Search

Exocytosis of myeloperoxidase (MPO) from activated neutrophils in the presence of the anionic polysaccharide heparin was studied. It was determined that the optimal concentration of heparin (0.1 u/ml), at which there is no additional activation of cells (absence of amplification of exocytosis of lysozyme contained in specific and azurophilic granules). It was found that after preincubation of cells with heparin (0.1 u/ml) the exocytosis of MPO from neutrophils activated by various stimulants (fMLP, PMA, plant lectins CABA and PHA-L) increased compared to that under the action of activators alone. In addition, it was shown that heparin in the range of concentrations 0.1-50 u/ml did not affect on the peroxidase activity of the MPO isolated from leukocytes. Thus, the use of heparin at a concentration of 0.1 u/ml avoids the artifact caused by the "loss" of MPO in a result of its binding to neutrophils, and increases the accuracy of the method of registration the degranulation of azurophilic granules of neutrophils based on determination of the concentration or peroxidase activity of MPO in cell supernatants.

A significant increase in the myeloperoxidase (MPO) activity has been found in plasma of patients with stable angina and with acute coronary syndrome (ACS) in comparison with the control group. MPO concentration was significantly increased in plasma of ACS patients. Reduced MPO activity in the treated ACS patients correlated with a favorable outcome of the disease. Generally, changes in plasma MPO concentration coincided with changes in lactoferrin concentration thus confirming the role of neutrophil degranulation in the increase of plasma concentrations of these proteins. The increase in MPO activity was obviously determined by modification of the MPO protein caused by reactive oxygen species and halogen in the molar ratio of 1 : 25 and 1 : 50. The decrease in plasma MPO activity may be associated with increased plasma concentrations of the physiological inhibitor of its activity, ceruloplasmin, and also with modification of the MPO protein with reactive oxygen species and halogen at their molar ratio of 1 : 100 and higher. Thus, MPO activity may be used for evaluation of effectiveness of the treatment of cardiovascular diseases.

Effects of water-soluble phenolic antioxidant sodium 3-(3'-tret-butyl-4'-hydroxyphenyl)-propyl thiosulfonate (TS-13), potassium 3,5-dimethyl-4-hydroxybenzyl thioetanoate (BEP-11-K) and potassium 3-(3',5'-ditretbutyl-4'-hydroxyphenyl)-propionate (potassium phenosan) on tumor cells proliferative activity and the role of redox-dependent and calcium-dependent signaling mechanisms in realization of tumor cell response to the antioxidant action were studied. Potassium phenosan and BEP-11-K were found to stimulate proliferation and ARE-inducing phenolic antioxidant TS-13 was found to inhibit tumor cell growth in culture. The tumor cell growth rate depended on the rate of intracellular reactive oxygen species production and was decreased by apocynin (a NADPH-oxidase inhibitor) and antimycin A (an ubiquinol-cytochrome c oxidoreductase inhibitor). TS-13 action on tumor cells was accompanied by a transient increase in intracellular reactive oxygen species production and the intracellular calcium concentration, whereas cell incubation with potassium phenosan and BEP-11-K did not influence the reactive oxygen species level and intracellular calcium ions. Cyclosporine A blocked the inhibitory effect of TS-13. Thus, it can be reasonably speculated that phenolic antioxidant TS-13 starts mitochondria-dependent apoptosis in tumor cells by the opening of permeability transition pores.

It is shown that human serum albumin, previously treated with HOCl (HSA-Cl), enhances luminol-dependent chemiluminescence of neutrophils activated by phorbol-12-myristate-13-acetate (PMA). The enzyme-linked immunosorbent assay revealed that addition of HSA-Cl to neutrophils promotes exocytosis of myeloperoxidase. Inhibitor of myeloperoxidase--4-aminobenzoic acid hydrazide, without any effect on lucigenin-dependent chemiluminescence of neutrophils stimulated with PMA, effectively suppressed luminol-dependent chemiluminescence (IC50 = 20 microM) under the same conditions. The transfer of the cells from medium with HSA-Cl and myeloperoxidase to fresh medium abolished an increase in PMA-induced luminol-dependent chemiluminescence, but not the ability of neutrophils to respond to re-addition of HSA-Cl. A direct and significant (r = 0.75, p) correlation was observed between the intensity of PMA stimulated neutrophil chemiluminescence response and myeloperoxidase activity in the cell-free media after chemiluminescence measurements. These results suggest the involvement of myeloperoxidase in the increase of neutrophil PMA-stimulated chemiluminescence response in the presence of HSA-Cl. A significant positive correlation was found between myeloperoxidase activity in blood plasma of children with severe burns and the enhancing effects of albumin fraction of the same plasma on luminol-dependent chemiluminescence of PMA-stimulated donor neutrophils. These results support a hypothesis that proteins modified in reactions involving myeloperoxidase under oxidative/halogenative stress, stimulate neutrophils, leading to exocytosis of myeloperoxidase, a key element of halogenative stress, and to closing a "vicious circle" of neutrophil activation at the inflammatory site.

Using previously developed spectro-photonmetrical method (Bioorg. Khim. 2009. V. 35. pp. 629-639), a significant increase of myeloperoxidase (MPO) activity was found in blood plasma of patients with type 2 diabetes mellitus (DM2) without of cardiovascular complications, as well as with ischemic heart disease (IHD). Plasma MPO concentration measured by an enzyme-linked immunosorbent assay was significantly higher only in blood plasma of patient with DM2 and IHD. A direct and significant correlation between MPO activity and MPO concentration was observed only in blood plasma samples from healthy donors. Increased MPO activity did not correlate with MPO concentration in blood plasma of patients with DM2 and DM2 with IHD. Taken together, these results highlight the necessity for studying of the MPO role in the development of pathological processes to determine both the amount of enzyme and its peroxidase activity in the blood. The proposed approach gives comprehensive information about the relationship between MPO activity and MPO concentration in patient blood. Since the high concentration of MPO is a diagnostically significant parameter in the prediction of endothelial dysfunction and cardiovascular disease development, the obtained results evidence the contribution of MPO-dependent reactions in cardiovascular complications associated with diabetes. MPO activity may serve as an additional diagnostic criterion for determination of risk of IHD in DM patients.

It is shown that in the presence of reduced glutathione at low concentrations (1-5 microM) the extent of platelet aggregation with neutrophils increases and the lag period of platelet aggregation induced by tumor cells decreases. At the same time in the presence of reduced glutathione at high concentration (3 mM) the extent of platelet aggregation with neutrophils decreases, and the lag period of platelet aggregation induced by tumor cells increases. It is established that glutathione-dependent regulation of the intercellular contact formation between platelets and neutrophils depends on the ratio of glutathione oxidized and reduced forms: at fixed total glutathione concentration of 5 microM, increase of glutathione redox potential from -175 mV to 0 mV led to reduction in platelet aggregation with neutrophils. Thus, it is shown for the first time, that GSH has priming effect on the platelet aggregation with neutrophils and tumor cells, which may contribute to the regulation of inflammatory diseases and cancer.

A model for the redox regulation of the functional state of the cell has been constructed on the basis of representation of electron transfer processes by equivalent electric circuits. The mechanism of action of redox active molecules on biosystems has been discussed in terms of circuit theory. A method for determining the parameters of cellular redox sensors has been proposed. It has been established that the concentration and redox potential of compounds entering the cell are the main regulatory parameters of redox signals for the cell. It has been experimentally shown that the calcium response to hydrogen peroxide in rat C6 glioma cells and human FL amnion cells depends on the redox buffer capacity of cells.

The effect of NO donors (sodium nitroprusside, S-nitrosoglutathione, dinitrosyl-iron complexes) on the functional and mechanical properties of human platelets and red blood cells has been investigated. It has been established by atomic force microscopy that NO donor-induced platelet disaggregation is accompanied by changes in the elastic properties of cells. It has been shown that, in the presence of NO donors, the detergent-induced hemolysis of red blood cells is delayed, and the elasticity modulus of these cells decreases. The results obtained indicate that NO donors regulate the structural and functional properties of platelets and red blood cells.

It has been shown by the method of fluorescent analysis that the rate of hydrogen peroxide generation in human U251 glioma cells under the effect of lipophilic (menadione) or hydrophilic (vikasol) analogues of vitamin K3 was different. Analyzing experimental data we can conclude that menadione underwent one- and two-electron reduction by intracellular reductases in glioma cells. Reduced forms of menadione interact with molecular oxygen leading to reactive oxygen species (ROS) generation. The theoretical model of ROS generation including two competitive processes of one- and two-electron reduction of menadione has been proposed. Rate constants of ROS generation mediated by one-electron reduction process have been estimated.

The parameters of the acid-base state and redox state of erythrocytes have been studied with the use of the fluorescent probes 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein and 2',7'-dichlorodihydrofluorescein. It has been found that the value of redox state parameters in erythrocytes depends both on the extracellular and intracellular concentrations of hydrogen ions. It has been shown that hydrogen peroxide induces a decrease in the value of the intracellular pH. The interrelation of cellular homeostasis parameters characterizing the acid-base and redox states of erythrocytes has been theoretically and experimentally substantiated.

A novel method for spectrometrical measurement of myeloperoxidase (MPO) activity in plasma with o-dianisidine (DA) as a substrate is proposed. We have determined the optimal conditions, including the pH and hydrogen peroxide concentration, under which MPO is the main contributor to DA oxidation in plasma. Specific MPO inhibitors, salicylhydroxamic acid or (4-aminobenzoyl)hydrazide, are added to measure the activity of other heme-containing peroxidases (mainly hemoglobin and its derivatives) and subtract their contribution from the total plasma peroxidase activity. Plasma MPO concentrations are quantified by a new enzyme-linked immunosorbent assay (ELISA) developed by us and based on the use of antibodies raised in rats and rabbits. The sensitivity of this ELISA is high: 0.2-250 ng/ml. A direct and significant (P < 0.0001) correlation was observed between the MPO activities measured spectrometrically and by ELISA in blood samples from 38 healthy donors. The proposed approaches to MPO measurement in plasma can be used to evaluate the enzyme activity and concentration, as well as the efficacy of mechanisms by which MPO is regulated under physiological conditions and against the background of various inflammatory diseases.

Effects of ascorbic acid on calcium homeostasis of human laryngeal carcinoma cells were studied. Intracellular concentration of free calcium and intracellular pH were measured by fluorescent analysis. Ascorbic acid in concentrations of 3-10 mM caused pH drop and sharply increased concentrations of free Ca ions in HEp-2 cells. Intracellular concentration of free Ca ions resulted from Ca ion release from the thapsigargin-sensitive Ca depots.

We investigated the influence ofneopterin and 7, 8-dihydroneopterin on the activity and secretory degranulation of myeloperoxidase in neutrophils and the ability of pteridines to interact with the main substrate of this enzyme, hydrogen peroxide, and with the intermediate product of halogenation cycle--hypochlorous acid. It was shown that neopterin and 7, 8-dihydroneopterin while being a redox-pair regulated the process of oxygen activation in neutrophils by functioning of myeloperoxidase. Depending on concentration, pteridines can influence the secretion of myeloperoxidase into intracellular medium and decrease the level of hydrogen peroxide and hypochlorous acid that are a substrate and an intermediate product of the enzyme respectively. It was shown that 7, 8-dihydroneopterin in micromolar concentration appeared to be noncompetitive inhibitor of myeloperoxidase. We suppose that myeloperoxidase assists 7, 8-dihydroneopterin oxidation by hypochlorous acid that leads to neopterin concentration increase. These changes modify the concentration of reactive oxygen species in intracellular and extracellular media.

The introduction of the parameters characterizing the redox state of the cell, such as the effective redox potential and the redox buffer capacity has been theoretically substantiated. A comparative study of the parameters of the redox state of erythrocytes from healthy donors and patients with diabetes and acute coronary syndrome has been performed. It was found that the redox buffer capacity in erythrocytes from patients with diabetes and acute coronary syndrome was reduced by 30-40% in comparison with the redox buffer capacity of erythrocytes from healthy donors. The largest change in the effective redox potential was observed for erythrocytes from patients with diabetes, which indicates a more expressed oxidative stress in this pathology.

Recent achievements of biophysics and physiology resulted to remarkable progress in the understanding of role of redox processes in cell vital function. It was established that oxidizing and reducing agents participate in processes of differentiation, proliferation and apoptosis. Representations about an existence of definite balance between oxidation and reduction processes in cells or a redox homeostasis were formed. Recent data about mechanisms of regulation of redox homeostasis were considered in the review. The correlation between redox homeostasis and metabolism were in detail analyzed. The special interest is given to a problem of quantitative description of redox phenomena in biological systems.

It has been demonstrated for the first time that GlcNAc-specific lectin from Solanum tuberosum induces the formation of haptenic sugar-resistant intercellular contacts (HSR-contacts) in platelet aggregation and does not induce stable neutrophil and lymphocyte aggregation. The formation of HSR-contacts in platelets was significantly impaired by the inhibitors of cAMP phosphodiesterase (papaverine) and arachidonic acid methabolism (indomethacin, aristolochic acid, and MK-886) as well as by the sulfhydryl reagent N-ethylmaleimide. The results obtained indicate that STA can be used to study the mechanisms of stable platelet aggregation, to screen drugs with potential antithrombotic activity, and to develop new cell engineering techniques.

The influence of H2O2 at concentrations of 10(-8)--10(-2) mol/l on neutrophil ability to generate the reactive oxygen and chlorine species (ROCS) and secrete myeloperoxidase (MPO) was studied, and H202 injurious effect on neutrophils was also investigated in this work. It was revealed that H2O2 at concentrations of 2 x 10(-3)--2 x 10(-2) mol/l induced disturbance of the neutrophil membrane barrier properties and lactate dehydrogenase release. The incubation of the neutrophils with the addition of 10(-4)--10(-7) mol/l H2O2 led to an increase in the cell ability to generate ROCS during phagocytosis and decreased neutrophil ability to secrete MPO and ROCS in extracellular medium during adhesion. The mechanisms of H2O2 effect are coupled with arachidonic acid metabolism. Inhibition of metabolic pathways of 5-lipoxygenase or cyclooxygenase increased the destructive effect of H2O2 on the cells. Five-lipoxygenase way prohibition led to cancellation of H2O2 influence on MPO and ROCS secretion and to enhancement of H2O2 effect on neutrophil ability to generate ROCS during phagocytosis. The data obtained testify to the high neutrophil resistance to destructive effect of H2O2 and confirm the regulatory role of H2O2 with respect to the neutrophil functions.

Effects of hydrogen peroxide on morphological characteristics, proliferation index, menadione-dependent lucigenin-enhanced chemiluminescence of C6 glioma cells were studied. It was established that H2O2 at 1 x 10(-8) - 5 x 10(-7) M concentrations acts as a regulator of morphological and functional properties of astrocytes by inducing their reactivation that is manifested as a cell body hypertrophy and an increase of proliferative activity and of menadione-dependent production of superoxide (O2- ). Cytodestructive action of hydrogen peroxide at a concentration higher than 1 microM on C6 glioma cells shows itself as a decrease of their proliferation index and the ability to generate O2- under menadione action. Using lipopolysaccharide B as a functional stimulator it has been shown that H2O2 modifies signaling pathways leading to the increase of mitotic activity of C6 glioma cells and decreases the yield of lucigenin-enhanced chemiluminescence of astrocytes under menadione action to the level of control values.

Processes of oxygen activation in monocytes stimulated with adhesion to glass were studied by methods of luminol-dependent and lucigen-independent chemiluminescence. It was shown that monocyte chemiluminescence was caused by cell adhesion to glass surface. Generation of reactive oxygen species at monocyte adhesion to glass was dependent on calcium ion concentration in the medium. The increase in the level of cytosolic calcium, as the extracellular calcium concentration elevated, was accompanied by the activation of phospholipase A2, 5-lypoxygenase and cycloxygenases. Magnesium ions exerted no influence on oxygen activation by cells. Incubation of cells in glucose-free medium, or the addition of glycolysis blocker (2-deoxy-D-glucose) to cell suspension led to a decrease in chemiluminescence intensity. By means of inhibitory analysis, it has been established that processes of oxygen activation are related to arachidonic acid metabolism, and depend on the activity of phospholipase A2.

The method for determination of intracellular oxidants content has been proposed. The method is based on the analysis of changes of 2,7-dichlorodihydrofluorescein oxidation. The theoretical model of hydrogen peroxide diffusion and its consumption in the cell and quantitative determination of hydrogen peroxide gradient through the cellular membrane has been built taking into account the experimental data. The coefficient for H2O2 permeability through the plasma membrane of human amnion cells is (2.1 +/- 0.1) x 10(-4) cm x c(-1).

Recent achievement of molecular biology and neurophysiology resulting in remarkable progress in the understanding of processes of neurodifferentiation, playing the relevant role both in an ontogenesis and working of adult brain. New representations have been formed about a role of cytoskeletal proteins in mechanisms of neurites growth. In the review we discussed recent data regarding mechanisms of neuronal differentiation regulation. The special interest is given to a problem of influence of electrical fields on neuronal growth and differentiation. The mechanisms of signal transduction from a cell surface to nuclear are in detail analyzed. The classification of the physical and chemical factors of growth neurites regulation is given.

The formation of two hemoglobin forms (methemoglobin and nitrite methemoglobin) in native human erythrocytes in the presence of sodium nitrite in suspension was shown. In normal erythrocytes, the interaction of intracellular oxyhemoglobin with nitrite ions results in the formation of methemoglobin, whereas in metabolically exhausted erythrocytes, this leads predominantly to the formation of nitrite methemoglobin. The nitrite methemoglobin reacts with hydrogen peroxide to form reactive intermediates (e.g. peroxynitrous acid) and the products of hemoglobin destruction. During the storage of erythrocyte suspensions containing methemoglobin and modified nitrite methemoglobin, differences in the forms of erythrocytes and the degree of their hemolysis were revealed. It is assumed that the formation of methemoglobin leads to the destruction of erythrocytes.

Cells of neuroglia--the astrocytes are of interest from the point of view of their participation in phagocytosis. Phagocyte ability to generate active oxygen forms (AOF) as used as the basic criterion of the estimation of their functional activity. For the purpose to clear up molecular and cellular mechanisms of phagocytosis a study of astrocyte redox-systems, participating in production of AOF, was undertaken. Registration of AOF in astrocytes was carried out using a method of luminol-dependent chemiluminescence. Primary culture of guinea pig astrocytes was used. Spontaneous chemiluminescence of low intensity was found for the astrocytes at the presence of luminol. The destruction of the cells was accompanied by a significant growth of the intensity of spontaneous chemiluminescence. Suspension of endocutosis inductors, particle of latex and phytohemagglutinin, added to astrocytes did not result in formation of AOF, characteristic for other cells, possessing phagocytosis. It was established, that addition of hydrogen peroxide destroys astrocytes at the presence of luminol and gives rise to the emission. Chemiluminescence was not observed in similar experiments with intact cells. A conclusion was made that inside astrocytes there are structures, which show peroxidase-like activity.

The content of circulating immune complexes (IC) interacting with plant lectins from Canavalia ensiformis (Con A), phaseolus vulgaris (PHA) and wheat germ (WGA) was examined in blood plasma of healthy donors and patients with ENT diseases (n = 36). It was found that the level of PHA- and WGA-reactive IC in patients with laryngeal carcinoma was significantly higher than in control, whereas only the level of WGA-reactive IC was elevated in patients with scleroma. The level of ConA-reactive IC was statistically uniform in control and both ENT diseases.

The effects of eight plant lectins on human neutrophils aggregation and H2O2 release were studied. Both processes were stimulated by lectins from Viscum album, Triticum vulgaris, Phaseolus vulgaris and Canavalia ensiformis while lectins from Solanum tuberosum, Sambucus nigra and Glycine mas displayed only aggregating activity and Arachis hypogaea lectin was not effective. Heating (46 degrees C) and UV-radiation during 4-5 min were found to suppress completely H2O2 release from neutrophils keeping the capacity of cells to be aggregated. The findings indicate that lectin-induced human neutrophils aggregation is necessary but not sufficient condition of respiratory burst of cells. The comparison of the aggregating activity of lectins with their ability to induce the H2O2 release is supposed to be the basis of screening of lectins with antitumor and immunomodulatory activity.

Hyperthermia (45 degrees C) has found to decrease the rate of rat thymocytes as well as splenocytes aggregation induced by lectins (concanavalin A, wheat germ agglutinin) and sulphated polysaccharides (heparin, dextran sulphate-500) and to increase those induced by cationic dye alcian blue. Shedding of plasma membrane glycoproteins and changes of surface charge are supposed to affect the aggregation.

It was shown that neutrophil aggregation is caused by all lectins with different specificity for carbohydrates but generation of active oxygen forms is induced only by some lectins. Polyspecific lectins-erythroagglutinin and phytohemagglutinins have the greatest activity in relation to both processes.

Human neutrophil aggregation induced by Escherichia coli K12 cells has been studied. D-mannose has been found to inhibit the process in the dose-dependent way causing the full blockage at 10 mM concentration. The process of disaggregation induced by the concentration is significantly higher at 10 degrees C than at 37 degrees C. The inhibitors of cellular oxygenation such as N-ethylmaleimide, nordihydroguaiaretic acid, trifluoperazine, and colchicine did not affect the aggregation process. Cellular aggregation was concomitant with production of H2O2, and this reaction was also blocked by D-mannose and depended on the number of bacteria present. The nordihydroguaiaretic acid and N-ethylmaleimide inhibited production of H2O2, while sodium azide enhanced the process. The results show the mannose-specific contact of bacteria and neutrophils to depend on the functional activity of phagocytes including their ability to produce H2O2.

As revealed by investigations made by the use of the method of luminol-dependent chemiluminescence and light dispersion, the addition of concanavalin A (ConA) to the suspension of human peripheral blood lymphocytes and subsequent incubation with recombinant interleukin-1 beta leads to a sharp increase in the yield of chemiluminescence and the rate of cell aggregation in comparison with similar parameters obtained for cells, not treated with interleukin-1 beta. The study revealed the potentiating action of recombinant interleukin-1 beta on ConA-induced proliferative response of thymocytes in the culture. The mechanisms of the priming action of recombinant interleukin-1 beta on ConA-induced reaction of lymphocytes are discussed.

The authors examined the interaction of 1,2-benzoquinone derivatives with HeLa cell cultures and ascite Ehrlich's cancer cells. 4-N (anilino)-5-methoxy-1,2-benzoquinone was found to produce a marked toxic effects against tumor cells. The cytotoxic effect is oxygen dependent and associated with the formation of oxygen radicals in quinone's redox cyclization reactions. The inhibitory analysis was used to show that the major mediators of the toxic action of the agent are superoxide radical-anion and hydrogen peroxide. It is concluded that the interaction of highly toxic oxygen radicals, which are generated in quinone redox cyclization, with plasma membrane cells is likely to be the mechanism responsible for cellular destruction when quinones act on tumor cells.

Arachidonate-induced aggregation and generalization of active oxygen forms (OAF) by peripheral blood neutrophils in donors were studied in donors and Hodgkin's disease patients. Leukocytes of the latter had incomplete ability to produce AOF in response to cell stimulation with arachidonic acid. The study of arachidonate-induced aggregation of neutrophils indicated no differences in the speed of the process in the patients and donors. AOF catchers did not act on the rate of leukocyte aggregation in the patients though accelerated the process in the donors. It is inferred that Hodgkin's disease is associated with dysfunction of oxygen activation by neutrophils. The findings suggest that defects in leukocytes ability to activate oxygen in Hodgkin's disease may entail deranged regulation of other processes essential for functional activity of polymorphonuclear leukocytes.

The kinetics of photoinduced damage sensitized by chlorine e6 and its derivatives was studied in Escherichia coli and Bacillus subtilis. The effectiveness of E. coli photoinactivation in the presence of chlorines was 100-200 times lower as compared with that of B. subtilis. The structural organisation of bacterial cell walls apparently played an essential role in the penetration of tetrapyrrole pigments into the cell and in their binding.

It has been shown that photosensitized action of chlorin e6 (Che6) on platelet-rich plasma leads to platelet aggregation inhibition. Che6-sensitized photoinactivation of platelets was intensified in the presence of singlet oxygen interceptor. The aggregation rate of light-irradiated platelets in the presence of Che6 in D2O buffer was higher than in H2O buffer. The participation of active oxygen forms in the determination of platelet function has been considered.

ADP-induced aggregation of platelets in the presence of chlorin E6 under the action of visible light depending on the pigment concentration and the time of light action was studied. It was shown that the photosensitized effect of chlorin E6 on the platelet-enriched plasma results in platelet aggregation inhibition.

The effects of culture media of various compositions on chemiluminescence developing in peroxidase oxidation of luminol with hydrogen peroxide were under study. The findings evidence that the presence of carbonate and bicarbonate ions in the medium results in a two-staged chemiluminescence kinetics and in more intensive chemiluminescence in the peroxidase-luminol-hydrogen peroxide system. This fact has brought the authors to a conclusion that carbonate and bicarbonate-containing media are more effective for the detection of low peroxidase concentrations by the chemiluminescence technique.

The method of luminol-dependent chemiluminescence was used to detect oxygen activation during interaction of HLA antibodies with HLA antigens expressed on lymphocyte surface. Introduction of specific anti-HLA serum into the lymphocyte suspension leads to a rapid decrease of intensity in chemiluminescence intensity follows. The maximum yield of induced chemiluminescence in case of using a specific antiserum is significantly lower than in the control. The method could be used for detection of HLA antigens on lymphocyte surface.

The effect of hydrogen peroxide on ADP-induced platelet aggregation in the presence of active oxygen species scavengers was studied. It was shown that the superoxide radical and singlet oxygen, alongside with hydrogen peroxide, may play a role in platelet interactions.

Arachidonate-induced aggregation of human peripheral blood neutrophils and the influence of certain interreceptors of oxygen active forms (OAF) on its rate were studied. The results obtained have shown that OAF play an important role in the processes of interaction between arachidonate and leucocytes. Regulation of the process of leucocyte aggregation may be realized during arachidonate-induced aggregation of neutrophils, due to interaction of cell-generated OAF with arachidonic acid and its lipoxygenase metabolites.

It has been shown that hyperthermia (45 degrees C, 20-120 min) produces a number of changes in HeLa cells at the plasma membrane level: shedding of membrane glycoproteins; changes in the lectin-induced cell agglutination; the outflux from cells of substances with the nonspecific esterase activity.

A study was made of the medium pH influence on structural states of platelets by optical methods. Within the pH range (6-8), two pH induced reversible changes of platelet state were observed. A conclusion is made that the structural rearrangements in platelets induced in the medium by changes in hydrogen ion concentration may involve some rearrangements in platelet proteins, and thus acting as a factor regulating platelet function.

The effect of different concentration of concanavalin A on intracellular pH of rat thymocytes has been studied by means of fluorescent probe 9-aminoacridine. It has been shown that the addition of mitogen to cells results in an increase of pH value inside the thymocytes. The value of pH changes depends on the concentration of concanavalin A.

Chemoluminescence induced by thrombin and aggregation of thrombocytes as well as effect of aspirin on the reactions were studied. Intensity of thrombin-induced chemoluminescence correlated with accumulation of malonic dialdehyde and hydroperoxides in cells. Chemoluminescence of thrombocytes appears to occur after the thrombin stimulation of cyclooxygenase pathway of arachidonic acid oxidation in cells.

4-N (p-sulfoaniline),5-methoxy,1,2-benzoquinon (1) is bound by hydrophobic regions of the native molecule of bovine serum albumin (BSA). In the temperature interval 0--65 degrees C the interaction characteristics such as energy, entropy and the average number of the binding sites on a BSA molecule were determined. Under experimental conditions BSA is found in at least in two equilibrium conformational states distinguished by quantity of hydrophobic regions capable of binding with 1. Below 17 degrees C no conformational changes of BSA was observed. With the increase of temperature from 17 to 47 degrees C the equilibrium is driven in the direction of protein form with the hydrophobic binding sites which are more available for the solvent. Heating above 47 degrees C produces "predenaturation" structural changes in the BSA molecule. Hydrophobic regions of the BSA have different thermal stability.

A substance inhibiting DNA synthesis in mouse leukemic cells was isolated from the regenerating calf spleen. When added to a suspension of leukaemic cells, this substance is adsorbed on their surface. The following changes in cell features being noticed: 1) a minute decrease in electrophoretic cell motility, 2) a decrease in esterase activity of the cells, 3) an increase in microviscosity of membrane lipids, 4) an increase in the intracellular pH values. With a longer contact with this substance, changes in nuclear chromatin structure were noticed, with special reference to weakened bonds between DNA and proteins. The data obtained are of significance for revealing molecular mechanisms of hematopoietic mediator action on target cells.