Relevance. The epidemiological situation in the world is characterized by an increase in the incidence of orthopoxvirus infections in humans and animals. In this regard, it is necessary to develop new safe vaccines against these infections.Aim. Conducting preclinical studies on the specific activity of the live vaccine against smallpox and other orthopoxvirus infections VACΔ6 based on the vaccinia virus (VACV) with six deleted virulence genes are presented.Matherials and methods. The studies were performed in accordance with the requirements of the Guidelines for conducting preclinical studies of drugs (immunobiological preparations), the State Pharmacopoeia XIII and the European Pharmacopoeia 7.0.Results and discussion. The vaccine strain VACΔ6 VACV showed significantly reduced neurovirulence in the model of intracerebral administration to suckling mice, and reduced inflammatorynecrotic activity in the model of intradermal administration to rabbits compared to the classical first­generation smallpox live vaccine approved for use in Russia. Preclinical studies of three series of the finished dosage form of the VAC∆6 vaccine showed its authenticity, thermal stability, non­pyrogenicity and safety. Double intradermal vaccination of rabbits at a dose of 106 PFU/animal, a 100% protective effect was provided against the intranasal infection of rabbits with VACV strain HB­-92 at a dose of 1995 LD50 and a double intradermal vaccination of mice at a dose of 106 PFU/animal, full protection was provided against the intranasal infection of mice with ectromelia virus strain K1 at a dose of 56 LD50.Conclusion. Based on the conducted complex of studies, it can be concluded that the created vaccine of the fourth generation VACΔ6 is safer compared to the live smallpox vaccine of the first generation and is not inferior to it in immunogenic and protective properties.

Due to cessation of mass smallpox vaccination in 1980, the collective immunity of humans against orthopoxvirus infections has virtually been lost. Therefore, the risk of spreading zoonotic human orthopoxvirus infections caused by monkeypox and cowpox viruses has increased in the world. First-generation smallpox vaccines based on Vaccinia virus (VAC) are reactogenic and therefore not suitable for mass vaccination under current conditions. This necessitates the development of modern safe live vaccines based on VAC using genetic engineering. We created the VACΔ6 strain by transient dominant selection. In the VACΔ6 genome, five virulence genes were intentionally deleted, and one gene was inactivated by inserting a synthetic DNA fragment. The virus was passaged 71 times in CV-1 cells to obtain the VACΔ6 strain from the VAC LIVP clonal variant. Such a long passage history might have led to additional off-target mutations in VACΔ6 compared to the original LIVP variant. To prevent this, we performed a genome-wide sequencing of VAC LIVP, VACΔ6, and five intermediate viral strains to assess possible off-target mutations. A comparative analysis of complete viral genomes showed that, in addition to target mutations, only two nucleotide substitutions occurred spontaneously when obtaining VACΔ4 from the VACΔ3 strain; the mutations persisting in the VACΔ5 and VACΔ6 genomes. Both nucleotide substitutions are located in intergenic regions (positions 1431 and 189738 relative to LIVP), which indicates an extremely rare occurrence of off-target mutations when using transient dominant selection to obtain recombinant VAC variants with multiple insertions/deletions. To assess the genome stability of the resulting attenuated vaccine strain, 15 consecutive cycles of cultivation of the industrial VACΔ6 strain were performed in 4647 cells certified for vaccine production in accordance with the “Guidelines for Clinical Trials of Medicinal Products”. PCR and sequencing analysis of six DNA fragments corresponding to the regions of disrupted genes in VACΔ6 showed that all viral DNA sequences remained unchanged after 15 passages in 4647 cells.

The article reports the results of introducing individual educational trajectories for preschool children into the educational process of preschool education organizations as a factor of children’s social and personality development. In conjunction with collective forms of work, these trajectories allow expanding the range of children’s ideas about different emotional states promoting feelings of empathy and sympathy. The introduction of individual educational trajectories explicated in trajectory maps of social and personality development contributes to improved communicability with peers and adults and reduces conflict in this process. The number of conflict situations in children’s communication with others is lowered. The introduction of individual educational trajectories promotes the development of each child’s readiness for independent goal-setting, action planning, and communication with children and adults. Children learn to evaluate the actions of peers and adults establishing themselves as social subjects and accounting for the social norms and regulations adopted in society, i.e. the norms of the human community.

The article dwells upon specific features and peculiar methods of teaching a foreign language to preschoolers which will contribute to their social communicative development. The authors emphasize a stage character of this process distinguishing the initial phonetic stage, the second lexical stage and the final evaluative and reflective stage. The enhancement of phonetic, lexical and speech skills stipulates such forms of classwork arrangements as individual, paired and choral, each with its advantages and functions. For this age group, an analytico-imitative method has been recognized as the most effective one: cognitive imitation and reproduction are the basic forms of learning. In this respect, the content of the material intended for cognitive imitation has to be valuable, informative and thought-provoking. Special genres have to be found and applied to motivate children to learn and take part in activities: songs, counting rhymes, poems, fairy-tales and games. Children should be constantly involved into the emotionally appealing canvas of the lesson. The results of the research show that emotional impact, exciting forms of presentation and the informative content of the material increase preschoolers' self-consciousness about their progress, create positive motivation and push forward their cognitive, communicative and social development.

The results of identifying the vaccinia virus with the use of nanowire biosensors manufactured on the basis of silicon-on-insulator (SOI) films were presented. In our experiments, the vaccinia virus, the LIVP strain from the collection of the State Research Center of Virology and Biotechnology VECTOR of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, and the rabbit blood serum containing specific polyclonal antibodies to the vaccinia virus were used. As shown by our studies, the polyvalent blood serum was electrically neutral at the sensor surface–viral suspension phase interface, the vaccinia virus was positively charged, and polyvalent blood serum–vaccinia virus vaccine complexes had a negative effective charge.

Mass vaccination has played a critical role in the global eradication of smallpox. Various vaccinia virus (VACV) strains, whose origin has not been clearly documented in most cases, have been used as live vaccines in different countries. These VACV strains differed in pathogenicity towards various laboratory animals and in reactogenicity exhibited upon vaccination of humans. In this work, we studied the development of humoral and cellular immune responses in BALB/c mice inoculated intranasally (i.n.) or intradermally (i.d.) with the VACV LIVP strain at a dose of 10(5) PFU/mouse, which was used in Russia as the first generation smallpox vaccine. Active synthesis of VACV-specific IgM in the mice occurred on day 7 after inoculation, reached a maximum on day 14, and decreased by day 29. Synthesis of virus-specific IgG was detected only from day 14, and the level increased significantly by day 29 after infection of the mice. Immunization (i.n.) resulted in significantly higher production of VACV-specific antibodies compared to that upon i.d. inoculation of LIVP. There were no significant differences in the levels of the T cell response in mice after i.n. or i.d. VACV administration at any time point. The maximum level of VACV-specific T-cells was detected on day 14. By day 29 of the experiment, the level of VACV-specific T-lymphocytes in the spleen of mice significantly decreased for both immunization procedures. On day 30 after immunization with LIVP, mice were infected with the cowpox virus at a dose of 46 LD50. The i.n. immunized mice were resistant to this infection, while 33% of i.d. immunized mice died. Our findings indicate that the level of the humoral immune response to vaccination may play a decisive role in protection of animals from orthopoxvirus reinfection.

This article contains the results of research on the topical problem of highly sensitive express registration of biological objects using field-effect transistors with the surface open for analyte access, which are made based on silicon-on-insulator (SOI) films. The possibilities of dielectrophoretic effects for controlling the concentration of the analyte in the area of sensory elements are considered on the example of the indication of viruses of nuclear polyhedrosis and vaccinia. It is shown that the use of the dielectrophoresis (DEPh) effect makes it possible to solve (1) the key tasks for creating sensor systems: increasing the detecting ability, as well as exrtacting and verifying the signal from the target particles; and (2) the fundamental task: determining the charge state of the analyte in solutions without modifying the sensors’ surface. The problems and prospects of the mass application of nanowire (NW) biosensors, including those with the dielectrophoretic effect, in biotechnology, virology, etc., are discussed.

Introduction. Currently, new directions in cancer therapy are actively developing, one of which is oncolytic immunotherapy. This approach would be to use of viruses as cancer specific cytolytic agents capable of stimulating both the tumor-specific and non-specific immune response.The objective paper was obtain a recombinant vaccinia virus containing genes encoding immunostimulating molecules and study oncolytic and immunostimulating properties of recombinant virus.Material and methods. MTT test, ELISA, methods of transient dominant selection.Results. The recombinant vaccinia virus (L-IVP_oncoB) were obtained with deletion of the gene encoding thymidine kinase and had an integrated gene encoding GM-CSF. Also the virus have deletion of the gene encoding viral growth factor and integrated genes encoding synthetic tumor-specific polyepitopic immunogens. It was shown that the modifications made to the viral genome did not affect the growth characteristics of the virus when cultured on CV-1 and 4647 cell cultures, and the cytopathogenic efficacy of the virus was determined in relation to cancer cultures of cells of various genesis. In in vivo experiment, it was revealed that the polyepitopic construct in the genome L-IVP_oncoB is able to initiate a change in the profile of cytokines.Discussion. The obtained data characterized L-IVP_oncoB as a promising cytopathogenic and immunostimulating agent and showed the need for further study of its properties as means of oncolytic immunotherapy. Conclusion. The basic experiments on the evaluation of the biological properties of the obtained L-IVP_oncoB, which are necessary for the characterization of the oncolytic virus, have been carried out.

We conducted a pedagogical experiment among students of the Faculty of Preschool Education of the "South Ural State Humanitarian Pedagogical University". After the experiment, it was dicovered how the students' motivation to learn a foreign language had changed. The percentage of motivated students in the experimental group increased by about 10%. According to the level of formation of the procedural component of interactive competence, the results in the experimental group changed as follows: the low level of formation of the procedural component of interactive competence changed from 31% to 15%, the number of students with an average level of formation of the procedural component of interactive competence increased from 54% to 70%, the percentage of students with a high level of formation of this component of interactive competence remained the same. We believe that the use of interactive teaching methods in a foreign language class contributes to the formation of interactive competence, the basis of which is the necessity to combine language competence, professional knowledge and skills, as well as the socio-behavioral context.

Tumor necrosis factor (TNF) is one among the key cytokines that mediate the immune system to protect humans against viral infections. Throughout evolution, anthropogenic Variola virus (VARV) has developed effective mechanisms to overcome human defense reactions. The viral genome encodes soluble proteins imitating the structure of cellular cytokine receptors. These proteins compete with cellular receptors for cytokine binding, thus blocking the antiviral immune response. In particular, the G2R gene of VARV encodes the TNF decoy receptor, VARV-CrmB protein. This protein consists of N-ended TNF-biding (TNF-BD) and C-ended chemokine binding (Ch-BD) domains. Recombinant VARV-CrmB protein has been produced in insect cells using molecular cloning methods and its TNF neutralizing activity has been shown in vitro and in vivo. To decrease the immunogenicity of this protein, a recombinant plasmid coding for shortened TNF-BD protein of VARV in Escherichia coli cells has been constructed. Using the method of immobilized metal affinity chromatography, recombinant TNF-BD protein corresponding to the TNF-biding domain of VARV-CrmB protein was purified from E. coli cells. The therapeutic potential of TNF-BD was studied using an experimental model of LPS-induced septic shock. After septic shock induction, several doses of recombinant TNF-BD were injected and the mortality of experimental animals was observed during 7 days. All mice not injected with TNF-BD had been dead by day 3 of the experiment, but 30, 40 and 60 % of the experimental animals, who received different TNF-BD doses, survived in a dose-dependent manner. Data obtained demonstrate that recombinant TNF-BD protein is pharmacologically active in the experimental model of LPS-induced septic shock.

Фактор некроза опухолей (TNF) – один из основных цитокинов, медиаторов иммунной системы, обеспечивающих защиту организма человека от вирусных инфекций. В процессе эволюции антропогенный вирус натуральной оспы (Variola virus, VARV) освоил эффективные механизмы преодоления иммунологических барьеров человека, кодируя в своем геноме белки, способные взаимодействовать с рецепторами цитокинов организма-хозяина, блокируя таким образом их активность. В частности, ген G2R этого вируса кодирует белок CrmB, который эффективно связывает TNF человека и мышей. При этом данный белок является двухдоменным и наряду с TNF-связывающим N-концевым доменом содержит С-концевой хемокин-связывающий домен. При использовании методологии молекулярного клонирования нами ранее получен рекомбинантный бакуловирус, продуцирующий в клетках насекомых рекомбинантный белок CrmB вируса натуральной оспы (VARV-CrmB), и показан его TNF-нейтрализующий потенциал в различных тестах in vitro и in vivo. С целью снижения иммуногенности этого вирусного белка при его многократном введении для терапии хронических воспалительных заболеваний получена рекомбинантная плазмида, направляющая в клетках Escherichia coli биосинтез укороченного однодоменного TNF-связывающего (TNF-BD) белка VARV. Методом металл-хелатной аффинной хроматографии из клеток выделен рекомбинантный белок TNF-BD. Терапевтический потенциал белка TNF-BD изучен в экспериментальной модели септического шока, индуцированного в организме мышей введением бактериального липополисахарида (ЛПС). После индукции септического шока животным вводили разные дозы рекомбинантного белка TNF-BD и регистрировали их гибель в течение 7 сут. Все мыши, не получавшие препарат белка TNF-BD после инъекции ЛПС, погибли через 3 сут, а в группах животных, которым вводили TNF-BD, в зависимости от дозы этого белка выжили 30, 40 или 60 % мышей. Результаты исследования демонстрируют наличие специфической фармакологической активности у рекомбинантного белка TNF-BD, синтезированного в бактериальных клетках, в мышиной модели ЛПС-индуцированного септического шока.

The modern approach to developing attenuated smallpox vaccines usually consists in targeted inactivation of vaccinia virus (VACV) virulence genes. In this work, we studied how an elevated production of extracellular enveloped virions (EEVs) and the route of mouse infection can influence the virulence and immunogenicity of VACV. The research subject was the LIVP strain, which is used in Russia for smallpox vaccination. Two point mutations causing an elevated production of EEVs compared with the parental LIVP strain were inserted into the sequence of the VACV A34R gene. The created mutant LIVP-A34R strain showed lower neurovirulence in an intracerebral injection test and elevated antibody production in the intradermal injection method. This VACV variant can be a promising platform for developing an attenuated, highly immunogenic vaccine against smallpox and other orthopoxvirus infections. It can also be used as a vector for designing live-attenuated recombinant polyvalent vaccines against various infectious diseases.

The article considers the history of Buddhism in Central Asia and in Russia. It outlines the main periods of development and special features of Buddhism in the region, its influence on the local culture. It explorers the contemporary state of the Buddhist sangha in Russia and Central Asian countries. Central Asia has played an important role in the development of Buddhism as a world religion. In I-III centuries A.D. missionaries from Central Asia carried out the sermon of the Buddhist teachings. The archeological findings illustrate the massive spread of Buddhism on the wide territories of the region which were part of the Kushan Kingdom. The second period of the flourishing of Buddhist teaching falls on the V – first part of the VIII centuries, when the geography on Buddhism in the region expanded, and it peacefully co-existed with other religions. By IX century, when the territories of the contemporary Uzbekistan, Kyrgyzstan, Turkmenistan and Tadjikistan stayed under the rule of Umayyad and Abbaside Caliphate, Islam eventually ousted Buddhism from these lands. The third period of rise of Buddhism in the region started with the appearance of Dzungars who aspired to take hold of the lands of Kazahstan. From 1690 to 1760 Central Asian region had become an area of struggle for the hegemony between the Buddhist Dzungarian khanate and China. The Dzungars promoted the spread of Buddhism in the Eastern part of Kazahstan and Northern part of Eastern Turkestan. The entry of Western Turkestan into the Russian Empire put an end to external threats and internal feudal strife. It gave the start to the process of consolidation of the Central Asian nations, which recognized their belonging to Muslim Ummah. In the absence of Dzungar and Chinese factors the influence of Buddhism in the region almost stopped. By the end of the XX century with the renaissance of religiosity on the post-Soviet space the interest to Buddhism slightly raised. However, at the present moment the number of the Buddhists in the region is insignificant. Among the followers of Buddhism the main place is taken by the Korean diaspora, residing in Central Asia since 1937. There also exist some single neo-Buddhist communities in the region. Buddhism made its contribution to the development of the unique socio-cultural identity of Russia as Eurasian by it’s nature. Buryatia, Kalmykia, Tuva, as well as several parts of Altai, Irkutsk and Chita regions represent historical areas of the spread of Buddhist teaching. At the present moment the Russian Buddhist sangha contains of the major independent centers in Buryatia, Kalmykia, Tuva, Moscow and St.Petersburg. Buddhism plays and important part in socio-cultural space of Russia, gradually moving far beyond the borders of the regions of its traditional location. Popularity of the Buddhist philosophy derives from the range of grounds, among which are the closeness of some of its principles to contemporary scientific ideas, first of all to cognitive sciences, as well as openness to dialogue with other cultural and religious traditions, humanism, ethics of non-violence and ideas of common responsibility.

El artículo presenta resultados del trabajo dirigido al desarrollo de la responsabilidad en preescolares mayores en el curso del proceso educativo en una institución de educación preescolar. Los autores han realizado un análisis del problema del desarrollo de la responsabilidad desde la perspectiva de diferentes ámbitos: filosofía, pedagogía y psicología. Con el fin de implementar el proceso de desarrollo de la responsabilidad en los niños, los autores han identificado tres etapas: formación del concepto y motivación para la actividad realizada (preparatoria), desarrollo de habilidades prácticas de comportamiento responsable (acompañamiento) y uso independiente consciente de lo acumulado, experiencia de comportamiento responsable en el curso de la propia actividad del niño (independiente). Aparte de eso, los autores han especificado las tecnologías pedagógicas aplicadas.