Milan Schara

Membrane fluidity characteristics of human lung cancer.

Membrane fluidity of non-cultured lung cancer tissue was studied by electron paramagnetic resonance (EPR). EPR spectra of a lipophilic spin probe in a tissue of resected tumor samples from 51 patients were compared with computer simulated spectra, which were superimpositions of spectra characterizing membrane domains with different fluidity. The membranes of tumor tissues were more fluid, than those of normal lungs; the most fluid domains were enlarged and their order parameter decreased in comparison to normal tissue. An empirical fluidity parameter (H13) was defined as the criterion to correlate EPR and clinical data. The histology of tumor, the quantitative presence of different tumor and non-tumor cells and the pathohisthological stage of the disease had no significant influence on fluidity.

Cell membrane fluidity and prognosis of lung cancer

BACKGROUND Membranes of tumor cells have been found to posses higher fluidity than membranes of non-tumor cells. Plasma membrane fluidity is significantly correlated with malignant potential of these cells. METHODS Seventy-five patients operated on for lung cancer were studied prospectively. During the operation, lung tumor samples were taken from the resected lung for evaluation by electron paramagnetic resonance. The fluidity variable H13, which is proportional to the plasma membrane fluidity, was determined from the electron paramagnetic resonance spectra. The association between H13 and survival was determined by survival analysis using Kaplan-Meier curves and Cox regression. RESULTS Pathologic TNM stage and the fluidity variable H13 were the only prognostic variables significantly associated with survival time in multivariate proportional hazards regression model. Thus, H13 was shown to be an independent prognostic variable for survival, which was also confirmed by a separate analysis relating the TNM stage and H13. Dividing the patients into two groups, one with an H13 value higher than the median and another with H13 below the median, resulted in significantly different survival curves (p = 0.01). CONCLUSIONS Patients with high plasma membrane fluidity, indicated by high H13 of the resected lung tumor tissue, seem to have poorer prognosis than those with less fluid membranes. We suggest that the fluidity variable could be used as an independent additional prognostic factor and a tool to identify patients who may be helped by adjuvant postoperative therapy.

Study of the Arrangement of Crystallites in y-Irradiated Human Enamel by Electron Paramagnetic Resonance

The arrangement of tooth enamel microcrystals has been studied on CO3 3- bound electrons by electron paramagnetic resonance. It was found that noncarious human maxillary central incisors have a greater degree of alignment of tooth enamel microcrystals than the carious ones. The outermost surface layer of enamel showed a greater crystallite degree of alignment than other parts.

Nitroxide reduction with ascorbic acid in spin labeled human plasma LDL and VLDL

The LDL and VLDL were spin labeled with Tempo which partitions both in the aqueous and lipid phase. The ESR spectra were measured in the equilibrium state as well as during the reduction of the spin label with ascorbic acid. The kinetics of the concentration decay curves was parametrized with two exponentials. The theoretical simulation of the experimental spectra revealed a drastic linewidth narrowing in the VLDL samples exposed to the ascorbic acid. Since the transport properties of the specific monolayer are reflected in the observed reaction rates, the analysis of the fatty acid composition of phospholipids, triglycerides and cholesterol esters in LDL and VLDL was performed. It is concluded that different lipid packing at the surface of LDL and VLDL might be the consequence of different intermolecular forces between phospholipids and cholesterol. This finding was connected to the experimentally detected different reaction kinetics in LDL and VLDL as well as their different susceptibility to the ESR linebroadening effects during the nonequilibrium conditions of the spin label reduction with ascorbic acid.

Lipid domains in the exoplasmic and cytoplasmic leaflet of the human erythrocyte membrane: a spin label approach.

Abstract The existence of different lipid domains in the monolayers of the human erythrocyte membrane was investigated at 4 °C by employing spin-labelled phospholipid analogues. Spectra of analogues located exclusively either in the exoplasmic or in the cytoplasmic leaflet of erythrocyte membranes were recorded. Spectra were simulated by variation of order parameter describing the average amplitude of motion of the long molecular axis of the nitrogen 2pπ orbital of the spin label and of the respective correlation times. For both leaflets at least three components were required to fit the experimental spectra, differing mainly in the order parameter. While the parameters of each component are not very different between both membrane halves, the relative contribution of each component to the spectrum is different between the exoplasmic and cytoplasmic leaflet. The order parameter of the most fluid component, presumably resembling the lipid bulk phase, is smaller in the cytoplasmic leaflet in comparison to the exoplasmic one. The lateral coexistence of different lipid domains in the human red blood cell membrane is concluded. The molecular nature of those domains is discussed.

Enhanced permeability of acid-etched or ground dental enamel

STATEMENT OF PROBLEM Acid etching creates retentive microcraters on enamel surfaces. Designing of a partial denture often involves reshaping the supporting and retentive teeth by grinding the enamel. Unfortunately, both these procedures damage the enamel surface. In vivo such surface damage takes several months to recover. PURPOSE This study evaluated the effect of 1-minute etching, prolonged etching, and grinding on the permeability of dental enamel for water-soluble molecules. MATERIAL AND METHODS With the electron paramagnetic resonance and a two-chamber diffusion cell, the influence of etching and grinding on the diffusion of spin label molecules through the enamel was studied quantitatively. The enamel permeability was measured in 30 sound enamel samples, of which 10 samples were exposed to 1-minute etching with 37% phosphoric acid, 10 samples were etched for 5 minutes, and 10 samples were ground with a diamond bur. RESULTS AND CONCLUSIONS All procedures significantly increased the permeability of dental enamel. These results demonstrate that in vivo the acid-etched and ground dental enamel surfaces are less protected and consequently, unless the tooth is properly protected, are more susceptible to carious lesions. Therefore ground or accidentally etched enamel should be protected.

The influence of jasmonic acid on biophysical properties of potato leaf protoplasts and roots.

SummaryJasmonic acid (JA) and its derivatives are a novel group of plant endogenous growth regulators. In our experiments some new data about the physiological effects of JA were obtained using electron paramagnetic resonance spectroscopy. Experiments were performed on potato plants (Solanum tuberosum L. cv. Vesna) grown in vitro. Different quantities of JA (0.1–10 μM) were added to the growth medium. Root samples of plants grown on media supplemented with JA showed a more rapid spin probe N-oxyl-2,2,6,6-tetramethyl piperidine reduction than the control plants, which is a possible indicator of altered root permeability. Samples of leaf protoplasts were probed with methyl ester of 5-doxyl-haxadecanoic acid. We observed a membrane fluidity decrease in protoplasts isolated from plants grown on higher concentrations of JA (1 and 10 μM).

Mirocrystal Arrangement in Human Deciduous Dental Enamel Studied by Electron Paramagnetic Resonance

The arrangement of microcrystallites in human deciduous enamel was studied by electron paramagnetic resonance using a stable CO33––radiation centre as a probe. Enamel

Diffusion of spin probes in tissues measured by field-gradient EPR

Abstract A linear magnetic field gradient was superimposed on a main static field to resolve the spatial distribution of diffusing paramagnetic molecules. The diffusion coefficient and distribution function were evaluated from the field-gradient electron paramagnetic spectra by comparison with spectra calculated from a corresponding model. Adipose tissue penetration of spin-labeled methyl ester of palmitic acid was measured.

Influence of aluminum on the membranes of mycorrhizal fungi

The influence of Al3+ on the membrane fluidity of the mycorrhizal fungus Lactarius piperatus (L.ex.Fr.) S. F. Gray was studied. When mycelia grew on media supplemented with 1 or 10 mM aluminum for 21 days, a significant increase in fluidity was detected, compared with the corresponding controls. The comparisons of the EPR spectra calculated by a model, with the experimental ones showed that the spectra are superimpositions of two components with different membrane ordering. The action of aluminum induced a relative enlargement of the less immobilized portion in the membrane. No significant alterations of fluidity were measured after 30-minute treatment, with Al3+.