Effect of the periodic properties of toxic stress on the oscillatory behaviour of glycolysis in yeast-evidence of a toxic effect frequency.

Starving and nondividing yeast cells induce changes in the electron donor nicotinamide adenine dinucleotide (NADH) levels in a cyclic and wave-like manner for over 90min. Yeast suspensions were used to examine the toxic effects of contaminants on the cyclic behaviour of metabolite changes during anaerobic glycolysis. The cyclic behaviour NADH levels in yeast cell suspensions starved for 2 to 5h was studied after the addition of 10mM glucose for 5min followed by 10mM KCN to block aerobic glycolysis. The effects of three toxic elements (CuSO, silver nanoparticles-nAg, and GdCl), known for their potential to alter glycolsysis, on NADH levels over time were examined during the 3-h starvation step. The data were analyzed using spectral analysis (Fourier transformation) to characterize the cyclic behaviour of NADH levels during anaerobic glycolysis. Increasing the starvation time by 3h increased the amplitude of changes in NADH levels with characteristic periods of 3 to 8min. Longer starvation times decreased the amplitude of oscillations during these periods, with the appearance of NADH changes at higher frequencies. Moreover, the amplitude changes in NADH were proportional to the starvation time. Exposure to the above chemicals during the 3-h starvation time led to the formation of higher frequencies with concentration-dependent amplitude changes. In comparison with nAg and Gd, Cu was the most toxic (decreased viability the most) and produce changes at higher frequencies as well. It is noteworthy that each element produced a characteristic change in the frequency profiles, which suggests different mechanisms of action in which the severity of toxicity shifted NADH changes at higher frequencies. In conclusion, the appearance of synchronized oscillations in dense yeast populations following synchronization stress could be induced by starvation and exposure to chemicals. However, synchronicity could be abolished when cells desynchronize as a result of loss of cell viability, which contributes to heterogeneity in yeast populations, translating into NADH changes at higher frequencies. This is the first report on the influence of environmental contaminants on the cyclic or wave-like behaviour of biochemical changes in cells.