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

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 irradiation with UV rays of various wave lengths. I. The expression of ABO and rhesus system antigens

UV irradiation (254 nm) in doses increasing erythrocyte haemolysis by 5, 10, 18 and 28 per cent was found to stimulate, by 2--16 times, the agglutination activity of ABO and Rh system antigens. The stimulation effect was the higher the lower the antigen activity before irradiation. In the Rh-negative (Rh-) erythrocytes, irradiation induced manifestation of the Rh0(D)-antigen specific activity suggesting that this antigen may be present in the Rh- erythrocyte membrane. The expression of Rh0(D)-antigen in Rh- erythrocytes, the stimulation of its activity in Rh-positive cells, and the activation of ABO system antigens may result from a photochemical destruction of the outer perimembraneous layer and release some of its components which stain in situ with alcian blue to be presumably glycoproteins. This effect is necessary to keep in mind when UV-irradiated blood transfusion is performed in therapeutic aims Rh- patients.     

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

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.     

Changes in the leukocyte phagocytic activity of donor blood after its UV irradiation. II. Simulation of the effect of the autotransfusion of UV-irradiated blood

The UV-irradiated blood of healthy adults was supplemented with non-irradiated blood in the ratio 1:10. The phagocytic activity (PhA) of monocytes and granulocytes was seen to increase markedly in the whole mixture of blood. In this case the rise of PhA was pronounced 1.4-1.7 times as much as in the case of the non-supplemented, directly UV-irradiated blood. The enhancement of PhA depends on its initial level and may occur simultaneously with structural changes of the cell surface components. It seems reasonable to propose that PhA stimulation may be one of the earliest mechanisms in immunocorrection by UV-irradiated blood therapy.     

Glycoprotein desorption from the surface of human peripheral blood lymphocytes after irradiation with short-wave UV rays

The vital quantitative method of the cell coat (outer perimembraneous layer-OPML) identification with alcian blue (AB), which was earlier developed for the rat hepatoma cells and human erythrocytes, has been adopted for human blood lymphocytes. AB is bind by glycoproteins, glycolipids and acid mucopolysaccharides of the cell surface. Under experimental condition to be used each lymphocyte adsorbed 1.1 X 10(-10) g of AB. Irradiation with non-lethal doses of UV light induced a decrease in AB sorption by 8-13%. At the same time, the release of substances took place, some properties of which are similar to those of glycoproteins. A conclusion is made that the lymphocyte OPML was destroyed by UV rays and its components released into the extracellular space. The role of this phenomenon is discussed in terms of the therapeutic effect of UV light.     

The effect of UV irradiation and of UV-irradiated autologous blood on the functional state of human peripheral blood lymphocytes

The effect of UV irradiation (UVI, 254 nm) and of UV-irradiated autologous blood on the spontaneous and mitogen-induced DNA-synthetic activity of intact lymphocytes has been studied. Lymphocytes were isolated from nonirradiated and irradiated blood, and from the mixture of UV-irradiated blood with the intact one in the volume ratio close to that in the blood stream during UV-irradiated blood autotransfusion (1:10, 1:40, 1:160). It has been shown that UVI of the whole blood caused in some donors the increase in spontaneous DNA synthesis, while in others the decrease or no statistically significant changes were observed. The analysis of the results obtained shows an inverse relation of the UVI effect to the initial level of spontaneous DNA synthesis (r = -0.68). In contrast to direct UVI effect, an addition of UV-irradiated blood to the autologous intact one resulted in an increase in spontaneous DNA synthesis in lymphocytes of all the samples examined. A 7-day cocultivation of lymphocytes, isolated from irradiated and nonirradiated blood samples, revealed a 1.8 times increase compared to the calculated value. The mitogen-induced DNA synthesis has a low sensitivity to UV rays, since the mitogens and the irradiation of optical range have presumably the common targets. It is assumed that photomodification of HLA-D/DR antigens can be a trigger mechanism for activation of immunocompetent cells by UVI.     

Structural-functional characteristics of the surface of the blood mononucleocytes in children suffering from chronic dermatoses. I. The correction of the glycocalyx structure by UV irradiation of the blood and in the course of treatment with an autotransfusion of UV-irradiated blood

In children suffering from chronic dermatoses (psoriasis and neurodermatitis), the glycocalix of blood mononuclears displays an Alcian blue dye sorption by 23-25% less than that in healthy children. The UV irradiation of their blood (254 nm), in addition to a course of UV-irradiated blood autotransfusion, resulted in an elevated sorption capacity of the mononuclear glycocalix up to the normal. A possible involvement of these changes in immunocompetent cell glycocalix in the pathogenesis of chronic dermatoses is discussed, as well as the significance of glycocalix normalization in the medicinal effect of UV-irradiated blood autotransfusion.     

Effect of UV radiation and the autotransfusion of UV-irradiated blood on the content of cationic proteins in the neutrophilic granulocytes of calves

A study was made of the influence of UV-irradiation (254 nm) of blood in vitro, of the autotransfusion of UV-irradiated blood (AUVIB), and of the mixture of UV-irradiated and intact blood in vitro on the content of bactericidal cation proteins (CP) in blood neutrophil of calves suffered from dyspepsia and broncho-pneumonia. Age differences were noticed in CP contents and their decrease in neutrophils following AUVIB in vivo and administration of the mixture of blood in vitro. The decrease in cell CP contents is presumably due to neutrophil degranulation and CP release into the blood plasma. Since the initial mechanisms of neutrophil degranulation are located on the cell surface, the CP release is supposed to result from a membranotropic effect of UV-irradiated blood on the intact autologous blood. This effect may explain the increase in nonspecific resistance of organism after the AUVIB, being one of the main therapeutic phenomena of the AUVIB-therapy.     

The effect of the UV microirradiation of the centrosome on cell behavior. III. The ultrastructure of the centrosome after irradiation]

One of the spindle poles of mitotic PK cells was irradiated with UV microbeam in metaphase or in anaphase. Electron microscopy showed that immediately after irradiation the microtubules around the centrosome were maintained, and that the ultrastructure of both irradiated and nonirradiated poles was similar. After microirradiation of the centrosome in metaphase, the mitotic halo around this centrosome was retained, but in due time the number of microtubules was getting less compared to that around the nonirradiated centrosome. When daughter cells with irradiated centrosomes are passing into the interphase, their centrioles are not separated from each other, no primary cilia are formed, and no replication of centrioles occurs. In the interphase cells with irradiated centrosomes, satellites are formed on the active centriole, but centrosome-attached microtubules are practically absent.     

Early changes in the lymphocyte surface membrane of rats after whole-body irradiation with neutrons and gamma rays

Biochemical changes in lymphocyte plasma membranes were studied 3 and 18 h after whole-body exposure of rats to neutrons and gamma-rays at doses from 2 to 6 Gy. It was shown that fast neutrons, with an average energy of 1.5-2.0 MeV, increased the rate of lipid peroxidation more markedly than gamma-rays did. In addition, there was an increase in the number of free aminogroups on the thymocyte surface. Dose- and time-dependent parameters of changes in the aminogroup content on the cellular surface were quantitatively different after the effect of radiation with different LET.     

Assessment of electrophoretic mobility changes of human erythrocytes by free flow electrophoresis after in vitro and in vivo irradiation

Single cell measurements of electrophoretic mobility showed a linear dose-response of human erythrocytes after exposure to ionizing radiation. To establish a biological indicator- or dosimeter-system for application to dose-estimations after accidental exposures, we tried to measure dose dependent EPM-changes by the free flow electrophoresis technique, a rapid and efficient method which supplies the capacity needed for application. The in vitro irradiated erythrocytes of most donors showed an oscillating dose-response but erythrocytes from several other individuals responded in a contradictory manner whereby the EPM of erythrocytes from radiotherapy patients increased linearly with the amount of administered dose. However, the wide interindividual EPM-range of unirradiated erythrocytes inhibits the application of this technique in biological dosimetry.     

Cation exchange in mammalian erythrocytes. III. The prolytic effect of x-rays on human cells

Freshly drawn heparinized human whole blood is exposed to x-rays in amounts up to 54,000 r in vitro and then equilibrated under a controlled atmosphere at 24 or 38°C. For as long as 26 hours following exposure, potassium is progressively lost from the cells and quantitatively replaced by sodium with little, if any, osmotic disturbance. The mean rate of loss at 20,000 r and 24°C. is about 0.4 per cent of the initial cell potassium per hour and approximately doubles for a 20,000 r increase. It is accentuated if blood is stored at low temperature (5°C.) following radiation exposure. Isotope experiments show that the rate of entrance of potassium into the cells is practically unaltered, the principal effect being an acceleration of the rate from cells to plasma. This suggests that radiation may have interfered with a mechanism of selective potassium accumulation based on preferential retention of the element. The sodium which enters the cells following irradiation contributes to the rapidly exchanging portion of the cellular sodium, suggesting that this fraction is ionic sodium.     

The use of phospholipase C to detect structural changes in the membranes of human erythrocytes aged by storage

Human blood was stored under blood transfusion conditions for up to 10 weeks. At various times samples were removed, erythrocytes isolated and the susceptibility of the erythrocyte membrane lipids to non-lytic concentrations of phospholipase C from either Bacillus cereus or Clostridium perfringens tested. The morphology of the cells at various times and the release of microvesicles from the erythrocytes were also assessed. Initially the cells were attacked very little by the phospholipases at the concentrations chosen, but their susceptibility increased markedly after about 2 weeks, stabilised until 5 weeks, and then increased again to approach a nearly stable value after 8–10 weeks. The first rise accompanied the conversion of most of the cells to crenated and echinocytic configurations and was reversed if cells were incubated in a ‘rejuvenating’ medium designed to restore their energy supplies. The second rise occurred during the period when the cells underwent extensive microvesiculation and eventually became spherocytes: this phase involved, in particular, an increase in availability of phosphatidylethanolamine for hydrolysis by phospholipase C and was not reversed by attempts at ‘rejuvenation’. When microvesicles released from the cells were harvested and their phospholipase susceptibility compared with that of the residual cells it was found that the microvesicles were the more susceptible. These changes in phospholipase susceptibility presumably reflect subtle changes in membrane organization that occur during storage and vesiculation of erythrocytes; the possible nature of such changes is discussed.     

The effect of pH and pO2 changes in the blood on the energy metabolism of human erythrocytes in old age

Changes in pH and pO2 of the blood have been studied for age peculiarities of their effect on the glycolysis rate and the content of ATP and 2,3-diphosphoglycerate (2,3-DPG) in erythrocytes (in vitro). The fresh venous blood of practically healthy young (aged 20-29) and old (aged 75-85) people was used. Acidosis was shown to induce inhibition of glycolysis and decrease of the ATP and 2.3-DPG concentrations in erythrocytes, while alkalosis and hypoxemia-an increase of the glycolysis rate and 2.3-DPG content. In the both cases changes in the indices studied were considerably lower in old people as compared to young ones.     

The indirect effect of hemoglobin on the electrophoretic mobility of human blood erythrocytes

Using the factor analysis, it was shown that the total content of hemoglobin in human blood plays a limited and indirect role in the regulation of average electrophoretic mobility of erythrocytes. In this case, it is not the only parameter governing this level. The statistical relationship between the total content of hemoglobin and erythrocyte mobility in electrical field is not stable and is determined by the dependence of both indices on their common factor of control of erythroid homeostasis.     

Effect of UV irradiation on the mitotic cycle of Chinese hamster cells with varying UV sensitivity. I. The time ratios after irradiation of the cells with UV light

The distribution of cells through the phases of the cell cycle by DNA flow cytofluorimetry was analysed to investigate the effects of UV irradiation on cell cycle progression in asynchronous Chinese hamster cells with different UV-sensitivity: cell line V79 (UV-resistant cells), and UV-sensitive clones: B6, CHS1, CHS2 and XII. The UV-irradiated cultures show a large accumulation of cells in S phase, the effect increasing with UV dose increase, which may point to an inhibition of the DNA chain elongation. UV-sensitive clones show a larger and more prolongated increase in the proportion of cells in S phase after irradiation with smaller dose than UV-resistant cells. Besides, the UV-sensitive clone XII shows an inhibition of movement of irradiated cells from G1 into S phase, that may testify to an inhibition of replicon initiation. These results suggest that there is a correlation in UV-irradiated Chinese hamster cells between alteration in cell cycle progression and UV-sensitivity of cells.     

Action of UV radiation on the surface of mammalian immunocompetent cells. III. Ultrastructural changes in the glycocalyx of mouse lymphocytes

The outer perimembrane layer of murine spleen lymphocytes were studied with electron microscopy. Mice were treated with short-wave (254 nm) and long-wave (365 nm) ultraviolet radiation in isoeffective lethal doses. The other perimembrane layers were stained with Alcian blue and Ruthenium red. The 254 nm UV treatment decreased the level of dye--sorption by these perimembrane layers, whereas no such effect was obvious after the 365 nm UV treatment.