卷 160, 编号 6 (2016)

Accumulation of c-Fos and Arc proteins in neurons in different regions of the hippocampus after single trial of contextual fear conditioning was studied by using immunohistochemical staining. We found that the dynamics of the c-Fos and Arc expression has a biphasic pattern: the first peak was observed in 15-30 min after learning and the second less pronounced peak in 1-3 h. Induction of Arc occurred earlier than c-Fos and the overall dynamics of the two waves slightly varied in the dentate gyrus and hippocampal CA1 and CA3 fields. The findings open the possibility of mapping the cognitive neural networks of the brain with higher temporal resolution and draw attention to fluctuations of hippocampal activity after a single brief episode of new experience.

Effects of nucleotide polyphosphate compounds (nicotinamide adenine dinucleotide, NAD⁺; diadenosine tetraphosphate, Ap4A) on the confi guration of action potentials were studied in isolated preparations of guinea pig sinoatrial node and right atrial appendage (auricle). In the working myocardium, NAD⁺ and Ap4A in concentrations of 10⁻⁵ and 10⁻⁴ M had no effect on resting potential, but significantly reduced the duration of action potentials; the most pronounced decrease was found at 25% repolarization. In the primary pacemaker of the sinoatrial node, both concentrations of NAD⁺ and Ap4A induced hyperpolarization and reduction in the rate of slow diastolic depolarization, but significant slowing of the sinus rhythm was produced by these substances only in the concentration of 10⁻⁴ M. Moreover, AP shortening and marked acceleration of AP upstroke were observed in the pacemaker myocardium after application of polyphosphates. Comparative analysis of the effects of NAD⁺ and Ap4A in the working and pacemaker myocardium drove us to a hypothesis on inhibitory effects of these substances on L-type calcium current accompanied by stimulation of one or several potassium currents, which induce enhancement of repolarization and hyperpolarization of membranes probably mediated by the activation of purine receptors.

The role of subtypes of vasopressin receptors in modulation of renal sodium reabsorption was studied in in vivo experiments on Wistar rats. Selective V_1a receptor agonist reduced sodium reabsorption in the kidneys and expression of these receptors increased by practically 100 times. This effect was similar to the effect of furosemide. Selective V₂ receptor agonist enhanced sodium reabsorption in the kidney and simultaneously increased reabsorption of solute-free water. Stimulation of V_1b receptors did not affect sodium transport. Our findings attest to the key role of V_1a receptors in the regulation of renal excretion of sodium ions.

Acetaminophen in a concentration of 5 mM increased the expression of JNK, HIF1A (hypoxiainduced factor), and CASP3, which indicated development of oxidative stress and apoptotic cell death. Acetaminophen in a concentration of 10 mM did not induce expression of HIF1A and CASP3, but reduced expression of chaperone HSP90, which attested to activation of a caspase-3-independent mechanism of cell death. The methods of preventing acetaminophen intoxication are discussed.

Сасо-2 cell line is widely used in in vitro studies of the intestinal wall permeability for drugs, but transport activity of these cells has not been thoroughly studied. We analyzed localization and expression of nucleoside transporters of the ENT family transferring a number of anticancer and antiviral drugs. Immunocytochemical staining revealed apical localization of ENT1 and integral expression of ENT2 in Caco-2 cells. Based on these data and on the value of hypoxanthine uptake by these cells, we created a mathematical model allowing quantification of transporter expression on the apical and basolateral membranes of Caco-2 cells. The discrepancy between gene and protein expression of the transporter complicating prediction of patient’s sensitivity to the drug upon individual therapy selection is discussed.

Differences in peritoneal macrophage polarization in mice of opposite lines CBA and C57Bl/6 and the effects of 60 kDa oxidized dextran were studied. Macrophages of C57Bl/6 mice demonstrated a phenotype close to M1, with increasing expression of CD86 costimulatory molecule and unchanged CD206 expression in response to activation. Macrophages of CBA mice demonstrated higher plasticity in response to activating agents; expression of the markers increased irrespectively on stimulated receptor (TLR-4 or mannose receptor) and both CD86 (classical activation) and CD206 (alternative activation) increased. Macrophage response to addition of oxidized dextran (60 kDa) to the culture medium could be characterized as potentiation of their alternative activation: expression of CD86 in CBA mice in response to LPS and LPS+IL-4 and in C57Bl/6 mice in response to IFN-γ and LPS+IFN-γ decreased, while expression of CD206 by intact macrophages of CBA mice and by macrophages stimulated by IFN-γ and IL-4 increased under the effect of 60 kDa oxidized dextran.

The levels of spontaneous and concanavalin A-induced production of some key cytokines (TNF-α, IL-1β, IL-4, Il-6, and IL-10) were assessed in the cultures of mononuclear cells derived from healthy volunteers examined for polymorphism of the promotor sites of the respective genes. The presence of certain allelic variants in the promoter regions of human cytokine genes mostly determines the rates of induced and spontaneous cytokine production. Allelic variant -31C of IL-1β gene is associated with high production rate of the related cytokine. In addition, polymorphism of allelic variant C-590T of IL-4 gene significantly affects the production rates of IL-1β, TNF-α, IL-6, and IL-10.

Molecule L1CAM is specific for nerve cells and tumors of various localizations. The expression of L1CAM is significantly higher in melanoma in comparison with benign nevi and correlates with the progress of melanoma and transition from radial to vertical growth. Monoclonal antibodies to L1CAM effectively and specifically attenuate melanoma growth, though stimulates the epithelial–mesenchymal transition. shRNA-mediated knock-down of L1CAM showed the involvement of L1CAM in regulation of activity of the canonical Wnt pathway and expression of genes of class I melanoma-associated antigens (MAGE).

IGF-1, IGF-2, and IGFBP-1,2,3 were assayed in blood serum of patients with malignant ovarian tumors (n=44), borderline ovarian tumors (n=11), and benign ovarian tumors (n=12) as well as in healthy women (n=33). In blood serum of patients with malignant ovarian tumors, the level of IGF-1 was lower and IGFBP-1 was higher than in other groups. In patients with malignant and borderline ovarian tumors, the level of IGFBP-2 was higher than in healthy women and in patients with benign ovarian tumors. There was no correlation between most examined parameters and the clinical and morphological peculiarities of ovarian tumors. The study revealed IGF/IGFBP imbalance in patients with malignant ovarian tumor and showed that IGFBP-2 proved to be a potential diagnostic serological marker w with 90% sensitivity and 90% specificity.

We studied the effect of officinal forms and nanoparticles of lithium carbonate and lithium citrate on proliferative activity of hepatoma-29 cells. Lithium carbonate nanoparticles suppressed proliferation of hepatoma-29 cells in lower concentrations than officinal form of this salt. The antiproliferative effect of lithium salts i activation of apoptosis and arrest of hepatoma-29 cells in the G₂/M phase of the cell cycle.

A complex morphological study of the auricle to determine the human age was performed by evaluating the metric sizes between fixed points in each auricle with axial guidelines. The auricular elastic cartilage in different age periods was characterized by thickening of the cartilaginous plate, different mature and immature cartilage zone ratio, variations in the volume density of the intercellular substance and elastic fibers, and change in the numerical density of individual chondrocytes and isogroups. Aggrecan content in the cartilage was shown to increase in different age periods. Age-related structural changes in the auricular cartilage expand the possibilities of forensic medical examination and hold much promise for the identification of personality.

3D cell cultures are extensively used to study in vitro toxic effect of xenobiotics. When using multicellular spheroids, the question about their optimal size should be solved: small spheroids are difficult to manipulate, while large size of spheroids impairs the transport of nutrients and oxygen into the center. Mathematical models describing the distribution of substances in multicellular spheroids numerical procedure for solving differential equation system, which complicates their use in laboratory practice. We proposed and experimentally evaluated a new mathematical model describing oxygen distribution in HepaRG cell spheroids. Markers of functional activity were studied in spheroids of different size. The maximum size of spheroids that can be maintained in culture for 9 days without necrosis was determined.

We developed a method for collection and processing of scintigraphic data to estimate myocardial reserve in a gamma-chamber with cadmium-zinc-telluride detectors. Dynamic single-photon emission computed tomography of the heart with ^99mTc-Technetril was performed in 16 coronary heart disease patients at rest and during pharmacological load. During data processing, regions of interest from the cavity and the myocardium of the left ventricle were formed and activity–time curves were constructed. The index of myocardial blood fl ow reserve was calculated as the difference between two ratios of the mean gamma-count from the myocardial area to the area under the left ventricle cavity curve (peak) during load and at rest. The mean indices of myocardial reserve in healthy volunteers and patients with coronary artery atherosclerosis were 1.86 (1.59; 2.20) and 1.39 (1.12; 1.69), respectively. The development of the method for studying myocardial reserve by single-photon emission computed tomography is an urgent problem and requires further investigations.

We studied the mechanisms of erythropoiesis stimulation and effectiveness of Poetam, a preparation containing release-active anti-erythropoietin antibodies as a supplement treatment for experimental iron deficiency anemia during gestation. The results confirmed potency of combined therapy to stimulate erythropoiesis, which was more efficacious in comparison with monotherapy as assessed by the count of erythrokaryocytes and erythroid progenitors in the hematopoietic tissue as well as by the content of erythrocytes and hemoglobin in the peripheral blood. Activation of erythropoiesis is related to the modulatory effect of Poetam on proliferative activity and differentiation of erythroid precursors, which in most aspects results from stimulatory action of Poetam on secretion of the hematopoietically active factors by adherent elements of the hematopoietic microenvironment.

We analyzed the effect of hemolysis on microRNA profi le of blood plasma. It was found that hemolysis of ~0.05% erythrocytes in a sample signifi cantly affected the concentration of 9 microRNA: hsa-miR-486-5p, hsa-miR-16-5p, hsa-miR-451a, hsa-miR-106a-5p, hsa-miR-17-5p, hsa-miR-93-5p, hsa-miR-20a-5p, hsa-miR-107, and hsa-miR-20b-5p. The effect of hemolysis on plasma content of miR-17 family microRNA was demonstrated.

Effect of a new antioxidant enoxifol exhibiting antiplatelet activity in vitro and in vivo on hemostasis parameters was assessed in laboratory rats with experimental diabetes mellitus. Gliclazide, a hypoglycemic agent with antiplatelet properties, and pentoxifylline, a preparation improving blood rheology, were used as the reference drugs. Enoxifol produced a pronounced inhibitory effect on platelet aggregation in rats with experimental diabetes comparable to the effect of gliclazide and decreased blood viscosity thus demonstrating a significant effect comparable to that of pentoxifylline. In view of the fact that oxidative stress is a pathogenetic components of vascular complications in diabetes, it can be assumed that improvement of hemostasis parameters under the effect of enoxifol is determined by its antiplatelet and antioxidant activities.

Toxicity of human blood serum for ciliate Tetrahymena pyriformis is determined by the complement system. When ciliate are dying after being exposed to blood serum, cell membrane permeability for low-molecular-weight compounds significantly increases, probably due to pore formation. Serine protease inhibitors or exposure to physical factors inducing complement inactivation (e.g., heating up to 56°C) completely prevented ciliate death under the effect of human serum. Activation of serum complement upon interaction with Tetrahymena cells occurred by the classical or lectin pathway, while the contribution of the alternative activation pathway was negligible.

The expression of immune response gene mRNA in the umbilical and venous blood were compared in newborns of the first day of life with and without signs of infection. The expression of il1b, il6, il8, il10, il12a, il15, il18, tnfa, tgfb1, tbx21, gata3, foxp3, rorc2, cd45, cd68, cd69, tlr2, tlr4, tlr9, and mmp8 mRNA was evaluated in umbilical and venous blood cells of newborns by reverse transcription real time PCR. In full-term newborns without signs of infection, the expression of il8, tlr2, tlr4, and mmp8 in venous blood was higher than in umbilical blood, while in preterm newborns, the levels of mmp8 transcript were elevated while the levels of tlr9, cd45, and gata3 were reduced. The expression of some markers differed in the umbilical and venous blood and in newborns with congenital infectious disease and without signs of infection.

We analyzed the dynamics of the expression of transcription factors, VEGF and its receptor VEGFR2, serine-threonine protein kinase mTOR and activity of proteasome and calpain in patients with metastatic renal cancer during therapy with tyrosine kinase inhibitor Votrient and mTOR blocker Afinitor. The expression of hypoxic nuclear factor HIF-1α in the tumor tissue decreased during therapy with the target preparations. The decrease of VEGF and its receptor VEGFR2 was observed only in patients treated with mTOR inhibitor. The increase in calpain activity in the tumor tissue was observed in both groups. These findings extend our understanding of the mechanism of action of target anticancer preparations as allow considering the studied markers as predictors in choosing optimal therapy.

The effects of water-dispersed gold nanoparticles (8.0±0.9 nm in diameter) on the rat small intestinal mucosa and Peyer plaques, liver, and spleen were studied by electron microscopy. Water-dispersed gold nanoparticles injected into isolated intestinal loop not only accumulated in the small intestinal mucosa and Peyer plaques, but also penetrated into other organs, e.g. liver and spleen. Ultrastructural changes in the cells (hyperplasia of endoplasmic reticulum) were detected in the studied organs.

Morphological changes in the mesenteric lymph nodes of male C57Bl/6 mice and subpopulation composition of lymphocytes in these nodes were studied in experimental acute and chronic ulcerative colitis induced by sodium dextran sulfate. Acute and chronic ulcerative colitis was associated with the development of reactive changes in the mesenteric lymph nodes. These changes were of mixed type and were characterized by follicular hyperplasia and sinus reaction. The content of CD19⁺ B cells in the mesenteric lymph nodes decreased in acute ulcerative colitis, while the content of CD3⁺CD8⁺ cytotoxic T cells increased, which presumably reflected activation of Th1 reactions. The increase in the count of CD4⁺CD25⁺FOXP3⁺ regulatory T cells and CD3⁺CD8⁺ cytotoxic T cells was due to intensive migration of lymphocytes from the thymus and the colonic compartment of the local immune system. Chronic ulcerative colitis was associated with higher levels of CD19⁺ B cells and CD3⁺CD4⁺ T helper cells in the mesenteric lymph nodes, which was characteristic of adoptive immunity reactions and chronization of the inflammatory process.

Using functional magnetic resonance imaging, we studied intracerebral dynamics during completion of Raven’s Progressive Matrices test. Solving the test organized in sets of progressively increasing difficulty cause changes in cerebellar activation functionally related to cognitive activities and operations. As the tasks became more complicated, we observed gradual suppression of the activity of default mode network (DMN). The most pronounced changes in cerebral activation patterns occurred the second set of the test and involved associative somatosensory area and Wernicke’s area that is known to play an important role in cognitive processes associated with synthesis and analysis of information.