Preanalytical features of the determination of circulating microRNAs as new specific biomarkers of the body’s response to physical activity

MicroRNAs are small non-coding single-stranded RNAs, 18 to 25 nucleotides long, they regulate gene expression at the post-transcriptional level through specific binding to the target mRNA, leading to its degradation. In recent decades, the development of technologies for determining the expression profiles of miRNAs has become an important part of research projects, and the role of miRNAs as potential highly informative molecular biomarkers of various physiological and pathological processes in the body is actively explored by the scientific community. In particular, physical activity is an important modifying factor for circulating miRNAs. Unlike classical blood biochemical parameters, which can change over time depending on the temperature and storage conditions of the sample, microRNAs remain stable during storage and even after multiple freeze-thaw cycles, which makes them an attractive and easily accessible target for detection. However, the determination of the microRNA expression profile in clinical practice is still difficult due to the high heterogeneity of analytical procedures used for testing. In sports medicine, the preanalytical stage is especially important, since often the conditions for sampling are not standardized and can affect the analysis result. This review shows the role of miRNAs as new sensitive biomarkers of the effectiveness of the training process and regulators of the body’s response to physical activity, and also discusses some preanalytical aspects of the analysis of miRNA expression profiles

The development of blockchain technology in Russia : outlook and trends

Purpose: The article addresses the issue of new scientific decisions shaping with respect to the study of problems, current trends and perspectives of blockchain technology usage in the Russian Federation. Design/methodology/approach: To achieve the objectives of this study the increasing interest to blockchain technology in Russia was discussed. Findings: The article determined main problems in blockchain technology which includes gaps in legislative regulation; the existence of a considerable number of projects that are undergoing the development stage and that have not proved own economic feasibility yet; incomplete understanding of the blockchain spheres’ implementation by state officials, society and business representatives as well as expected outcomes according to the amount and time of their receiving; disputes on cryptocurrencies turnover in the country’s territory and their influence in the national economy. Practical implications: The study has demonstrated the interest growth mainly by businesses to the usage of blockchain technology in order to improve own competitiveness and to obtain additional benefits, including the form of their profits. Originality/value: The research has also determined the area of further key studies in blockchain technology usage in Russia and the world.peer-reviewe

Comparison of microRNA expression profiles of athletes involved in endurance sports and non-athletic volunteers using a hypoxia signaling pathway panel

In recent years, interest in the search for new potential molecular biomarkers to assess the effectiveness of the training process in elite sports, various pathologies caused by overload, and abuse of prohibited substances has reached an unprecedented level. MicroRNAs that regulate the processes of metabolism, differentiation, proliferation, and apoptosis of cells at the post-transcriptional level are the most valuable candidates for use in this regard. They are found in all biological fluids of the human body, are stable during long-term storage and resistant to changes in environmental conditions. It is known that changes in the miRNA expression profile are closely associated with physical activity, as well as with the administration of recombinant hormones, erythropoiesis-stimulating agents, and other substances and methods that improve sports performance, which are prohibited by the World Anti-Doping Agency (WADA).Objective: In this study, plasma samples from athletes involved in endurance sports (race walking) and non-athletic volunteers were analyzed using a hypoxia signaling pathway panel to identify hypoxia candidate markers.Materials and methods: Expression profiles of plasma circulating miRNAs were assessed by reverse transcription followed by real-time quantitative polymerase chain reaction (RT Q-PCR). The obtained data were subjected to statistical processing using the CFX Manager Software v3.1 program.Results: Three potential microRNA markers were identified: hsa-miR-210-3p, hsa-miR-320a and hsa-miR-935 (increased in athletes by 61.6 times, 51.8 and 41.0 times, respectively) — of physiological response to the training, load, which may be associated with the emergence of hypoxia during endurance training.Conclusion: We have obtained preliminary data on differences in the expression profiles of circulating miRNAs in healthy non-athletic volunteers and professional athletes in the out-of-competition period. In the future, it is planned to expand the sample of studied blood plasma samples and compare the expression profiles of circulating microRNAs in athletes in competition and out-of-competition periods, as well as to compare the expression profiles of circulating microRNAs during aerobic and anaerobic loads. In addition, it is of interest to analyze changes in the expression levels of circulating miRNAs when using hypoxia mimetics applied to improve sports performance

The thylakoid carbonic anhydrase associated with photosystem II is the component of inorganic carbon accumulating system in cells of halo- and alkaliphilic cyanobacterium Rhabdoderma lineare

AbstractThe organization of carbonic anhydrase (CA) system in halo- and alkaliphilic cyanobacterium Rhabdoderma lineare was studied by Western blot analysis and immunocytochemical electron microscopy. The presence of putative extracellular α-CA of 60 kDa in the glycocalyx, forming a tight sheath around the cell, and of two intracellular β-CA is reported. We show for the first time that the β-CA of 60 kDa is expressed constitutively and associated with polypeptides of photosystem II (β-CA-PS II). Another soluble β-CA of 25 kDa was induced in low-bicarbonate medium. Induction of synthesis of the latter β-CA was accompanied by an increase in the intracellular pool of inorganic carbon, which suggests an important role of this enzyme in the functioning of a CO2-concentrating mechanism

INCREASE OF DEMAND FOR GRADUATES OF HIGHER EDUCATION INSTITUTIONS

Abstract. The aim of the research is identification of the objective and subjective factors influencing professional self-realization of graduates of the field of study «Public and Municipal Administration». Methods. The authors have conducted anonymous opinion survey of prestigious economic universities undergraduates (bachelor and master degree course) and graduates specialized in this sphere. The questions were about correspondence of gained academic qualification and actual occupation, the importance of degree certificate, satisfaction with education quality, attitude to different aspects of studying etc. In order to make relevant conclusions, the authors apply comparative, statistical, eneralization and induction methods. Results. Informants’ answers helped to count out the percentage of graduates whose occupation is connected with public administration, understand the reasons of choosing relevant master course, assess the significance of acquired competence and qualification for job placement at governmental organizations. The main scientific result of the research is working out a system of recommendations aimed at increasing demand for graduates specialized in the sphere of civil service, and, consequently, improving the efficiency of investment in management training. It is specified that the most important issues in this regard are forethought of career choice, students’ vision of academic planning, and approach to teaching, practice-oriented type of education. Scientific novelty. Having organized the data and compared it with previous surveys’ results, the authors reveal the factors essential to professional success and application of knowledge as well as correlation of these factors which means meeting the objective of the research.Practical significance. The presented materials can be used for the further modernization of the process of educational planning and taken into notice while developing academic programs for the direction «Public and Municipal Administration». Исследование выполнено в рамках проекта – Грантового соглашения (Договора) №16-27-01001/16 от 30.05.2016 г. в РГНФ. Международный конкурс РГНФ – Белорусский республиканский фонд фундаментальных исследований (БРФФИ) 2016 г. «Разработка Концепции стратегического развития межстрановой интеграции национальных инновационных систем Союзного государства до 2030 года»: http:// grant.rfh.ru/taskИсследование выполнено в рамках проекта – Грантового соглашения (Договора) №16-27-01001/16 от 30.05.2016 г. в РГНФ. Международный конкурс РГНФ – Белорусский республиканский фонд фундаментальных исследований (БРФФИ) 2016 г. «Разработка Концепции стратегического развития межстрановой интеграции национальных инновационных систем Союзного государства до 2030 года»: http:// grant.rfh.ru/tas

Определение возможных микроРНК-маркеров злоупотребления препаратами кобальта методом ПЦР в реальном времени с использованием панелей сигнального пути гипоксии

Objectives. Cobalt mimics the state of hypoxia to prevent degradation of the alpha subunit of hypoxia-inducible factor, resulting in an increase in blood oxygen capacity and endurance. Athletes can use this property to gain competitive advantage. Nowadays, direct methods of inductively coupled plasma mass spectrometry and liquid chromatography-tandem mass spectrometry are used to determine total cobalt levels in the body. However, the World Anti-Doping Agency is yet to establish a maximum allowable threshold concentration of this element in biofluids. The lack of clear identification criteria complicates the interpretation of the obtained results for the purposes of doping control. In this regard, the present work proposes a new approach for the indirect determination of possible cobalt abuse based on changes in the expression levels of miRNAs involved in the regulation of hypoxia signaling pathways. Here, the aim is to identify possible microRNA markers whose expression does not depend on exercise-induced hypoxia, but changes markedly when taking cobalt preparations.Methods. MicroRNA isolation was performed from blood plasma samples using the PAXgene Blood miRNA Kit. Quantitative real-time polymerase chain reaction (PCR) was performed on CFX96 Bio-Rad (USA) analyzer using miScript® SYBR® Green PCR Kits and panels for studying the expression profiles of mature microRNAs of the hypoxia signaling pathway miScript® miRNA PCR Array.Results. Based on the statistical analysis of the data, it was found that the expression of hsa-miR-15b-5p in the blood plasma of the subjects does not depend on physical activity, but increases when taking cobalt preparations.Conclusions. The difference in expression levels during anaerobic exercise-induced hypoxia and cobalt-induced hypoxia makes hsa-miR-15b-5p a potential candidate to be a marker of erythropoiesis-stimulating agent abuse.Цели. Кобальт имитирует состояние гипоксии, препятствуя деградации альфасубъединицы гипоксия-индуцируемого фактора, что приводит к увеличению кислородной емкости крови и может использоваться спортсменами в качестве допинга для получения конкурентных преимуществ. На сегодняшний момент для определения общего кобальта в организме используют прямые методы масс-спектрометрии с индуктивно связанной плазмой, жидкостной хроматографиитандемной масс-спектрометрии, однако Всемирным антидопинговым агентством не установлена максимально допустимая пороговая концентрация этого элемента в биожидкостях. Отсутствие четких критериев идентификации осложняет интерпретацию полученных результатов. В связи с этим, в данной статье впервые предлагается подход по косвенному определению возможных злоупотреблений кобальтом для целей допинг-контроля, основанный на изменении уровней экспрессии микроРНК, задействованных в регуляции сигнального пути гипоксии. Цель исследования заключалась в поиске возможных микроРНК-маркеров, экспрессия которых не зависит от гипоксии, вызванной физическими нагрузками, но заметно изменяется при приеме препаратов кобальта.Методы. Выделение микроРНК из образцов плазмы крови проводили при помощи набора PAXgene Blood miRNA Kit. Количественную полимеразную цепную реакцию в реальном времени проводили на амплификаторе CFX96 Bio-Rad (США) с помощью наборов miScript® SYBR® Green PCR Kit и панелей для исследования профилей экспрессии зрелых микроРНК сигнального пути гипоксии Hypoxia Signaling Pathway miScript® miRNA PCR Array.Результаты. На основании статистического анализа данных было установлено, что экспрессия hsa-miR-15b-5p в плазме крови испытуемых не зависит от физической нагрузки, но возрастает при приеме препаратов кобальта.Выводы. Разница в уровнях экспрессии при гипоксии, вызванной анаэробной физической нагрузкой, и имитацией гипоксии за счет применения кобальта делает hsa-miR-15b-5p потенциальным претендентом на роль маркера злоупотребления данным эритропоэзстимулирующим агентом

Spontaneous symmetry breaking in a two-lane model for bidirectional overtaking traffic

First we consider a unidirectional flux \omega_bar of vehicles each of which is characterized by its `natural' velocity v drawn from a distribution P(v). The traffic flow is modeled as a collection of straight `world lines' in the time-space plane, with overtaking events represented by a fixed queuing time tau imposed on the overtaking vehicle. This geometrical model exhibits platoon formation and allows, among many other things, for the calculation of the effective average velocity w=\phi(v) of a vehicle of natural velocity v. Secondly, we extend the model to two opposite lanes, A and B. We argue that the queuing time \tau in one lane is determined by the traffic density in the opposite lane. On the basis of reasonable additional assumptions we establish a set of equations that couple the two lanes and can be solved numerically. It appears that above a critical value \omega_bar_c of the control parameter \omega_bar the symmetry between the lanes is spontaneously broken: there is a slow lane where long platoons form behind the slowest vehicles, and a fast lane where overtaking is easy due to the wide spacing between the platoons in the opposite direction. A variant of the model is studied in which the spatial vehicle density \rho_bar rather than the flux \omega_bar is the control parameter. Unequal fluxes \omega_bar_A and \omega_bar_B in the two lanes are also considered. The symmetry breaking phenomenon exhibited by this model, even though no doubt hard to observe in pure form in real-life traffic, nevertheless indicates a tendency of such traffic.Comment: 50 pages, 16 figures; extra references adde

Tumor suppressor function of the SEMA3B gene in human lung and renal cancers

The SEMA3B gene is located in the 3p21.3 LUCA region, which is frequently affected in different types of cancer. The objective of our study was to expand our knowledge of the SEMA3B gene as a tumor suppressor and the mechanisms of its inactivation. In this study, several experimental approaches were used: tumor growth analyses and apoptosis assays in vitro and in SCID mice, expression and methylation assays and other. With the use of the small cell lung cancer cell line U2020 we confirmed the function of SEMA3B as a tumor suppressor, and showed that the suppression can be realized through the induction of apoptosis and, possibly, associated with the inhibition of angiogenesis. In addition, for the first time, high methylation frequencies have been observed in both intronic (32-39%) and promoter (44-52%) CpG-islands in 38 non-small cell lung carcinomas, including 16 squamous cell carcinomas (SCC) and 22 adenocarcinomas (ADC), and in 83 clear cell renal cell carcinomas (ccRCC). Correlations between the methylation frequencies of the promoter and the intronic CpG-islands of SEMA3B with tumor stage and grade have been revealed for SCC, ADC and ccRCC. The association between the decrease of the SEMA3B mRNA level and hypermethylation of the promoter and the intronic CpG-islands has been estimated in renal primary tumors (P < 0.01). Using qPCR, we observed on the average 10- and 14-fold decrease of the SEMA3B mRNA level in SCC and ADC, respectively, and a 4-fold decrease in ccRCC. The frequency of this effect was high in both lung (92-95%) and renal (84%) tumor samples. Moreover, we showed a clear difference (P < 0.05) of the SEMA3B relative mRNA levels in ADC with and without lymph node metastases. We conclude that aberrant expression and methylation of SEMA3B could be suggested as markers of lung and renal cancer progression