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The theoretical and methodological foundation for studying the problems of sustainable development today is insufficiently formed. The issues of knowledge in the field of sustainable development at the state and regional levels remain open. The object of the study is the Ural macroregion, which includes seven constituent entities of the Russian Federation (Sverdlovsk, Chelyabinsk, Kurgan, Orenburg regions, Perm Territory, the Republic of Udmurtia and the Republic of Bashkortostan). The set of industries (types of economic activity) that are part of the military-industrial complex is important in ensuring the sustainability for the socio-economic development of the Urals. The findings and conclusions of scientists on the problems of ensuring sustainable development, as well as the functioning of the military-industrial complex, were adopted as the methodological basis for the study. The research methodology is based on formalization and systematization approaches, bibliometric and content analysis of scientific literature devoted to the theoretical and methodological knowledge of sustainable development, as well as the development of territories where the military-industrial complex dominates in the economic space. The main results of the study are: putting forward and proving a scientific hypothesis confirming the influence of the functioning and development of the military-industrial complex on ensuring sustainable socio-economic development in the Ural macroregion. The results obtained during the presented research can be used in the development of strategic documents aimed at forming sustainable socio-economic development of territories at various levels.

The theory, methodology and practice of research of socioeconomic and theoretical-methodological aspects of sustainable development are at the stage of comprehension. These provisions are rather poorly elaborated at the level of regional economy. However, the negative events in the economic, political, environmental and social spheres, which we are witnessing, confirm the high significance of this scientific agenda. The authors see the strategic vector of socio-economic development of the regions of the Russian Federation in the context of reducing spatial disproportions, which will ensure sustainable socio-economic development of the entire state, as well as its constituent regions. The research presented in this article is aimed at substantiating the most important factor of sustainable development dynamics of the Russian Federation, namely, overcoming spatial disproportions between the regions that make up the state. The methodological basis of the study is based on the findings and conclusions of scientists who study the problems of sustainable socioeconomic development, as well as those working in the field of regional economics. The research methodology is based on the approaches of formalization and systematization, bibliometric and content analysis, as well as statistical, analytical, comparative, economic and mathematical methods. Scientific publications of domestic and foreign scientists, normative, analytical, program and strategic materials of the Government of the Russian Federation, state regional authorities formed the information base. The main results of the study are: the nomination and proof of the scientific hypothesis, confirming the importance of the reduction of spatial disparities in the system of factors to ensure sustainable development of the state. The results of the study contribute to the development of theoretical and methodological provisions of regional economics, state management of economic and social processes in order to achieve sustainable development.

The theory, methodology and practice of studying the functioning and development of sanatorium and resort activities in the context of ensuring stable regional dynamics and sustainable development of territories are at the stage of comprehension. The strategic vector of socio-economic development of a new subject of the Russian Federation, the Republic of Crimea, is seen by the authors in close connection with the development of sanatorium and resort activities. The study is aimed at substantiating the most important factor in the sustainable dynamics of the Republic of Crimea, namely the stable functioning and development of sanatorium and resort activities. The methodological basis of the study was the findings and conclusions of scientists studying the issues of sustainable functioning of sanatorium and resort activities and working in the subject field of regional economics, health economics, the concept of human capital, the concept of the quality of life of the population, and the theory of services. The research methodology is based on formalization and systematization approaches, bibliometric and content analysis, and the method of analogies and comparative analytical methods are also used. The information base consisted of scientific publications of domestic and foreign scientists, regulatory, analytical, program and strategic materials of the Government of the Russian Federation, state regional authorities of the Republic of Crimea. The main results of the study are: putting forward and proving a scientific hypothesis confirming the importance of the functioning and evolution of sanatorium and resort activities and medical tourism in the system of factors for ensuring sustainable development of the Republic of Crimea. The results of the study contribute to the development of theoretical and methodological principles of regional economics, public management of economic and social processes in order to achieve sustainable regional development.

The basis for development of an optimal organizational and economic mechanism for land tenure and land use adapted to the conditions of the digital economy is based on the principles and patterns of its regulation through the identification and application of effective areas of administrative influence at the federal level through use of innovative tools. The analysis of the distribution of the land fund of the Russian Federation by land categories for 2016-2020 is carried out and the need to solve some of the fundamental problems of formation of the mechanism under consideration through use of the identified development patterns formed by the regulatory, economic and digital environment for its regulation, considering the dynamics of changes in the category of agricultural land within the study period is confirmed. It has been established that interaction of subjects of land relations depends on the degree of practical and scientific substantiation of the organizational and economic mechanism of land tenure and land use, the functional state of the digital and institutional environment.

An automated technology is developed that combines the processes of describing of the use case (precedent) and compiling of test cases sets. For this purpose, a mathematical model of the precedent is proposed, representing it in the form of an oriented graph. Each vertex of the graph corresponds to a precedent item, and each edge defines the transition conditions and data that define these conditions. The classification of the test case scenarios was used, which made it possible to distinguish 7 types of items. For each type of test case item, a separate test case template has been developed. Templates have 3 sections: data entered in this item; data previously received in the system; and the results of the implementation of the item. An algorithm for identify-ing independent paths and a procedure for determining the completion of traversal is developed. Software tools that support the proposed technology of drawing test cases have been developed. The tests showed a significant reduction in the time when this technology was used in comparison with existing solutions which provide the separate processes for describing of use cases and drawing up of test cases.

The production of the initial liquid metal with minimal departures from the regulated chemical composition is one of the main conditions for the production of high-quality castings with predetermined physical, mechanical and operational properties. An important factor is also the optimization of the composition of the furnace charge to reduce production costs. The program complexes to automate the management of the entire melting process, incl. calculation of furnace charge and refining the liquid metal to the predetermined chemical composition during the melting process cannot be claimed by small and medium businesses because of the high cost, complexity and increased requirements for the qualification of personnel. Aim: The aim of the research is to automate the calculation of the furnace charge for obtaining a melt with a given set of physical, mechanical and operational properties by developing and implementing appropriate algorithms to obtain a minimum cost composition of the furnace charge, taking into account the limitations of the components. Materials and Methods: Proposed method for calculating the mass of components for obtaining the melt of predetermined chemical composition using the model of initial and output data. The model takes into account the relationships between the required melt composition in the form of a multitude of chemical elements and a multitude of components, in turn the component is described as a set of chemical elements. Algorithms for checking the correctness of the initial data on the range of the number of components and chemical composition, determining the output set of components with subsequent correction for determining the minimum total cost, an algorithm for eliminating the excess of the chemical element in the melt are described. Results: The proposed method allows identifying at the initial stage of calculation the unacceptable values in the initial data. Algorithms are implemented in the form of a software product that can be used by small and medium business.

The article is devoted to the current amendments to the Russian legislation on targeted training aimed at meeting the employers’ needs in specialists with secondary vocational or higher education. It raises the issue of the legal nature of a contract on targeted training, which is compared with an apprenticeship contract. It is proved that the contract on targeted training is in the field of labour law, therefore, the relevant legal relations should be regulated by labour-law norms

In the article presented, the authors prove the objective need for the development of the electricity industry of the Republic of Crimea on the basis of the active use of the renewable energy sources, caused by the world processes of the formation of the “green” economy. The object of the study is the territory of the Republic of Crimea - a bright representative of the energy deficit region. The development of the electricity industry of the Republic of Crimea on the basis of the use of the renewable energy sources serves as the basis for ensuring regional energy security, the fragility of which was determined by the political events of 2015. The authors studied the research papers of leading domestic and foreign scientists devoted to the analysis of the renewable energy sources in order to justify the further active development of a wind energy, a solar energy as well as bioenergy and landfill gas energy in the Republic of Crimea. The development of the Republic of Crimea’s electricity industry through the use of the renewable energy sources and local fuels is an essential area of modern active regional policy. It is determined by the fact that this direction, firstly, is considered as environmentally friendly generation, and secondly, contributes to a qualitative increase in regional energy security level.

The article focuses on the need to study the processes of a new high-tech industrialization (neo-industrialization) at the macroregion level. The object of the study is the economic region of the Russian Federation (the macroregion) as a relatively integral spatially localized and complexly structured socio-economic entity, which includes four Ural regions – Kurgan region, Orenburg region, Sverdlovsk region and Chelyabinsk region, Perm krai and two republics – Republic of Bashkortostan and Republic of Udmurtia, which corresponds to the borders of the Ural economic region according to the all-Russian classifier of economic regions. The purpose of this study is to prove the relationship between the level of industrial production and the level of the pawnshop market development in the Ural macroregion. The territory of the Ural macroregion is analyzed from the perspective of the relationship between the level of industrial production and the level of the pawnshop market development. The Authors proved the existence of a direct relationship between the level of industrial production and the level of pawnshops territorial concentration, and also identified the territories that are leaders in the pawnshops concentration. The Authors put forward a scientific hypothesis about the most effective model of pawnshop activity. The conclusion is made about mutually beneficial cooperation between precious metal processing enterprises and pawnshops, which contributes to the development of both the pawnshop sector and this type of industrial activity.

Due to the global world transformations caused by the spread of the new coronavirus infection, the regional development policy requires a radical rethinking. The world economic community is busy looking for new factors to accelerate economic dynamics. The use of space resources, the advantages of polarized development is the basis for the formation and development of growth poles. However, the problems of assessing the effectiveness of this territorial entity remain poorly developed. The purpose of this article is to propose a methodological basis for assessing the effectiveness of growth poles in the economic space of the region.

The practical application of the growth pole concept is one of the possible sources of accelerating economic dynamics. The purpose of the study is to identify growth poles in the Ural macroregion. The scientific article proposes to consider the theoretical basis of the growth pole concept, as well as practical application in various forms of the growth pole concept in the territory of the Russian Federation, including in the Ural macroregion. In addition, the authors will apply theoretical and empirical methods in this study. The novelty of the authors' research lies in the hypothesis on the classification of growth poles in Russian practice, in identifying the number of growth poles in the territory of the Russian Federation and the Ural macroregion. This article will be of interest to executive authorities, the scientific community, teachers and students of higher educational institutions.

The article is devoted to the issues of organization and prospects of audit development in terms of digitalization. The state and the measures needed to improve audit effectiveness in the context of automation are investigated and analyzed. The factors that negatively influence the audit digitization process and the possible directions of development of audit activity in the computerized environment are identified. The use of automation helps auditors process more data and focus on identifying risks, allowing for higher quality audits. Classification of information technology of audit, conditions for the development of audit software, factors that impede audit automation are given. Audit digitization is a new level in the audit field that is rapidly evolving and rapidly expanding in enterprises, especially those that use an automated accounting method. The software currently used by auditors needs to be refined to meet the demands of today's world. Digitization will become relevant for all audit firms seeking to gain competitive advantage and take a place in the audit services market.

During the coronavirus pandemic the problems of sustainability of the Russian economy, as well as the economy of regions that form it, sharply worsened. The protracted, global course of the COVID-19 epidemic has given the exceptional urgency to the problem of the sustainable development of regions. There is a lack of knowledge of the global, national, regional dynamics in the conditions of the epidemic, despite the world scale of the problem. The sustainability and integrity of the country depend on the sustainability of the regional social and economic system. The purpose of this research is to identify trends in the development of the regional consumer market in the conditions of the pandemic, to reveal its significance in forming the region’s sustainable development. To conduct the research the authors used the first statistic data about the functioning of the national and regional consumer market in the conditions of the pandemic. The object of the study is one of the Russian regions –the Sverdlovsk region. The theoretical basis of the research is the concept of the sustainable development of the social and economic system.

In article the authors argue that the industrial future is the most important factor in ensuring sustainable development of a traditional industrial region. The conclusions are based on the study of the first results of socio-economic development in the context of the coronavirus pandemic, a prominent representative of traditionally industrial regions - the Sverdlovsk region. The authors base their own research on the conceptual guidelines of the scientific paradigm of sustainable development. Operative statistical data on the Sverdlovsk region are presented. In the study the conclusions were reached about the immediate prospects for the development of the Sverdlovsk region economy in the context of a new coronavirus infection. The processes that are an approximate scenario for the development of events at the regional level were identified. The importance of maintaining industrial specialization in the new realities of the economy as an important factor in sustainable development not only at the national level, but also at the regional level was revealed.

System research of gnosiological nature of economic interests is conducted in the article. Certainly essence of economic interests, and the features of going are selected near the analysis of this question. It was concluded that the origin and formation of the content of economic interest is determined by the fact of his involvement in social media combined process of production.В статье проводится системное исследование гносеологической природы экономических интересов. Определена сущность экономических интересов, и выделены особенности подходов к анализу этого вопроса. Сделан вывод, что возникновение и формирование содержания экономического интереса детерминировано фактом привлечения его носителя в общественно комбинированный процесс производства.

Project management offices (or PMOs) are becoming a very well known concept and widely used organizational unit which is organized to support and develop project management systems and methodologies in companies in various sectors of economy. PMOs are being organized in more and more Russian engineering companies. However, PMOs have different forms and functions in different organizational context. It is very useful for theory and practice to analyze the real practice and define which characteristics and forms of PMOs are more accepted. The research presented in the article shows that PMOs are very popular among Russian engineering companies and the trends in their development are in line with the trends in other countries and industries.

The real educational system must be modified because of global changing in social life. It is necessary to use the innovative technologies in the educational process. The modern labour market takes the special claiming to the graduates. It is important to study the competitiveness of students in the culture sphere. Thus, the article is devoted to the study of the competitiveness of students studying in educational institutions in the culture and art spheres; problems of modern information technologies introduction in educational process of such educational institutions.

Introduction: Cytomegalovirus (CMV) is a DNA-containing virus that is widespread worldwide and is of great importance in infectious pathology of children and adults. The aim of this study is to evaluate the prevalence of CMV among children and immunocompromised patients in the Nizhny Novgorod region (central Russia) and to perform a phylogenetic analysis of the identified strains., Materials and Methods: DNA samples of CMV detected in frequently ill children and adult recipients of solid organs were studied. The genetic diversity of CMV was assessed for two variable genes: UL55 (gB) and UL7 3(gN), using NGS technology on the Illumina platform. Phylogenetic trees were constructed in the MEGA X program, the reliability of the cluster topology on the trees was confirmed using the rapid bootstrap method based on the generation of 500 pseudo-replicas., Results: Circulation of 5 CMV genotypes by gene UL55 (gB) and 5 genotypes by gene UL73 (gN) was established in the territory of the Nizhny Novgorod region. While genotypes gB1 and gB2 dominated both in children and in adults, genotype gB5 was detected only in children. The spectrum of gN genotypes was more diverse: genotypes gN4a and gN3b prevailed in children, and gN1 and gN4b genotypes were predominant in adults. The obtained results allowed us to establish the similarity of the landscape of CMV genotypes circulating in Russia (Nizhny Novgorod region), Brazil, China and the USA., Conclusion: The obtained data indicate the similarity of the landscape of CMV genotypes circulating in Russia (Nizhny Novgorod region), Brazil, China and the USA: in children, the predominant genotypes are gB1 (40.0%), gB2 (33.3%), gN4a (42.8%), and gN3b (28.6%), while in adults (recipients of solid organs) genotypes gB1 (37.5%), gB2 (37.5%), gN1 (26.3%), and gN4b (26.3%) are prevailed.

Introduction: Rotavirus infection is the major cause of severe dehydrating diarrhea requiring hospitalization in young children worldwide. Due to their segmented genome, rotaviruses are capable of gene reassortment, which makes the emergence and spread of genetically novel strains possible. The purpose of this study was to search for unusual rotaviruses circulating in Nizhny Novgorod in 2021‒2023 and their molecular genetic characterization based on all genome segments., Materials and Methods: Rotavirus-positive stool samples of children were examined by PCR genotyping and electrophoresis in PAAG. cDNA fragments of each of the 11 genes ( VP1‒VP4 , VP6 , VP7 , NSP1‒NSP5 ), 570 to 850 nucleotide pairs in length were sequenced for the selected strains. The phylogenetic analysis was performed in the MEGA X program., Results: In the study period 2021‒2023, 11 G[P] combinations with a predominance of G3P[8] (59.5%) were identified. Six atypical Rotavirus А (RVA) strains were identified: 2 strains of the G2P[4] genotype (G2-P[4]-I2-R2-C2-M2-A3-N2-T3-E2-H3, G2-P[4]-I2-R2-C2-M2-A3-N2-T3-E3-H2) and 4 G3P[9] strains (all strains had the genotype G3-P[9]-I2-R2-C2-M2-A3-N2-T3-E3-H3). Phylogenetic analysis based on all genes showed an evolutionary relationship between rotaviruses similar to rotaviruses of cats and dogs (BA222-like) and unusual strains of the G2P[4] genotype, for which a mixed combination of genotypes was identified and characterized for the first time., Discussion: The results obtained expand the understanding of the diversity of reassortant RVAs, as well as complement the data on the genotypic structure of the rotavirus population in Nizhny Novgorod., Conclusion: The wide genetic diversity of reassortant RVA can help rotaviruses overcome the immunological pressure provided by natural and vaccine-induced immunity. In this regard, to control the emergence of new variants and assess changes in the virulence of rotaviruses after reassortment processes, continuous molecular monitoring for circulating RVA is necessary.

Enteroviruses are one of the most common causative agents of infectious diseases of the central nervous system. They are characterized by genetic variability, the ability to infect a wide range of cells, including brain microglial cells and astrocytes, and persist in the central nervous system tissue, causing delayed and chronic diseases. The review presents data on the basis of neurovirulence of non-polio enteroviruses and the most common pathogens causing enteroviral neuroinfections.

Since 2013, there has been an increase in reports of the spread of a double intergroup reassortant strain of rotavirus type A (RVA) with the genotype G3P[8] and other genes belonging to the second genogroup I2-R2-C2-M2-A2-N2-T2-E2-H2. In our study, we provide a molecular genetic characterization of rotaviruses with genotype G3P[8]-I2 isolated in Nizhny Novgorod. In our study, we used RT-PCR, Sanger sequencing, RNA-PAGE methods. Phylogenetic and phylodynamic analysis were performed using the Bayesian approach. According to our study, there was a significant increase in the proportion of G3P[8] from 15% during the period of 2020-2021 to 53% during the period of 2021-2022 in Nizhny Novgorod, Russia. Phylogenetic analysis based on the VP4 gene revealed that DS-1-like RVAs isolated in Nizhny Novgorod belong to different clusters of the P[8]-3.1 lineage, with a level of variation ranging from 1.1% to 1.3%. Based on the VP6 gene, the equine-like RVAs identified by us carry genetic variants belonging to three distinct clusters of the lineage I2-V, with a variation level ranging from 2.0% to 4.5%. These data indicate the genotypic diversity of circulating DS-1-like G3 RVAs. Phylogenetic analysis of the VP7 gene allowed us to assign the isolates identified in our study to the G3-1 lineage. We estimated that the circulation of the most recent common ancestor of the spreading strains dates back to 2002. Additionally, we determined the typical level of mutations in the VP7 gene, which amounted to 2.14*10-3 substitutions/per site/per year., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Aim      To determine the effect of sodium-glucose cotransporter-2 inhibitors (SGLT2i) on kidney function in acute decompensated heart failure (ADHF).Material and methods  A controlled randomized study on the dapagliflozin treatment in ADHF was performed. Patients were randomized to a main group (standard therapy supplemented with dapagliflozin) or a control group (standard therapy for ADHF). The primary endpoint was the development of acute kidney injury (AKI). 200 patients were included (mean age, 74±12 years; 51% men). 31% of patients had type 2 diabetes mellitus (DM2). Mean left ventricular ejection fraction (LV EF) was 47±14 %; in 44.5% of patients, LV EF was less than 45%. Median concentration of N-terminal pro-brain natriuretic peptide (NT-proBNP) was 5225 [3120; 9743] pg / ml, glomerular filtration rate (GFR) was 51 [38; 64] ml / min / 1.73 m2.Results In-hospital mortality was 6.5%. Analysis of the dynamics of body weight loss showed significant differences (4200 [2925; 6300] g vs. 3000 [1113; 4850] g; p=0.011) in favor of the dapagliflozin group. The requirement for increasing the daily dose of furosemide and adding an another class diuretic (thiazide or acetazolamide) did not differ between the groups. However, median furosemide dose during the stay in the hospital was lower in the dapagliflozin group (80 [67; 120] mg vs. 102 [43; 120] mg; p=0.016). At 48 hours after randomization, GFR significantly decreased in the dapagliflozin group (-5.5 [-11; 3] ml/min/ 1.73 m2) compared to the control group (-0.3 [-4; 5] ml / min/1.73 m2, р=0.012). Despite this, GFR did not differ between the groups at discharge (51 [41; 66] ml/min/1.73 m2 and 49 [38; 67] ml/min/1.73 m2, respectively; p = 0.84). In the dapagliflozin group, frequency of AKI episodes was not increased compared to the control group (13 and 9.4%, respectively; p = 0.45).Conclusion      The dapagliflozin treatment in ADHF is associated with more pronounced body weight loss and lower average doses of loop diuretics during the period of stay in the hospital, with no associated clinically significant impairment of renal function.

Reassortant DS-1-like rotavirus A strains have been shown to circulate widely in many countries around the world. In Russia, the prevalence of such strains remains unclear due to the preferred use of the traditional binary classification system. In this work, we obtained partial sequence data from all 11 genome segments and determined the full-genotype constellations of rare and reassortant rotaviruses circulating in Nizhny Novgorod in 2016-2019. DS-1-like G3P[8] and G8P[8] strains were found, reflecting the global trend. Most likely, these strains were introduced into the territory of Russia from other countries but subsequently underwent further evolutionary changes locally. G3P[8], G9P[8], and G12P[8] Wa-like strains of subgenotypic lineages that are unusual for the territory of Russia were also identified. Reassortant G2P[8], G4P[4], and G9P[4] strains with one Wa-like gene (VP4 or VP7) on a DS-1-like backbone were found, and these apparently had a local origin. Feline-like G3P[9] and G6P[9] strains were found to be phylogenetically close to BA222 isolated from a cat in Italy but carried some traces of reassortment with human strains from Russia and other countries. Thus, full-genotype determination of rotavirus A strains in Nizhny Novgorod has clarified some questions related to their origin and evolution., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Circulating serum miRNA are increasingly used as biomarkers and potential treatment targets in several clinical scenarios, including cardiovascular diseases. However, the current data on circulating miRNA in thoracic aorta aneurism (TAA) patients are inconclusive. The aim of the present study is to compare the levels of several circulating miRNA in patients with degenerative TAA, coronary artery disease (CAD), and controls for special profile identification. We have identified several candidates for the role of new biomarkers: miR-143-3p, miR-181-5p, miR-126-3p, miR-126-5p, miR-145-5p, miR-150-5p, and miR-195-5p., Materials and Methods: Serum samples of 100 patients were analyzed, including 388 TAA patients scheduled for elective surgery, 67 patients with stable CAD and 17 controls, were used for miRNA isolation and identification., Results: More specific for TAA with very high predictive ability in ROC analysis was an increase in the levels of miR-21-5p, miR-29b-5p, miR-126-5p/-3p, miR-181b-5p, and miR-92a-3p, with the latter microRNA being investigated as a novel potential marker of TAA for the first time., Conclusion: TAA and CAD patients demonstrated a significant increase in the levels of circulating miR-126-5p/-3p, miR-181b-5p, and miR-29b-3p. More specific for TAA with very high predictive ability in ROC analysis was an increase in the levels of miR-21-5p, -29b-5p, -126-5p/-3p, 181b-5p, and -92a-3p, with the latter microRNA being investigated as a potential marker of TAA for the first time.

The picobirnaviruses (Picobirnaviridae, Picobirnavirus, PBVs) are currently thought to be animal viruses, as they are usually found in animal stool samples. However, no animal model or cell culture for their propagation has yet been found. In 2018, a hypothetical assumption about PBVs belonging to prokaryotic viruses was put forward and experimentally substantiated. This hypothesis is based on the presence of Shine-Dalgarno sequences in the genome of all PBVs before three reading frames (ORF) at the ribosomal binding site, with which the prokaryotic genome is saturated, while in the eukaryotic genome such regions occur with low frequency. The genome saturation with the Shine-Dalgarno sequences, as well as the preservation of this saturation in the progeny, according to scientists, allows us to attribute PBVs to prokaryotic viruses. On the other hand, there is a possibility that PBVs belong to viruses of eukaryotic hosts - fungi or invertebrates, since PBV-like sequences similar to the genome of fungal viruses from the families of mitoviruses and partitiviruses have been identified. In this regard, the idea arose that, in terms of reproduction mode, PBVs resemble fungal viruses. The divergence of views on the true PBV host(s) has sparked discussions among scientists and required further research to elucidate their nature. The review highlights the results of the search for a PBV host. The reasons for the occurrence of atypical sequences among the PBV genome sequences that use an alternative mitochondrial code of lower eukaryotes (fungi and invertebrates) for the translation of viral RNA-dependent RNA polymerase (RdRp) instead of the standard genetic code are analyzed. The purpose of the review was to collect arguments in support of the hypothesis about the phage nature of PBVs and to find the most realistic explanation of the reasons for identifying non-standard genomic sequences for PBVs. Based on the hypothesis about the genealogical relationship of PBVs with RNA viruses from other families with similar segmented genomes, such as Reoviridae, Cystoviridae, Totiviridae and Partitiviridae, virologists support the assumption of a decisive role in the origin of atypical PBV-like reassortment strains between PBVs and viruses of the listed families. The collected arguments given in this review indicate a high probability of a phage nature of PBVs. The data presented in the review show that the belonging of PBV-like progeny to prokaryotic or eukaryotic viruses is determined not only by its genome saturation level with a prokaryotic motif, standard or mitochondrial genetic code. The primary structure of the gene encoding the viral capsid protein responsible for the presence or absence of specific proteolytic properties of the virus that determine its ability for independent horizontal transmission into new cells may also be a decisive factor., Competing Interests: The authors declare no conflict of interest., (Copyright © AUTHORS.)

Hand-foot-and-mouth disease (HFMD) is the most common enteroviral infection in South-East Asia. When evaluating the role of enterovirus 71 (EVA71) as an etiological agent of infectious disease in South Vietnam, we revealed a high proportion of EVA71 among identified species A enteroviruses found in 3542 samples from HFMD cases; 125 samples from cases of enteroviral meningitis; and 130 samples from acute flaccid paralysis (AFP) cases. These represent 50%, 54.8%, and 51.5%, respectively. According to molecular analysis, 90% of EVA71 were attributed to genotype C4 and 10% were attributed to genotype B5. The predominance of EVA71 circulation among the population proves the need to strengthen surveillance (with monitoring of enterovirus circulation for facilitation of HFMD outbreak prediction) and to increase the effectiveness of preventative measures by the implementation of vaccination against EVA71-associated infections. A phase III trial of a Taiwanese vaccine (EV71vac) in Taiwan and South Vietnam showed its safety, tolerability, and efficacy in children aged 2-71 months. This B4 genotype-based vaccine, which features cross-protection against B5 and C4 genotypes, and other existing EV71 vaccines can serve as a good approach to solving the HFMD problem, which is so important for Vietnam.

Introduction: In Russia, rotavirus A is the main cause of severe viral gastroenteritis in young children. The molecular features that allow a rotavirus of a particular genotype to gain an evolutionary advantage remain unclear, therefore, the study of the genetic diversity of rotaviruses based on genes encoding nonstructural proteins (NSPs) responsible for the reproduction of the virus in the cell is an urgent task., Objective: To study the genetic diversity of rotaviruses of genotype G9P[8], which dominated Nizhny Novgorod in 20112020, based on genes encoding nonstructural proteins., Materials and Methods: Rotavirus-positive samples were subjected to PCR-genotyping and sequencing of NSP1 NSP5 genes. Phylogenetic analysis was carried out in the MEGA X program., Results: In the period 20112020, G9P[8] rotaviruses with four variants of the NSP2 gene were co-circulating in Nizhny Novgorod. New alleles were noted in 2012 (N1-a-III), 2016 (N1-a-IV) and in 2019 (N1-a-II). The appearance of new variants of other genes occurred in 2014 (E1-3, NSP4), 2018 (T1-a3-III, NSP3) and in 2019 (A1-b-II, NSP1). NSP2 gene had the most variable amino acid sequence (16 substitutions), 2 to 7 substitutions were observed in NSP1, NSP3 and NSP4, NSP5 was conservative., Discussion: The results obtained are consistent with the literature data and indicate the participation of NSP genes in maintaining the heterogeneity of the rotavirus population., Conclusion: Until 2018, the genetic diversity of rotaviruses in Nizhny Novgorod was determined by the circulation of strains carrying several alleles of the NSP2 gene and conservative genes NSP1, NSP3NSP5. By the end of the study period, new variants of the genotype G9P[8] were formed in the population, carrying previously unknown combinations of alleles of nonstructural genes.

Modern molecular genetic methods, massive parallel sequencing in particular, allow for genotyping of various pathogens with the aim of their epidemiological marking and improvement of molecular epidemiological surveillance of actual infections, including cytomegalovirus infection. The aim of the study is to evaluate the next-generation sequencing (NGS) technology for genotyping clinical isolates of cytomegalovirus (CMV)., Materials and Methods: The object of the study were samples of biological substrates (leukocyte mass, saliva, urine) taken from patients who underwent liver and kidney transplantation. Detection of CMV DNA was carried out by a real-time PCR using commercial diagnostic AmpliSense CMV-FL test systems (Central Research Institute for Epidemiology, Moscow, Russia). DNA extraction was performed using DNA-sorb AM and DNA-sorb V kits (Central Research Institute for Epidemiology) in accordance with manufacturer's manual. The quality of the prepared DNA library for sequencing was assessed by means of the QIAxcel Advanced System capillary gel electrophoresis system (QIAGEN, Germany). Alignment and assembly of nucleotide sequences were carried out using CLC Genomics Workbench 5.5 software (CLC bio, USA). The sequencing results were analyzed using BLAST of NCBI server., Results: CMV DNA samples were selected for genotyping. The two variable genes, UL55 (gB) and UL73 (gN), were used for CMV genotype determination, which was performed using NGS technology MiSeq sequencer (Illumina, USA). Based on the exploratory studies and analysis of literature sources, primers for genotyping on the UL55 (gB) and UL73 (gN) genes have been selected and the optimal conditions for the PCR reaction have been defined. The results of sequencing the UL55 (gB) and UL73 (gN) gene fragments of CMV clinical isolates from recipients of solid organs made it possible to determine the virus genotypes, among which gB2, gN4c, and gN4b were dominant. In some cases, association of two and three CMV genotypes has been revealed., Conclusion: The application of the NGS technology for genotyping cytomegalovirus strains can become one of the main methods of CMV infection molecular epidemiology, as it allows for obtaining reliable results with a significant reduction in research time., Competing Interests: Conflicts of interest. The authors have no conflicts of interest to declare.

Noroviruses are important etiological agents causing acute intestinal infection in humans. In the last decades, the most common norovirus genotype was GII.4 despite a significant genetic diversity among strains, while the active circulation of noroviruses with other genotypes was observed periodically. This study shows an increase in the detection rate of recombinant GII.3[P12] norovirus in Nizhny Novgorod, Russia, from 6.8% in 2018-2019 to 34.9% in 2020-2021. We performed a phylogenetic analysis based on the nucleotide sequences of noroviruses possessing this genotype obtained in this work, as well as presented in the GenBank database. It has been shown that the circulation of GII.3[P12] noroviruses in the study area was the result of several independent introductions, either directly from the Western Pacific region, or through the Asian part of Russia. The polyphyletic origin, the geographical expansion, and the growth of the epidemic significance of the recombinant GII.3[P12] noroviruses were noted., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: The novel coronavirus infection COVID-19 is a major public health problem worldwide. Several publications show the presence of gastrointestinal (GI) symptoms (nausea, vomiting, and diarrhea) in addition to respiratory disorders.The aim of this study was the monitoring of RNA of COVID-19 pathogen, coronavirus SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) in children hospitalized with acute intestinal infection (AII), with following molecular-genetic characterization of detected strains., Material and Methods: Fecal samples of children with AII hospitalized in infectious hospital of Nizhny Novgorod (Russia) in the period from 01.07.2020 to 31.10.2021 were used as material for the study. Viral RNA detection was performed by real-time polymerase chain reaction (RT-PCR). The nucleotide sequence of S-protein gene fragment was determined by Sanger sequencing., Results and Discussion: SARS-CoV-2 genetic material was detected in 45 out of 2476 fecal samples. The maximum number of samples containing RNA of the virus occurred in November 2020 (detection rate of 12.2%). In 20.0% of cases, SARS-CoV-2 RNA was detected in combination with rota-, noro-, and adenoviruses. 28 nucleotide sequences of S-protein gene fragment complementary DNA (cDNA) were determined. Phylogenetic analysis showed that the studied SARS-CoV-2 strains belonged to two variants. Analysis of the S-protein amino acid sequence of the strains studied showed the absence of the N501Y mutation in the 2020 samples, which is a marker for variants with a high epidemic potential, called variants of concern (VOC) according to the World Health Organization (WHO) definition (lines Alpha B.1.1.7, Beta B.1.351, Gamma P.1). Delta line variant B.1.617.2 was identified in two samples isolated in September 2021., Conclusion: The detection of SARS-CoV-2 RNA in the fecal samples of children with AII, suggesting that the fecal-oral mechanism of pathogen transmission may exist, determines the necessity to optimize its monitoring and to develop an algorithm of actions with patients with signs of AII under the conditions of a novel coronavirus infection pandemic.

Surveillance for acute flaccid paralysis syndrome (AFP) in children under 15 is the backbone of the Global Polio Eradication Initiative. Laboratory examination of stool samples from AFP cases allows the detection of, along with polioviruses, a variety of non-polio enteroviruses (NPEV). The etiological significance of these viruses in the occurrence of AFP cases has been definitively established only for enteroviruses A71 and D68. Enterovirus Coxsackie A2 (CVA2) is most often associated with vesicular pharyngitis and hand, foot and mouth disease. Among 7280 AFP cases registered in Russia over 20 years (2001-2020), CVA2 was isolated only from five cases. However, these included three children aged 3 to 4 years, without overt immune deficiency, immunized with 4-5 doses of poliovirus vaccine in accordance with the National Vaccination Schedule. The disease resulted in persistent residual paralysis. Clinical and laboratory data corresponded to poliomyelitis developing during poliovirus infection. These findings are compatible with CVA2 being the cause of AFP. Molecular analysis of CVA2 from these patients and a number of AFP cases in other countries did not reveal association with a specific phylogenetic group, suggesting that virus genetics is unlikely to explain the pathogenic profile. The overall results highlight the value of AFP surveillance not just for polio control but for studies of uncommon AFP agents.

Introduction: Enterovirus (nonpolio) infection is widespread all over the world, registered as sporadic cases and large-scale outbreaks and can cause severe lesions such as serous meningitis. Epidemiological studies have shown that enterovirus (Picornaviridae; Enterovirus) variant Echovirus 30 (E30) is the most frequently detected variant in patients with enterovirus meningitis in the Russian Federation. However, no vaccines to prevent the disease caused by this pathogen have been developed so far. One of the promising modern trends in terms of creating vaccine preparations is the use of virus-like particles (VLPs), including chimeric ones containing the biological structures of viruses belonging to different species.The aim of this work was to obtain norovirus (Caliciviridae; Norovirus) VLPs displaying enterovirus Echovirus E30 full-length VP1 on the surface., Material and Methods: The nucleotide sequences of VP1 protein of norovirus genotype GII.4 and VP1 E30 of genotype h circulating in Russia were used. The SN-VP1E30 protein was constructed, in which the shell (S) and the hinge regions of the norovirus VP1 are fused into one molecule with the full-length VP1 of the E30 virus. The protein was expressed in E. coli, purified using affinity chromatography, and characterized by polyacrylamide gel electrophoresis (PAGE) and immunoblotting. VLPs were visualized by electron microscopy., Results: The S N-VP1E30 protein expressed in E. coli as insoluble form, so the conditions for SN-VP1E30 solublisation were defined. Sucrose has been shown to significantly increase the efficiency of renaturation. Electrophoretic mobility comparison of denatured and non-denatured SN-VP1E30 demonstrated that most monomers form high molecular weight compounds. Electron microscopy showed that renatured SN-VP1E30 spontaneously forms empty virus-like particles about 50 nm in diameter., Conclusion: Chimeric protein SN-VP1E30 self-assemble into VLPs displaying the VP1 protein of E30 variant that is highly prevalent in Russia. Further immunological research is necessary to characterize VLPs potential for development of the vaccine for enteroviral meningitis prevention.

Introduction: The pentavalent rotavirus vaccine has been registered in Russia, however, the vaccination coverage remains low, and an annual increase in the incidence of rotavirus infection is unavoidable. In this regard, molecular monitoring of rotaviruses in order to search for new variants possessing epidemic potential is an urgent task., Material and Methods: PCR genotyping and VP4 and VP7 genes sequencing were used to characterize rotaviruses circulating in Nizhny Novgorod in 2012-2020. The phylogenetic analysis of the strains was carried out using the BEAST software package., Results: The spectrum included 17 genotypes with predominance of G9P[8] (37,4%). Detected in this study genotypes G1P[4], G1P[9], G2P[8], G4P[4], G4P[6], G8P[8], and G9P[4] were not previously identified in Nizhny Novgorod. The circulation of DS-1-like strains possessing genotypes G1P[8], G3P[8], G8P[8], or G9P[8] and a short RNA pattern had been shown. Rotaviruses of the common genotypes were genetically heterogeneous and belonged to different phylogenetic lineages and/or sublineages (P[4]-IV-a; P[4]-IV-b; P[8]-3.1; P[8]-3.3; P[8]-3.4 and P[8]-3.6; G1-I; G1-II; G2-IVa-1; G2-IVa-3; G3-1; G3-3; G4-I-c; G9-III; G9-VI)., Discussion: These results extend the available data on the genotypic structure of rotavirus populations in Russia and show the genetic diversity of viral strains. G3P[8] DS-1-like viruses were representatives of the G3-1 lineage, new for the territory of Russia, and had the largest number of amino acid substitutions in the VP7 antigenic epitopes., Conclusion: The emergence and spread of strains with new genetic features may allow rotavirus to overcome the immunological pressure formed by natural and vaccine-induced immunity, and maintain or increase the incidence of rotavirus infection.

Introduction: Rotavirus A is one of the leading causes of acute gastroenteritis in children in the first years of life. Rotavirus infection is currently classified as a preventable infection. The most abundant rotavirion protein is VP6., Material and Methods: Phylogenetic analysis and calculation of phylodynamic characteristics were carried out for 262 nucleotide sequences of the VP6 gene of rotavirus species A, isolated in Russia, using the BEAST v.1.10.4 software package. The derivation and analysis of amino acid sequences was performed using the MEGAX program., Results: This study provides phylodynamic characteristics of the rotaviruses in Russia based on the sequences coding VP6 protein. Bayesian analysis showed the circulation of rotaviruses of three sublineages of genotype I1 and three sublineages of genotype I2 in Russia. The level of accumulation of mutations was established, which turned out to be similar for genotypes I1 and I2 and amounted to 7.732E-4 and 1.008E-3 nucleotides/site/year, respectively. The effective population sizes based on nucleotide sequences of the VP6 I1 and I2 genotypes are relatively stable while after the 2000s there is a tendency of its decreasing. Comparative analysis of the amino acid sequences in the region of the intracellular neutralization sites A (231-260 aa) and B (265-292 aa) made it possible to reveal a mutation in position V252I in a proportion of Russian strains of genotype I1 some strains of genotypes I1 and I2 had mutation I281V. These substitutions were not associated with any sublineages to which the strains belong. The analysis of three T-cell epitopes revealed four amino acid differences (in aa positions 305, 315, 342, 348) that were associated with the first or second genogroup., Conclusion: Based on the phylodynamic characteristics and amino acid composition of antigenic determinants, it was concluded that the VP6 protein is highly stable and could potentially be a good model for development of a rotavirus vaccine.

Rotavirus infection is one of the leading causes of acute gastroenteritis in children in their first years of life. We studied the genotypic diversity of rotavirus A (RVA) strains in Nizhny Novgorod, Russia, during the period 2016-19. In total, 4714 samples of faeces from children admitted to the Nizhny Novgorod Hospital for Infectious Diseases with acute gastroenteritis were examined. The share of rotavirus-positive samples was 31.5% in 2016-17. It decreased to 21.6% in 2018-19. In Nizhny Novgorod, all six global types of RVA were detected (G1P[8], G2P[4], G3P[8], G4P[8], G9P[8] and G12P[8]), as well as sporadic samples with genotypes G9P[4], G3P[9], G9P[9], G8P[8], G2P[8], G4P[4], G3P[9]. The fraction of strains with genotype G2P[4] gradually increased from 5.9% in 2016-17 to 39.1% in 2018-19. Simultaneously, the proportion of G9P[8] strains decreased from 63.2% to 27.7% in the same period. Phylogenetic analysis showed that rotaviruses with the G2P[4] genotype carried ubiquitous alleles of the VP7 and VP4 genes during the period of their prevalence: G2-IVa-1 and G2-IVa-3; P[4]-IVa and P[4]-IVb. As rotavirus vaccination is not widely used in the region because it is not included in the national vaccination calendar in Russia so far, the increase in the number of G2P[4] RVA is likely due to natural strain fluctuations.

Currently, the full-genome-based classification is widely used to investigate rotavirus A (RVA) strains found in different countries around the world. However, the information on the full genotypes of rotaviruses circulating in Russia is limited. Using partial sequencing, this study determined the full genotype constellations of 15 RVA strains in total commonly detected in Nizhny Novgorod (European part of Russia) in 2017-2018, three from each of the following genotypes G1P[8], G4P[8], and G9P[8] and six from G2P[4]. There were two intergenogroup mono-reassortants possessing an identical genotype constellation of G4-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1 with the DS-1-like NSP4 gene of probably local origin. A variety of subgenotype lineages and their combinations of Wa-like rotaviruses and genetic heterogeneity among G9P[8] and G1P[8] strains were shown on the basis of phylogenetic analysis of each gene. Moreover, two distinct co-circulating variants that differed in all 11 genome segments were found among DS-1-like rotaviruses., (Copyright © 2020 Elsevier B.V. All rights reserved.)