Vol 164, No 1 (2017)

Mechanisms of amnesia caused by impairment of consolidation or reconsolidation of conditioned food aversion memory with protein synthesis inhibitor cycloheximide were studied in Helix lucorum. Cycloheximide injection during training or memory reconsolidation in trained snails produced amnesia. In both cases, repeated training 10 days after amnesia induction led to short-term memory formation, while long-term memory was not formed, despite the fact that the number of conditioned and reinforcing stimuli combinations was higher than during initial training. The possibility of formation of short-term memory not transforming into long-term memory is one of the key characteristics of anterograde amnesia. Our findings data and experimental model can be used for analysis of specific molecular mechanisms of anterograde amnesia.

We studied the role of calcium-regulating structures of slow- (m. soleus, SOL) and fast-twitch (m. extensor digitorum longus, EDL) skeletal muscles of rats during adaptation to exhausting physical activity and the possibility of modulating this adaptation with decaffeinated green tea extract. It was established that EDL adaptation is mainly aimed at Са²⁺ elimination from the sarcoplasm by Са-ATPase and its retention in the reticulum by calsequestrin. Administration of green tea extract increased endurance due to involvement of slow-twitch muscles whose adaptation is associated with enhanced expression of all the studied genes responsible for the regulation of Ca²⁺ balance.

Intravenous injection of nonselective antagonists of opioid receptors (OR) naltrexone (5 mg/kg) and naloxone methiodide (5 mg/kg), selective δ₁-OR antagonist BNTX (0.7 mg/kg), selective δ₂-OR blocker naltriben (0.3 mg/kg), selective κ-OR antagonist norbinaltorphimine (2 mg/kg), and selective blocker of ORL1 opioid receptors JTC-801 (0.1 mg/kg) produced no effect on reperfusion injury to the heart in rats narcotized with α-chloralose. In contrast, selective μ-OR antagonist CTAP (1 mg/kg) limited the infarct size, although this effect was not observed at a lower CTAP concentration of 0.1 mg/kg. Probably, the myocardial infarct size-limiting effect of CTAP was associated with activation of the non-opioid receptors. It was hypothesized that endogenous OR agonists did not affect heart resistance to reperfusion injury in unadapted rats.

Mexicor treatment (8 mg/kg body weight per day) during the posttraumatic period after concomitant traumatic brain injury and acute blood loss in rats increased electrophoretic mobility and concentration of 2,3-diphosphoglycerate, and reduced malondialdehyde content in erythrocytes. These changes improved hemodynamics and oxygen-transporting function of the blood. The most pronounced effects of Mexicor were observed at the early stages of posttraumatic period.

We studied the balance of fatty acids and their correlation with parameters of lipid metabolism and inflammation markers in men with coronary atherosclerosis. In the blood of patients and healthy men, the content of myristic (C14:0), pentadecanoic (C15:0), palmitic (C16:0), stearic (C18:0), arachidic (C20:0) and docosanic (C22:0), TNFα, IL-1β, IL-6, IL-8, C-reactive protein, cholesterol, triglycerides, HDL, and LPO intensity were measured. In patients with coronary atherosclerosis, increased concentrations of saturated fatty acids, triglycerides, LPO products, IL-6, IL-8, and C-reactive protein were found. Correlation analysis revealed multiple correlations between the studied fatty acids, correlation of C16:0, C18:0, and C20:0 with inflammation markers, C14:0, C16:0, and C18:0 with triglyceride level, C16:0, C18:0, and C20:0 with cholesterol. The contents of C14:0, C15:0, C16:0, and C18:0 correlated with the presence of coronary atherosclerosis. Lipid metabolism disturbances in coronary atherosclerosis were accompanied by changes in not only lipid spectrum and inflammation markers, but also fatty acids balance.

Effect of alkoxyamines on normal and tumor cells was studied in vitro and in vivo. In vitro experiments showed that alkoxyamines produce a dose-dependent toxic effect on cells of human breast tumor MCF7 line. Transplantation of Krebs-2 ascites carcinoma cells preincubated with alkoxyamines to mice did not induce tumor growth. An opposite effect was observed in normal mouse cells: functional activity of peritoneal macrophages increased. The possibility of using alkoxyamines as theranostic agents is discussed.

Experimental model for in vitro evaluation of Campylobacter genus bacteria growth kinetics, inhibition, or inactivation is proposed. The model allows quantitative evaluation of the sensitivity to various types of stress exposure and promotes detection of the regularities of their transformation into uncultivable forms. The model implies the use of 96-well plates for parallel culturing of several subpopulations of the test strain in media with various parameters. The proposed algorithm includes evaluation of the proportion of viable CFU to total level of planktonic and uncultivable cells in the population, which is estimated by the content of genomic DNA in the samples by quantitative PCR (or real-time PCR) with ciaB, cdtB, or 16S rRNA primers. The presence of biofilm matrix is detected by the intensity of staining of polystyrene plates. This model can be used for evaluation of the most significant types of exposure, including low-dose antibacterial treatment, promoting the formation of stable microorganism variants. The model has been used to study the effects of culturing conditions on the characteristics of C. jejuni populations. The most characteristic feature of C. jejuni is reduction of the count of viable cells up to complete disappearance of cultivable forms under favorable conditions of growth. The level of viable cells in the populations decreased 10-fold and more, on average, after 48-h incubation. Not all strains exhibit this property, some strains retain their viability, which is detected by the culturing method, and contributes to biofilm formation.

In vivo modifying effects of bone marrow mesenchymal stromal cells of humans and laboratory mice on ROS production by mouse blood mononuclears are studied by luminol-dependent zymosan-induced chemiluminescence after syngeneic and xenogeneic transplantation into systemic blood flow. The chemiluminescent activity of mouse blood mononuclears has increased early (1 day) after syngeneic (mouse mesenchymal stromal cells) and xenogeneic (human mesenchymal stromal cells) transplantation. Later, 7-21 days after syngeneic and xenogeneic transplantation, the chemiluminescent activity of mouse mononuclears is suppressed. The probable mechanisms of involvement of the transplanted mesenchymal stromal cells in reprogramming of the blood mononuclear phagocytes from proinflammatory (M1) to anti-inflammatory (M2) phenotype under conditions of their in vivo interactions are discussed; a frequent manifestation of this reprogramming is transition of the phase of activation into inhibition of ROS-producing activity of macrophages.

Stable red fluorescing line of human ovarian epithelial cancer cells SK-OV-3ip-red was generated expressing gene coding for protein TurboFP635 (Katushka) fluorescing in the far-red spectrum region with excitation and emission peaks at 588 and 635 nm, respectively. Fluorescence of SK-OV-3ip-red line remained high during long-term cell culturing and after cryogenic freezing. The obtained cell line SK-OV-3ip-red can serve a basis for a model of a scattered tumor with numerous/extended metastases and used both for testing anticancer drugs inhibiting metastasis growth and for non-invasive monitoring of the growth dynamics with high precision.

Despite promising vista of the use of microRNA in molecular diagnosis of bladder cancer, there are few data on their expression profiles, which impedes assessment of diagnostic value of these marker molecules. In this study, suppression subtractive hybridization, on-chip hybridization, and high-throughput deep sequencing focused on profiling microRNA and assessing the diagnostic value of revealed marker molecules.

A cross-sectional study of universe sample of patients (N=135; mean age —49.7±0.8 years) with metabolic syndrome yielded the incidence of cardiac autonomic neuropathy of 37.5%. The pathogenetic peculiarities were revealed for the onset and development of this neuropathy. At the early stages, the progress of cardiac autonomic neuropathy closely correlated with elevation of blood glucose, while endothelial dysfunction progressing at the later period against the background of persistent hyperglycemia is viewed as an extra factor contributing to the development of this disease.

The polypeptide complex of the epiphysis and the peptide AEDG, constructed on the basis of its amino acid analysis, exert similar biological effects. Both bioregulators normalize melatonin synthesis in the pineal gland, functioning of the brain, eye retina, cardiovascular, endocrine, and immune systems; they also act as antioxidants, stress-protectors, and geroprotectors. Within the epiphysis polypeptide complex, free amino acids (3.26%), dipeptides (23.19%), tripeptides (50.72%), tetrapeptides (22.10%), and pentapeptides (0.72%) were revealed by mass spectrometry and HPLC. Peptide AEDG was detected among the tetrapeptides of the epiphysis polypeptide complex by selective reaction monitoring method. The biological effects of the epiphysis polypeptide complex are determined by the effect of its component AEDG.

We compared the effect of xymedon (100 mg/kg) and Mexidol (50 mg/kg) on morphometric parameters of erythrocytes and oxygen-transporting function of hemoglobin in rats with Walker-256 carcinoma treated with docetaxel (15 mg/kg once intraperitoneally on day 11 after tumor cells transplantation). Xymedon and Mexidol were injected intramuscularly for 10 days starting from day 11 of the experiment. The studied parameters were evaluated on experimental days 14 and 22. Similar to Mexidol, xymedon prevented changes in the erythrocyte geometric parameters induced by docetaxel and neoplastic process, but increased hemoglobin packing density in erythrocytes (by 32%) more effectively than Mexidol. Optimization of oxygen-transporting function of hemoglobin and normalization of its structural and functional parameters changed by docetaxel treatment (content of oxyhemoglobin, hemoglobin affinity for oxygen and its ability to bind and release oxygen, intensity of symmetric and asymmetric vibrations of pyrrole rings) in rats receiving xymedon took longer time than in case of Mexidol treatment, but the effects were similar.

We developed a multiplexed DNA microarray-based assay allowing identification of 12 causative agents of reproductive tract infections with the simultaneous detection of 47 genetic determinants of resistance to antimicrobial substances. The microarray was tested on 93 isolates of Neisseria gonorrhoeae, 32 isolates of Treponema pallidum and 29 samples of Ureaplasma spp./Mycoplasma spp. The N. gonorrhoeae isolates had multiple mutations in the penA, ponA, rpsJ, gyrA, parC, and mtrR genes; their prognostic value significantly increased when combinations of mutations were detected. In the analyzed T. pallidum isolates, single A2058G substitution in the 23S rRNA gene responsible for macrolide resistance was found. DNA sequences of Ureaplasma spp./Mycoplasma spp. were determined as wild type, which was not fully consistent with the results of analysis of their antimicrobial susceptibility

Influenza virus hemagglutinins are surface proteins responsible for fusion of the viral and cellular membranes. Their capacity to mediate membrane fusion (fusogenic activity) is studied by various methods, including the syncytium formation and pseudovirus transduction methods. We constructed plasmids coding for genes of three H1 and one H5 hemagglutinins and compared their fusogenic activities. Hemagglutinin capacity to induce syncytium formation did not always correlate with the transduction activity of the respective pseudoviruses. Hemagglutinin H5 exhibited high fusogenic activity in studies by both methods, however, two of the studied H1 hemagglutinins induced the formation of syncytia, but did not mediate pseudovirus transduction. This could be due to different capsid sizes of influenza virus and vesicular stomatitis virus, which determines their different permeability through the fusion pore.

A real-time PCR with hybridization and fluorescent detection was used to analyze the distribution of p53 G215C, p21A1026G, and G369C gene polymorphisms in patients with stomach cancer and healthy subjects. It was found that allele C, genotypes of CC and GC of G215C p53, and G369C p21 polymorphisms and allele A and AA and GA genotypes of A1026G p21 polymorphism are significantly associated with the risk of stomach cancer development.

DAI-1 receptor (DNA-dependent activator of IFN-regulatory factors; DLM-1/ZBP-1) is an innate immunity cytoplasmic receptor of the DNA-recognition receptor class of antiviral immunity. DAI-1 expression reflects the severity of the inflammatory response that plays the key role in the pathogenesis of pregnancy complications. We studied DAI-1 receptor expression in the placental villi in early- and late-onset preeclampsia. In case of early-onset preeclampsia DAI-1 staining intensity was lower (p=0.01), and in case of late preeclampsia — significantly higher (p<0.005) than in the reference groups at the corresponding gestational age. There was revealed a correlation between the decrease in DAI-1 receptor expression and the severity of disease progression.