United States v. F/V Taiyo Maru. 395 F. Supp. 413

Abstract

We have recently described the construction of a dual-cell, nonlinear dielectric spectrometer, and its application to the study of cell suspensions of S. cereuisiae (A.M. Woodward and D.B. Kell, Bioelectro-them. Bioenerg., 24 (1990) 83). Substantial, odd harmonics were generated by these cells when stimulated by very modest sinusoidal electrical fields, within fairly sharp voltage-and frequency windows (ca. 0.8-2.5 V cm-', l-50 Hz). Resting cells were found to generate only odd-numbered harmonics. In the present work, we have simultaneously applied two sinusoidal frequencies which were indiuidually of unsuitable frequency and/or amplitude for the generation of harmonics when applied to suspensions of S. cerevisiae. Strong "sidebands" or "beat frequencies" were observed which were the (odd-numbered) sums and differences of the exciting frequencies (viz. fi f 2f2, f2 +2f,). The generation of these beat frequencies was strongly inhibited by low concentrations of sodium metavanadate, suggesting that they may be ascribed largely to the H+-ATPase present in the plasma membranes of these cells. We show that the ability of dc fields to inhibit the manifestation of nonlinear dielectric behaviour by these cells is explicable in terms of their ability to act as a field of zero Hz, forcing the excitation out of the amplitude window. When the cells were allowed to glycolyse, beat frequencies of even order (fi f f*, fi f 3fi) were observed. The present approach provides a novel and powerful approach to the registration of nonlinear dielectric spectra, which, due to the greater precision inherent in the discrimination of frequencies rather than voltages may be expected to provide a more sensitive means of detecting nonlinear dielectric properties in biological systems. If the transduction of exogenous electrical field energy recorded by this method is representative of the natural turnover of the H+-ATPase in viva, it may be calculated that the efficiency of the capture of electric field energy by this enzyme is some 3%.

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