Nucleotide Sequence and Phylogenetic Analysis of Glycoprotein-G of the Russian Fixed Rabies Virus Strain “Moscow 3253”

Fully sequenced have been glycoprotein-G, sha-psi region, as well as H-end site of the L-gene in the rabies virus strain “Moscow 3253”. Compared are amino acid sequences of proteins of “Moscow 3253” strain and other fixed strains of the virus. Established is 98 % DNA homology with RV-97, and 91% homology with PV (Pasteur virus) strain. Constructed has been phylogenetic tree of the strain under study alongside with various groups of fixed rabies virus. It is revealed that “Moscow 3253” strain has closer genetic relations with Japanese group of strains, than with PV strain. Put forward is an assumption that PV strain does not derive from the virus isolated by Pasteur, but relates to the American group of strains

Signatures of the slow solar wind streams from active regions in the inner corona

Some of local sources of the slow solar wind can be associated with spectroscopically detected plasma outflows at edges of active regions accompanied with specific signatures in the inner corona. The EUV telescopes (e.g. SPIRIT/CORONAS-F, TESIS/CORONAS-Photon and SWAP/PROBA2) sometimes observed extended ray-like structures seen at the limb above active regions in 1MK iron emission lines and described as "coronal rays". To verify the relationship between coronal rays and plasma outflows, we analyze an isolated active region (AR) adjacent to small coronal hole (CH) observed by different EUV instruments in the end of July - beginning of August 2009. On August 1 EIS revealed in the AR two compact outflows with the Doppler velocities V =10-30 km/s accompanied with fan loops diverging from their regions. At the limb the ARCH interface region produced coronal rays observed by EUVI/STEREO-A on July 31 as well as by TESIS on August 7. The rays were co-aligned with open magnetic field lines expanded to the streamer stalks. Using the DEM analysis, it was found that the fan loops diverged from the outflow regions had the dominant temperature of ~1 MK, which is similar to that of the outgoing plasma streams. Parameters of the solar wind measured by STEREO-B, ACE, WIND, STEREO-A were conformed with identification of the ARCH as a source region at the Wang-Sheeley-Arge map of derived coronal holes for CR 2086. The results of the study support the suggestion that coronal rays can represent signatures of outflows from ARs propagating in the inner corona along open field lines into the heliosphere.Comment: Accepted for publication in Solar Physics; 31 Pages; 13 Figure

ДНК- и РНК-вакцины: современное состояние, требования к качеству и особенности проведения доклинических исследований

This review focuses on DNA and RNA vaccines whose potential use was first considered at the end of the 20th century. However, not a single bacterial plasmid-based or mRNA vaccine has been used since that time in public healthcare for the prevention of infectious diseases. Nevertheless, vaccines containing recombinant nucleic acids as the active ingredient still attract interest due to the possibility of rapid development, low-cost production, safety of the technology and the potential to activate cellular and humoral immunity. Recent technological advances have largely overcome the problems of low immunogenicity, instability, and difficulties with the delivery of DNA and RNA vaccines in humans. The aim of this review was to present the main strategies of development of DNA and RNA vaccines designed to prevent infectious diseases, and to summarise requirements for the quality control and preclinical studies. The article examines the general principles of creation of plasmid vectors encoding protective antigens. It describes new technologies used in the creation of DNA vaccines with plasmids encoding an attenuated virus genome (iDNA and PPLAV), and RNA vaccines based on mRNA and self-amplifying RNAs. The article presents current regulatory requirements for the choice of quality parameters to be tested and the general principles of preclinical studies of DNA and RNA vaccines.Обзор посвящен ДНК- и РНК-вакцинам, возможность использования которых была показана еще в конце XX века. При этом до сих пор ни одна вакцина, основанная на использовании бактериальных плазмид и мРНК, не нашла применения в практике здравоохранения для профилактики инфекционных заболеваний. Но, несмотря на это, интерес к вакцинам, действующим веществом которых являются рекомбинантные нуклеиновые кислоты, сохраняется из-за возможности их быстрой разработки, малозатратного производства, безопасности технологии и возможности активации клеточного и гуморального иммунитета. Последние технологические достижения в значительной степени преодолели проблемы низкой иммуногенности, нестабильности и трудности доставки при применении ДНК- и РНК-вакцин у человека. Цель работы — изложение основных стратегий создания ДНК- и РНК-вакцин, предназначенных для профилактики инфекционных заболеваний, обобщение требований к оценке их качества и проведению доклинических исследований. Представлены общие принципы создания плазмидных векторов, кодирующих протективные антигены. Описаны новые технологии создания ДНК-вакцин, плазмиды которых кодируют геном аттенуированного вируса (iDNA и PPLAV). Приведены стратегии создания РНК-вакцин на основе мРНК и самоамплифицирующихся РНК. Представлены современные регуляторные требования к выбору необходимых показателей качества и общим принципам проведения доклинических исследований ДНК- и РНК-вакцин

Международный опыт нормативно-правового регулирования препаратов, содержащих жизнеспособные клетки человека

The intensive development of cellular technologies stipulates the introduction at the global level of medicinal products based on viable human cells, which in most countries are referred to as biomedical cell products. The authors conducted a comparative analysis of the regulatory framework in different countries and determined special aspects of regulation of cell therapy products (analogues of biomedical cell products). Some countries have mechanisms for priority review of cell therapy products for marketing authorization, such as accelerated assessment, accelerated approval, or conditional marketing authorisation. These mechanisms are currently absent in Russia, because of the novelty of the regulatory framework, and the biological properties of innovative cell products. Biomedical cell products are regarded as a separate class of medicinal products in Russia, they are not treated as biologicals and are regulated by the Federal Law No. 180-FZ «On Biomedical Cell Products» of June 23, 2016. The main difference in regulation of cell-based products in the Russian Federation is the principle of unified requirements for marketing authorisation of autologous, allogeneic, and combined biomedical cellular products, and the absence of the «hospital exemptions» mechanism that exists in many countries. This mechanism allows prescription and use of personalised autologous medicines produced in the laboratory of a medical institution for a particular patient.Интенсивное развитие клеточных технологий обусловливает внедрение в мировую медицинскую практику препаратов на основе жизнеспособных клеток человека, которые в большинстве стран определяются как биологические лекарственные препараты. Авторами проведен сравнительный анализ нормативно-правовой базы разных стран мира и определены особенности регулирования препаратов для клеточной терапии (аналогов биомедицинских клеточных продуктов). В некоторых странах существуют механизмы приоритетного рассмотрения препаратов для клеточной терапии для вывода на рынок, например процедуры ускоренного рассмотрения, ускоренного утверждения, условной регистрации. Учитывая новизну нормативной базы и биологические особенности инновационных препаратов — биомедицинских клеточных продуктов, в Российской Федерации подобные механизмы в настоящее время отсутствуют. Биомедицинские клеточные продукты в России являются отдельным классом медицинских средств, отличным от биологических лекарственных препаратов, и регулируются Федеральным законом № 180-ФЗ «О биомедицинских клеточных продуктах» от 23 июня 2016 г. Основным отличием регулирования клеточных препаратов в России является принцип единых требований вывода на рынок аутологичных, аллогенных и комбинированных биомедицинских клеточных продуктов и отсутствие механизма «исключения для больничного производства» (hospital exemptions), действующего во многих странах и заключающегося в допущении применения персонифицированного аутологичного препарата, произведенного в конкретной лаборатории при медицинской организации для определенного пациента по назначению конкретного врача

Features of developing SARS-CoV-2 nucleocapsid protein population-based seroprevalence during the first wave of the COVID-19 epidemic in the Russian Federation

The novel coronavirus (SARS-CoV-2) pandemic, dubbed COVID-19, has become one of the most serious challenges for human populations in the vast majority of countries worldwide. Rapid spreading and increased mortality related to it required new approaches to manage epidemic processes on a global scale. One of such approaches was based on analyzing SARS-CoV-2 seroprevalence associated with COVID-19. Our aim was to summarize the results on assessing seroprevalence to the SARS-CoV-2 nucleocapsid antigen (Nc) in residents from 26 regions of the Russian Federation, carried out during the first wave of the COVID-19 epidemic.Materials and methods. Seroprevalence distribution was examined in 26 model regions of the Russian Federation according to the unified method developed by the Rospotrebnadzor with the participation of the Federal State Institution Saint Petersburg Pasteur Research Institute of Epidemiology and Microbiology. Such approach implied formation of a group of volunteer subjects in model geographic region who were tested by ELISA for anti-Nc serum antibody level in peripheral blood. Analyzed primary data obtained in separate regions were either accepted for publication or released.Results. The current paper finalizes the data obtained in all 26 regions of the Russian Federation. The total SARS-CoV-2 seroprevalence was 19.5 (10.0–25.6)% with the maximum and minimum value found in the Kaliningrad Region and the Republic of Crimea, respectively (50.2% vs. 4.3%). A pattern of age-related seroprevalence distribution indicates insignificant predominance of seroprevalence among subjects of 1–17 years old: 22.1 (13.1–31.8)%. Among COVID-19 convalescents positive for SARS-CoV Nc antibodies it reached 60.0 (40.0–73.3)%. The number of contact persons comprised 6285 subjects or 8.5% of total volunteer cohort, with the level of seroprevalence reaching up to 25.3 (17.95–35.8)%. A direct correlation was revealed between levels of seroprevalence in convalescent and contact volunteers. In addition, the reproductive number for SARS-CoV was calculated comprising 5.8 (4.3–8.5) suggesting that one convalescent subject can infect at least 4 healthy individuals. A high level of asymptomatic forms of COVID-19 among seropositive subjects was confirmed empirically comprising up to 93.6 (87.1–94.9)%.Conclusion. A single cross-sectional study performed during 2020 June–August timeframe allowed to assess pattern of sex- and agerelated COVID-19 seroprevalence for general population in 26 Russian Federation regions. The data obtained may serve as a basis for the longitudinal cohort investigation with serial subject sampling. The timing and duration of study will be determined by dynamics of ongoing COVID-19 epidemic

Altered Variants of Cholera Agent Isolated in the Territory of the Russian Federation

Carried out was the retrospective genetic analysis of cholera vibrio strains which caused the outbreaks and sporadic cases of cholera in the territory of the Russian Federation from 1993 to 2006. It was elucidated that beginning from 1993 the disease had been caused by the altered strains of V. cholerae El Tor containing ctxB gene of classical type, encoding B subunit of cholera toxin, in their CTXφ prophage genome. In addition, identified were two strains whose prophage genome contained classical type rstR gene

DNA and RNA Vaccines: Current Status, Quality Requirements and Specific Aspects of Preclinical Studies

This review focuses on DNA and RNA vaccines whose potential use was first considered at the end of the 20th century. However, not a single bacterial plasmid-based or mRNA vaccine has been used since that time in public healthcare for the prevention of infectious diseases. Nevertheless, vaccines containing recombinant nucleic acids as the active ingredient still attract interest due to the possibility of rapid development, low-cost production, safety of the technology and the potential to activate cellular and humoral immunity. Recent technological advances have largely overcome the problems of low immunogenicity, instability, and difficulties with the delivery of DNA and RNA vaccines in humans. The aim of this review was to present the main strategies of development of DNA and RNA vaccines designed to prevent infectious diseases, and to summarise requirements for the quality control and preclinical studies. The article examines the general principles of creation of plasmid vectors encoding protective antigens. It describes new technologies used in the creation of DNA vaccines with plasmids encoding an attenuated virus genome (iDNA and PPLAV), and RNA vaccines based on mRNA and self-amplifying RNAs. The article presents current regulatory requirements for the choice of quality parameters to be tested and the general principles of preclinical studies of DNA and RNA vaccines