[Cell-to-cell interactions in microgravity: experiments in vitro].

The review summarizes the results of studying the space and simulated microgravity effects on cell-to-cell interactions in three models: 1) interaction of a human lymphocytes culture with a subinoculated suspension culture of tumor myeloblasts K-562; 2) formation of embryoid bodies and differentiation of mice embryonic stem cells (ESCs); 3) gene differentiation and expression in multipotent mesenchymal cells (MCSs) obtained from the human marrow. It was shown that microgravity modifies the cell-to-cell interactions without cell contact inhibition; however, micro-g causes reversible changes in expression of genes responsible for regulation of the cytoskeleton, focal adhesion proteins, and some cytokines. Prolonged exposure of progenitor cells in simulated microgravity inhibits the differentiation processes of and reprofiles significantly expression of the genes that control various cell activities, including cell-to-cell interactions.

Cryoglobulin composition in patients with myocardial infarction.

The formation of an ischemic focus in the myocardium is paralleled by autoimmune processes involving immunoglobulins with abnormal thermal solubility, cryoglobulins. Serum levels of cryoglobulins and their complexes were measured in 25 patients with Q-forming myocardial infarction on days 1, 2, 3, 7, and 14 of the disease. Cryoprotein levels were measured by spectrophotometry, concentrations of IgM, IgA, IgG, and fibronectin by ELISA. Cryocomplexes in patients with myocardial infarction consist of polyclonal immunoglobulins of the main classes IgG and IgM, less often IgA, while cryoglobulinemia in this patient population is referred to type III according to the commonly used classification by Brouet. Fibronectin was detected in the cryocomplexes throughout the entire period of observation, its dynamics being opposite to that of the cryocomplexes level.

Relationship between apoptosis and expression of heat shock proteins in peripheral blood lymphocytes of patients with myocardial infarction.

We studied activity and dynamics of apoptosis of peripheral blood lymphocytes in patients with myocardial infarction and analyzed the relationship of these processes with expression of heat shock proteins with a molecular weight of 70 kDa playing an essential role in preventing cell death. Thus, we first demonstrated activation of apoptosis in peripheral blood cells of patients with myocardial infarction compared to the control (healthy individuals) and revealed the expected negative correlation between the expression of heat shock proteins with a molecular weight of 70 kDa by lymphocytes and intensity of their death. The observed dynamics of mononuclear cell apoptosis in the peripheral blood of patients with myocardial infarction can reflect activity of programmed cardiomyocyte death in the focus of ischemic injury.

[Clinostatting effects on neuronal differentiation of embryonic stem cells from mice strain R1].

The model of embryonic stem cells from R1 mice at the stage of embryoid bodies was used to study effects of slow clinostatting on neuronal differentiation with the help of two markers--beta-III tubulin (early differentiation) and MAP2 (late differentiation). As compared with the control, the number of beta-III tubulin-positive neurons was found increased and of MAP2-positive neurons--decreased. As regards MAP2- positive neurons, it is concluded that the gravity factors have a specific effect on EB. The beta-III tubulin staining makes possible determination of the total number of neuronal cells at different stages of development. The observed increase in the number of beta-III tubulin-positive neurons may evidence a nonspecific mechanic effect of clinostatting at the EB stage. It was shown that EB cells are particularly sensitive to clinostatting.

[Cryoglobulins in patients with ischemic stroke].

The composition of circulating immune complexes with cryo-properties (cryo-IC) were determined in 70 patients on days 1, 2, 3, 7, 14 and 21 after ischemic carotid stroke. The quantitative and qualitative assessments of cryo-IC content were conducted on a spectrophotometer and on a vertical electrophoresis, respectively. Concentrations of immunoglobulins (Ig) M, A and C, fibronectin, percentage of IgM with rheumatoid factor activity, IgG isotypes (IgG1, IgG2, IgG3, IgG4) were measured with ELISA. The control group included 15 healthy age- and sex-matched people. Compared to the controls, the cryo-IC content was higher in 100% of patients studied, it was higher in atherothrombotic variant than in cardioembolic one. The cryo-IC level was directly correlated with the clinical state severity (p < 0.01). IgG, IgM and fibronectin were observed in 100%, IgA--in 11% and IgM with rheumatoid factor activity--in 6% of patients. The percentage of IgG1 and IgG2 was not less than 90% of the total amount, with the IgG1 percentage about 50%. Cryo-IC changing in the time in dependence on ischemic variant were formed on different stages of development of acute brain ischemia. Dependence of concentrations of fibronectin and IgG, IgA, IgM on the time of observation was similar for both variants of ischemic stroke, but with a time axis shift, running faster and with a smaller range of changes of parameters studied in cardioembolic than in atherothrombotic stroke.

[Genosystematics and new insight on phylogeny and taxonomy of liverworts].

The paper covers a current state of molecular studies of liverworts including new data from authors. The traditional conceptions of liverwort phylogeny and systematics were changed greatly in a result of recent molecular researches. The phylogenetic inferences from studies of different DNA loci and species sampling are mainly congruent. The phylogeny and systematics of one of the largest and taxonomically difficult suborder Jungermaniineae was discussed based on analyses of internal transcribed spacer ITS1-2 of nuclear rDNA and chloroplast trnL-F of large species number.

[Influence of clinorotation on embryoid bodies morphology].

The compensative effects of gravitation at the first stages of embryonic development stages have been investigated using slow clinorotation of embryoid bodies from mouse R1 stem cell lines. Semithin sections (1-2 microm) analysis and electron microscopy study of embryonic bodies cells allowed to reveal morphological features of cells at different maturation stages. Significant decrease in the number of embryonic stem cells undergoing apoptosis as well as noticeable reduction of "lacunas" relative areas were found in clinorotated embryonic bodies compared to control. We propose that large cyst absence may be caused by initial differentiation and morphogenesis stages delay associated with autophagy processes in embryonic bodies.

[Dynamics of blood fibronectin and its correlation with the phenomenon of cryoglobulinemia in patients with ischemic stroke].

Seventy patients with carotid ischemic stroke (38 with atherothrombotic and 32 with cardioembolic variants) were examined. The dynamics of fibronectin level and its correlation with blood cryoglobulins concentrations were evaluated in the 1st, 2nd, 3rd, 7th, 14th and 21st days after disease onset. The quantitative assessment of cryoglobulins content was carried out using spectrophotometric method; fibronectin concentration was measured by ELISA. In patients with atherothrombotic variant of stroke, the level of fibronectin in the blood serum and in the cryocomplex was higher compared to patients with cardioembolic variant in the 1st day (p < 0.001) and during all the period of the study (p < 0.05). The concentration of cryocomplexes in the blood serum of patients with ischemic stroke was in reverse correlation with percentage of fibronectin in their content and with the pathogenetic variant of ischemia (r = -0.94; p < 0.05 and r = -0.87; p < 0.05 or atherothrombotic and cardioembolic variants, respectively).

[Possible causes of temperature-dependent disorders of regional circulation in cryoglobulinemia].

We studied effects of changed blood theology in cryoglobulinemia on temperature-dependent disorders of peripheral circulation. The rheological status was assessed by aggregation characteristics of blood at temperature 37 degrees C and 4 degrees C at nephelometry of reverse light diffusion. Blood cryoglobulins concentration was determined spectrophotometrically. Turbidimetry estimated the size of cryocomplexes at different temperatures and energetic characteristics of their formation. It was found that blood of healthy donors in cooling does not change its rheological properties significantly. Blood of patients with cryoglobulinemia cooled to 4 degrees accelerated formation of linear erythrocyte aggregates and increased their strength. The results agreed with estimated values of molecular connection energy in cryocomplexes. The greatest difference between the patients' and donors' blood was in strength of the largest aggregates. Sharp strengthening of both large and small aggregates in blood cooling makes vascular red cell desaggregation impossible. This may entail microcirculatory arrest.

The effects of prolonged gravity vector randomization on differentiation of precursour cells in vitro.

It was shown that proliferative rate of human mesenchymal stem cells (hMSCs) decreased during gravity vector randomization (clinorotation). Clinorotated hMSCs exhibited changes in expression of some external cell markers and adhesion molecules. Marked suppression of osteogenic differentiation of hMSC as to formation of bone nodules and calcium deposits was observed. F-actin stress fibers of cytoskeleton were altered in clinorotated cultures. Prolonged clinorotation of embryoid bodies (EBs) resulted in a delay of embryonic stem cells (ESCs) differentiation into cardiomyocytes. Percentage of beating cardiomyocytes in each EB was significantly reduced. Thus, long-term gravity vector randomization decreases the differentiation processes of precursor cells in vitro.

[Phylogeny of the genus Lophozia (Dumort.) Dumort. s. str. inferred from nuclear and chloroplast sequences ITS1-2 and TRNL-F].

Maximum parsimony and maximum likelihood phylogenetic trees were constructed for 21 taxa of Lophozia s. str. and the related genera, Schistochilopsis (5 species), Protolophozia elongate, and Obtusifolium obtusum based on pooled nuclear ITS 1-2 and chloroplast trnL-F DNA sequences. The trees were characterized by similar topology. It was demonstrated that the genus Lophozia s. str. was monophyletic, excluding L. sudetica, which deserved isolation into a distinct cryptic genus. The species distribution among the clades disagreed with the sections distinguished based on anatomical and morphological data. The relationships within the genus Schistochilopsis were consistent with the sectioning of the genus, based on morphological characters. Analysis of molecular data provided more precise definition of the systematic position of a number of taxa. Small genetic divergence of geographically distant forms was demonstrated.

[Studies of the primary effects of chronic clinostatting of embryonal mice stem cells in vitro].

Effects of randomized gravity vector (clinostatting) on embryonal stem cells (ESC) of mice were evaluated in vitro with respect to proliferation, proliferative potential, and differentiability. Colony formation remained normal up to hour 72 of clinostatting; however, further exposure led to fusion of the ESC colonies. No reliable shifts in the proliferative activity were found, whereas morphometric analysis showed different dynamics of the ESC colonies size in specific periods of the experiment comparing with the control. Evaluation of the ESC proliferative potential after the experiment revealed a trend upward in the number of colonies when compared with the dynamic and static controls. However, the number of resulted EBs in the control tended upward contrary to EBs formed under the conditions of clinostatting and continuous agitation pointing to the importance of local medium conditioning at the beginning of ESC differentiation.

Effect of routine therapy and selective plasmapheresis on electrical properties of cryoglobulins during ischemic stroke.

The charge properties of cryoglobulins were examined during the first 21 days of ischemic stroke (atherothrombotic and cardioembolic). Routine drug therapy produced no effect on the charge of cryoglobulins. Plasmapheresis significantly modified the electrokinetic parameters of cryoglobulins during atherothrombotic stroke and produced less pronounced effect on these proteins during cardioembolic stroke.

The use of statistical methods for predicting the status of patients with acute cerebral and coronary ischemia.

The most significant informative clinical laboratory parameters were determined and the exceptional value of cryoglobulin concentrations for predicting the clinical status was demonstrated. The efficiency of the proposed protocol of selective plasmapheresis for patients with atherothrombotic variant of stroke is confirmed, mathematical models describing and predicting the patient status are plotted.

[The effect of water with varying isotopic composition on the proliferative activity of endothelial cell in vitro].

Effects of water with varying isotopic composition (high and low deuterium content) were studied on the model of cultivated human endothelial cells. In laboratory conditions, changes in the isotopic content of water in nutrient substrate did not affect significantly the cell proliferative activity. In cells cultivated in no-serum medium increased deuterium concentration inhibited growth with a consequent of cell death. Both decreased and increased concentration of deuterium activated proliferation in originally low-density cell cultures supplemented with serum. It is hypothesized that changed isotopic composition of water has an effect only when combined with some other cell cultivation factors.

Cryoglobulin level in patients with ischemic stroke and effect of selective plasmapheresis on this parameter.

Serum concentrations of cryoglobulins were measured in patients with ischemic stroke; inadequate changes in these concentrations over the course of the entire acute period of the disease were shown. Positive effect of selective plasmapheresis on the decrease in cryoglobulin concentration and recovery of patients was confirmed.

[Involvement of autoimmune mechanisms in development of ischemic brain damage].

Autoimmune processes take place during an acute phase of brain injury as well as during chronic vascular diseases. Cryoglobulinemia is a nonspecific marker of long-term stimulation of the immune system which reflects duration of autoimmune sensitization. It is characterized by production of abnormal temperature-dependent immunoglobulins named cryoglobulins. The highest cryoglobulinemia was found in patients with atherothrombotic stroke that corresponds to the results of previous studies reporting the greatest level of neurospecific antibodies during slow atherothrombosis development. It is shown that cryoglobulinemia is an important factor in pathogenesis of stroke which contributes to microcirculatory and hemorheological disorders, increases blood viscosity and results in the development of immunocomplex endovasculitis of small brain blood vessels in the ischemic lesion.

[Crioglobulinemia in cerebral and coronal atherothrombosis].

Forty-seven patients: group A--17 patients with acute ischemic atherothrombotic stroke and group B--30 patients with acute Q-formation myocardial infarction. The levels of dynamics and comparative clinical relevance of crioglobulinemia in patients of the both groups have been evaluated. Crioglobulines (CG) in the peripheral blood serum were determined at 1, 2, 3, 7th and 14th days after the disease development. On the 1st day, the elevated CG level similar in the both groups as well as similarity of crioglobulinemia dynamics in whole was observed. In the group A, but not in the group B, the CG level was directly correlated with disease severity (p<0,05). In the group B, there was a trend, which depended on the blood CG level on the 1st day of the disease and on the presence of previous stenocardia (p<0,1).

Determination of optimal conditions for studies of electrophoretic mobility of human erythrocytes loaded with cryoglobulins.

For studies of the charge characteristics of human erythrocytes loaded with cryoglobulins the cells should be stored in Alsever solution with 0.1% sodium azide no longer than 1 week after blood collection. Erythrocytes should be loaded with cryoglobulins by incubation during at least 4 h, the final concentration of cryoproteins in the solution being no lower than 30 microg/ml.

[Cryoglobulinemia and extracorporal plasma therapy in severe ischemic stroke]. / Krioglobulinemiia i ee korrektsiia pri ostrom ishemicheskom insul'te.

Forty-five patients with carotid ischemic stroke have been examined within the first 24 hours from the disease onset. The Orgogozo and NIH scales were used to objectify a severity of the patient's state and the Barthel scale--to evaluate a degree of functional rehabilitation. Cryoglobulins (CG) were measured in blood serum by spectrophotometric method on the 1st, 2nd, 3rd, 7th and 21st days. Selective plasmapheresis was included in the therapeutic complex for 22 randomly selected patients. In both groups, a level of CG positively correlated with a severity of the patient's state on the 1st day of the monitoring (r=-0.5; p<0.01). The study of hemostasis indicated the presence of a direct correlation between an increase of platelet aggregation, blood hyperviscosity and CG concentration (r=0.46; p<0.05 and r=0.74; p<0.01, respectively). Higher scoring on the Orgogozo scale and Barthel scale prevailed in the group treated with plasmapheresis that significantly correlated with a decrease in CG concentration in blood serum (r=-0.6; p<0.05 and r=0.5; p<0.05 respectively). An evaluation of platelet aggregation, blood viscosity and a coagulogram on the 21st day of the disease revealed a significant improvement of blood rheology in the group of patients switched to efferent therapy.