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F W Kleinhans

Publications and source records attributed to F W Kleinhans.

39 records · Page 3Linked to original sources

ESR determinations of membrane permeability in a yeast sterol mutant.

Yeast sterol mutants were subjected to ESR analysis in an attempt to elucidate how altered sterol composition correlates with membrane permeability. The technique requires spin labeling the intact yeast cells with a small, water-soluble nitroxide probe (2,2,5,5 tetramethyl-3-pyrrolin-1-oxyl-3-carboxylic acid, PCA), suspending cells in a NiCl2 solution, and measuring the extent of Ni2+ entry through the membrane by its magnetic dipolar line broadening effect on the PCA signal. The wild type, A184D, was found to be impermeable to Ni2+ during all growth phases while the sterol mutant erg 6/2 was readily permeable to Ni2+. Other sources of line broadening such as increased rotational correlation time and cell nonviability are shown to be neglibible. Internal Ni2+ concentrations for erg 6/2 and kinetics of Ni2+ entry were determined.

Cell Membrane Permeability↗

Membrane fluidity alterations in a cytochrome P-450-deficient mutant of Candida albicans.

A cytochrome P450-deficient mutant of the pathogenic fungus, Candida albicans, which accumulates exclusively 14 alpha-methylsterols in place of the normal end product sterol, ergosterol, was examined for alterations in membrane fluidity by electron paramagnetic resonance. The results using four nitroxyl spin labels indicated that exponential phase cultures of the mutant strain, D10, had a uniformly more rigid membrane than similarly grown wild type. Since D10 shows a sterol spectrum similar to that of wild type cells treated with imidazole and triazole antifungal agents, many of the physiological effects reported as the result of azole application may be the result of alterations in membrane fluidity.

Animals↗

Measurement of human sperm intracellular water volume by electron spin resonance.

An electron spin resonance technique using the spin label tempone and the broadening agent potassium chromium oxalate was used to measure the water volume of human sperm. The toxicity of tempone (5 mmol/L) and potassium chromium oxalate (50 mmol/L) to sperm was measured over a time span of 120 minutes using computer-assisted semen analysis. Tempone had no effect on any computer-assisted semen analysis parameters, including motility. Potassium chromium oxalate reduced sperm motility by an average of 24% during the first 30 minutes of exposure. After selection by swim-up and correction for the presence of dead cells and cytoplasmic droplets, a water volume of 20.0 +/- 2.9 microns3 was obtained. This yields a total volume of 33.9 microns3 if a water compartment of 59% by volume is assumed. These results are consistent with other shape-independent techniques for measuring volume, but larger than the generally accepted optical and electronic particle counter sizes.

Blood Viscosity↗