G1/G0 phases. The combined use of HK-T5 with TA synchronized the cells at the phase slightly proceeding from G1 to S phase without toxicity. On the other hand, the combined use with aspirin made the inhibitory effect of HK-T5 inactive. Hence, the HK-T5 is attractive as a drug for the extension of cell lifespan, and anticancer therapy.
傅立葉轉換於奈米螢光鑽石超微量偵測之研究
螢光奈米鑽石(Fluorescent Nanodiamond, FND)主要應用於生物定位，其結構中與一個氮原子相鄰的晶格缺陷部分(Nitrogen-Vacancy, NV^-)照射波長532nm的雷射會發出637nm的螢光，對FND施加磁場會使螢光強度減弱。由於在低濃度溶液中螢光訊號會被溶液的背景雜訊掩蓋而難以偵測，因此設計實驗對FND施加穩定變換的磁場，此動作能夠使螢光強度也進行相同週期的變化。針對此週期進行快速傅立葉轉換(Fast Fourier Transformation, FFT)得出的數值會與螢光強度呈正比，進而推知FND濃度，有效排除不隨磁場變動的背景雜訊。研究結果顯示，施加磁場並使用FFT能夠成功排除牛血清蛋白(Bovine Serum Albumin, BSA)、碘化丙啶(Propidium Iodide, PI)、水、血液的背景輻射，且FND在高離子濃度溶液中會沉澱，在表層包覆BSA則可以有效地改善此現象。FND不易受到血液的背景輻射干擾螢光測定。