[Some aspects of structural alterations of erythrocyte membranes under the effect of uranyl chloride at low concentrations].

The influence of nanomolar concentrations of the uranyl ion on the parameters of some membrane structures of rodent erythrocytes (laboratory mice and tundra voles--classical objects of radioecological monitoring) was investigated in vitro. A high sensitivity of the tundra vole red blood cells to the uranyl influence was shown. This fact may be determined by the cross-species difference in the membrane structures of erythrocytes--the low sphingomyelin content in tundra voles. Investigation into the phospholipid composition of the erythrocytes incubated in vitro with uranyl ions demonstrates the absence of the membrane lipid component reactions "typical" for the cells circulating in blood and also the changes pointing to the initial stages of eryptosis. Latent alterations in the membrane structure of red blood cells of both species induced by a short time contact with uranyl ions were confirmed by the increase in their sensitivity to nonionic detergent Triton X-100 and indicate the changes in orderliness of the membrane lipid phase.

Use of red blood cell membranes to evaluate the antioxidant potential of plant extracts.

Antioxidant phytochemicals in fruits and vegetables of a vegetarian diet may account for the reduced risk of aging and stress oxidative associated diseases. In this study, a simple, rapid and accurate new bioassay for the determination of the antioxidant activity of purified or crude plant extracts and thier interactions is described, based on the fluorimetric determination of thiobarbituric acid reactive substances (TBARS) released by UV-B radiated red blood cell (RBC) ghosts. Pure resveratrol, white and red wine and pomegranate juice (PJ) were used as antioxidant source to test the biological method. TBARS production is a function of radiation time, the number of RBC ghosts in the radiated sample and the loaded antioxidant. The antioxidant activity of resveratrol was detected at a submicromolar concentration range [0.02 µg/mL-0.1 µmol/L]. The activity of red wine was almost 10 times higher than that of white wine, and PJ juice had the highest activity. Submaximal protective effects of PJ and red wine were additive.

[The influence of uranyl in nanomolar concentrations on erythrocyte sensitivity (in vitro) to factors inducing acute oxidative stress].

The influence of nanomolar concentrations of UO2Cl2 on the erythrocyte sensitivity (in vitro) to the factors inducing acute oxidative stress was investigated. It was shown that even a short-run exposure of uranyl ions resulted in the changes of the physico-chemical properties of the membrane. It can affectnot only the survival of cells but significantly modify their reaction on the effect of damaging factors, particularly on the impact of oxidative stress inductors. The character of modification depends on a radical source and results from the mechanisms of their effects on the cell and the ability of uranyl to catalyze ROS-forming processes. The detailed investigation into the mechanisms of the effect of uranyl ions at low concentration on cells is required.

Penetration of laser light through red blood cell ghosts.

Hemoglobin is the main absorber of visible light in blood and blood-perfused tissues. However, hemoglobin is released from a red blood cell (RBC) during hemolysis. Hemolysis may be caused by a large number of medical conditions, including photodynamic therapy (PDT) and this subsequently can affect passage of light through the treated biological structures. The purpose of the present study was to determine the penetration of a laser beam through a suspension of hemoglobin-free human red blood cells (RBCs) - ghosts. Although hemoglobin has been efficiently removed from the samples used in our experiments, our measurements show that the samples still effectively attenuate the radiant power of penetrating laser light. We established penetration depths of 12.6mm and 15.4mm for two different laser light wavelengths, 532nm and 630nm, respectively. The penetration depth of laser light was about one order of magnitude higher for hemoglobin-free RBC ghosts as compared to intact RBCs [8,10,12]. These results can be important in case of phototherapy or biostimulation, since all photons that penetrate in a biological object may interact with it and evoke biological response.

Modulatory action of α-tocopherol on erythrocyte membrane adenosine triphosphatase against radiation damage in oral cancer.

To investigate the possible effects of α-tocopherol on erythrocyte membrane adenosine triphosphatases against radiation damage in oral cancer patients. Adenosine triphosphatase activities were analysed in oral cancer patients before and after radiotherapy (at a dosage of 6000 cGY in five fractions per week for a period of six weeks) and after supplemented with α-tocopherol (400 IU per day for entire period of radiotherapy). The membrane bound enzymes such as Na(+)/K(+)-ATPase, Ca(2+)-ATPase, Mg(2+)-ATPase and some trace elements were altered in oral cancer patients before and after radiotherapy. Supplemented with α-tocopherol modulates the erythrocyte membrane which is damaged by radiotherapy which suggests that α-tocopherol protects the erythrocyte membrane from radiation damage in oral cancer patients.

Effect of low power laser irradiation on disconnecting the membrane-attached hemoglobin from erythrocyte membrane.

In our previous study we found that low power laser irradiation improved the erythrocyte deformability, but the mechanism is unclear. The membrane-attached hemoglobin (Hbm) may be one of the determining factors for the erythrocyte deformability. We report here for the first time, that laser irradiation can reduce the Hbm contents in pig's erythrocytes, providing the explanation for the improvement of erythrocyte deformability. The decrease of the Hbm was proportional to the irradiation dose, but the relative change of Hbm was saturated around 35%. The 532 nm laser was more efficient at lowering Hbm than the 632.8 nm laser, consistent with the absorption spectrum of Hbm.

Effect of low-intensity (3.75-25 J/cm2) near-infrared (810 nm) laser radiation on red blood cell ATPase activities and membrane structure.

OBJECTIVE: The biostimulation and therapeutic effects of low-power laser radiation of different wavelengths and light doses are well known, but the exact mechanism of action of the laser radiation with living cells is not yet understood. The aim of the present work was to investigate the effect of laser radiation (810 nm, radiant exposure 3.75-25 J/cm(2)) on the structure of protein and lipid components of red blood cell membranes and it functional properties. The role of membrane ATPases as possible targets of laser irradiation was analyzed. BACKGROUND DATA: A variety of studies both in vivo and in vitro showed significant influence of laser irradiation on cell functional state. At the same time another group of works found no detectable effects of light exposure. Some different explanations based on the light absorption by primary endogenous chromophores (mitochondrial enzymes, cytochromes, flavins, porphyrins) have been proposed to describe biological effects of laser light. It was suggested that optimization of the structural-functional organization of the erythrocyte membrane as a result of laser irradiation may be the basis for improving the cardiac function in patients under a course of laser therapy. MATERIALS AND METHODS: Human red blood cells or isolated cell membranes were irradiated with low-intensity laser light (810 nm) at different radiant exposures (3.75-25 J/cm(2)) and light powers (fluence rate; 10-400 mW) at 37 degrees C. As the parameters characterizing the structural and functional changes of cell membranes the activities of Na(+)-, K(+)-, and Mg(2+)-ATPases, tryptophan fluorescence of membrane proteins and fluorescence of pyrene incorporated into membrane lipid bilayer were used. RESULTS: It was found that near-infrared low-intensity laser radiation changes the ATPase activities of the membrane ion pumps in the dose- and fluence rate-dependent manner. At the same time no changes of such integral parameters as cell stability, membrane lipid peroxidation level, intracellular reduced glutathione or oxyhaemoglobin level were observed. At laser power of 10 mW, an increase of the ATPase activity was observed with maximal effect at 12-15 J/cm(2) of light dose (18-26% for the total ATPase activity). At laser power of 400 mW (fluence rate significantly increased), inhibition of ATPases activities mainly due to the inhibition of Na(+)-, K(+)-ATPase was observed with maximal effect at the same light dose of 12-15 J/cm(2) (18-23% for the total ATPase activity). Fractionation of the light dose significantly changed the membrane response to laser radiation. Changes in tryptophan fluorescent parameters of erythrocyte membrane proteins and the increase in lipid bilayer fluidity measured by pyrene monomer/excimer fluorescence ratio were observed. CONCLUSIONS: Near-infrared laser light radiation (810 nm) induced long-term conformational transitions of red blood cell membrane which were related to the changes in the structural states of both erythrocyte membrane proteins and lipid bilayer and which manifested themselves as changes in fluorescent parameters of erythrocyte membranes and lipid bilayer fluidity. This resulted in the modulation of membrane functional properties: changes in the activity of membrane ion pumps and, thus, changes in membrane ion flows.

[Effect of membrane-stabilizing magnetolaser therapy on cardiodynamics in patients with ischemic heart disease]. / Vliianie membranostabiliziruiushchei magnitolazernoi terapii na kardiodinamiku u bol'nykh ishemicheskoi bolezn'iu serdtsa.

AIM: To evaluate cardiodynamic changes in response to magnetolaser therapy (MLT) and these changes links with lipid shifts in cell membrane. MATERIAL AND METHODS: The study enrolled 50 patients with effort angina (functional class II-III). Of them, 37 patients were exposed to 10-day courses of MLT, 13 patients were exposed to sham procedures. Before the treatment and 3 months after it measurements were made of lipid peroxidation (LPO) products, structure of erythrocytic membrane and cardiodynamic parameters. RESULTS: MLT resulted in a significant reduction of LPO products, stabilization of cell membrane structure and positive shifts in cardiodynamics. Correlation was found between the above parameters. CONCLUSION: Improvement of inotropic, diastolic functions of the myocardium and abatement of cardiac remodeling in coronary heart disease patients in response to MLT is realized primarily due to structural stabilization of cell membrane lipid biolayer.

Low-intensity near-infrared laser radiation-induced changes of acetylcholinesterase activity of human erythrocytes.

OBJECTIVE: The aim of the present study was to investigate the transformations of red blood cells produced by low-intensity infrared laser radiation (810 nm). BACKGROUND DATA: Low-intensity (the output power of a laser device in the milliwatt range) laser radiation as a local phototherapeutic modality is characterized by its ability to induce non-thermic, nondestructive photobiological processes in cells and tissues. However, the exact theory concerning the therapeutic effects of laser biostimulation has not been developed. MATERIALS AND METHODS: The suspensions of human erythrocytes in PBS (10% hematocrit) were irradiated with near-infrared (810 nm) therapy laser at different light doses (0-20 J) and light power (fluence rate; 200 or 400 mW) at 37 degrees C. As the parameters characterizing the cell structural and functional changes membrane acetylcholinesterase (AchEase) activity, the membrane potential, the level of intracellular glutathione, the level of products of membrane lipid peroxidation, and the cell osmotic stability were measured. RESULTS: It was found that near-infrared low-intensity laser radiation produced complex biphasic dose-dependent changes of the parameters of AchEase reaction in the dose-dependent manner: at smaller doses of radiation (6 J) the maximal reaction rate and Michaelis-Menten constant value decreased, and at higher radiation doses these parameters increased. No significant changes of erythrocyte stability, cellular redox state (reduced glutathione or lipid peroxidation product levels), or cell membrane electrochemical potential were observed. CONCLUSION: Low-intensity near-infrared laser radiation (810 mn) produced AchEase activity changes, reflecting the effect of light on the enzyme due to energy absorption. Protein molecule conformational transitions and enzyme activity modifications in cells have been suggested as laser radiation-induced events.

[The mechanisms of action of extracorporeal helium-neon laser irradiation in acute exogenous poisonings]. / Nekotorye mekhanizmy deistviia ékstrakorporal'nogo gelii-neonovogo lazernogo oblucheniia pri ostrykh ékzogennykh otravleniiakh.

Extracorporeal exposure to LG-79 He-Ne laser of 12 mWt power was used in 57 patients hospitalized at the intoxication reanimation department with acute poisonings with psychotropic drugs. The clinical result was a decrease of the incidence of pneumonia in the patients with x-ray signs of venous congestion from 52% among those administered to physiochemotherapy to 24% after this treatment modality. Laser hemotherapy brought about a temporary normalization of the erythrocyte membrane permeability, which was changed biophysically by means of a diffractometer. Red cell aggregation was approximating the norm, decreasing by 20%, and platelet aggregation decreased by 17%. Analysis of the results brought as to a conclusion that He-Ne laser exposure is an effective source of singlet stimulation of molecular O2 evenly dissolved in the blood, which causes resonance oscillations of water difields. This leads to membrane depolarization, which is probably responsible for purification of polarized membranes from toxic agents fixed by them.

Photophysical alterations of erythrocyte membrane by 4,5',8-trimethyl psoralen: membrane response towards psoralen phototherapy.

Despite the wide and versatile use of psoralen phototherapy in various dermal diseases including psoriasis and leukoderma, the proposed mechanism of action of psoralen is disputed. This study launches an original documentation of free-radical resulting in erythrocyte membrane damage studied by spin-labeling using the probes malnet and 5 doxyl stearate as protein and lipid domains, respectively. Both Type I and Type II photodynamics appear plausible and the therapeutic aspects of this innovation are being verified.

[The effect of the naphthenic hydrocarbons of naphthalane on the change in the permeability of biological membranes after ionizing radiation]. / Vliianie naftenovykh uglevodorodov naftalana na izmenenie pronitsaemosti biologicheskikh membran posle ioniziruiushchego oblucheniia.

Nephthene hydrocarbons, a naphthalan active component, were tried for ability to change permeability of erythrocytic and vascular membranes for dyes. It was established that this preparation can return to normal hyperpermeability of biological membranes induced by radiation. Its mechanism of action is suggested to be participation in biosynthesis of adrenal steroid hormones and activation of some enzymatic systems.

Modification of membrane composition, eicosanoid metabolism, and immunoresponsiveness by dietary omega-3 and omega-6 fatty acid sources, modulators of ultraviolet-carcinogenesis.

Dietary sources of lipids containing predominantly n-3 or n-6 fatty acids (FA) have been examined for effect upon several potential pathophysiologic parameters. Epidermal, plasma, and red blood cell (RBC) membrane FA composition exhibited marked differences between animals fed the respective dietary lipid sources. Reduced levels of 18:1, 20:3 and 20:4 occurred in the n-3 FA fed animals which exhibited significantly higher levels of 20:5 and 22:6. Approximately equal levels of 18:2 were present in animals fed either diet. Despite marked differences in RBC membrane FA composition, only marginal effect upon osmotic fragility occurred. Lower levels of 20:3 and 20:4 found in n-3 fed animals could result from a deficit of elongase and/or delta 5-desaturase activity. Whether lower 20:4 levels in n-3 fed animals could rate-limit eicosanoid metabolism is unknown, but epidermal capacity to metabolise arachidonic acid in these animals was found to be closely related to n-6 FA intake. Animals fed n-3 FA exhibited markedly lower levels of plasma PGE2, even when the diet was supplemented with n-6 FA. In addition, UV-radiated animals receiving the n-3 FA source demonstrated a reduced (approximately 30%) response to inflammatory stimulus and a greater (4.5-fold) delayed hypersensitivity (DH) to dinitrochlorobenzene than animals fed the n-6 FA source. These data demonstrate that dietary lipid strongly influences tissue FA composition, eicosanoid metabolism, and, in the case of DH, at least one type of T-cell mediated immune response in UV-irradiated animals.

[The relationship of the potassium transport function of human erythrocyte membranes to the structural state of the glycocalyx]. / O sviazi kaliitransportnoi funktsii membran éritrotsitov cheloveka so strukturnym sostoianiem glikokaliksa.

The two structural-functional state parameters of erythrocyte membrane in healthy donors and ischemic heart disease patients have been compared--the potassium exchange rate constant (x), and the quantity of Alcian blue sorption capacity of glycocalyx (y). The connection between the first parameter--x--and the other one--y--has been observed in cases, when the value of the latter experienced both the seasonal oscillations (the coefficient of lineal correlation was: r = -0.87, P less than 0.01) and the changes in the cause of ischemic heart disease (r = -0.72, P less than 0.01) and its treatment (the coefficients of correlation attitude of each of parameter changes were: eta xy = 0.83, P less than 0.01; eta yx = 0.52, P less than 0.05). The above connection may indicate the participation of membrane glycoproteins and glycolipids in a regulation of the cation transport function of the erythrocyte membrane.

[Trigger mechanisms of the therapeutic effects of autologous transfusion of UV-irradiated blood (membranotropic effect on erythrocytes and thrombocytes)]. / Puskovye mekhanizmy lechebnykh éffektov autotransfuzii UF-obluchennoi krovi (membranotropnoe deistvie na éritrotsity i trombotsity).

The article sums up numerous investigations of the authors on studying different parameters of membrane-dependent properties of erythrocytes and thrombocytes after UV irradiation of blood samples, mixing of irradiated and nonirradiated blood, transfusions of UV-irradiated auto blood. It was shown that membranotropic action of UV-irradiated blood and transfusion of UV-irradiated blood are responsible for rheological and hemostatic properties of blood.

[The activation of the immunosorptive properties of blood during its UV irradiation at therapeutic doses]. / Aktivatsiia immunosorbtsionnykh svoistv krovi pri ee UF-obluchenii v terapevtichekikh dozakh.

UV irradiation of donor rhesus-positive blood in apparatus, applied in Soviet hospitals for autotransfusion of UV-irradiated blood produces a 2-fold increase of the blood capacity to bind antirhesus antibodies in blood or serum from sensibilized women. The above data can be used for increase in therapeutic effect of blood exchange transfusion in children with rhesus-conflict hemolytic disease.

[Functional and structural changes in the surface of human erythrocytes following irradiation with ultraviolet rays of various wave lengths. V. Modification of the glycocalyx in autotransfusions of UV-irradiated blood]. / Funktsionalnye i strukturnye izmeneniia poverkhnosti éritrotsitov cheloveka posle oblucheniia UF luchami raznoi dliny volny. V. Mofifikatsiia glikokaliksa pri autotransfuziiakh UF-obluchennoi krovi.

A study was made of the structural and functional state of the erythrocyte surface components of patients with ischemic heart disease, and of patients with ulcer disease during the treatment with UV-irradiated blood autotransfusions. The cytochemical and isoserological methods employed showed some structural disturbances in the state of erythrocyte, glycocalyx and its modification in the course of treatment. The clinical results of treatment correlated with these changes.

[Functional and structural changes in the surface of human erythrocytes after UV irradiation with rays of various wavelengths. IV. Changes in the physicochemical properties of autotransfused UV-irradiated blood]. / Funktsional'nye i strukturnye izmeneniia poverkhnosti éritrotsitov cheloveka posle oblucheniia UF luchami raznoi dliny volny. IV. Izmeneniia fiziko-khimicheskikh svoistv pri autotransfuziiakh UF-obluchennoi krovi.

Properties of erythrocyte surface were investigated for patients with ischemic heart disease in the course of treatment with the UV-irradiated blood autotransfusion (UVIBA). Application of methods of light-scattering, photometry and cytochemistry revealed rapid and significant changes in deformability and aggregation properties of the erythrocytes immediately following each UVIBA procedure, which was accompanied by considerable blood viscosity decrease.

[Clinico-biochemical parallels against a background of traditional treatment and laser therapy of patients with ischemic heart disease]. / Kliniko-biokhimicheskie paralleli na fone traditsionnogo lecheniia i lazeroterapii bol'nykh ishemicheskoi bolezn'iu serdtsa.

The paper is devoted to analysis of clinico-biochemical indices and a quest for additional pathologico-chemical criteria at the cell membrane level of the efficacy of helium-neon laser therapy in coronary heart disease (CHD). Another task was to decipher a number of metabolic signs of the phenomenon of CHD "exacerbation" in laser therapy (during 4-6 sessions). Positive clinical results were obtained in 98% of the patients. A favorable time course was noted in serum lipoprotein spectrum indices and in lipid and phospholipid structure of erythrocyte membranes. Laser therapy caused mobilization of the cell membrane antioxidant defense. The 4th-6th session of irradiation was marked by temporary clinical deterioration--"exacerbation" of disease accompanied by an increase in the level of erythrocyte total phospholipids (at the expense of PTEA), a decrease in free fatty acid deficiency against a background of advancing alpha-tocopherol deficiency and a rise of the blood level of lipid peroxidation primary products (diene conjugates). A conclusion was that membrane protective drugs and drugs enhancing cell energy and antioxidant defense resources promoting the "deactivation" of influence on the membrane of lipid peroxidation metabolites (tocopherol, Essentiale, retinol, etc.) should be incorporated in therapy of CHD in order to prevent the phenomenon of its "exacerbation".

[Functional and structural changes in the surface of human erythrocytes after irradiation by different wave lengths of UV rays. III. The immediate effect of the autotransfusion of UV-irradiated blood]. / Funktsional'nye i strukturnye izmeneniia poverkhnosti éritrotsitov cheloveka posle oblucheniia UF luchami raznoi dliny volny. III. Nemedlennyi éffekt autotransfuzii UF-obluchennoi krovi.

The autotransfusion of UV-irradiated blood (70-200 ml) results in the structural modification of cell surface in all the circulating erythrocytes of cardiological patients. The effect is registered within 1 hour after transfusion and involves some decrease in the distribution coefficient of erythrocytes registered in two-phase polymer system dextran-poly(ethylene glycol), which depends on membrane surface properties other than charge. This effect is suggested to be responsible for the main peculiarities of the therapeutic effect of UV-irradiated blood autotransfusion--high rate of appearance, prolongation and wide spectrum of the therapeutic action.