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Aim: The search for etiopathogenetic agents to prevent the development of severe and extremely severe COVID-19 remains relevant. A placebo-controlled randomized clinical trial was conducted to evaluate the efficacy and safety of the antibody-based biological drug (Raphamin)., Materials and Methods: 785 outpatients 18-75 y.o. with laboratory confirmed mild COVID-19 were included within 24 hours from the disease onset. 771 patients were randomized to the group Raphamin (n=382) and the Placebo group (n=389). The study drug/placebo was prescribed for 5 days. The rate of progression to a more severe degree of COVID-19 by day 28 as well as the time to sustained clinical recovery and the frequency of hospitalization were evaluated. Safety was assessed taking into account adverse events, vital signs and laboratory parameters., Results: The number of cases of progression to a more severe degree of COVID-19 in participants receiving Raphamin was 59 (15.5%) [52 (14.6%)] versus placebo - 89 (22.9%) [85 (23.7%)], ITT and [PP] analysis data are presented. The odds ratio between groups was OR=0.6157 [OR=0.5494], 95% confidence interval 0.4276-0.8866 [0.3750-0.8048], which meant a reduction in the chance of progression to a more severe degree by 38.4% [45.1%] or 1.48 [1.62] times; p=0.0088 [p=0.0019]. The time to sustained recovery in the Raphamin group was 4.5±2.4 [4.6±2.4] days, versus placebo - 5.8±4.7 [6.0±4.8] days; p=0.0025 [p=0.0036]. No adverse events with a certain relationship were registered., Conclusion: Raphamin reduces the risk of progression to a more severe degree of the COVID-19 and significantly shortens the duration of clinical symptoms.

The effects of bone marrow multipotent mesenchymal stromal cells and their secretion products on the subpopulation composition of thymic and splenic lymphocytes were studied in female Wistar rats with experimental chronic inflammatory process in the internal genitals. Stromal cells and medium conditioned by these cells in different administration routes (intravenous or lymphotropic injection) produces different modulating effect on blood leukocyte count and on subpopulation composition of the splenic and thymic lymphocytes. The most manifest anti-inflammatory effect was observed after lymphotropic injection of multipotent mesenchymal stromal cells creating a high concentration and long persistence of the factors produced by these cells in the focus of inflammation.

We studied the effect of a biomedical cell product (bone marrow multipotent mesenchymal stromal cells and products secreted by these cells in conditioned medium) on subpopulation composition of lymphoid cells of central and peripheral lymphoid organs of Wistar rats under normal conditions. Changes in the subpopulation composition of lymphoid organs depended on the route of administration of biomedical cell product.

We studied the effect of dyslipidemia induced by poloxamer 407 (300 mg/kg twice a week for 30 days) on cellular composition of the spleen and splenocyte lysosomes in mice. Changes in blood lipid profile included elevated concentrations of total cholesterol, aterogenic LDL, and triglycerides most pronounced in 24 h after the last poloxamer 407 injection; gradual normalization of lipid profile was observed in 4 days (except triglycerides) and 10 days. The most pronounced changes in the spleen (increase in organ weight and number of cells, inhibition in apoptosis, and reduced accumulation of vital dye acridine orange in lysosomes) were detected on day 4; on day 10, the indices returned to normal. Cathepsin D activity in the spleen also increased at these terms. The relationship between changes in the cellular composition of the spleen and dynamics of serum lipid profile in mice in dyslipidemia caused by repeated administrations of relatively low doses of poloxamer 407 is discussed.

We studied the response of the pineal gland and organs of the immune system to melatonin treatment in Wistar rats kept under conditions of abnormal illumination regimen. The animals were kept under natural light regimen or continuous illumination for 14 days and then received daily injections of melatonin (once a day in the evening) for 7 days. Administration of melatonin to rats kept at natural light cycle was followed by a decrease in percent ratio of CD4+8+ splenocytes and CD4-8+ thymocytes. In 24-h light with the following melatonin injections were accompanied by an increase in percent rate and absolute amount of CD4+8+ cells in the spleen, and a decrease in percent rate of CD11b/c and CD4-8+ splenocytes. In the thymus amount of CD4-8+ cells increased, and absolute number of CD4+25+ cells reduced. Melatonin significantly decreased lipofuscin concentration in the pineal gland during continuous light. Direction and intensity of effects of melatonin on parameters of cell immunity and state of the pineal gland were different under normal and continuous light conditions. It should be taken into account during using of this hormone for correction of immune and endocrine impairments developing during change in light/dark rhythm.

We compared the effectiveness of immunomodulators used in the treatment of patients with chronic salpingitis and oophoritis with or without changes in succinate dehydrogenase (SDH) activity in blood lymphocytes at incubation with the drug. Diurnal variations in individual reaction of SDH in blood lymphocytes to thymalin or ridostin were revealed. In the groups of women receiving ridostin or thymalin during the reaction of lymphocyte SDH to it, improvement of clinical laboratory and immunological parameters was observed in the majority of the patients and no effect was found in a lesser group of patients than in the groups treated with drugs during the absence of lymphocyte SDH reaction thereto. The timing of the presence of SDH reaction to drugs in the immunocompetent cells makes it possible to set the optimal daily regime of their application and to select a drug that would be most effective in each particular case.

We found the peculiarities of the effects of dipeptidyl peptidase IV inhibitor diprotin A on immunological parameters of lymphocytes in lymphoid organs in intact animals and in animals with experimental autoimmune process. In intact animals, diprotin A increased cellularity of the thymus and spleen, number of CD4+, CD8+, and CD4+8+ thymocytes and CD3+ splenocytes and reduced cellularity of lymph nodes and the number of antibody-forming cells in the spleen. Diprotin A administered against the background of autoimmune process, reduced thymus cellularity, number of antibody-forming cells in the spleen, and number of CD4 and CD4+8+ thymocytes in comparison with the corresponding parameters of autoimmune process.

We studied the chronodependent effect of IL-2 in the experimental model of immunodeficiency, cyclophosphamide-induced immunosuppression in mice. IL-2 in a dose of 100 U/ mouse was administered at 10.00 and 16.00 for 3 days after injection of cyclophosphamide. In contrast to the morning treatment with the cytokine, evening administration produced antiapoptotic effect on splenocytes and stimulated proliferation to a greater extent. This was accompanied by an increase in the number of CD4(+), CD25(+) and CD4(+)25(+) cells in the spleen to a level of intact mice. More pronounced effect of the evening mode of IL-2 administration on the proliferation and subpopulation composition of mouse spleen cells in the studied model can be associated with high blood level of CD25(+) cells at this time of the day.

We studied the effects of melatonin on the status of immune organs and parameters of lipid metabolism in rats with alimentary obesity and parameters of lipid metabolism and immune status in Wistar rats kept on high-fat diet and receiving melatonin solution per os. Melatonin leveled the changes in blood and liver parameters of lipid metabolism, which was paralleled by normalization of cellular composition of immune organs. We conclude that melatonin can be a promising agent for the treatment of lipid metabolism and immune status disorders in alimentary obesity.

The effects of dipeptidyl peptidase IV inhibitor sitagliptin on immunological parameters were studied in animals with experimental autoimmune process. The effects of the drug administered in preventive (before manifestation of autoimmune processes) and therapeutic (after manifestation of autoimmune process) modes were studied.

It has been shown that psychoneurological disorders are accompanied by different disturbances of immunity. Paper aimed to study the effects of repeated experience of aggression in daily agonistic interactions leading to the development of behavioral psychopathology on the parameters of cellular immunity in the thymus and spleen. There were no found the changes in the weight indexes, the number of cells in the thymus, spleen and blood in aggressive mice. In the spleen of aggressive mice percent of B-lymphocytes--CD19+ and CD16/32+, as well as T-lymphocytes CD4+8-, CD4-8+, and CD4+25(hi) decreased and percent of CD4-25+ increased in comparison with the controls. In the thymus percent of CD4-25+ cells are decreased without changes of other types of lymphocytes. Flow cytometric analysis revealed decreased percentage of apoptotic (A(0)) and resting (G0/G1) cells and increased percentage of proliferating cells in phase S+G2/M in the spleen of aggressive male mice in comparison with the control. The percentage of apoptotic thymocytes is increased and the percentage of thymocytes in S+G2/M phase is decreased under the repeated experience of aggression. Data suggest the possible development of an autoimmune procceses in male mice under the influence of repeated experience of aggression.

Neuroprotective effect of erythropoietin administered before ischemia has been previously demonstrated. The efficiency of erythropoietin administration after ischemia was not studied, though in case of success these protocols would be applied in clinical neurology. In our experiments on the model of transitory focal ischemia, erythropoietin was injected intraperitoneally during the early and delayed postischemic period (3 and 12 h). The size of the necrotic zone, neurological deficit, and the severity of brain edema were evaluated in 48 h. Injection of erythropoietin in 3 and 12 h after ischemia significantly reduced the size of necrosis (p = 0.0007 and p=0.0016, respectively), neurological deficit (p=0.0013 and p=0.0062, respectively), and brain edema (p=0.02 and p=0.0186, respectively). Injection of erythropoietin after transitory focal cerebral ischemia produced a pronounced neuroprotective effect.

Unlabelled: Chronic social defeat stress leads to the development of mixed anxiety-depression state, which accompanied by immune deficiency in male mice. Paper aimed to study effects of ronkoleukin on the parameters of cellular immunity in the thymus and spleen and psychoemotional state in these animals., Methods: Mixed anxiety/depression state was produced by chronic social defeat stress during 20 days in male mice. Roncoleukin (5000 ME/kg, i/p) and saline were chronically injected to depressive mice during 2 weeks without agonistic interactions. After this period subpopulations of lymphocytes in the thymus and spleen were studied in male mice. The animals were also studied in behavioral tests estimating the levels of communicativeness, anxiety and depressiveness., Results: Roncoleukin decreases the number of lymphocytes in the thymus and spleen, and increased the number of lymphocytes in blood and thymus index. Medication increased per cent of CD4+8+ lymphocytes in the thymus and per cent of CD8+ and CD3+25- lymphocytes in the spleen. Roncoleukin induced anxiogenic, stimulative and antidepressive effects., Conclusion: Roncoleukin has small efficacy for treatment of immune suppression induced by chronic social defeat stress and has anxiogenic, stimulating and weak antidepressive effects.

The objective of this investigation was to determine the complex response of the pineal gland (PG) and of the organs of the immune system in Wistar rats in response to a violation of the illumination regime in the experiment. Animals were kept under natural light regime and continuous illumination for 14 days. After that rats were sacrificed and the mass of the body, PG, gonads, thymus and spleen was measured. Thymus and spleen cell subpopulations were determined by flow cytometry. The lipofuscin content in PG was determined by measuring an autofluorescence intensity in frozen tissue sections in the wavelength range of 505-545 nm using a confocal laser scanning microscope LSM 510 META (Carl Zeiss). The correlation analysis showed an increase in the amount and the change of sign and direction of relations between the indices of the state of PG and the immune system. This indicates the up-regulation of the intensity of inter-system relationships and the change of migration and differentiation vector of immunocompetent cells.

Changes in the cellular and humoral immunity parameters were revealed in animals receiving thyroxin and mercazolyl per os for 2 weeks. Specific features of effects of the hormone and its inhibitor on the parameters of immune system were found.

We analyzed activities of lysosomal cystein cathepsins B and L in mouse LS lymphosarcoma and its drug-resistant RLS 40 strain and their correlations with the dynamics of the percentage of cells with fragmented DNA and CD14 (+) phagocytes over 3 days after cyclophosphamide injection. LS regression and inhibition of RLS 40 growth after cyclophosphamide injection were paralleled by an increase in cathepsins B and L activities in tumor tissues. The antitumor effect of cyclophosphamide associated with apoptosis intensity and protease activities were significantly higher in LS. Positive correlations between activities of cathepsins B and L and the LS tissue content of cells with fragmented DNA and CD14 (+) phagocytes and negative correlations thereof with tumor weight were detected. It seems that the increase in cathepsins B and L activities in LS tissues was caused by cyclophosphamide induction of apoptosis and depended on the level of tumor cell infiltration with mononuclear phagocytes.

The levels of CD19 (+) and CD20(+) microvesicles were estimated in the blood of patients with B-cell chronic lymphocytic leukemia and indolent non-Hodgkin lymphoma by flow cytometry method. It was found that the number of B cell microvesicles is several times higher in patients than in volunteers. The level of CD20 (+) microvesicles directly correlated with the number of CD20(+) lymphocytes in patients with chronic lymphoproliferative diseases. Extramedullary tumors cells can be a source of CD19 (+) microvesicles.

We studied the effects of IL-2 administration at different times of day to female B6D2F1 mice with experimental immunopathology, graft-versus-host chronic reaction induced by the administration of lymphoid cells from parental female DBA/2 mice. Recombinant murine IL-2 was injected subcutaneously at 10.00 (group 1) or 16.00 (group 2) after the first cell transfer. Evening administration of IL-2 in contrast to its morning injection prevented cell proliferation in the thymus and spleen, decreased the number of apoptotic splenocytes, significantly stimulated differentiation processes in the thymus, increased the amount of CD4 (+) 25 (high) thymocytes, and decreased the number of CD4 (+) 27 (low) splenocytes. These findings can serve as a prerequisite for the development of chronotherapeutic schemes for immunocorrection.

We studied the effect of bezafibrate on hepatic PPARα activity and immune parameters in hypertensive ISIAH rats in comparison with normotensive WAG rats under conditions of LPS treatment. Bezafibrate increased activity of PPARα in WAG rats, but not in ISIAH rats. As differentiated from WAG rats, bezafibrate produced a potent effect on the content of T cell subpopulations in the thymus and spleen of ISIAH rats. Administration of LPS after injection of bezafibrate caused death of 50% ISIAH animals (but not WAG rats), which was associated with low level of HDL cholesterol and increased triglyceride content. Our results suggest that the hypolipidemic treatment (e.g., bezafibrate) can increase the severity of complications in patients with infectious and inflammatory diseases in association with low level of HDL.

The external and internal control samples are used in case of polymerase chain reaction application to ensure validity of results. The external control samples are used to establish operability of reagents included into diagnostic kit. The internal control samples make it possible to check not only all the stages of reaction but also the process of amplification in each individual test tube with reaction mixture.

The study examined the effect of long-term hypothermic (4°C) storage of mouse carcasses on motility, cell membrane damage, in vitro survival and capacitation of epididymal spermatozoa before and after cryopreservation. It was shown that the number of spermatozoa with rectilinear forward motion decreased with increasing storage time. There were no significant changes in the total sperm motility and integrity of their plasmalemma. Pronounced effects of hypothermia and long-term storage of the mouse carcasses on cryocapacitation of spermatozoa during cryoconservation were demonstrated.

We hypothesize that early ischemic preconditioning (IPC) can afford protection against focal brief and prolonged cerebral ischemia with subsequent reperfusion as well as permanent brain ischemia in rats by amelioration of regional cerebral blood flow. Adult male Wistar rats (n=97) were subjected to transient (30 and 60 minutes) and permanent middle cerebral artery (MCA) occlusion. IPC protocol consisted of two episodes of 5-min common carotid artery occlusion + 5-min reperfusion prior to test ischemia either followed by 48 hours of reperfusion or not. Triphenyltetrazolium chloride and Evans blue were used for delineation of infarct size and anatomical area at risk (comprises ischemic penumbra and ischemic core), respectively. Blood flow in the MCA vascular bed was measured with use of Doppler ultrasound. The IPC resulted in significant infarct size limitation in both transient and permanent MCA occlusion. Importantly, IPC caused significant reduction of area at risk after 30 min of focal ischemia as compared to controls [med(min-max) 11.4% (3.59-2 0.35%) vs. 2.47% (0.8-9.31%), p = 0.018] but it failed to influence area at risk after 5 min of ischemia [med(min-max) 7.61% (6.32-10.87%) vs. 8.2% (4.87-9.65%), p > 0.05]. No differences in blood flow were found between IPC and control groups using Doppler ultrasound. This is suggestive of the fact that IPC does not really influence blood flow in the large cerebral arteries such as MCA but it might have some effect on smaller arteries. It seems that, along with well established cytoprotective effects of IPC, IPC-mediated reduction of area at risk by means of improvement in local cerebral blood flow may contribute to infarct size limitation after focal transient and permanent brain ischemia in rats.

Background: the present study was aimed at investigation of neuroprotective effects of alpha-lipoic acid (LA) and superoxide dismutase (SOD) in brain ischemia in rats., Methods: two models were used to produce brain ischemia: focal ischemia (permanent left middle cerebral artery occlusion) and permanent ligation of both CCA without subsequent reperfusion. Cu/Zn-SOD at a dose 5 mg/kg, i.v. and LA at a dose of 20 mg/kg, i.p. were injected 30 minutes prior or 5 minutes after onset of ischemia. The end-points of the study were histochemically determined: infarction size, ultrastructural changes in the cerebral tissue, and survival rate., Results: LA administration 30 minutes prior to ischemia dramatically reduced infarction size (p < 0.001). Injection of LA 5 minutes after beginning of ischemia did not affect the infarction size. Besides, infarction ion size was unchanged after injection of SOD 30 minutes prior to ischemia., Conclusion: the LA treatment regimen used in this study resulted in significant cerebral protection against ischemia. In contrast, SOD did not show any protective effects in focal and forebrain ischemia.

The review is devoted to the secondary prevention of ischemic events after ischemic stroke. Recommendations for antithrombotic therapy and prolonged indirect anticoagulant therapy with INR monitoring based on evidence-based medicine of the secondary prevention of ischemic stroke and TIA are presented. Patients who do not need anticoagulants should receive antiplatelet therapy. Due to the increasing number of patients resistant to antiaggregant therapy, the rapid and adequate evaluation of platelet function is needed. Current laboratory diagnostic methods have some shortcomings and do not meet criteria of evidence-based medicine. It is necessary to improve the laboratory diagnosis with the elaboration of reliable standardized methods.

Normotensive and hypertensive male Wistar rats were subjected to the 3 h bilateral carotid artery occlusion followed by 72 h of reperfusion. The selective cycloxygenase-2 inhibitor meloxicam was administered intramuscularly after the cerebral reperfusion at a daily dose of 15 mg/kg. Doppler ultrasound was used to evaluate the cerebral blood flow during the reperfusion phase. The level of malondialdehyde (MDA) within the brain tissue homogenate was determined using spectrophotometry. The presence of arterial hypertension caused the altered response of brain tissue to ischemia/reperfusion. The meloxicam treatment significantly decreased the MDA level in normotensive rats subjected to ischemia-reperfusion. The protective effect of meloxicam against cerebral ischemia/reperfusion injury was more pronounced in hypertensive rats as compared to normotensive animals.

The present study was designed to determine experimentally the effect of a biologically active phytocomposition (BAP) on the gastric mucosa structural components during the formation and healing of ulcer induced by a single intraperitoneal injection of epinephrine. Experiments were conducted using 160 Wistar albino male rats. The fundic mucosa was studied 2, 5 and 10 days after the beginning of the experiment. The integrative assessment of a gastric mucosa was given by means of a statistical matrix method under the condition of phytocorrection. BAP was shown to change the rates of cell renewal that resulted in the restoration of the dimensions of mucosal compartments. Administration of BAP decreased the degree of mucosal damage and increased the initial size of mucosal compartments by 14-15%, as compared to those at similar time intervals in animals receiving no BAP. The number of neck mucous cells in the fundic glands was stably increased. Administration of BAP provided high parameters of the general normalization index for an organ. These changes collectively favored faster restoration of the gastric mucosa integrity after an ulcer. BAP was shown to have a protective and stimulating effect on the organ structures.

Cancer of stomach is currently regarded as the final result of a staged multifactor process during which the microenvironment affects cells and causes their changes. One of the main triggering factors is Hp infection. Adenocarcinoma of stomach develops via stages of gastritis, precancerous changes, and cancer. The possibility to prevent cancer ensues from the potential irreversibility of premalignant processes in gastric mucosa, in the first place its atrophy; hence, the importance of its early diagnosis. The state of the endoscopic service in this country is inadequate for mass screening of patients with symptoms of dyspepsia. "GastroPanel", a new serological test for the diagnosis of gastric pathology provides information about histological and functional characteristics of gastric mucosa in the antral and fundal regions of the stomach. The method determines serum gastrin-17, pepsinogen-1, and IgG expressed in response to Hp infection. Our results demonstrate high diagnostic efficiency of "GastroPanel" as a screening technique for atrophic gastritis and assessment of stomach cancer risk.

Luminescence and absorption stains specific for DNA (acridine orange, ethidium bromide), proteins (silver nitrate), and lipids (Sudan III) were used to study the distribution of DNA, proteins, and lipids in sections of the olfactory bulb in rats, studies being performed after fixation of brains with paraformaldehyde. DNA was found to be more abundant in the glomerular cell layer than the mitral cell layer. Higher quantities of DNA were present in the granular layer, located beneath the mitral layer. The characteristics of cell layers in the olfactory bulb were studied in rats aged two days and one month. There were differences between the layers of rats of different ages in terms of the content and distribution of DNA, though there were no differences in the total protein or lipid contents. Glomeruli were immature in two-day-old rats.

The influence of continuous ultrasound on the embryonic development of grass frog Rana temporaria has been investigated. Intact embryos at the blastula stage were treated by ultrasound of different frequency (0.88 and 2.64 MHz), intensity (0.05-1.0 W/cm2), and duration (1-15 min). The treatment with ultrasound of frequency 0.88 MHz and intensity 0.05 W/cm2 for 1-5 min tended to increase the proportion of normally developing embryos up to hatch (10-25% of control). Increasing the intensity of ultrasound (0.88 MHz) to 0.7-1.0 W/cm2 and the duration of its action to 5-15 min induced the death of almost all of treated embryos. No significant differences were found between the development of control embryos and embryos treated with ultrasound of middle intensity (0.2-0.7 W/cm2) for 1-5 min. The exposure of amphibian embryos to ultrasound of frequency 2.64 MHz and intensity 0.05-0.7 W/cm2 for 1-5 min did not change their survival. Increasing the intensity of ultrasound (2.64 MHz) to 1.0 W/cm2 and the duration of its action to 5 min decreased the number of normal developing embryos (by 35%).

It was shown by fluorescent and electron microscopy that the physiological state of ground squirrels subjected to ionizing radiation at different phases of the torpor-normothermia cycle plays a determining role in the alteration of the conditions of the protein-synthesizing system in neurons of hippocampus fields CA1 and CA3 and sensomotor area of the brain. In ground squirrels irradiated under normothermia, the neurons were less radioresistant and restored slower compared with torpor animal cells, the distinctions being most expressed in CA1 field neurons. The effect of irradiation was minimum during the entrance into torpor and maximum during arousal. It was supposed that the inhibition of protein synthesis in the latter case occurred at the elongation stage when heavy polyribosomes were formed in neuron cytoplasm.

It was determined whether high-frequency ultra sound can change the permeability of gray toad Bufo bufo and grass frog Rana temporaria under the action of high-frequency ultrasound. The changes in the permeability of embryonic envelope were assessed by using slowly penetrating fluorochromes ANS, FDA, and fluorescein. It was found that the ultrasound of 0.88 MHz and 0.4 - 0.7 W/cm2 intensity increased the permeability of amphibian embryonic envelope for ANS and FDA, whereas the ultrasound of 2.64 MHz and the same intensity increased that for fluorescein with the retention of low permeability for FDA. Embryos continued the normal development after treatment with ultrasound under these conditions.

Experiments on (CBA x C57Bl)F1 mice showed that constant (day and night) illumination for 2 weeks led to the development of internal desynchronosis, which significantly modulated the reaction of the immune system to benz(a)pyrene compared to those in intact animals. Presumably, the differences were associated with changed circadian rhythms of the immune parameters in experimental animals determining different ratios and, presumably, functions of immunocompetent cells during benz(a)pyrene exposure in intact controls and animals with desynchronosis.

New technologies of diagnostics of post-operative period complications in newborns (measure of procalcitonin level, serum wedge-shaped dehydration method), as well as introduction of discrete plasmopheresis and fiber-optic bronchoscopy via the laringeal mask, present necessery additions to traditinally used methods of treatment in a children's intensive care unit, improving the results of its work.

The paper is based on the results obtained during examination and intensive care in 175 neonatal infants operated on for various malformations who were treated in the pediatric intensive care unit, Moscow Regional Research Institute of Clinical Studies, in 1998 to 2000. Analyzing the results has indicated that 24% of the patients have signs of early sepsis just within the first 24 postoperative hours. The marker procalcitonin was determined in 75 neonatal infants. The studies performed lead to the conclusion that procalcitonin is a highly sensitive and specific marker of sepsis by which postoperative septic complications can be diagnosed in the early periods. The early diagnosis of sepsis, by using the marker procalcitonin permitted timely rational correction of antibiotic therapy, which improved the basic parameters of treatment and reduced deaths from septic complications from 48 to 36%.

Subpopulation spectrum of T lymphocytes (CD3+, CD4+, CD8+, CD25+, and CD3(+)25+) in thymus, spleen and inguinal lymphatic nodes have been studied in male mice after 20 days of psychoemotional stress produced by social defeats in daily agonistic confrontations. A reduction of total number of cells, of absolute numbers of all researched subpopulations of lymphocytes and % CD3+ cells in thymus of submissive mice was shown in comparison with intact animals. Reduction of total number of splenocytes and absolute numbers of CD4+ and CD8+ lymphocytes has been observed in a spleen of submissive mice. Besides, % CD3+, CD25+, CD4+ and CD25+ cells were increased in these animals in comparison with intact mice. The absolute number of cells with CD8 phenotype was increased in inguinal lymphatic nodes. The data obtained suggest that the chronic psychoemotional stress is accompanied by serious changes of the cellular link of immunity. The effect of chronic emotional social stress on mutual interaction of the central and peripheral links of immunity has been discussed.

The cryotolerance of totipotent cells from dissociated embryos of amphibian (grass frog Rana temporaria and grey toad Bufo bufo) was studied. Cell integrity and preservation of the cell barrier function were evaluated by fluorescent analysis. It was shown that the best cryopreservation of the cells was achieved by using the cryoprotective agent 10% dimethyl sulfoxide and 10% saccharose. These cells were successfully used for the homotransplantation of nuclei into enucleated eggs. The development of reconstructed eggs to the blastula stage was noted.

The autofluorescence of horsetail Equisetum arvense spores excited with UV-light of 360-380 nm was studied by microspectrofluorimetry during their development from an individual cell to the formation of a multicellular thallus with the generative organs. The investigation involved the registration of the fluorescence spectra of individual intact developing cells and the measurement of the ratio of cell fluorescence intensities in the blue and red regions of the spectrum. Dry blue-fluorescing microspores showed the maxima at 460 and 530 nm and a small maximum at 680 nm. Thirty minutes after moistening in water, red-fluorescing cells arose among blue-fluorescing microspores, indicating the onset of development. Red fluorescence with a maximum at 680 nm enhanced as cells put off their cover, which brightly fluoresced in the blue region of the spectrum with the main maximum at 460 nm. By estimating the ratio of autofluorescence intensities in the blue region of the spectrum to red lightening of microspores at the first stages of development up to 24 h (in particular, their first division, the formation of nonfluorescencing rhizoid, etc.), nonviable (only blue-lightening) cells were distinguished from viable cells, in which red fluorescence began to prevail. After 25-40 days of development, the gametophyte fluorescing mainly at 680 nm formed male organs, antheridia, with blue-green-fluorescing spermatozoids. Then female generative organs archegonia with the egg cell appeared, which fluoresced blue, whereas the surrounding cells fluoresced red. It was supposed that the lightening in the blue and green regions of the spectrum is due to the presence of phenols, terpenoids, and azulenes, whereas the emission in the red region is associated with the presence of chlorophyll and azulenes. The observation of autofluorescence makes it possible to easily distinguish generative cells without additional staining.

The work was carried out on the pollen of Hippeastrum hibridum, whose size (ca 55 mcm) permits to follow both the pollen tube formation and development of the vegetative nucleus and generative cell within the first two hours before the growing point arising. Using fluorescent dye Hoechst 33342 chromatin state alterations accompanying changes in the pollen physiological state were investigated. The maximum fluorescent intensity was observed in 30 min of staining and reflected the maximum chromatin functional activity. Histograms of pollen distribution according to fluorescent intensity differ considerably with doses of irradiation (500, 1000, 2000, 3000 and 4000 Gy). Besides, a germinative index of fluorescence intensity was calculated. A comparative analysis of these data has shown that the lowest decrease in fluorescent intensity observed at 500, 3000 and 4000 Gy, was accompanied by different germinative indices. At 500 Gy, the index was 2.5 times lower than in the control, but at 4000 G no germination was not noted. The process of pollen grain development is supposed to be more intensive and faster at 500 Gy, than in the control. At 4000 Gy, a decrease in the functional pollen activity is accompanied by decrease or inhibition of chromatin functional activity.

It is well known that artificial increase in expression of growth factors and their receptors can lead to tumorigenic transformation of cells. Additionally, multiple data on the increased expression of growth factors in many human and animal tumorigenic cells have been published. Nevertheless description of the functional role of endogenous growth factors in maintenance of tumorigenic phenotype remains obscure. Previously, we described a new model for studying neoplastic transformation and dormant metastasis. This model consists of cognate tumorigenic and nontumorigenic cell clones, the latter being obtained as a result of spontaneous reversion of tumorigenic ones. All revertant clones demonstrate the three well known features of normal cells: monolayer growth on a plastic, incapability to grow in soft agar with regular culture media with 10% FCS, and incapability to form tumors in syngeneic animals. Using RT-PCR, we measured expression ratios of main growth factors, commonly believed to be associated with malignization, in tumorigenic and revertant clones of our model. For some of these clones, we also measured an activity of growth factors in conditioned media. The data obtained argue that the levels of growth factor expression, measured by both the methods, are distributed between tumorigenic and revertant clones in a sporadic manner and do not correlate with cell tumorigenicity. Thus, our experimental observations enable us to consider the variability of growth factor expression as insignificant events in the reversion of tumorigenic cells to a nontumorigenic phenotype.

A fluorescent technique with Hoechst-33258 and acridine orange staining was used to assess changes in chromatin state induced by radiation. Fluorochromes with different modes of binding to DNA were chosen. In T lymphocytes chronic irradiation caused a rearrangement of binding between nonhiston proteins and lipids accompanied by conformational changes in DNA, resulting in chromatin condensation. The decrease in fluorescence probably resulted from a reduction in the number of sites accessible for dye binding. After acute irradiation, the fluorescence intensity decreases predominantly due to double-strand breaks.

Levels of the major regulatory subpopulations of lymphocytes in the thymus and spleen and blood lymphocyte dehydrogenase levels were measured in male mice with aggressive and submissive patterns of behavior formed over 10 or 20 aggressive confrontations leading to repeated experience of victory or defeat. The results showed that repeated experience of social confrontation non-specifically increased the proportion of segmented neutrophils and lactate dehydrogenase activity in both participants in aggressive encounters, and decreased the numbers of CD4+ and CD8+ T-lymphocytes in the spleen. Succinate dehydrogenase specifically decreased in the lymphocytes of aggressive mice and increased in the lymphocytes of submissive mice. The proportion of CD4+ T-lymphocytes in victims' thymuses also decreased. Changes in metabolic measures and percentage ratios of lymphocyte contents were dynamic and depended on the duration of confrontational interactions.

We studied the effects of amitriptyline (preparation stimulating melatonin synthesis in the pineal gland) on daily variations of cell composition in immune organs of Wistar rats with experimental desynchronosis. Amitriptyline administered in the evening restored the daily dynamics of the number of thymus cells.

Three major groups can be distinguished among children hospitalized in resuscitation and intensive care wards in a state of unconsciousness: patients with bacterial meningitis and meningoencephalitis, viral meningoencephalitis, and noninfectious involvement of the NCS. Common strategy of treatment of these patients is characterized by some specific features: etiotropic antibiotic therapy in group 1 patients, antiviral drugs in group 2, and mainly symptomatic measures in group 3. Immunotherapy used in each of these groups is substitute in group 1, immunomodulating and immunostimulating in group 2 and even more so in group 3. Better results in group 1 are due to highly effective etiotropic antibiotics and drugs.

Recurrent experience of a social confrontation was shown to increase the per cent of cleavage nucleus neutrophiles in both mice-participants, activation of the lymphocyte dehydrogenase, and reduction of number of the CDA+ and CD8+ T-lymphocytes. Dynamic character of changes in the metabolism parameters and the number of lymphocytes is obviously dependent on duration of the confrontations in mice.