New concept for a toxicity assay based on multiple indexes from the wave shape of damped metabolic oscillation induced in living yeast cells (part I): characterization of the phenomenon.
The damped glycolytic oscillation phenomenon occurring in starved cells of the yeast Saccharomyces cerevisiae (NBRC 0565) was characterization for application to a toxicity bioassay. S. cerevisiae was grown under semi-anaerobic conditions. The transient oscillations were observed photometrically as the time course of the fluorescent intensity of reduced pyridine nucleotide resulting from instantaneous addition of glucose to a cell suspension. In this study, simple and reproducible conditions inducing damped oscillations were obtained by modifying a literature method. For estimation of the wave shapes of the damped oscillations we used six indexes. To investigate the total reproducibility as the averaged relative standard deviation (RSD(av)) for the six indexes obtained from the wave shapes, the damped oscillations were induced under the optimum conditions and the RSD(av) values were calculated as 14% in a buffer cell suspension (n = 62) and 22% in a water cell suspension (n = 78). Finally, the effects of glucose concentration on the six indexes were examined, and all the indexes changed when the glucose concentration was changed. Excellent correlations were obtained between the index of oscillation-state time and the concentration of glucose in a buffer cell suspension (r = 0.9985, 0.5-250 mmol L(-1), 10 points) and in a water cell suspension (r = 0.9989, 2.5 micromol L(-1)-250 mmol L(-1), 12 points), respectively.
Nakamura
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H
School of Bionics, Graduate School of Bionics, Computer and Media Sciences, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo, Japan. nakamura@bs.teu.ac.jp
Suzuki
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M
D002455:Cell Division / Q000187:drug effects; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D009249:NADP / Q000378:metabolism; D009991:Oscillometry; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature; D018675:Toxicity Tests / Q000379:methods*
D009249:NADP; D005947:Glucose