Vol 167, No 4 (2019)

A new phenomenon (or paradox) consisting in higher statistical stability of samples, for example, tremorograms, in a group of different subjects than in the same subject in unchanged homeostasis. At the same time, it turns out that 15 different subjects are more similar (statistically) to each other than one person to himself in 15 repetitions of tremorogram recording (in unchanged homeostasis). This poses serious problems for physiology and medicine, and in the context of transition to individualized medicine, this raises the question of how different people can be compared if they are statistically more similar than one person to himself.

The study examined the effects of hyperpolarization-activated funny current (If) on HR and coronary flow in Langendorff-isolated hearts from newborn rats. Blockade of If current with ZD7288 changed the examined cardiac parameters. The blocker in a concentration of 10^—9 M decreased HR by 26.8% (p≤0.05). In concentrations 10^—8, 10^—7, 10^—6, and 10^—5 M ZD7288 produced minor differently directed effects. In a concentration of 10^—5 M, ZD7288 reduced coronary flow in the isolated heart (p≤0.01). In other concentrations, the blocker produced no significant effects on coronary flow.

Cardiac activity parameters were analyzed in rat fetuses with preserved placental circulation on gestation days 17-20 after administration of muscarinic receptor blocker atropine either directly to the fetuses, or to the females. The obtained results attested to the absence of chronotropic effect from the cholinergic system in rats during the fetal period. At the same time, changes in heart rate variability observed after atropine injection showed that cholinergic receptors are involved into heart rhythm regulation in rat fetuses and that their role increases from the 17th to 20th day of the embryonic period.

We studied the peculiarities of oxidative and substrate phosphorylation in isolated cardiomyocytes and myocardial mitochondria in 2- and 15-month-old rats. It was shown that age factor reduces the degree of oxidation and phosphorylation coupling both in cardiomyocytes and in mitochondria, which was associated with the decrease in the rate of consumption of energy resources.

The anticonvulsant effect of ethosuximide (T-type calcium channel blocker) was evaluated in Krushinsky–Molodkina rats predisposed to audiogenic epilepsy. Ethosuximide given with drinking water (300 mg/kg/day) over 45 days slightly reduced proneness to audiogenic epilepsy and increased locomotor activity of the animals at the periphery of the open field. Neonatal administration of ethosuximide (3-4 mg per animal, from 2 to 10 days of life) insignificantly modulated the parameters of audiogenic epilepsy in these animals at the age of 1.5 months and reduced manifestation of audiogenic myoclonic convulsion that developed after long daily sound presentation started at the age of 3 months. The findings attested to a weak anticonvulsant effect of ethosuximide on tonic convulsions with its predominant effect on convulsions with forebrain focus location.

We studied the effect of sulfated glycosaminoglycan on the infection properties of high-virulence Dal’negorsk strain and low-virulence Primorye-437 of tick-borne encephalitis virus. Differences in reproductive activity of these strains and their tropism to the target cells were revealed. Glycosaminoglycan reduced pathogenetic activity of high-virulence strain in vitro, but had no effect on low-virulence strain. The interaction of imperfect virus particles of non-pathogen strain with the glycosaminoglycan led to their accumulation in cell, but in the culture medium of SPEV cells infected with experimental and control samples, accumulation of virus particles did not differ. The results on activity of glycosaminoglycan binding with strains differing by their biological and molecular genetic characteristics can be used to assess their pathogenic potential.

We studied antibacterial properties of organo-inorganic hybrid coatings on the AMg2 aluminum alloy including superhydrophilic and superhydrophobic nanotextured metal substrates with applied bacteriophage particles. Bactericidal activity of surfaces after artificial contamination with a bacterial suspension was evaluated. To increase bactericidal effect of the plates, bacteriophage was sorbed on their surface. In the experiments simulating possible spreading of HAI pathogens, higher bactericidal activity of superhydrophilic surfaces in comparison with superhydrophobic ones. Application of bacteriophage particles did not prevent primary colonization of textured metal surfaces by strains used in the experiment, but in some cases increases their bactericidal activity.

The parameters of sperm head in Wistar male rats orally treated with antifungal agent for 48 days during spermatogenesis were studied by the method of image analysis. The degree of roundness (roughness) of sperm head was calculated. Significant differences in morphometric parameters of sperm head, such as length, width, head angle, and roundness were revealed between the treatment and control group. The index of deformation of sperm head in the treatment group rats was 4.93 arb. units. These data indicated microcephaly accompanied by the enlargement of the head, transition of an acute angle to a right angle, and acquiring of a round form. Potential gonadotoxicity was confirmed by the analysis of the functional activity of spermatozoids of male rats (increased count of spermatozoa with head pathology), fertilization ability (enhanced fertility index), and genotoxicity (increased number of chromosomal aberrations polychromatophilic erythrocytes of murine bone marrow). These changes can be responsible for reduced fertility.

We studied proliferative activity of colorectal cancer cells with different expression level of CD133 molecule associated with cancer stem cells phenotype. Analysis of BrdU incorporation into Caco-2 and HT-29 cell lines showed that the percentage of cells in the DNA synthesis phase in the CD133^+/high population is higher than in CD133^—/low population. The expression of proliferation marker Ki-67 and the percentage of Ki-67⁺ cells were also higher in the CD133^+/high population. Colorimetric analysis with crystal violet dye showed that the number of cells after 10-days culturing was higher in the CD133^+/high population in both cell lines. These findings suggest that cells with high level of CD133 expression are characterized by higher proliferative activity, which can contribute to the tumor progression.

Morphological changes in the allograft of rat anterior cerebral vesicle at the early stages after transplantation into the peripheral nerve of an adult rat were studied by immunohistochemical methods. Immunohistochemical reaction to bromodeoxyuridine showed that the delay of mitotic division in neural stem/progenitor cells in the grafts occurred during S/G2 stage. In transplants of rat embryonic neocortex (E13), changes in the cell cycle of neural stem/progenitor cells in 3 h after transplantation into the nerve correlated with abnormal distribution of adherens junctions and interkinetic nuclear migration.

We analyzed the expression of transcriptional factor Oct4 in rat adrenal cortical cells during postnatal development. It was found that Oct4 is expressed by typical cortical cells of the zona glomerulosa, zona fasciculata, and zona reticularis in pubertal and postpubertal periods. The maximum number of Oct4⁺ cells was found in the zona glomerulosa. An inverse correlation between the number of Oct4⁺ glomerulosa cells and serum level of aldosterone both in pubertal and postpubertal periods was revealed. After puberty, the number of Oct4⁺ glomerulosa cells directly correlated with the number of Ki-67⁺ cells. A hypothesis was put forward that Oct4 is involved in postnatal morphogenesis, regeneration, and functioning of the adrenal cortex.

We analyzed changes in multipotent mesenchymal stromal cells of patients with chronic myeloid leukemia before discontinuation of tyrosine kinase inhibitors. Withdrawal syndrome was significantly more common in patients who have been taking tyrosine kinase inhibitors for a longer time and in patients of older age and with lower body weight. In patients with withdrawal syndrome, the total production of mesenchymal stromal cells and expression of FGFR2 and MMP2 genes were significantly lower; loss of deep molecular response was also less frequent in this group of patients. At the same time, the expression of genes important for the maintenance of stem cells (SOX9, PDGFRa, and LIF) was significantly lower in the mesenchymal stromal cells of patients with withdrawal syndrome and loss of deep molecular response. We observed a clear-cut relationship between the development of withdrawal syndrome and the loss of deep molecular response. The decrease in the expression of FGFR2 and MMP2 genes in the mesenchymal stromal cells of patients with chronic myeloid leukemia before discontinuation of treatment can be a predictor of withdrawal syndrome, while simultaneous decrease in the expression of SOX9, PDGFRa, and LIF in these cells attests to undesirability of therapy discontinuation at the moment.

The effect of porosity on diffusion characteristics of scaffolds and invasive capacity of MCF-7 and PC-3 tumor cells was studied for gelatin hydrogels. According to MTS test results, the efficiency of population of a macroporous cryogel by cells applied by different techniques increased in the following order: migration from the monolayer

We studied the effects of neonatal administration of non-opioid leu-enkephalin analogue peptide NALE) on the morphological parameters of the liver and redox status of 60-dayold albino rats subjected to antenatal hypoxia. In animals subjected to antenatal hypoxia, a decrease body weight and reduction of the area of hepatocytes and total area of their nucleoli were observed; these changes were accompanied by activation of free-radical processes and impairment of antioxidant defense in the liver and blood serum. Intraperitoneal administration of NALE (100 μg/kg, daily) during postnatal days 2-6 normalized body weight, hepatocyte area, and total area of their nucleoli, significantly reduced the intensity of ROS generation, and improved antioxidant protection in the liver and blood serum in 60-day-old animals subjected to antenatal hypoxia.

A system for detection of malignantly transformed cells, including follicular lymphoma Bcells, was developed and experimentally validated. The system is based on the use of bacteriophages carrying exposed ligands for pathogenic B-cell receptors. The efficiency of binding to target cells is several times higher than in systems with chemically synthesized biotinylated peptides. The new method is proposed as a noninvasive diagnostic test for mapping B-cell lymphoma and for determining the specificity of B-cell receptors and high-throughput combinatorial selection of various repertories of B cells.

Focal unilateral traumatic brain injury in the sensorimotor cortical region disturbed the functions of contralateral limbs controlled by the damaged hemisphere. A single intravenous injection of methylene blue (1 mg/kg) immediately before or 30 min after the injury significantly weakened functional disorders in the affected extremities. In vitro experiments showed that methylene blue effectively reduced death of cultured neurons provoked by paraquat or zinc ions producing the toxic effects on mitochondrias.

We studied the role of native α-fetoprotein preparation in the regulation of proliferation and functional activity of naïve T cells and immune memory T cells in vitro. The study was carried out on separated fractions of naïve T cells (CD45RA⁺) and immune memory T cells (CD45R0⁺) incubated with α-fetoprotein under conditions of TCR activation. At the level of naïve T cells, α-fetoprotein in a concentration of 100 U/ml reduced the expression of CD28, but increased the expression of CD25, while at the level of immune memory T cells α-fetoprotein (50 and 100 U/ml) only suppressed the expression of CD25. No effects of α-fetoprotein on the proliferative status of the studied lymphocyte subpopulations and on the expression of CD71 (proliferation marker) by these cells were detected. Addition of α-fetoprotein in a concentration of 100 U/ml increased the level of IL-2 in naïve T cell culture supernatants, while production of IL-2 by memory T cells remained unchanged. These data demonstrated the priority aspects of regulation of the functional activities of naïve T cells and immune memory T cells.

A total of 100 women with early pregnancy loss were recruited and further classified into two subgroups: sporadic pregnancy loss and recurrent pregnancy loss; each subgroup consisted of 50 women. The control group included 56 women with normal pregnancies. Genotyping was performed by PCR with restriction fragment length polymorphism analysis. A statistically significant increase in the frequencies of TT genotype and T allele for DNMT3B rs2424913 polymorphism was found in the total patient group and in both patient subgroups in comparison with the control. Moreover, homozygous TT genotype was associated with increased risk of early pregnancy loss (both sporadic and recurrent). DNMT3B rs2424913 gene polymorphism in women can be used a marker of predisposition to early pregnancy loss and recurrent pregnancy loss.

Antiviral activity of a Russian drug flakozid towards infection caused by a cytopathogenic variant of hepatitis C virus in SPEV cells is studied. Flakozid is an individual natural flavonoid glycoside 7-O-β-D-glucopyranoside-8-(methyl-but)-2-enyl)-5,4’-dioxyflavanolol), isolated from the leaves of Phellodendron amurense Rupr. and Ph. amurense var. lavallei (Dode) Sprague, Rutaceae family. High antiviral activity of flakozid manifests in response to its addition into the monolayer of SPEV cells infected by hepatitis C virus during all periods of experiment. Flakozid exhibits weak cytotoxicity and no viricidal activity towards hepatitis C virus, which is comparable to activity of ribavirin (antiviral drug). Chemotherapeutic index of flakozid is 15-25 times higher than that of the reference drug ribavirin.

The study focused on activation of lymphocytes derived from healthy donors and cancer patients in the medium supplemented with IL-2 and IL-15 at low concentrations. We studied morphological features of cultured cells, their viability, and proliferative and functional activities during culturing for up to 14 days. In comparison with lymphocytes derived from cancer patients, white blood cells derived from healthy donors demonstrated a number of advantages under the given culturing conditions such as higher viability, greater proliferative activity, and the potency for earlier activation; moreover, during activation they secreted the anti-inflammatory cytokines IL-4 and IL-10 at greater concentrations.

We analyzed biocompatibility, elastic-strength properties, and biointegration potential of a new biomaterial made of xenopericardium for reconstructive cardiovascular surgery. The biomaterial manufactured by the proposed technology demonstrated high biocompatibility and biointegration potential and its elastic-strength properties 2-4-fold surpassed that of native pericardium. The obtained results attested to good prospects of using the proposed technology for preparing biomaterials for reconstructive cardiovascular surgery.

We studied the expression of ARID1A, prostaglandin E2 synthase, and prostaglandin E2 receptor in the endometrium and ovarian, peritoneal, and intestinal endometrioid heterotopies in women with endometriosis of young and middle reproductive age. ARID1A protein is a tumor suppressor, its expression reduced in different types of cancer. Prostaglandin E2 synthase and prostaglandin E2 receptor are involved in the signaling cascade of inflammatory reactions presumably underlying the development of endometriosis. In endometrioid heterotopies, expression of ARID1A was reduced in 1.2-4.0 times, the expression of prostaglandin E2 synthase and prostaglandin E2 receptor was reduced in 2.9-5.2 times These findings suggest that ARID1A, prostaglandin E2 synthase, and prostaglandin E2 receptor can be used as predictors of malignant transformation of endometrioid heterotopies in women with endometriosis.

We studied neuronal death in the sensorimotor cortex, hippocampus, and supraoptic and paraventricular nuclei of the hypothalamus and dynamics of HER-2/neu expression in late ontogenesis in young and old transgenic HER-2/neu mice. Wild-type FVB/N mice served as the control. The intensity of apoptosis (TUNEL) and HER-2/neu expression (Western blotting) in the same brain regions were measured. HER-2/neu was detected in the cortex, hippocampus, and hypothalamus of transgenic and wild-type mice, and its expression increased with age. The effect of HER-2/neu on the intensity of cell death in various brain regions depended on the stage of ontogenesis and animal genotype. Enhanced expression of HER-2/neu determines low rate of cell death in the studied brain regions during pathological ageing.

The effects of environmental factors (flavors, different ethanol concentration, alcoholic deprivation, and food reinforcement) on the formation of alcohol motivation was studied in rhesus macaques (Macaca mulatta, n=6). Motivation for alcohol intake was induced in two stages: initiation (sessions 1-160) and formation of motivation (sessions 161-516). Monkeys preferred multifruit flavor and 4% ethanol solution, while ethanol deprivation did not stimulate alcohol consumption. The pronounced individual differences in the pattern of alcohol motivation were revealed: the intake of 4% ethanol solution ranged from 0.21±0.03 to 0.43±0.06 g/kg without food reinforcement and increased from 0.78±0.03 to 1.22±0.09 g/kg with food reinforcement. The results suggest that the proposed method is valid and can be used as an experimental model of alcohol dependence in non-human primates.

The effects of inhalation exposure to a mixture containing acetone, acetaldehyde, and ethanol in concentrations typical of closed environment in space vehicles on the structure of mesenteric lymph nodes in F1 male mice were studied by histological technique. The long-term exposure to modeled atmosphere led to pronounced structural changes in these nodes that were clearly seen on day 22 and increased by day 36 of the experiment. The thickness of the capsule and trabeculae of mesenteric lymph nodes as well as diameter of lymphatic sinuses did not differ from the control values up to day 8, but then increased on days 22-70. Starting from day 22, the thickness of the medullary cords decreased and attained the minimum to the end of the experiments, which can indicate depletion of immunocytopoiesis and inhibition of humoral immunity. The present data are important for the space medicine, because they indicate structural changes in the peripheral lymphoid organs, the key elements of immune system.

A model of moderate hyperhomocysteinemia associated with mechanical injury of the musculoskeletal system was developed and experimentally substantiated. The adequacy of this model for studies of morphological and functional regularities is verified. This model can be used for the development of a new concept of evaluation of thrombotic complications of mechanical injury.

Olfactory ensheathing cells showed significant effects on the regeneration of the spinal cord in experimental models and in clinical trials. However, the use of these cells in the therapy of posttraumatic cysts of the spinal cord has not been studied. Cultures of human and rat olfactory mucosa were obtained according to the protocols developed by us. Passage 3-4 cultures are most enriched with olfactory ensheathing cells and are preferable for transplantation. We performed transplantation of 750,000 olfactory ensheathing cells into the region of modeled cysts. The therapeutic effect of human cells was more pronounced. The positive dynamics of recovery of motor activity in the hind limbs of rats can reflect regenerative processes in the spinal cord after transplantation of olfactory ensheathing cells into the region of posttraumatic cysts.