[Indicators of spontaneous behavior of rhesus monkeys with short-term course alcohol self-administration under free choice]. / Pokazateli spontannogo povedeniya makak-rezus pri subkhronicheskom potreblenii alkogolya v modeli so svobodnym vyborom.

OBJECTIVE: To analyze spontaneous behavior in 36 male rhesus macaques during formation of alcoholic motivation under free choice. MATERIAL AND METHODS: The model composed the initiation stage with the provision of an aqueous ethanol solution of increasing concentration from 1 to 4% with a sweetener and restricted access to the tap water supply and the subsequent stage of the formation of alcoholic motivation with a free choice between 4% ethanol solution without sweetener and water. The behavior was recorded by the «One-Zero¼ method with ethogram compiled in accordance with the behavioral peculiarities of rhesus monkeys when housed individually. Three subgroups of high, medium and low-level ethanol consumption were distinguished. RESULTS: In the subgroup of high ethanol consumption median consumption was 1.70 g/kg/day at initiation (p<0.05, compared to other subgroups) and 1.79 g/kg/day (p<0.05) at free choice stage. Animals of high consumption subgroup had significantly higher frequency being at the cage bottom and in a sitting posture. We observed significant changes in a number of indicators of spontaneous behavior depending on the level of ethanol consumption, which included displacement behavior, stereotypic behavior and posture of animals. In the high consumption subgroup, there was a significant increase in the frequency of stereotypic behavior, atypical behavior, being on four legs, as well as a decrease in the frequency of being at the back side of the cage and of displacement behavior. At the same time, the inhibitory effect of ethanol on the hypothalamic-pituitary-adrenal system was revealed. CONCLUSION: In the model of free choice alcohol self-administration rhesus monkeys demonstrate a significant change in a number of indicators of spontaneous behavior depending on the level of ethanol consumption, which includes displacement, stereotypic behaviors and animal posture.

Genotyping of Macaque Population on the Brain-Derived Neurotrophic Factor Gene Polymorphism by Mismatch Amplification Mutation Assay (MAMA)-PCR.

Human rs6265 (196G>A) polymorphism in the BDNF gene is associated with many clinically significant phenotypic manifestations. Rhesus monkey (Macaca mulatta) has a functionally significant rs309950446 ( 136G>A) polymorphism. To determine this polymorphism in macaques, we used mismatch amplification mutation assay (MAMA)-PCR method with non-complementary nucleotide to the template chain at the 3rd position from the 3'-end of the allele-specific primers (mismatch primers), which allowed the best discrimination of the alleles. Genotyping of male rhesus monkeys (n=178) and cynomolgus monkeys (Macaca fascicularis) (n=90) was carried out. The A/A, G/G, and G/A genotypes were found in 16, 34, and 50% rhesus macaques, respectively. In the cynomolgus macaques, the mutant polymorphic allele was not detected. The study results allow considering rhesus macaques as a potential biological model for assessment of the gen-environment interaction of the BDNF gene polymorphism.

[Effect of 6-month S-methylmethionine intake on the quality of life and dyspepsia symptoms in patients with chronic gastritis].

S-methylmethionine (methylmethionine sulfonium chloride), better known as vitamin U, is a metabolic substrate that affects many metabolic processes in the human organism. Since its discovery, a large number of studies has been produced demonstrating its safety and effectiveness in various diseases, especially in diseases of the gastrointestinal tract. The purpose of the study was to evaluate the effect of methylmethionine sulfonium chloride (vitamin U) intake on the symptoms of dyspepsia and the quality of life of patients with chronic gastritis. Material and methods. The study included 37 patients (21 men and 16 women) aged 35-60 years with chronic gastritis of various etiologies. After inclusion in the study, all patients were prescribed S-methylmethionine at a dose of 300 mg per day. Clinical manifestations of dyspepsia were assessed using the GSRS questionnaire (Gastrointestinal Symptom Rating Scale), quality of life was assessed using the SF 36 questionnaire. The survey was conducted before the start of the therapy, after 3 and 6 months of complex diet therapy. Results. The most pronounced manifestations were dyspeptic (from 3 to 9 points) and diarrheal syndromes (from 2 to 5 points). Other indicators of the GSRS scale did not exceed 4 points. The total score was 15 points. By the 3rd month of therapy, there was a statistically significant decrease in the total score to 9 points (p<0.05). By the 6th month of therapy, the total GSRS score averaged 5.5 points (p<0.05). According to the SF 36 questionnaire, by the end of the 3rd month of therapy, indicators such as PF - physical functioning, BP - Bodily pain and SF - social functioning improved. By the end of the 6th month of therapy, several other indicators also improved (RP - role-physical functioning, GH - general perception of health, VT - viability, RE - Role-Emotional; MH - mental health) (p<0.05). Conclusion. The study showed that the appointment of dietary supplements containing methylmethionine sulfonium chloride at a dose of 300 mg per day helps to reduce the severity of dyspeptic symptoms in patients with chronic gastritis and their quality of life.

[Features of CD68+ and CD163+ expression in the urinary bladder as a marker of cancer in age aspects.]

The study presents a study of the expression of CD68+ and CD163+ in the bladder as a marker of cancer in age-related aspects. As part of the work performed, 35 patients with bladder cancer with histological verification of the disease aged 36 to 70 years were studied. The role of CD68+ and CD163+ cells in bladder reactions can be interpreted as an indicator of diagnostic parameters of the activity of tissue macrophages and endothelial cells in cancer. Their progression during oncogenesis is especially pronounced in the group of middle-aged patients. In old age, there is a decrease in these parameters, which indicates in favor of a decrease in the activity of the development of oncological processes with age. The combination of morphological and immunological atypism leads to a violation of the functional specialization of the organ.

[Changes in telomere length in leukocytes of male rhesus macaques of different ages.]

Telomeres are specialized terminal sections of chromosomes that ensure the stability of the latter. DNA duplication during cell division is associated with telomere shortening due to the phenomenon of terminal underreplication. As cells divide, shortening of telomere length is considered to be one of the most important causes of cell aging. Estimation of telomere length still remains the subject of scientific research in gerontology and it is not used in clinical practice. Most often, rodents are used as a model object for studying the aging process, but the neuroendocrine mechanisms that influence, among other things, the regulation of the aging process differ in rodents and humans. The model objects closest in phylogenetic relation to humans are monkeys. In particular, Rhesus macaques is one of the representatives of the Old World most often used in biomedical research. However, data on age-related changes in telomere length in monkeys are extremely scarce. We studied the absolute average length of telomeres in DNA from blood leukocytes of 29 clinically healthy male rhesus monkeys aged from 4 to 24 years using quantitative PCR-method. The data obtained did not correspond to the normal distribution and the correlation analysis showed the absence of a significant dependence of telomere length on the age of the animals (rs=0,27; p>0,05). Thus, our study does not confirm the dependence of changes in the average length of telomeres of blood leukocytes with age.

[Identification regulatory noncoding RNAs of human papilloma virus type 16 (Papillomaviridae: Alphapapillomavirus: Human papillomavirus) in cervical tumors].

INTRODUCTION: High carcinogenic-risk human papillomaviruses (hrHPVs) are recognized as etiological agents of cervical cancer. Constant expression of the viral oncoproteins, E6 and E7, is required for maintenance of the malignant phenotype of tumor cells. The exact mechanism of regulation of viral oncogenes expression in tumor cells is not fully elucidated. THE PURPOSE: identification of viral noncoding RNAs (ncRNAs) in HPV16-positve cervical cancer. MATERIALS AND METHODS: The reverse transcription polymerase chain reactions were used to detect viral ncRNAs in HPV16-positve primary cervical squamous cell carcinomas and SiHa and CasKi cell lines. The knockdown technique with oligonucleotides complementary to ncRNAs was used to elucidate their functions. RESULTS: We have identified ncRNAs transcribed in the upstream regulatory region of HPV16 in the cervical carcinoma cell lines and in 32 out 32 cervical squamous cell carcinomas with episomal or integrated forms of HPV16 DNA. Knockdown of sense or antisense strains of ncRNAs by oligonucleotides results in a decrease or increase of the E6 and E7 oncogenes mRNA levels in cells, respectively. These changes of oncogenes mRNA levels are accompanied by the modulation of the levels of the p53 protein, the main target of the E6 oncoprotein. CONCLUSION: The presence of regulatory ncRNAs in all examined tumors and cell lines revealed for the first time indicates their necessity for maintenance of constant expression of E6 and E7 oncogenes in them. The findings can be useful for understanding of the fundamental aspects of the viral expression regulation in HPV16-positive tumors.

Experimental Study of the Efficacy of Sodium Deoxyribonucleate Used in Combination with Co-Transplantation of Mesenchymal and Hematopoietic Stem Cells after Exposure to γ-Radiation.

We studied the possibility of using sodium deoxyribonucleate (Derinat) for improving the efficiency of co-transplantation of mesenchymal (MSC) and hematopoietic stem cells (HSC) to female F1(CBA×C57BL/6) mice with bone marrow aplasia caused by exposure to γ-radiation. It was found that immunomodulator Derinat enhanced the effect of co-transplantation, in particular, triple post-irradiation administration of Derinat accelerated hematopoiesis recovery judging from the parameters of peripheral blood, total cellularity of the bone marrow and spleen, and animal survival. Single or double administration of Derinat prior to irradiation was ineffective. The optimal result was obtained when the following scheme was applied: MSC→HSC with an interval of 48 h starting during the first hours after irradiation and triple administration of Derinat (in 10-15 min, 3 and 7 days after irradiation) in a dose of 3 mg/mouse.

Evaluation of New Immunohistochemical Approaches for the Study of Kidney Tumors in Geriatric.

Kidney malignancies are among the most deadly genitourinary tumors. It is more common in males and is often seen in people aged 60-70 years old. The incidence rate of kidney cancer seems to be increasing. One reason for this may be the fact that imaging techniques, such as computed tomography scans are more commonly used. These tests may lead to the accidental detection of more kidney cancers. Fortunately, kidney cancer is often detected in the early stages, when the tumor is small and confined to the kidney. The objective of this study was the development of new diagnostic immunohistochemical methods. Clinical examination material of 134 people, including 94 (70%) males and 40 (30%) females, were used in this study. Immunohistochemical staining of tryptase was carried out in compliance with the requirements using Anti-Mast Cell Tryptase antibodies. Goat anti-mouse antibodies #AS-M1-HRP were used as secondary antibodies, visualized with ImmPACTTM DAB Peroxidase Substrate Kit (#SK-4105) according to the instructions of the manufacturer. The nuclei were counterstained with Mayer's hematoxylin, and the sections were embedded in a permanent mounting medium. The immunohistochemical study showed an increase in both tryptase- and chymase-positive mast cells in the renal parenchyma, compared with the control group. The number of mast cells with tryptase expression directly in the tumor was significantly less than the peritumoral localization. A similar pattern was observed for chymase-positive mast cells as the content of the tumor was more than 10 times higher than the intratumoral arrangement. The histological and immunological characteristics did not differ in different age groups. The immunohistochemical method of research in the diagnosis of renal tumors plays an important diagnostic and prognostic value. It can assist pathologists in difficult and ambiguous cases to correctly diagnose renal tumors. This will make it possible to prescribe the correct treatment and predict the course of malignant tumor growth in patients.

[Comparative effectiveness of ursodeoxycholic acid preparations in the treatment of biliary sludge].

In the clinical classification of cholelithiasis, biliary sludge (BS) is distinguished as the pre-stone stage. Ursodeoxycholic acid (UDCA) is a drug with an evidence base for effective and safe effects on BS. The therapeutic equivalence of various UDCA drugs remains an important issue for clinical practice. AIM: To conduct a comparative analysis of the effectiveness of the use of UDCA: Ursofalk with other UDCA drugs for the treatment of BS in a fixed dose of 10 mg/kg of body weight. MATERIAL AND METHODS: The observation group consisted of 225 patients with various types of BS. In randomized groups, the comparison of the effectiveness of UDCA drugs in the dissolution of BS was determined by the data of ultrasound of the gallbladder. Dynamic ultrasound cholecystography using a standardized technique was performed to study the effect of the compared drugs on the contractile function of the liver. When analyzing the impact of ursotherapy on the clinical manifestations of BS, the dynamics of biliary pain syndrome and dyspeptic disorders were evaluated. RESULTS: A comparative analysis of the effectiveness of UDCA drugs for BS lysis with a high degree of confidence established the advantage of Ursofalk: after 3 months in 80%, and after 6 months of therapy in 95.65% of patients, a regression of BS was detected. In the group of patients who took other UDCA, the corresponding indicators were 46.36% and 67.27%. The greatest effectiveness of ursotherapy was demonstrated in the BS variant by the type of suspension of hyperechogenic particles, the lowest in the variant by the type of putty-like bile in the form of mobile or fixed clots. In the last variant of BS, the relationship between the success of lysis with the extension of the UDCA intake period to 6 months and the preferred choice of Ursofalk was traced. Normalization of the contractile function of the gallbladder was noted in patients who took Ursofalk. CONCLUSION: From the standpoint of therapeutic effectiveness, the drug of choice for the treatment of BS, regardless of its type, is Ursofalk, which has proven the best dynamics of BS litolysis, recovery of the contractile function of the gastrointestinal tract, and relief of clinical symptoms.

[PHARMACOGENETIC ASPECTS OF TREATMENT OF PATIENTS WITH CHRONIC HEART FAILURE (REVIEW)].

This article reviews data for pharmacogenetics findings for treatment for chronic heart insufficiency of heart failure. Also, it discusses connections of genetic polymorphism with a risk of developing heart failure and how it affects the choice of a treatments medicine. The article investigates genetically determined factors of the risk of developing glycoside toxicity. One of the causes is polymorphism of the MDR1 gene encoding glycoprotein P, a transporter, that is involved in the absorption of drugs in the intestines and excretion by the kidneys. Also, the genetic characteristics of ACE in patients with heart failure are discussed. The data is presented from the study of the role of genetic liver metabolism polymorphism regarding efficacy and safety of loop diuretic torasemide. Data for genetic polymorphism of the metabolism of the main beta blocker, used in chronic heart failure metoprolosuccinate is also discussed. Data for roll ß1-AP polymorphism in the clinical use of ß-AB in patients with heart failure. Further observations are needed from longer studies, that will evaluate long-term efficacy and safety outcomes for medicines related to genetic traits.

[CLINICAL DIAGNOSTIC VALUE OF OXIDATIVE STRESS MARKERS IN PATIENTS WITH CHRONIC HEART FAILURE. OPPORTUNITIES OF THEIR PHARMACOLOGICAL CORRECTION BY ETOXIDOL].

Purpose - to evaluate the effectiveness of a fundamentally new antioxidant drug Amoxidal malate in patients with chronic heart failure (CHF) and its effect on marker of oxidative stress 2,3-diphosphoglycerate -2,3-DPG regulating the dissociation of oxyhemoglobin into hemoglobin and oxygen depending on the partial pressure of oxygen in the lungs and effect on other markers of oxidative stress. Clinical study of etoxazole was conducted in the city hospital N 23. 32 people were examined. At the age of 55 to 76 years (men and women) with coronary heart disease, stable angina, who had a myocardial infarction with a diagnosis of chronic heart failure II-IU FC according to the NYHA classification. Hypertension was diagnosed in 15 patients and type 2 diabetes mellitus in 8 patients. The study included patients with an ejection fraction of less than 40%. Permanent atrial fibrillation was diagnosed in 6 patients. Patients were divided into 2 groups: 1 main group, 22 patients who were added to the standard therapy with intravenous infusions of Ethylmethylhydroxypyridine malate, 2 group - control group, 10 people who received standard pharmacotherapy of CHF. Indicators of oxidative status, especially 2,3-DPH, were evaluated. and also, the voltage of oxygen (pO2), pCO2, pH, concentration of superoxide dismutase (SOD), malondialdehyde (MDA), the concentration of total peroxides in the blood of these patients. Etilmetilgidroksipiridinamaalat restores oxygenation of the blood in patients who are intravenously introduced Amoxidal compared with patients not receiving this treatment with CHF IV FK. Analysis comparative assessment of treatment results shows that the addition of Ethoxide to standard therapy is most beneficial effect on patients> IU F. K. the Inclusion of the new Patriotic antioxidant drug Amoxidal in standard therapy of patients with CHF is pathogenetically justified and promising.

[CLINICAL CASE: THE SIDE EFFECTS OF STATINS].

The clinical studies, conducted in recent years, suggest that statins increase the activity of telomerase and by that decelerate speed of telomerase shortening. Thus, on one hand, it reduces a risk of cardiovascular diseases development, decelerate aging, but on the other hand, increasing the activity of telomerase, lead to expression rising of gene hTERT, that can make prerequisites for malignancy. That's why, it's necessary to study the subject and develop reliable criteria for safety use of activators-telomerase.

The Mechanism of Action of Ethoxidol on Oxidative Stress Indices in Heart Failure and Hypotension.

The aim of the investigation was to study the effect of 2-ethyl-6-methyl-3-hydroxypyridine malate (Ethoxidol) on the concentration of oxidative stress metabolites in patients with chronic heart failure (CHF) and hypertension. MATERIALS AND METHODS: 126 patients with FC I-III CHF have been examined. In addition to their individual therapy these patients received intravenous infusions of Ethoxidol. Blood content of 2,3-diphosphoglycerate (2,3-DPG), oxygen tension (рО2), pH, concentration of total peroxides, lactate, and aldosterone were identified. 2,3-DPG levels (g/L erythrocytes) in whole blood samples were determined by an enzyme assay using the reagent kit (Rosh, Germany), values of рО2, рСО2, рН, lactate in the venous blood were measured using gas analyzer Stat Profil pHOx Ultra (Nova Biomedical, USA). Indices of oxidative stress, i.e. the concentration of plasma total peroxides, were investigated by ELISA using OxyStat kit (Biomedica, Austria). Peripheral venous blood samples were collected from all patients before and 6 days after the daily intravenous Ethoxidol infusion. RESULTS: In patients with FC I, II, III CHF, on day 7 after intravenous Ethoxidol infusion at a dose of 100 mg/day, statistically significant growth (p=0.0002) of PaO2 level by 15.7, 17.4, and 22.8%, respectively, was noted. In patients with FC I, II, III CHF in the group receiving standard therapy, statistically significant (p=0.002) reduction of 2,3-DPG level by 2.7, 2.4, and 4.0%, respectively, was registered. On day 7 after the infusion of Ethoxidol at a dose of 100 mg/day, its decrease by 5.7, 10.5, and 26.2%, respectively (p<0.0001), was also observed. CONCLUSION: The increased concentrations of active oxygen forms have been established to negatively affect various bodily functions and adversely influence the pathophysiology of numerous diseases. Application of antioxidants, including Ethoxidol presented by us in this article, may become a clue to the development of preventive measures for many serious diseases.

Formation of Alcohol Motivation in Monkeys (Macaca mulatta) Kept in Isolation.

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.

Effect of Different Regimens of Co-Transplantation of Hemopoietic and Mesenchymal Hemopoietic Stem Cells on the Rates of Hemopoietic Recovery in Laboratory Mice Treated with Cyclophosphamide in Sublethal Doses.

We studied the effect of mesenchymal stem cells and hemopoietic stem cells co-transplanted in different regimens (sequence and intervals between the injections within 0-48 h) on the rate of the bone marrow hemopoiesis recovery in 468 inbred mice with cytostatic aplasia caused by a single injection of cyclophosphamide in a dose of 500 mg/kg. The efficacy of stem cells was evaluated by animal survival and general physical condition, body weight dynamics, peripheral blood counts (leucocytes, platelets, and reticulocytes), total cellularity and the presence of hemopoietic stem cells in the bone marrow on day 9 after cyclophosphamide injection. The relative content of hemopoietic stem cells in the total myelokaryocyte pool was assessed by flow cytofluorometry using specific monoclonal antibodies to CD117 and CD184. The stimulating effect of co-transplantation of mesenchymal stem cells and hemopoietic stem cells on hemopoiesis was demonstrated by the indices of total cellularity and the relative content of CD117+ hemopoietic stem cells. The schemes with injection of mesenchymal stem cells 24-48 h prior to injection of hemopoietic stem cells were most effective.

First Finding of Representatives of the Eccrinida Order in the Digestive Tract of King Crab Specie from the Barents Sea.

This is the first report on the finding of large intra-intestinal symbionts belonging to the order Eccrinida in crab-like decapod crustaceans of the family Lithodidae, Paralithodes camtschaticus and Lithodes maja, inhabiting the Barents Sea. Studies have been regularly conducted since the early 2000s in Kola Bay and in one of the inlets of the Eastern Murman Coast. Since 2005, Eccrinida representatives (presumably, a new species of the genus Arundinula) have been found in the guts of the red king crab. A brief description of the endosymbionts and data on their frequency of occurrence are presented. The possible reasons for the distribution of eccrinids in the crab-like decapod crustaceans of the Barents Sea are discussed.

Investigation of the new sorption preconcentration systems for determination of noble metals in rocks by inductively coupled plasma-mass spectrometry.

The reversible sorption preconcentration of noble metals (NMs) prior to their determination by inductively coupled plasma-mass spectrometry (ICP-MS) was investigated. Six new hypercrosslinked polystyrene sorbents were tested. The dependence of the degree of NMs sorption on the average degree of polymer network crosslinking and pore diameters was investigated. It was found that sorbents HP-100/6, HP-300/6 and HP-500/6 have low efficiency of NMs chlorocomplexes extraction. Among Stirosorb sorbents (Stirosorb-2, Stirosorb-514 and Stirosorb-584) the highest efficiency of the extraction of NMs' chlorocomplexes has Stirosorb-514. Tributylamine (TBA), N-methylbenzylamine (MBA), N,N-dimethylbenzylamine (DMBA), N,N-dibenzylmetylamine (DBMA) were studied as the reagents for extraction of Ru, Rh, Pd, Ir, Pt and Au chlorocomplexes from hydrochloric acid solutions in the form of ion associates by reversed-phase mechanism. The reversible quantitative extraction of Ru, Pd, Pt and Au in system Stirosorb-514 - TBA - 1M HCl in ethanol as eluent was achieved. It was found that resulting eluates do not contain matrix components which may cause spectral interferences on the stage of NMs determination by ICP-MS. The found scheme of NMs reversible sorption was validated by the analysis of certified reference materials of basic and ultrabasic rocks GPt-5, GPt-6 and SARM-7. Good agreement between the measured NMs concentrations and the certified values was demonstrated. The achieved limits of detection for Ru, Pd, Pt and Au vary within 10(-8)-10(-7)wt% range.

Ration of the red king crab on coastal shoals of the Barents Sea.

In different habitats of the Kola Bay (Western Murman) and Dalnezelenetskaya and Yarnyshnaya bays (Eastern Murman), the size and structure of ecological rations (foraging of benthos) of the red king crab, which is an alien species in the Barents Sea, was established. The material for the study was collected in 2000-2009. In the Kola Bay, significant variability in time of this nutrition was detected for individuals of the same size category, which was associated with the depletion of food resources due to the high abundance of invaders in the area. The stable values of the ration and its structures in Eastern Murman bays indicated the prosperous state of benthic communities and an insignificant impact of crabs on these communities.

[Comparative efficiency of algorithms based on support vector machines for binary classification].

Methods of construction of support vector machines require no further a priori infoimation and provide big data processing, what is especially important for various problems in computational biology. The question of the quality of learning algorithms is considered. The main algorithms of support vector machines for binary classification are reviewed and they were comparatively explored for their efficiencies. The critical analysis of the results of this study revealed the most effective support-vector-classifiers. The description of the recommended algorithms, sufficient for their practical implementation, is presented.

[Comparative Efficiency of Algorithms Based on Support Vector Machines for Regression].

Methods of construction of support vector machines do not require additional a priori information and can be used to process large scale data set. It is especially important for various problems in computational biology. The main set of algorithms of support vector machines for regression is presented. The comparative efficiency of a number of support-vector-algorithms for regression is investigated. A thorough analysis of the study results found the most efficient support vector algorithms for regression. The description of the presented algorithms, sufficient for their practical implementation is given.