Vol 165, No 2 (2018)

Changes in the frequency characteristics of EEG alpha rhythm in during falling asleep were studied in three healthy individuals under conditions of long-term isolation (MARS-500 project). Falling asleep was preceded by enhanced alpha rhythm frequency. An inverse correlation between the duration of falling asleep and prevailing alpha rhythm frequency during active and relaxed wakefulness was revealed in the left hemisphere. These results demonstrate the principal possibility of predicting the duration of falling asleep by using alpha rhythm spectral analysis. It is assumed that the frequency of the alpha range spectral peak can be a marker of drowsiness and reflect the current need for sleep.

We studied the effects of social isolation for 1, 2 or 3 weeks on behavioral reactions of male rats. As social isolation is a common model for inducing depressive-like state in rodents, classical tests for depressive-like behavioral features were conducted: Porsolt forced swimming test and tests for anhedonia and social interest. None of the experimental groups showed statistically significant disorders in comparison with the control group kept under standard conditions. Thus, social isolation for up to 3 weeks did not cause behavioral abnormalities in male rats. Single housing can be used in other models of induction of depressive-like states, but the use of this paradigm as an independent model for the development of depressive-like behavior requires longer period of social isolation.

The parallel placebo-controlled study examined the therapeutic effects of dual-target repetitive transcranial magnetic stimulation (rTMS) of the motor cortex (bilaterally) and the left prefrontal cortex (dorsolaterally) on spontaneous and mitogen-stimulating synthesis of pro- and anti-inflammatory cytokines by the blood cells and the level of brain-derived neurotrophic factor (BDNF) in blood serum of patients with Parkinson’s disease. The significantly steeper positive clinical dynamics (assessed by UPRSD scale) observed in rTMS group in comparison with the placebo group was accompanied by a significant drop in spontaneous production of proinflammatory cytokines IFNγ and IL-17A. rTMS produced no significant effect on serum BDNF. The possible mechanisms of rTMS therapeutic action on the level of cytokines associated with neuroinflammation in patients with Parkinson’s disease are discussed.

The study examined the effect of bone marrow multipotent mesenchymal stromal cells and their products secreted into the conditioned medium on uterine microcirculatory bed in Wistar rats during chronic inflammation. The parameters of blood and lymphatic microcirculation in the uterus changed in various directions in relation to the administration routes of biomedical cell product, which is important when the cell therapy is employed in the treatment of inflammatory-degenerative alterations in the lesser pelvis organs.

А cross-sectional study was performed on a continuous sample of patients (N=52) with the gastroesophageal reflux disease. Specific features of this disease in patients with overweight were revealed. Regurgitation predominated in the structure of complaints. Fibrogastroduodenoscopy revealed endoscopically negative form of the diseases in 38% of cases. According to 24-h pH monitoring results, alkaline reflux pH>7 took more than 25% time in supine position at high DeMeester score, which limits the usage of aggressive acid-lowering therapy.

Cells of E. coli isolates from the gut of healthy volunteers (N=5) and patients with Crohn’s disease (N=5) and laboratory E. coli strain DH5α bound mucin in vitro in similar amounts ranging from 0.02 to 0.12 mg/mg of bacterial dry weight. Binding was evaluated by the decrease in optical absorption of mucin solution at 214 nm after incubation with bacteria. Detailed analysis of mucin binding by one of isolates showed that during incubation of 0.09 mg/ml bacteria in 0.15 M NaCl containing 0.1 mg/ml mucin at 25^oC, maximum binding was reached in 30 min, while in the presence of 14 mM α-methyl mannoside, mucin binding decreased by 46% (p<0.05). Confocal microscopy revealed intensive binding of FITC-labeled mucin to the surface of a small number of bacterial cells. Mucin binding did not significantly affect zeta potential of bacteria and their energetic status assessed by ATP content; at the same time, ATP content in the extracellular environment slightly increased.

Antiviral efficiency of oxidized dextrans (OD) with different molecular weights and oxidation degree (OD40min, OD70min, OD40max, and OD70 max) was studied in vitro and in vivo. Dextrans OD40max and OD70max prevented the development of the cytopathic effect of influenza A(H1N1)pdm09 virus in more than 50% MDCK cells vs. control (no OD). Four intranasal doses of OD40min, OD40max, and OD70min and one intranasal dose of OD70max before infection of BALB/c mice with A(H1N1)pdm09 influenza virus significantly reduced mortality and prolonged life span in comparison with controls receiving saline. These and our previous data attest to clear-cut preventive effect of OD in influenza infection.

The major salivary glands of rats release into the saliva and blood a wide spectrum of bioactive substances, essential for many organs, including the testes. Sialoadenectomy leads to the development of degenerative changes in the cells of the twisted testicular tubules. However, the effects of bioactive factors released by the major salivary glands on the morphology and function of Leydig cells remain little studied. Sialoadenectomy in adult rats led (in 1-4 weeks) to a decrease in the nuclear and cytoplasmatic areas of Leydig cells, violation of the plasmalemma integrity, dilatation of perinuclear space and agranular endoplasmatic reticulum vesicles, and to destruction of the mitochondria. Ultrastructural changes caused by sialoadenectomy completely resolved by week 6 of the experiment at the expense of compensatory activation of the synthesis of the major salivary gland factors by other sources in the organism of rats.

A method of measuring of glutathione peroxidase activity using H₂O₂ was adapted for homogenates of murine brains. If the amount of reduced glutathione was at the constant level of 0.55 mM, the concentration of H₂O₂ of 0.192 mM was saturating for glutathione peroxidase of murine brain and was selected as an optimal concentration for the estimation of enzyme activity in tris-HCl buffer with addition of NaN₃ and EDTA (pH 8.5) at the incubation temperature of 37°C. The homogenates were dissolved by the reaction mixture by 10.4 times. The duration of incubation did not exceed 60 sec, if 13% homogenate was used. The experiment based on this method showed increased activity of glutathione peroxidase in the brain of mice treated with a derivative of acetaldehyde ammonia during long-term intermittent normobaric hypoxia. These data might reflect activation of glutathione peroxidase.

Oxidative kidney injury was compared in newborn and adult rats under conditions of ischemia/reperfusion and in experimental model of systemic inflammation induced by endotoxin (LPS of bacterial cell wall) administration. Oxidative stress in the kidney accompanied both experimental models, but despite similar oxidative tissue damage, kidney dysfunction in neonates was less pronounced than in adult animals. It was found that neonatal kidney has a more potent regenerative potential with higher level of cell proliferation than adult kidney, where the level proliferating cell antigen (PCNA) increased only on day 2 after ischemia/reperfusion. The pathological process in the neonatal kidney developed against the background of active cell proliferation, and, as a result, proliferating cells could almost immediately replace the damaged structures. In the adult kidney, regeneration of the renal tissue was activated only after significant loss of functional nephrons and impairment of renal function.

We analyzed changes in angiotensin-converting enzyme activity in rat aorta at the early terms after irradiation in doses equal to one fraction dose used in tumor radiotherapy. Male Wistar rats were exposed to whole body or local (chest) X-ray irradiation (200 kV, 1-7.5 Gy). The activity of the enzyme in aorta segments was measured in 1-48 h after irradiation by hydrolysis of hippuryl-histidine-leucine. Activity of angiotensin-converting enzyme in rat aorta was increased 1-24 h after whole body irradiation in a dose of 2.5 Gy with a peak in 2 h after exposure. After local exposure, enzyme activity also increased in 2 h, but returned to the control level in 24 h. In 2 h after whole-body irradiation in doses >2.5 Gy, the increase in enzyme activity was less pronounced and after exposure to 7.5 Gy, it did not differ from the control. During local exposure, the effect did not decrease with increasing the irradiation dose. The fraction of blood monocytes adherent to plastic in rats subjected to whole body irradiation decreases with increasing the dose. In rats subjected to local irradiation in a dose of 7.5 Gy, monocyte adhesion to plastic did not differ from the control. These data suggest that the increase in activity of angiotensin-converting enzyme in the aorta after irradiation is determined by monocyte adhesion to the endothelium; the decrease in this effects with increasing the dose can be explained by radiation damage of monocytes.

We studied combined effect of β-hairpin antimicrobial peptide tachyplesin I and cytotoxic agent cisplatin on tumor and normal human cell lines. MTT assay and flow cytometry showed that tachyplesin I selectively sensitized cancer cells to cisplatin in specified concentration ratios. In vitro experiments demonstrated that combined use of tachyplesin I and cisplatin allows decreasing the effective dose of the cytostatic thus reducing nonspecific toxicity.

We studied the effect of hormones estriol, ghrelin, kisspeptin, and chorionic gonadotropin in concentrations corresponding to their content in the peripheral blood in each trimester of pregnancy on the expression of membrane molecules on myeloid and plasmacytoid dendritic cells of the thymus. It was found that thymic myeloid dendritic cells are sensitive to the action of estriol and kisspeptin. Estriol in a concentration of the first trimester of pregnancy reduces the number of myeloid dendritic cells expressing receptor for thymic stromal lymphopoietin (CD11c+TSLP-R⁺) and inhibitory molecule B7-H3 (CD11c⁺CD276⁺). In contrast to estriol, kisspeptin regulates the processes of differentiation of thymic myeloid dendritic cells in concentrations typical of the second-third trimesters and reduced their total number (CD11c⁺) and the number of cells expressing TSLP-R (CD11c⁺TSLP-R⁺). Estriol and kisspeptin do not affect the total number of plasmacytoid dendritic cells (CD303⁺) and expression of TSLP-R and CD276 by these cells. Ghrelin and chorionic gonadotropin in the studied concentrations had no significant effect on the total number of thymic myeloid and plasmacytoid dendritic cells and on the expression of membrane molecules of TSLP-R and CD276.

We developed a pyrosequencing protocol for monitoring of stability of attenuating mutations in the genome of vaccine reassortants based on master donor virus of Russian live attenuated influenza vaccine B/USSR/60/69. The developed protocol allows rapid and accurate assessment of mutations and can be used for analysis of genetic stability of reassortants during vaccine strain development and manufacturing, as well as genetic stability of vaccine isolates of influenza B virus during pre-clinical and clinical trials.

To determine the role of NMDA receptors in the functional regulation of immunocompetent cells, comparative assay was carried out for genes expressed in the mononuclears in peripheral blood of healthy persons under normal conditions and after blockade of these receptors. The genes, whose expression changed in response to blockade of NMDA receptors in mononuclears, encode the products involved in regulation of the major functions of immune cells, such as proliferation (IL4, VCAM1, and CDKN2A), apoptosis (BAX, MYC, CDKN2A, HSPB1, and CADD45A), activation (IL4R, IL4, VCAM1, and CDKN2A), and differentiation (IL4, VCAM1, and BAX).

We analyzed the expression of galectin-1 and galectin-3 in tumor tissue in stomach and colorectal cancer with and without tissue eosinophilia. Low expression of galectin-3 was detected in all patients with malignant gastrointestinal tumors irrespective of the presence of eosinophilia. Low expression of galectin-1 was detected only in patients with gastrointestinal cancer associated with eosinophilia. Association of galectin-1 expression with eosinophilic infiltration of the tumor tissue in stomach and colorectal cancer was detected.

We have previously developed a polycaprolactone (PCL) vascular graft with incorporated vascular endothelial growth factor (VEGF). Functioning of the PCL/VEGF graft in rat circulatory system over 1, 3 and 6 months after implantation into abdominal aorta was tested. Graft patency and formation of vascular wall elements were assessed histologically and by immunofluorescence staining for von Willebrand factor, CD31, CD34, and collagens I and IV and DAPI staining. Local application of VEGF promoted endothelialization and improved patency of the graft. The wall of the PCL/VEGF graft underwent remodeling due to active cellular infiltration and the extracellular matrix deposition.

Changes in the biochemical composition of the tear film is a critical risk factor for the development of chronic perioperative dry eye syndrome, because increasing the duration of general anesthesia did not affect the dynamics of tear secretion recovery, but slowed down normalization of its structure and antioxidant activity in the post-anesthetic period.

Store-operated channels activated in response to intracellular calcium store depletion represent the main pathway of calcium entry from the extracellular space in nonelectroexcitable cells. Adapter proteins organize the components of this system into integral complex. We studied the influence of adapter proteins of the Homer family on endogenous store-operated calcium Imin channels in A431 cells. Monomeric Homer 1a proteins increase activity of Imin channels, but did not modulate their electrophysiological properties. Recombinant Homer 1c protein did not block the induced calcium currents.

The numerical density of secretory granules dense-core vesicles (DCV) in the cytoplasm of pinealocytes of the pineal gland was estimated by transmission electron microscopy in male white mice and Wistar rats. The 3-day biorhythm and lunaphase changes in the DCV content in the perikaryon and the processes of pinealocytes, which are manifested significantly in different seasons of the year, are established. The three-day biorhythm in adult male mice in comparison with younger male rats is not expressed uniformly in different phases of the moon. The in-phase manifestation of infradian biorhythms in different species of animals during the year with an unchanged daily photophase indicates the existence of common external synchronizers for mammals of these biorhythms that are not associated with the light/dark cycle.

We propose an optimized single-step assay of total nitrite and nitrate concentrations based on serum deproteinization by ultrafiltration. After deproteinization, the residual protein concentration was 0.0800±0.0007 mg/ml, which secured necessary transparency of the specimens in contrast to methanol deproteinization. The optimized assay for serum nitrite and nitrate ions is characterized by high reproducibility and accuracy (variation coefficient <7%) and can be recommended for routine use in clinical practice.

The second author’s name should read A. Yu. Romanov.