RESUMO
Although fluorescent proteins are widely used as biomarkers (Yin), no study focuses on their influence on the microbial stress response. Here, the Green Fluorescent Protein (GFP) was fused to two proteins of interest in Saccharomyces cerevisiae. Pab1p and Sur7p, respectively involved in stress granules structure and in Can1 membrane domains. These were chosen since questions remain regarding the understanding of the behavior of S. cerevisiae facing different heat kinetics or oxidative stresses. The main results showed that Pab1p-GFP fluorescent mutant displayed a higher resistance than that of the wild type under a heat shock. Moreover, fluorescent mutants exposed to oxidative stresses displayed changes in the cultivability compared to the wild type strain. In silico approaches showed that the presence of the GFP did not influence the structure and so the functionality of the tagged proteins meaning that changes in yeast resistance were certainly related to GFP ROS-scavenging ability (Yang).
Assuntos
Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Yin-Yang , Estresse Oxidativo/fisiologiaRESUMO
Steady state (SS) and transient temperature-rise in tissue from radiofrequency exposure forms the underlying basis for limits in international exposure guidelines. Periodically pulsed or intermittent exposures form a special case of having both peak and average levels, producing temperature-rise oscillations in the SS. Presented here is a method for determining tissue temperature-rise for periodic specific absorption rate (SAR) modulation having arbitrary waveform. It involves the finite difference solution of a form of the Pennes Bioheat Transfer equation (BHTE) and uses the concept of the transfer function and the Fast Fourier Transform (FFT). The time-dependent BHTE is converted to a SS harmonic version by assuming that the time-dependent SAR waveform and tissue temperature can both be represented by Fourier series. The transfer function is obtained from solutions of the harmonic BHTE for an assumed SAR waveform consisting of periodic impulses. The temperature versus time response for an arbitrary periodic SAR waveform is obtained from the inverse FFT of the product of the transfer function and the FFT of the actual SAR waveform. This method takes advantage of existing FFT algorithms on most computational platforms and the ability to store the transfer function for later re-use. The transfer function varies slowly with harmonic number, allowing interpolation and extrapolation to reduce the computational effort. The method is highly efficient for the case where repeated temperature-rise calculations for parameter variations in the SAR waveform are sought. Examples are given for a narrow, circularly symmetric beam incident on a planar skin/fat/muscle model with rectangular, triangular and cosine-pulsed SAR modulation waveforms. Calculations of temperature-rise crest factor as a function of rectangular pulse duty factor and pulse repetition frequency for the same exposure/tissue model are also presented as an example of the versatility of the method.