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Biomedical subjects

W Leyko

Publications and source records attributed to W Leyko.

At least 37 records · Page 2Linked to original sources

Hyperthermic modification of bleomycin--DNA interaction detected by electron spin resonance.

Electron spin resonance spectra of DNA labeled with each of four spin-labeling compounds have been studied to detect interaction between the antibiotic bleomycin and DNA. Only one of these labels, compound IV, resulted in a modified spectrum when bound to DNA and the latter was subjected to bleomycin. This property has been used to monitor DNA-bleomycin interactions under physiological and hyperthermic conditions. Bleomycin produced an increase in rotational correlation time of the residue bound to DNA at 37 degrees C and a significantly higher increase at 43 degrees C. Some effect was still detected with bleomycin at 37 degrees C after preheating at 43 degrees C. Parallel studies have revealed enhanced binding of 59Fe-bleomycin to DNA during and after hyperthermic treatment.

Bleomycin↗

The crypto-OH radical in the damage of DNA by bleomycin-Fe2+?

1. Effects of various OH scavengers, superoxide dismutase and catalase on the formation of malondialdehyde-like products from DNA by bleomycin-Fe2+ were studied. In no case was a protective effect observed. 2. These results can be interpreted on the basis that a crypto-OH radical mediates the damage to DNA by bleomycin-Fe2+.

Bleomycin↗

Effect of .OH scavengers on radiation damage to the erythrocyte membrane.

Thiourea, and .OH scavenger, reduced the gamma-radiation-induced changes in the rigidity of erythrocyte membrane lipids, the state of membrane proteins, and lipid peroxidation. Several .OH scavengers, but not superoxide dismutase, also inhibited the radiation-induced acceleration of the transport of a hydrophilic non-electrolyte spin label TEMPOL across the erythrocyte membrane. The effects of scavengers were usually biphasic with a maximum of the protective effect. Higher concentrations of the scavengers were less effective, as in the case of radioprotection of erythrocytes to haemolysis studied previously.

Animals↗

Effect of hyperthermia and ionizing radiation on the erythrocyte membrane.

Spin-label studies of the effects of hyperthermia on the erythrocyte membrane revealed a decrease in the fluidity of lipids and changes in the state of membrane proteins. The rate of haemolysis in iso-osmotic glycerol solution was increased. Changes of most of the parameters studied when plotted in Arrhenius coordinates revealed a discontinuity (critical hyperthermic transition in the membrane) between 46 and 50 degrees C. Studies of the combined action of ionizing radiation (100 Gy) and hyperthermia (43 degrees C) showed the same direction of changes for (Na-K-Mg)-ATPase activity and spectra of membrane-bound maleimide spin label for both agents, but the additivity of changes depended on the parameter studied.

Animals↗

Interaction of local anaesthetics with model phospholipid membranes. The effect of pH and phospholipid composition studied by quenching of an intramembrane fluorescent probe.

The interaction of local anaesthetics, tetracaine and procaine, with model phospholipid membranes has been examined by measurement of drug-induced quenching of the fluorescence of a membrane incorporated probe, 12-(9-anthroyl)stearic acid. The pH dependence of quenching curves obtained for neutral phosphatidylcholine and acidic phosphatidylserine bilayers indicates that, contrary to previous suggestions, both charged and uncharged forms of amine local anaesthetics may be incorporated into lipid membranes.

Anesthetics, Local↗

Interaction of propranolol with model phospholipid membranes. Monolayer, spin label and fluorescent spectroscopy studies.

The interaction of propranolol with model phospholipid membranes was studied using various experimental techniques. The partition coefficient of propranolol in the negatively charged membranes of vesicles prepared from phosphatidylserine and phosphatidic acid was found to be more than 20-times higher than in neutral phosphatidylcholine membranes. Preferential interaction of propranolol with acidic phospholipid membranes was confirmed using the monolayer compression isotherm technique and the spin-labelling method. Phosphatidylserine monolayers were markedly expanded even at a relatively low drug concentration (5 . 10(-6) M). In contrast, the effect of propranolol on phosphatidylcholine monolayers was much smaller, being detectable only at a higher concentration of the drug (1 . 10(-4) M). Spin-labeling experiments show that propranolol exerts marked ordering effect on bilayers prepared from acidic phospholipids and does not change the order parameter of phosphatidylcholine membranes. The dependence of the propranolol fluorescence spectrum on the polarity of the solvent allowed us to identify the intercalation region of the drug in the membrane. The fluorophore moiety of propranolol was found to be localized in the lipid polar head groups region of the bilayer. The role of electrostatic and hydrophobic effects in propranolol-membrane interaction is discussed and the effect of propranolol on the ordering of phospholipid bilayers is compared with the effects of other anesthetic-like molecules.

Electron Spin Resonance Spectroscopy↗

Effect of gamma radiation on the transport of spin-labeled compounds across the erythrocyte membrane.

The effect of ionizing radiation on the non-electrolyte, anion and cation permeability of the erythrocyte membrane was studied by measurement of the reduction rate of appropriate nitroxyl derivatives. Irradiation of bovine erythrocytes in the dose-range of 2-50 krad resulted in a regular dose-dependent increase in the reduction rates of a cation (TEMPO-choline) and a hydrophobic non-electrolyte (TEMPO), and non-regular changes in the reduction rate of a hydrophilic non-electrolyte (TEMPOL). The permeation constant for TEMPO-choline also showed a non-regular response to radiation, similar to the response pattern of other red blood cell parameters. These results also demonstrate that the effects of radiation on the transport of various solutes can be used as a means of distinguishing between different channels of membrane transport.

Animals↗

Changes in the tryptophan fluorescence in gamma-irradiated human haemoglobin.

The effect of gamma-radiation on human haemoglobin was studied by means of fluorescence spectroscopy. Irradiation with doses in the range of 0-2.4 Mrad led to an increase in fluorescence intensity and a red shift of the fluorescence maximum. Such spectral changes should be attributed mainly to a more polar environment of tryptophan residues in irradiation haemoglobin caused by protein unfolding and to a concomitant increase in separation between tryptophan residues and haem groups. Differences in fluorescence intensity but not spectral distribution between control and irradiated samples denatured in guanidinium hydrochloride indicate a radiation-induced decrease in tryptophan content. Fluorescence quenching by caesium ions was observed in irradiated haemoglobin but not in native haemoglobin. The Stern-Volmer constant calculated for caesium quenching indicates that the fraction of tryptophan residues accessible to the quencher increases after irradiation.

Cesium↗

Radioprotection of bovine erythrocytes to haemolysis.

The effect of several free-radical scavengers on the radiation-induced haemolysis of bovine erythrocytes (50 krad, 60Co, cell concentration: 0.05 v/v) was studied. A two-phase effect of the additives was found with a maximum of the protective action in the millimolar concentration range. A correlation was established between the degree of protection at a fixed concentration (5mM) and the rate constants of reactions of respective compounds with OH. A radiosensitizing effect of diethyldithiocarbamate, an inhibitor of superoxide dismutase (SOD) was not confirmed and no specific protective effect of SOD was seen.

Animals↗

Ultrastructural modifications of the erythrocyte membrane in Down's syndrome.

Spin label studies of erythrocyte membranes from patients with Down's syndrome showed no differences in the rigidity of membrane lipids with respect to normal subjects and a tendency for alterations in the state of membrane proteins. Electron microscopic studies demonstrated the occurrence of ultrastructural defects in membranes from trisomics which can be due to accelerated red cell aging as similar alterations were found in the fraction of oldest (most dense) cells in normal subjects.

Adolescent↗

Osmotic fragility and lipid peroxidation of irradiated erythrocytes in the presence of radioprotectors.

Gamma radiation causes a decrease in the osmotic fragility of porcine erythrocytes and induces peroxidation of membrane lipids, as reflected by an increase in malondialdehyde content. With respect to osmotic fragility all the protectors used have the radioprotective effect. Malondialdehyde content was reduced in the presence of catalase, cysteine and glutathione, whereas SOD had no protective effect.

Animals↗