Vol 168, No 1 (2019)

Functional peculiarities of paralemniscal subdivision of the thalamocortical system were examined in normal Wistar and in WAG/Rij rats genetically prone to absence epilepsy. In 6-7-month-old WAG/Rij characterized by developed epileptic activity, the response of cortical somatosensory neurons to single electrical stimulation of the posterior thalamic nucleus was phasic, whereas in normal Wistar rats, similar reaction was tonic. The study views this phasic response as neural equivalent of spike-wave discharges known as typical EEG symptom of absence epilepsy.

The study examined stochastic stability of EEG samples recorded in succession from the same healthy volunteer at unchanged state. The absence of statistical repetitions not only for EEG samples, but also for the corresponding spectral densities was proven. A method to calculate the pairwise comparison matrices of ECG samples and quasi-attractors was proposed. The diagnostic effectiveness of these calculations was demonstrated during photostimulation, when successive EEG samples chaotically changed under overall homeostatic conditions.

We studied the structure of rhythms of BP, HR (by telemetric monitoring), electrolyte excretion (by capillary electrophoresis), and products of epiphyseal melatonin (by the urinary concentration of 6-sulfatoxymelatonin measured by ELISA) in normotensive Wistar-Kyoto rats and spontaneously hypertensive SHR rats maintained at 16/8 h and 20/4 h light-dark regimes. In Wister-Kyoto rats exposed to prolonged daylight, we observed changes in the amplitude, rhythm power (% of rhythm), and range of oscillations of systolic BP; HR mezor decreased. In SHR rats, mezor of HR also decreased, but other parameters of rhythms remained unchanged. Changes in electrolyte excretion were opposite in normo- and hypertensive rats. Under conditions of 20/4 h light-dark regime, daytime melatonin production tended to increase in normotensive rats and significantly increased in SHR rats. At the same time, nighttime melatonin production did not change in both normotensive and hypertensive animals. As the secretion of melatonin has similar features in animals of both lines, we can say that the epiphyseal component of the “biological clock” is not the only component of the functional system that determines the response of the studied rhythms to an increase in the duration of light exposure.

Male C57Bl/6J mice were exposed to daily 24-h illumination over 14 days and daily intragastrically received melatonin (1 mg/kg) or water (placebo). Controls were kept under standard day/night (14/10 h) conditions. Melatonin prevented the development of anemia in mice exposed to continuous illumination, which was proven by higher blood hemoglobin levels by the end of the experiment in melatonin-treated animals in comparison with the placebo group. Studies by the low-field NMR spectrometry detected lower lean body mass, total body water, and especially, fat content (by ~13%) in animals receiving placebo. Melatonin treatment led to an increase in the lean body mass and total body water on day 7 (in comparison with the placebo group) without affecting fat mass. On day 14 of continuous illumination, lean body mass increased in comparison with the corresponding parameter in the control and placebo groups. Melatonin had no effect on the physical endurance of mice exposed to continuous illumination (assessed by the grid hanging test).

In the presence of Fe³⁺ cation, S-nitrosoglutathione (GSNO) loses the potency to inhibit catalase in the system containing hemoglobin (an NO trap) with iron chelator or -SH inhibitor (a “sulfhydric poison” Hg²⁺). In the absence of hemoglobin, the inhibitory potency is retained in both cases. These properties are characteristic of dinitrosyl-iron complexes containing ferrous iron and thiols (DNIC/RSH). Since the potency to inhibit catalase results from the presence of -NO group, its loss in the presence of hemoglobin relates probably to transfer of this group to hemoglobin. The nitrosothiols are relatively stable compounds, so their ability to release NO under the action of iron chelators, which is characteristic of DNIC/RSH, can have important physiological implications, because the role of such chelators can be played by some endogenous agents as well. Thus, release of NO from the donor compounds can be controlled and regulated. Probably, the agents such as nitrosothiol+Fe³⁺ are the major constituents in the pool of nitroso compounds.

We studied antidepressant and antiparkinsonian properties of N-(5-hydroxynicotinoyl)-Lglutamic acid calcium salt (Ampasse) in rodents. It was found that Ampasse in a dose of 30 mg/kg exhibited antidepressant activity in the forced swimming test in mice and in a dose of 0.1 mg/kg maximally alleviates the symptoms of parkinsonian syndrome induced by systemic administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in C57Bl/6 mice, and haloperidol-induced catalepsy in rats.

We studied the effect of endomorphin-1 on isolated mesenteric lymphatic vessels in rats. It was found that endomorphin-1 caused a dose-dependent increase in the contractile activity of lymphangions, which was associated with stimulation of intracellular calcium depots. The observed effect of endomorphin-1 in isolated lymphatic vessels has a complex mechanism; it depends on the concentration of the applied peptide and is probably determined by its interaction with non-opioid receptors

We studied the effect of the organoselenium compound 2,6-dipyridinium-9-selenium-bicyclo[ 3,3,1]nonan dibromide (974zh) on the severity of pathological changes in the organs of experimental animals immunized with live tularemia and brucellosis vaccines. It was found that 974zh reduced reactogenicity of vaccines for experimental animals. Our findings indicate the prospects for further studies of the effects of 974zh on the functional state of experimental animals.

The effects of metabolites of Bacillus sp. strain M3 and Bacillus megaterium strain 8/75-1 isolated from permafrost formations, as well as a medicinal Bacillus cereus strain IP5832 on the secretion of TNFα, IL-1β, and IL-10 by mononuclear cells of human peripheral blood depending on the temperature of bacteria culturing (-16oC, -5oC, and 42oC) were studied. Metabolites of 8/75-1 and M3 strains cultured at 42oC produced more potent stimulating effect on TNFα and IL-10 synthesis than metabolites from these bacteria cultured at -16oC and -5oC (p<0.01). Temperature did not affect the ability of metabolites to modulate IL-1β synthesis in all bacterial strains and the ability of IP5832 strain metabolites to modulate TNFα production. IL-10 secretion was minimum under the effect of metabolites of IP5832 strain cultured at 42oC. The TNFα/IL-10 and IL-1β/IL-10 ratios indicated that metabolites of permafrost bacteria cultured at 42oC can shift the balance towards the synthesis of proinflammatory cytokines.

The role of the VNTR polymorphism of the AS3MT gene in determining the clinical features of schizophrenia and schizophrenic spectrum disorders was studied. The analysis included 670 individuals. We found no differences in PANSS scores for positive, negative, and common psychopathological symptoms between the carriers of different genotypes. The interaction of the studied polymorphism and obstetrical complications as an environmental factor was found. The genotype-environment interactions were identified for one of the characteristics reflecting the severity of schizophrenia: the level of negative symptoms. Women with the V2/V2 genotype, who have obstetrical complications, showed significantly higher negative symptoms scores, which was associated with a poor prognosis of the disease.

Studies by flow cytometry, scanning electron microscopy, and X-ray microanalysis have shown that fixation/internalization of quantum dots differ in lymphocytes and neutrophilic granulocytes. Punctate fixation of quantum dots to cell surface is characteristic of lymphocytes, while phagocytosis with perinuclear location of quantum dot aggregations is characteristic of neutrophils. The presence of complex functional groups on the surface of quantum dots can inhibit significantly the fixation/internalization of quantum dots by blood cells.

We evaluated the content of cell-free fetal DNA in maternal blood and expression of ZBP-1 receptors in the placental tissue of women with uncomplicated pregnancy, preeclampsia, and preterm labor. The study included 16 women with preeclampsia (early and late-onset preeclampsia, 8 cases each), 16 women with preterm labor, and 21 women with uncomplicated pregnancy. The concentration of cell-free fetal DNA was measured by PCR by detecting hypermethylated region of the RASSF1A gene. Immunohistochemistry was performed on paraffin-embedded sections of the placenta samples using primary polyclonal antibodies to ZBP-1. Significant increase in the level of cell-free fetal DNA was found in women with preeclampsia (both early and late-onset form) in comparison with uncomplicated pregnancy. The concentration of cell-free fetal DNA in preterm labor group did not differ from the control group; however, it was significantly lower than in early-onset preeclampsia, but not late preeclampsia. Immunohistochemical study showed higher expression of ZBP-1 in the villus syncytiotrophoblast in early-onset preeclampsia in comparison with that in preterm labor group (p=0.006). Fragments of damaged placental cells, predominantly trophoblast, enter maternal circulation and are the source of cell-free fetal DNA and a potential ligand for ZBP-1, which leads to further cell damage and the formation of a vicious circle. The increase in the content of cell-free fetal DNA in maternal blood and ZBP-1 expression in the syncytiotrophoblast in preeclampsia are interrelated processes reflecting impaired morphofunctional state of the placenta.

Prognostic significance of the proportion of cancer stem cells in cervical scrapings from 38 patients with uterine cervical cancer before treatment and after irradiation in a total dose of 10 Gy was assessed for immediate results of radio- and combined chemoradiotherapy evaluated by the degree of tumor regression in 3-6 months after the treatment. Cancer stem cells were detected as cells with CD44+CD24low immunophenotype by flow cytometry. The proportion of cancer stem cells in patients with the complete tumor regression decreased by on average 2.2±1.1% after irradiation, while in patients with partial regression this indicator increased by on average 3.3±2.3% (p=0.03). Multiple regression analysis revealed two independent indicators affecting tumor regression: the stage of the disease (which is quite expected) and change in the proportion of cancer stem cells after the first irradiation sessions (R=0.60, p<0.002 for the model in the whole). The proportion of cancer stem cells before the treatment did not have prognostic significance.

We studied the effects of intravenous and lymphotropic administration of bone marrow multipotent mesenchymal stromal cells and products secreted by these cells into conditioned medium on the blood and lymph circulation in the uterus and ovaries, as well as on folliculogenesis in female Wistar rats. It was shown that stromal cells and conditioned media of these cells administered via both routes lead to an increase in the number and diameter of blood vessels in the uterine wall and in the cortical layer of the ovaries. Neither mesenchymal stromal cells, not conditioned media affected the ovarian follicular apparatus.

We studied the effect of a bioactive peptide complex isolated from bovine serum on the proliferative potential and migration rate of mesenchymal stromal cells in vitro, as well as on the healing of modeled bone defects in rats. This bioregulatory preparation stimulated proliferation of mesenchymal stromal cells from deciduous tooth pulp in vitro, but did not affect the rate of their migration in two-dimensional cultures. In vivo experiments showed that application of this preparation in combination with hydroxyapatite and chitosan gel accelerated bone tissue regeneration, thus ensuring restoration of morphologically normal bone matrix. Thus, cattle blood serum is an available source for the production of bioregulatory preparations for medical purposes.

Elevated plus-maze test was used to reveal the role of excitation and inhibition mechanisms in the CNS in the organization of the behavior of male Wistar rats under normal conditions and during modeling experimental post-traumatic stress disorder (with or without administration of low doses of high-molecular-weight heparin). The data about insufficiency in the system of excitation mechanisms and pronounced excess in the system of inhibition mechanisms in post-traumatic stress disorder are prioritized. Heparin seems to be a promising and effective preparation to cope with extreme psychotraumatic abnormalities in CNS.

A translational rat model of chronic heart failure was employed to examine the cardioprotective effect of fabomotizole hydrochloride. Fabomotizole therapy for 28 days (15 mg/kg/day intraperitoneally) restored inotropic function of the left ventricle and increased ejection fraction from 54±3 to 65±3% (p=0.001). The inotropic function returned to normal against the background of significantly reduced myocardial expression of angiotensin (p=0.01) and glucocorticoid (p=0.03) receptors and significant increased expression of sigma-1 receptors (p=0.04). Inhibition of abnormal expression of angiotensin and glucocorticoid receptors responsible for activation of the pathological cascades underlying the postinfarction remodeling of the left ventricle as well as activation of the expression of cytoprotective sigma-1 receptors are viewed as the key features of the cardioprotective action of fabomotizole hydrochloride.

We studied antibacterial activity of a hybrid polymeric construction consisting of continuous and porous layers of ultrahigh-molecular-weight polyethylene reinforced by titanium. Experimental samples were impregnated with amoxycillin in subcritical Freon R22. The contact of bacterial culture with hybrid polymeric constructions saturated with amoxycillin suppressed the growth of microorganisms and the formation of their colonies. These results attest to the presence of a bactericidal effect of hybrid scaffold samples impregnated with an antibacterial component.

The risk of essential arterial hypertension was assessed in carriers of the NOS2 gene variants (rs1800482 (-954G>C), rs3730017 (C>T)). In subjects carrying C allele (rs1800482), the risk for essential arterial hypertension developing was higher by 1.7 times (OR=1.712, 95%CI 1.07-2.74), while the presence of T-allele (rs3730017) had a protective effect (OR=0.304, 95%CI 0.192-0.482). In patients with essential arterial hypertension, the presence of the C allele (rs1800482) was associated with a higher content of NO metabolites in the blood plasma. A positive correlation was found between the plasma content of nitrites and nitrates and the level of transcripts of VCAM1, ICAM1 genes in peripheral blood leukocytes. We found the influence of the C allele carriership on the expression VCAM1 and ICAM1 genes in patients with essential hypertension. It was hypothesized that this polymorphic site in the NOS2 gene can be involved in the development of endothelial dysfunction and essential arterial hypertension through modulation of NO level under condition of inflammation.

The presence of virus DNA integration into the cell genome was studied for 47 primary HPV16-positive patients with morphologically verified stage III cervical cancer. By using ROC analysis, the patients were divided into two groups: with and without HPV DNA integration into the host cell genome. The differences between the groups by the histological type, degree of tumor differentiation, and primary response to therapy were statistically insignificant. Virus DNA integration more than 7-fold reduced 5-year relapse-free survival and 1.7-fold reduced overall survival rate in comparison with patients without HPV DNA integration (p=0.0002 and p=0.05, respectively). The relative risk of adverse outcome of the disease in patients with the presence of HPV16 DNA integration increases by 4 times over a period of less than 3 years (р=0.0006) at high AUC level. The probability of earlier progression of the disease in patients with of HPV DNA integration calculated according to the Cox proportional hazards model was 85.5% (hazard ratio 5.96; p=0.002). Thus, the results suggest that the presence of HPV16 DNA integration into the cell genome is an independent factor in predicting clinical outcome of advanced cervical cancer and can serve as an effective criterion for the individual choice of treatment tactics for the patients.

Combined chronic treatment of Ehrlich solid carcinoma (EC) with an NOS inhibitor 1-isobutanoyl-2-isopropylisothiourea hydrobromide (T1023) and a PDK1 inhibitor dichloroacetate was accompanied by statistically significant synergetic antitumor effects manifested in a significant and stable suppression of neoplasm growth (by 55-65%). Separate treatment with T1023 and dichloroacetate induced moderate short-term inhibition of tumor growth (by 30-35%) followed by weakening of tumor sensitivity to these substances. These results attest to synergetic antitumor effects NOS inhibitor T1023 and PDK1 inhibitor dichloroacetate producing antiangiogenic and hypoxia-targeted cytotoxic effects, during their combined administration, which allows overcoming the adaptive potential of the tumors.

The effects of composite fibroin-gelatin microparticles (100-250 μ) on the rate of wound healing and regeneration under conditions of contraction prevention were studied on the model of splinted full-thickness skin wound in a mouse. Subcutaneous injection of these particles into the defect area accelerated wound healing and promoted re-epithelialization and recovery of normal structure of the epidermis. In addition, the composite microparticles promoted the formation of connective tissue of characteristic structure, replacing the derma over the entire defect, and stimulated regeneration of subcutaneous muscle (panniculus carnosus) and skin appendages (sebaceous glands and hair follicles).

The effects of a fluroquinolone levofloxacin on apoptosis of peripheral blood lymphocytes from patients with infiltrative pulmonary tuberculosis were studied in vitro. It was found that levofloxacin stimulated apoptotic cell death in tuberculosis. Addition of levofloxacin to cell suspension from patients with drug-susceptible form of tuberculosis led to an increase in the number of CD95+ and AnnV+ lymphocytes. In patients with drug-resistant form of tuberculosis, only the number of apoptotic lymphocytes, but not the count of CD95+ cells increased under these conditions.

We performed a comprehensive clinical and morphological examination of 70 menopausal patients aged 42-62 years with dysuria and chronic pelvic pain. Assessment of the bladder and vaginal microcirculation by laser Doppler flowmetry in menopausal patients with genitourinary syndrome revealed reduced (by 12-65%) microcirculation parameters. Analysis of the quantitative and qualitative composition of the bacterial microflora of the urethra and vagina revealed abnormalities of microbiota of varying severity, which can be the cause of infectiousinflammatory processes in the pelvic organs leading to chronic pelvic pain syndrome and incontinence. During menopause, the genitourinary syndrome in women is associated with the combined development of atrophic changes in the bladder and uterine mucosa that by their morphological characteristics differ from age-related involutive changes. Atrophy (hypoplasia) of the endometrium and bladder mucosa develops against the background of pronounced fibrosis and is accompanied by hyperemia and hemorrhages.

We studied the effect of IL-1β on the permeability of brain capillaries in healthy mice. Intravital microscopy demonstrated that parenteral administration of IL-1β was followed by an increase in vascular permeability ensuring passage of free Alexa488 fluorescent label through the capillary walls, but not sufficient for penetration of liposomes. In addition, experiments on mice with intracranial M6 glioma showed penetration of liposomes through the walls of tumor capillaries after parenteral administration of IL-1β in a concentration of 2 μg/ml. Thus, the use of proinflammatory cytokine IL-1β in the therapy of brain tumors can significantly increase the therapeutic efficacy of drug delivery systems, in particular, for drugs poorly crossing the blood—brain barrier.

We studied the effect of short-term hypoxic stress on the phenotypic polarization of monocyte-derived macrophages and their secretory activity during interaction with mesenchymal stromal cells. In the presence of mesenchymal stromal cells, monocyte-derived macrophages exhibited the signs of M2 polarization, which was evidenced by increased expression of CD206 and CD163 markers, as well as increased transcription and translation of IL-6. Short-term hypoxic stress promoted a shift of macrophage phenotype from inflammatory M1 towards anti-inflammatory M2 in monoculture and co-culture with mesenchymal stromal cells. In addition to the immunoregulatory action, mesenchymal stromal cells demonstrated stromal activity and maintained high viability of monocyte-derived macrophages.