Expression Level of Membrane Markers CD5, CD19, and CD20 in B Cells after UV-Irradiation and Incubation in the Presence of Autologous Plasma.

The effect of UV-light (240-390 nm) in doses of 151 and 755 J/m2 on the expression of membrane markers CD5, CD19, CD20 in human peripheral blood B cells was studied by flow cytometry. In 24 h after exposure to UV light, we observed activation of processes accompanied by structural rearrangements of B-cell membranes leading to changes in the expression of receptor molecules: the content of of CD19 and CD20 increased due to activation of the synthesis of these proteins, while the content of CD5 decreased. The percentage of CD5+ cells decreased over 24 h after UV-irradiation of lymphocytes, while addition of autologous plasma to the incubation medium produced a photoprotective effect on CD5+ cells.

[The Influence of UV-Light on the Sub-Populational Composition and Expression of Membrane Markers of Lymphocytes of Donor Blood].

The influence of UV-light (240-390 nm) at dozes of 151 and 755 J/m2 on the content of membrane markers of lymphocytes using the method of flow cytometry was investigated. It was demonstrated that during incubation of UV-irradiated lymphocytes the change of their populational and sub-populational composition occurs. Expression of complexes of CD3, CD 19,.CD8, CD 16, CD25 and CD95 increased. This increase was caused mainly by de novo synthesis. UV-light had immunostimulating effect on CD8+ T-lymphocyte population. Together with the increase of cytotoxic cells and NK-cells, activation of lymphocytes (increased amount of CD25+ and CD95+ cells) took place. Amount of cells undergone apoptosis or necrosis increased proportionally to the dosage. These changes were more expressed during incubation of lymphocytes in nutrition medium without autological blood serum, e.g. under deficiency of growth factors and antioxidants.

[Apoptosis and necrosis of lymphocytes induced by UV-radiation in the presence of autological plasma].

The influence of UV-light (240-390 nm) in doses 151-3020 J/m2 on the nature of the death of lymphocytes cells of donor's blood (using markers of apoptotic and necrotic death of cells) and on the level of expression of the marker of apoptotic pre-preparation--CD95-receptor has been investigated. We have shown that UV-radiation increases expression of CD95-receptors which is caused mainly by de novo synthesis of the receptors. It has been revealed that during daily incubation of photo-modified lymphocytes (151 and 755 J/m2) without autological blood cell death occurs by receptor-involved apoptosis. Exposure to high doses of radiation (1510 and 3020 J/m2) causes massive necrotic death of immunocytes. The use of autologous blood plasma during incubation of photo-modified lymphocytes allows decreasing the number of both apoptotic and necrotic cells.

[Influence of UV-light on the profile of receptors of lymphocytes of donated blood].

The influence of UV-light (240-390 nm) at the dozes of 151 and 755 J/m2 on the maintenance of membrane markers of lymphocytes: CD3, CD19, CD4, CD8, CD25, CD95 using the method of flow cytometry was investigated. It was demonstrated that UV-irradiation of lymphocytes induced the decrease in the number of CD3, CD4 and CD19 markers together with a simultaneous increase of the amount of CD8, CD25 receptors during the daily incubation of cells in a nutrient medium without plasma. When autologous plasma is used during incubation of both native and UV-irradiated lymphocytes, the percentage of cells with the specified markers is not different from the corresponding values in the cells immediately after their purification. This finding points at the important role of antioxidants of the blood plasma and growth factors for the maintenance of the sub-population structure of lymphocytes. However, the level of expression of CD95, one of the markers of apoptosis, increases in the photomodified cells incubated both in the presence and in the absence of plasma. This increase was revealed to be caused mainly by the synthesis of the protein de novo under the influence of apoptosis-inducing factors.

[Spectral and functional properties of ceruloplasmin under UV irradiation].

During oxydase reaction spectral characteristics of ceruloplasmin at absorption of copper ions and protein part of the molecule are shown to change. It has been ascertained, that when irradiating ceruloplasmin by UV-light the functioning of intramolecular electron transport chain is broken, the degree of positive cooperativity (a Hill's constant) on substrate decreases. It is supposed, that these changes are caused by disturbance of interdomain interactions in a protein molecule.

[The protective effect of autologous plasma against the development of oxidative stress in UV-irradiated lymphocytes of peripheral blood of donors].

The influence of UV-light (240-390 nm) at the dozes of 151 and 755 J/m2 on the intensity of processes of the lipid peroxidation, activity of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), cytochrome c oxidase and the level of the energy supply of donors' blood lymphocytes in the absence and presence of autologous plasma was investigated. It was shown that during the incubation of native and UV-irradiated lymphocytes, autologous plasma reduces the intensity of lipid peroxidation, thus protecting cells from oxidative stress. As a result, the endocellular level of ATP is restored in UV-irradiated lymphocytes (during the daily incubation), which reflects the intensification of the adaptive ability of cells in the presence of autologous plasma.

[Peculiarities of metabolism of UV-irradiated lymphocytes].

The influence of UV-light (240-390 nm) at a dose of 151 and 755 J/m2 on the functional properties of lymphocyte metabolism key enzymes from donors' human blood: lactate dehydrogenase (LDH), cytochrome c oxidase, succinate dehydrogenase (SDH), Ca2(+)-ATPase of plasma membranes has been investigated. It has been revealed that photoinactivation of enzymes immediately after UV-irradiation which leads to the decrease of the ATP content in lymphocytes is replaced by the increased activity of the enzymes under investigation during daily incubation of lymphocytes. As a result, the level of ATP in photo-modified lymphocytes does not differ from that in native cells before incubation. This indicates the normalization of biochemical processes in lymphocytes influenced by UV-light applied in autotransfusion of UV-irradiated blood.

Effect of UV irradiation on functional activity of donor blood neutrophils.

Activation of NADPH-oxidase enzymatic complex was observed after UV irradiation of the blood and neutrophil suspension in a dose of 151 J/m(2). UV irradiation in doses of 75.5 and 151.0 J/m(2) corrected myeloperoxidase activity and intensity of LPO processes in donor blood.

[Influence of UV-light and dexamethasone on functional properties of lymphocytes and neutrophils].

UV-light and dexamethasone influence on functional properties of lymphocytes and neutrophils of peripherical donors' blood was studied. An increase of phagocytic activity of neutrophils was observed after their incubation with photomodified lymphocytes. It was found that UV-irradiation of lymphocytes activated synthesis of interleukines 1beta and 2. Dexamethasone presence in lymphocyte suspension inhibited the synthesis of the studied cytokines, especially by the incubation with photomodified cells. It was shown by the method of fluorescent labels that UV-irradiation improved interaction between dexamethasone and cell membrane.

[Thermal denaturation of oligomeric proteins: structural and functional modifications, the sequence of steps]. / Teplovaia denaturatsiia oligomernykh belkov: strukturno-funktsional'nye izmeneniia, posledovatel'nost' stadii.

On the basis of literature and own experimental data changes in the structural and functional properties of some oligomeric proteins of the blood system (hemoglobin, lactate dehydrogenase, superoxide dismutase, catalase) exposed to the influence of temperature in a broad range were analyzed. The many-phase character of the temperature modification of protein molecules with different values of functional and kinetic parameters for each of revealed stages was discovered. At a critical temperature and at higher values, the dissociation of oligomeric proteins into separate subunits was shown to occur along with the typical "loosening" of the protein globule. It was shown that low-molecular components (subunits) can subsequently associate with one another and with oligometic proteins, which leads to irreversible denaturation and to the unusual physicochemical behavior of protein molecules. Schemes of processes underlying the temperature modifications of the proteins studied were elaborated.

[Effect of temperature on structure and functional properties of horseradish peroxidase]. / Struktura i funktsional'nye svoistva peroksidazy khrena pri temperaturnom vozdeistvii.

The dynamics of structural and functional changes proceeding in a peroxidase molecule under the effect of temperature was studied. It was shown that peroxidase thermoinactivation proceeds in two consequent stages. Based on the analysis of enzyme peroxidase and oxidase activity and peroxidase spectral and buffer characteristics, it was established that at temperatures from 20 to 55 degrees C reversible conformation there occur changes of the hemoprotein molecule related to consequent unfolding and folding of the protein globule. The influence of temperature of 60 degrees C and above induces the protein globule unfolding and loosing of peroxidase activity.

[Effect of quamatel on neutrophil properties, parameters of lipid peroxidation and the antioxidant defense system in blood of patients with duodenal ulcer and pancreatitis]. / Vliianie kvamatela na svoistva neitrofilov, pokazateli peroksidnogo okisleniia lipidov i antioksidantnoi zashchity v krovi bol'nykh iazvennoi bolezn'iu dvenadtsatiperstnoi kishki i pankreatitom.

The properties of neutrophils, the parameters of lipid peroxidation, and the characteristics of antioxidant protection (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase activity) were studied in the blood of patients with duodenal ulcer and pancreatitis in the course of quamatel administration. The pattern of changes induced by quamatel shows evidence of the antioxidant activity of the drug.

[Study of the effects of laser irradiation (540 nm) on some components of blood enzyme antioxidant system]. / Izuchenie vliianiia lazernogo izlucheniia (540 nm) na otdel'nye zven'ia fermentativnoi antioksidantnoi sistemy krovi.

The influence of blood laser irradiation (lambda = 540 nm) in doses range 0.3-1.2 J/cm2 on the neutrophyles superoxide-dismutase and katalase activity has been studied. Correcting effect of laser light (0.6; 1.2 J/cm2) on the functional activity of studied enzymes has been found. The high degree of correlation (r = 0.6) between neutrophil superoxide dismutase and neutrophil catalase activity of the irradiated blood has been discovered. The close correlation between components of the antioxidant system and processes of active oxygen metabolite generation has been revealed. The scheme of free radical mechanisms of the laser irradiation influence on the several parts of the enzyme antioxidant system has been suggested.

[Effects of low intensity laser (632.8 nm) and ultraviolet radiation on spectral properties and functional activity of human ceruloplasmin]. / Deistvie nizkointensivnogo lazernogo (632.8 nm) i ul'trafioletovogo izluchenii na spektral'nye svoistva i funktsional'nuiu aktivnost' tseruloplazmina cheloveka.

Photoinduced changes of human ceruloplasmin (hCp) under laser and ultraviolet (UV) radiation have been studied. A complex character of hCp spectral modifications points to a different degree of the protein molecule folding. After the influence of UV radiation (240-390 nm) in a dose range 906-4530 J/m2 the decrease of oxidase hCp activity was registered. These changes are thought to be connected with modification of the conformation of hCp active centre or copper reduction that was confirmed by the decrease of optical density at 610 nm. An inactivation constant (kf = 3.8 x 10(-4) 1/s) and a photoinactivation quantum yield (1.4 x 10(-4)) were calculated. Kinetic dependencies of o-phenilen-diamine oxidation by intact and UV-modified glycoprotein do not correspond to Michaelis-Menten equation. UV radiation in a dose range 151-4530 J/m2 induced increasing of ceruloplasmin superoxiddismutase activity. The influence of He-Ne laser radiation (1.3 mW; lambda = 632.8 nm, exposure time from 1 up to 30 min) leads to the raise of the hCp functional activity; superoxiddismutase activity changes more significantly. Possible photophysical and photochemical processes in hCp molecule, leading to photomodification and changes of protein functional activity have been represented on the scheme.

Effect of laser irradiation on functional activity of human neutrophils: activation of myeloperoxidase in the presence of hematoporphyrin.

Laser irradiation (lambda=540 nm) of the blood increased myeloperoxidase activity in polymorphonuclear leukocytes. We revealed photoinduced activation of myeloperoxidase in irradiated neutrophils in the presence of hematoporphyrin. The photodynamic effect was most pronounced, when the modifier was incorporated into the cell membrane or sorbed on it.

[Oligomeric proteins: modulation of UV-sensitivity and characteristics of photochemical transformations]. / Oligomernye belki: moduliatsiia UF-chuvstvitel'nosti i zakonomernosti fotokhimicheskikh prevrashchenii.

The results of our research devoted to the study of photophysical and photochemical transformations of some compound proteins of oxidative, antioxidant, transport and immune systems of an organism under conditions of different microenvironment (presence of chemical modificators, the change of pH, temperature, doses and spectral composition of ultraviolet irradiation) have been analyzed. Methods and the degree of modulation of UV-sensitivity of proteins of the mentioned systems have been discussed. Conditions, on which the used chemical agents show photoprotective and photosensitizing effect on protein molecules, have been found.

Effect of helium-neon laser on activity and optical properties of catalase.

The effects of laser (632.8 nm) on functional and spectral properties of catalase at pH 6.0-7.4 were studied. Laser irradiation led to photoactivation of the enzyme at pH 7.1-7.4. Changes in the spectral properties of photomodified hemoprotein were found in the absorption spectrum of the protein component: apoenzyme displayed protective effects in relation to ferroporphyrin. Structural modifications of catalase induced by helium-neon laser irradiation correlated with its functional properties. These results can be used in clinical practice to design the individual management program.

[Kinetic regularities of thermal inactivation of superoxide dismutase]. / Kineticheskie zakonomernosti termoinaktivatsii superoksiddismutazy.

An interest to the study of superoxide dismutase (SOD) is growing as it has become known that this enzyme may be used as a medical preparation and in the biochemical research. The aim of the work presented is to investigate the influence of temperature and UV radiation on the structural-functional properties of SOD. Copper- and zinc-containing SOD from bovine erythrocytes with the light of a DRT-400 lamp (dose of irradiation 1.5 x 10(3) J/m2) through an UFS-1 light filter (240-390 nm). The influence of temperature (20-85 degrees C) on the functional properties of native and UV modified SOD has been studied. The process of thermodenaturation of the enzyme proceeds at least two consecutive stages. SOD is stable within the temperature interval 20-70 degrees C. Kinetics of thermodenaturation of protein has been studied. It was established that UV light increases enzyme thermostability. Changes found in the kinetic characteristics of the UV-irradiated SOD are apparently due to its conformational rearrangements.

[Activation of superoxide dismutase molecules under the effect of ultraviolet radiation]. / Aktivatsiia molekul superoksiddismutazy pod vliianiem ul'trafioletovogooblucheniia.

The effect of UV light (240-390 nm) with dose range 1.51-30.2 x 10(2) J/m2 on the functional properties of superoxide dismutase was investigated at various pH values. Photoactivation was shown to be maximum at pH values extreme for the display of superoxide dismutase activity.