Kira Shalepo Shalepo, T. Khusnutdinova, O. Budilovskaya, A. Krysanova, K. V. Sapozhnikov, A. Savicheva, I. Kogan
Introduction. Colonization of the reproductive organs of pregnant women with group B streptococci (GBS; Streptococcus agalactiae) can lead to severe perinatal and neonatal pathology. In modern conditions, aside from the prevention of antenatal infection of the fetus during childbirth using antibacterial drugs, vaccination is also necessary. In this regard, surveillance of GBS genotypes is an important task. Objective. To determine the molecular genetic determinants of virulence of Streptococcus agalactiae isolated from pregnant women and newborns, and to monitor the distribution of capsular polysaccharides types and pili profiles in GBS clinical isolates. Materials and methods. The study used clinical isolates of GBS (n = 420) isolated from pregnant women and newborns in 2010–2023.The bacteriological method was used for isolation of S. agalactiae. PCR method was used to determine the types of capsular polysaccharides, pili, and whether the strains belonged to the hypervirulent sequence type ST-17. Results. During 13 years of observation, the predominance of Ia, III and V genotypes of GBS capsular polysaccharides was noted both in pregnant women and in newborns. The frequency of occurrence of genotype Ib increased from 0.7 to 6.7%, genotype V from 12.1 to 24.4%, and the prevalence of genotype III decreased significantly from 41.1 to 21.1%. Hypervirulent sequence type ST-17 was detected in 6 pregnant women and 2 newborns. However, there were no signs of neonatal infection in these children. More than half of all clinical isolates of S. agalactiae had pili genotypes PI-1 + PI-2a, as well as pili genotypes PI-2a and PI-1 + PI-2b. The distribution of pili types did not change over 13 years of the surveillance period. Conclusion. Surveillance of the GBS capsular polysaccharides and pili genotypes is necessary for the development of effective preventive vaccines.
{"title":"Molecular genetic determinants of virulence of Streptococcus agalactiae isolated from pregnant women and newborns in St. Petersburg and the Leningrad region in 2010–2023","authors":"Kira Shalepo Shalepo, T. Khusnutdinova, O. Budilovskaya, A. Krysanova, K. V. Sapozhnikov, A. Savicheva, I. Kogan","doi":"10.36233/0372-9311-501","DOIUrl":"https://doi.org/10.36233/0372-9311-501","url":null,"abstract":"Introduction. Colonization of the reproductive organs of pregnant women with group B streptococci (GBS; Streptococcus agalactiae) can lead to severe perinatal and neonatal pathology. In modern conditions, aside from the prevention of antenatal infection of the fetus during childbirth using antibacterial drugs, vaccination is also necessary. In this regard, surveillance of GBS genotypes is an important task. \u0000Objective. To determine the molecular genetic determinants of virulence of Streptococcus agalactiae isolated from pregnant women and newborns, and to monitor the distribution of capsular polysaccharides types and pili profiles in GBS clinical isolates. \u0000Materials and methods. The study used clinical isolates of GBS (n = 420) isolated from pregnant women and newborns in 2010–2023.The bacteriological method was used for isolation of S. agalactiae. PCR method was used to determine the types of capsular polysaccharides, pili, and whether the strains belonged to the hypervirulent sequence type ST-17. \u0000Results. During 13 years of observation, the predominance of Ia, III and V genotypes of GBS capsular polysaccharides was noted both in pregnant women and in newborns. The frequency of occurrence of genotype Ib increased from 0.7 to 6.7%, genotype V from 12.1 to 24.4%, and the prevalence of genotype III decreased significantly from 41.1 to 21.1%. Hypervirulent sequence type ST-17 was detected in 6 pregnant women and 2 newborns. However, there were no signs of neonatal infection in these children. More than half of all clinical isolates of S. agalactiae had pili genotypes PI-1 + PI-2a, as well as pili genotypes PI-2a and PI-1 + PI-2b. The distribution of pili types did not change over 13 years of the surveillance period. \u0000Conclusion. Surveillance of the GBS capsular polysaccharides and pili genotypes is necessary for the development of effective preventive vaccines.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":" 14","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140990295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. Akimkin, Tatyana A. Semenenko, S. Ugleva, D. Dubodelov, Elena N. Kolosovskaya
The topic of epidemiologic surveillance is one of the basic concepts in the theory and practice of epidemiologic science. In Russia, generalization of the accumulated factual material and theoretical developments have allowed us to formulate a number of provisions on the nature of the epidemic process. The pandemic of a new coronavirus infection has forced adjustments in all spheres of society, including the activities of the infectious disease epidemiological surveillance system, requiring the development and implementation of innovative solutions. Based on the experience of prompt response to the tasks set by the COVID-19 pandemic, the authors raised the problem of development and implementation of a system of molecular genetic monitoring for pathogens of emerging and re-emerging infections as a priority vector of epidemiological surveillance development.The introduction of modern molecular biological technologies for the identification of pathogens with epidemic potential, taking into account their genetic diversity, into the system of epidemiologic surveillance has been substantiated based on the experience of using platform solutions created by the Central Research Institute of Epidemiology of Rospotrebnadzor. The strategy of genomic epidemiologic surveillance as a powerful tool to ensure readiness for response measures and management of the epidemic process by implementing and adjusting preventive and anti-epidemic measures was developed.The Russian platform for aggregation of information on virus genomes (VGARus) developed at the Central Research Institute of Epidemiology of Rospotrebnadzor as a technological, scientific, organizational and infrastructural base of genomic epidemiological surveillance, acting as an interdepartmental consortium, has been introduced into practice. The efficiency of VGARus was shown to assess the mutational variability of SARS-CoV-2, the influence of evolutionary development of circulating pathogens on the characteristics of the epidemic process, the implementation of operational and retrospective analysis of morbidity and prediction of the spread of genetic variants of pathogens.
{"title":"Genomic surveillance strategy. Problems and perspectives","authors":"V. Akimkin, Tatyana A. Semenenko, S. Ugleva, D. Dubodelov, Elena N. Kolosovskaya","doi":"10.36233/0372-9311-507","DOIUrl":"https://doi.org/10.36233/0372-9311-507","url":null,"abstract":"The topic of epidemiologic surveillance is one of the basic concepts in the theory and practice of epidemiologic science. In Russia, generalization of the accumulated factual material and theoretical developments have allowed us to formulate a number of provisions on the nature of the epidemic process. The pandemic of a new coronavirus infection has forced adjustments in all spheres of society, including the activities of the infectious disease epidemiological surveillance system, requiring the development and implementation of innovative solutions. Based on the experience of prompt response to the tasks set by the COVID-19 pandemic, the authors raised the problem of development and implementation of a system of molecular genetic monitoring for pathogens of emerging and re-emerging infections as a priority vector of epidemiological surveillance development.The introduction of modern molecular biological technologies for the identification of pathogens with epidemic potential, taking into account their genetic diversity, into the system of epidemiologic surveillance has been substantiated based on the experience of using platform solutions created by the Central Research Institute of Epidemiology of Rospotrebnadzor. The strategy of genomic epidemiologic surveillance as a powerful tool to ensure readiness for response measures and management of the epidemic process by implementing and adjusting preventive and anti-epidemic measures was developed.The Russian platform for aggregation of information on virus genomes (VGARus) developed at the Central Research Institute of Epidemiology of Rospotrebnadzor as a technological, scientific, organizational and infrastructural base of genomic epidemiological surveillance, acting as an interdepartmental consortium, has been introduced into practice. The efficiency of VGARus was shown to assess the mutational variability of SARS-CoV-2, the influence of evolutionary development of circulating pathogens on the characteristics of the epidemic process, the implementation of operational and retrospective analysis of morbidity and prediction of the spread of genetic variants of pathogens.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140683899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Mironov, A. Titkov, K. Kuleshov, A. E. Platonov
Introduction. Borrelia miyamotoi is a pathogen of erythema-free ixodid tick-borne borreliosis (ITBB), a disease widespread in Russia. To date, there are no generally accepted methods for B. miyamotoi genotyping. The multilocus sequencing typing (MLST) scheme of Borrelia was originally developed for B. burgdorferi, and does not have the required discrimination power for monitoring the ITBB pathogens. The objective of this study is to develop the MLST scheme for B. miyamotoi. Materials and Methods. The whole genome sequences of 10 reference strains (GenBank) were analyzed for the selection of the house-keeping loci. The MLST scheme development was based on principles published by the authors of the method. For this experiment, 81 B. miyamotoi strains and positive clinical samples were used to test the MLST scheme. Results. After analyzing the genomic data, 8 house-keeping loci were chosen for MLST, for which the PCR and sequencing primers were designed. Each MLST loci was represented by several alleles (from 4 to 7) which form 15 sequence types. The genetic diversity of pathogens isolated from ITBB patients and ticks were characterized. Discussion. Based on pairwise distances between allelic profiles, the sequence types can be classified into four groups. The first two groups are clonal complexes; the other two groups are formed by once identified sequence types. The first clonal complex unites 11 sequence types (80 or 88% of the characterized B. miyamotoi), the second consists of 2 sequence types (9 or 9.8%). The genetic differences between B. miyamotoi are associated with the sources of strains and biological isolates. The MLST based classification confirms the previously described genetic heterogeneity of B. miyamotoi populations associated with ecologically unrelated vectors of ITBB pathogens. Conclusion. The proposed MLST scheme is an appropriate tool for ITBB pathogen classification and evolutionary change characterization within clonal complexes.
{"title":"Multilocus sequence-typing scheme for Borrelia miyamotoi — the erythema-free ixodid tick-borne borreliosis pathogens","authors":"K. Mironov, A. Titkov, K. Kuleshov, A. E. Platonov","doi":"10.36233/0372-9311-419","DOIUrl":"https://doi.org/10.36233/0372-9311-419","url":null,"abstract":"Introduction. Borrelia miyamotoi is a pathogen of erythema-free ixodid tick-borne borreliosis (ITBB), a disease widespread in Russia. To date, there are no generally accepted methods for B. miyamotoi genotyping. The multilocus sequencing typing (MLST) scheme of Borrelia was originally developed for B. burgdorferi, and does not have the required discrimination power for monitoring the ITBB pathogens. \u0000The objective of this study is to develop the MLST scheme for B. miyamotoi. \u0000Materials and Methods. The whole genome sequences of 10 reference strains (GenBank) were analyzed for the selection of the house-keeping loci. The MLST scheme development was based on principles published by the authors of the method. For this experiment, 81 B. miyamotoi strains and positive clinical samples were used to test the MLST scheme. \u0000Results. After analyzing the genomic data, 8 house-keeping loci were chosen for MLST, for which the PCR and sequencing primers were designed. Each MLST loci was represented by several alleles (from 4 to 7) which form 15 sequence types. The genetic diversity of pathogens isolated from ITBB patients and ticks were characterized. \u0000Discussion. Based on pairwise distances between allelic profiles, the sequence types can be classified into four groups. The first two groups are clonal complexes; the other two groups are formed by once identified sequence types. The first clonal complex unites 11 sequence types (80 or 88% of the characterized B. miyamotoi), the second consists of 2 sequence types (9 or 9.8%). The genetic differences between B. miyamotoi are associated with the sources of strains and biological isolates. The MLST based classification confirms the previously described genetic heterogeneity of B. miyamotoi populations associated with ecologically unrelated vectors of ITBB pathogens. \u0000Conclusion. The proposed MLST scheme is an appropriate tool for ITBB pathogen classification and evolutionary change characterization within clonal complexes.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140245397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Piterskiy, O. A. Khodakov, Tatyana V. Mikheeva, Natalia V. Bilalova, Alena B. Konkova-Reidman, Yuliya A. Zakharova, A. V. Semenov
Background. Due to the wide coverage with antiretroviral therapy, the life expectancy of HIV infected people has significantly increased. Against the background of a decrease in mortality from HIV infection, HIV-associated neurocognitive disorders, which develop even during effective treatment, are of high importance. The overall prevalence of this pathology among HIV-infected people reaches 42.6%. The objective of the study was to research the genetic features and phylogenetic position of HIV-1 persisting in the central nervous system. Materials and methods. The clinical study group consisted of 38 patients with severe neurocognitive disorders against the background of HIV infection in stage 4B. The viral load of HIV-1 in blood plasma and cerebrospinal fluid (CSF) was measured using the "AmpliSens HIV Monitor-FRT" reagents kit. Sanger sequencing was performed using the AmpliSens HIV-Resist-Seq assay kit on an Applied Biosystems 3500 analyzer. Phylogenetic analysis of the pol gene fragments of HIV-1 strains (the site encoding the viral protease and part of the reverse transcriptase) was carried out using maximum likelihood method with the GTR+G nucleotide substitution model. Comparisons of the tertiary structure of viral proteins were performed according to three-dimensional models of the protease and p51 and p66 reverse transcriptase subunits obtained by homologous reconstruction using the SWISS-MODEL tools. Results. The viral load in the sample of patients with severe CNS lesions in blood plasma was 6.27 times higher than in CSF and amounted to 4.67 and 3.87 lg copies/ml respectively by median (p = 0,004). Phylogenetic analysis with the use of all available HIV-1 genomes from GenBank, which differed from the studied ones by less than 5% showed close genetic relations of viruses circulating in Chelyabinsk region, apart from strains circulating in Russian Federation, with viruses circulating in neighboring countries, in most abundance — from Ukraine and Kyrgyzstan, slightly less — from Belarus, Tajikistan, Kazakhstan and Armenia and also with strains from certain foreign countries: Poland and Germany. Phylogenetic analysis of 38 HIV-1 genomes revealed significant genetic distances between HIV isolates from blood plasma and CSF in 5 patients, 4 of whom were PWID, which may indicate an event of superinfection. The amount of independent amino acid substitutions in protease in isolates from blood plasma ranged from 1 to 3, in isolates from CSF — from 1 to 2. An amount of such substitutions in a fragment of reverse transcriptase in isolates from blood plasma ranged from 1 to 6, while in isolates from CSF, it ranged from 1 to 7. HIV isolates from blood plasma and CSF from 5 patients had differences in the tertiary structure of HIV-1 reverse transcriptase p51 subunit in amino acid positions 16–20 and 210–235. Isolates from 3 other patients differed in the tertiary structure only in amino acid positons 210–235. Isolates from 3 patients differed in t
{"title":"Phylogenetic position and genetic features of HIV-1 in CNS","authors":"M. Piterskiy, O. A. Khodakov, Tatyana V. Mikheeva, Natalia V. Bilalova, Alena B. Konkova-Reidman, Yuliya A. Zakharova, A. V. Semenov","doi":"10.36233/0372-9311-442","DOIUrl":"https://doi.org/10.36233/0372-9311-442","url":null,"abstract":"Background. Due to the wide coverage with antiretroviral therapy, the life expectancy of HIV infected people has significantly increased. Against the background of a decrease in mortality from HIV infection, HIV-associated neurocognitive disorders, which develop even during effective treatment, are of high importance. The overall prevalence of this pathology among HIV-infected people reaches 42.6%. \u0000The objective of the study was to research the genetic features and phylogenetic position of HIV-1 persisting in the central nervous system. \u0000Materials and methods. The clinical study group consisted of 38 patients with severe neurocognitive disorders against the background of HIV infection in stage 4B. The viral load of HIV-1 in blood plasma and cerebrospinal fluid (CSF) was measured using the \"AmpliSens HIV Monitor-FRT\" reagents kit. Sanger sequencing was performed using the AmpliSens HIV-Resist-Seq assay kit on an Applied Biosystems 3500 analyzer. Phylogenetic analysis of the pol gene fragments of HIV-1 strains (the site encoding the viral protease and part of the reverse transcriptase) was carried out using maximum likelihood method with the GTR+G nucleotide substitution model. Comparisons of the tertiary structure of viral proteins were performed according to three-dimensional models of the protease and p51 and p66 reverse transcriptase subunits obtained by homologous reconstruction using the SWISS-MODEL tools. \u0000Results. The viral load in the sample of patients with severe CNS lesions in blood plasma was 6.27 times higher than in CSF and amounted to 4.67 and 3.87 lg copies/ml respectively by median (p = 0,004). \u0000Phylogenetic analysis with the use of all available HIV-1 genomes from GenBank, which differed from the studied ones by less than 5% showed close genetic relations of viruses circulating in Chelyabinsk region, apart from strains circulating in Russian Federation, with viruses circulating in neighboring countries, in most abundance — from Ukraine and Kyrgyzstan, slightly less — from Belarus, Tajikistan, Kazakhstan and Armenia and also with strains from certain foreign countries: Poland and Germany. Phylogenetic analysis of 38 HIV-1 genomes revealed significant genetic distances between HIV isolates from blood plasma and CSF in 5 patients, 4 of whom were PWID, which may indicate an event of superinfection. \u0000The amount of independent amino acid substitutions in protease in isolates from blood plasma ranged from 1 to 3, in isolates from CSF — from 1 to 2. An amount of such substitutions in a fragment of reverse transcriptase in isolates from blood plasma ranged from 1 to 6, while in isolates from CSF, it ranged from 1 to 7. HIV isolates from blood plasma and CSF from 5 patients had differences in the tertiary structure of HIV-1 reverse transcriptase p51 subunit in amino acid positions 16–20 and 210–235. Isolates from 3 other patients differed in the tertiary structure only in amino acid positons 210–235. Isolates from 3 patients differed in t","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"13 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140244967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Asya S. Levina, M. Repkova, O.YU. Mazurkov, E. V. Makarevich, N. A. Mazurkova, V. F. Zarytova
Relevance. Studies on model systems have confirmed the effectiveness of antisense oligonucleotides, including those that contain photoactive groups, for the modification of nucleic acids. However, this strategy has not yet found wide application due to the lack of successful methods for the cellular delivery. The development of effective preparations capable of acting on target nucleic acids in cells is an urgent task. The objective of the work is to create nanocomposites consisting of TiO2 nanoparticles, antisense oligonucleotides, and photoactive groups and to study their effect on target nucleic acids by the example of inhibition of influenza A virus replication in the cellular system. Materials and methods. Influenza virus A/Aichi/2/68 (A/H3N2), N-succinimide ether of p-azidotetrafluorobenzoic acid, TiO2 nanoparticles, and oligodeoxyribonucleotides have been used in the work. The antiviral activity of the proposed nanocomposites has been studied on the MDCK cells infected with the A/H3N2 virus. Results and discussion. Unique nanocomposites have been created, which consist of three functional components, i.e., titanium dioxide nanoparticles, antisense oligonucleotides, and the photoactive tetrafluoroarylazide group, respectively, providing penetration into cells, selective interaction with target nucleic acids, and photomodification of the targets. A significant antiviral site-specific action of the nanocomposites has been demonstrated against the influenza A virus in the cellular system, which exceeds the effect of the analogous samples that contain no photoactive groups. Conclusion. The biological activity of the created nanocomposites has been demonstrated by the example of highly effective suppression of influenza A virus replication in the cellular system. The results indicate the prospects of using the proposed drugs to affect target nucleic acids inside cells.
{"title":"Nanocomposites consisting of titanium dioxide nanoparticles, antisense oligonucleotides, and photoactive groups as agents for effective action on nucleic acids","authors":"Asya S. Levina, M. Repkova, O.YU. Mazurkov, E. V. Makarevich, N. A. Mazurkova, V. F. Zarytova","doi":"10.36233/0372-9311-456","DOIUrl":"https://doi.org/10.36233/0372-9311-456","url":null,"abstract":"Relevance. Studies on model systems have confirmed the effectiveness of antisense oligonucleotides, including those that contain photoactive groups, for the modification of nucleic acids. However, this strategy has not yet found wide application due to the lack of successful methods for the cellular delivery. The development of effective preparations capable of acting on target nucleic acids in cells is an urgent task. \u0000The objective of the work is to create nanocomposites consisting of TiO2 nanoparticles, antisense oligonucleotides, and photoactive groups and to study their effect on target nucleic acids by the example of inhibition of influenza A virus replication in the cellular system. \u0000Materials and methods. Influenza virus A/Aichi/2/68 (A/H3N2), N-succinimide ether of p-azidotetrafluorobenzoic acid, TiO2 nanoparticles, and oligodeoxyribonucleotides have been used in the work. The antiviral activity of the proposed nanocomposites has been studied on the MDCK cells infected with the A/H3N2 virus. \u0000Results and discussion. Unique nanocomposites have been created, which consist of three functional components, i.e., titanium dioxide nanoparticles, antisense oligonucleotides, and the photoactive tetrafluoroarylazide group, respectively, providing penetration into cells, selective interaction with target nucleic acids, and photomodification of the targets. A significant antiviral site-specific action of the nanocomposites has been demonstrated against the influenza A virus in the cellular system, which exceeds the effect of the analogous samples that contain no photoactive groups. \u0000Conclusion. The biological activity of the created nanocomposites has been demonstrated by the example of highly effective suppression of influenza A virus replication in the cellular system. The results indicate the prospects of using the proposed drugs to affect target nucleic acids inside cells.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"259 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140247352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. V. Toporkov, E. Putintseva, S. K. Udovichenko, Natalya V. Boroday, E. Molchanova, Olga S. Bondareva, AS Antonov
Introduction. The relevance of the study is due to the need to assess the real expansion of West Nile fever (WNF) in Russia, analyze the manifestations of epizootic and epidemic processes and study the population structure of West Nile virus (WNV). Aim. To obtain objective data on the intensity of WNV circulation in certain territories of Russia and study the properties of the isolated strains of the pathogen. Materials and methods. 4564 samples of field material from 23 subjects of the Russian Federation and 1547 samples of clinical material from 12 subjects of the Russian Federation were examined. A set of laboratory diagnostic methods was used: ELISA, RT-PCR, virological, sequencing. Results. In 20 out of 1331 (1.5%) examined febrile patients, IgM antibodies to WNV were detected, including positive cases identified for the first time in the Karachay-Cherkess Republic, Tver and Vladimir regions. The presence of immunity to WNV was found on average in 8.6% of 11 federal subjects' population with high seroprevalence rates in Zaporozhye (24,5%), Tula (15,4%) and Kursk (11,1%) regions. Low-avidity IgG antibodies were detected in 44 (33,1%) samples from a population of 9 federal subjects. Intensive circulation of WNV lineage 2 in the epizootic cycle in the southern part of European Russia was confirmed. 12 WNV isolates were obtained, including those isolated for the first time in the Republic of Crimea, the Karachay-Cherkess Republic and the Stavropol Territory. The heterogeneity of circulating WNF causative agent's strains related to the two clades of the WNV lineage 2 formed in 2021 and 2022 was established. Conclusion. In the course of comprehensive monitoring studies, a new northern border of the WNF range was established, which as of 2022 passes through the territory of the Tver region. Data from seroepidemiological studies, including the detection of low-avidity IgG antibodies, confirm fairly intense, but undiagnosed contact of the population of the European part of Russia with the WNF pathogen. In the southern region of the European part of Russia, WNV genotype 2 predominantly circulates, belonging to at least two clades formed in 2021 and 2022.
{"title":"Study of the circulation and properties of the West Nile virus in Russia in 2022","authors":"A. V. Toporkov, E. Putintseva, S. K. Udovichenko, Natalya V. Boroday, E. Molchanova, Olga S. Bondareva, AS Antonov","doi":"10.36233/0372-9311-432","DOIUrl":"https://doi.org/10.36233/0372-9311-432","url":null,"abstract":"Introduction. The relevance of the study is due to the need to assess the real expansion of West Nile fever (WNF) in Russia, analyze the manifestations of epizootic and epidemic processes and study the population structure of West Nile virus (WNV). \u0000Aim. To obtain objective data on the intensity of WNV circulation in certain territories of Russia and study the properties of the isolated strains of the pathogen. \u0000Materials and methods. 4564 samples of field material from 23 subjects of the Russian Federation and 1547 samples of clinical material from 12 subjects of the Russian Federation were examined. A set of laboratory diagnostic methods was used: ELISA, RT-PCR, virological, sequencing. \u0000Results. In 20 out of 1331 (1.5%) examined febrile patients, IgM antibodies to WNV were detected, including positive cases identified for the first time in the Karachay-Cherkess Republic, Tver and Vladimir regions. The presence of immunity to WNV was found on average in 8.6% of 11 federal subjects' population with high seroprevalence rates in Zaporozhye (24,5%), Tula (15,4%) and Kursk (11,1%) regions. Low-avidity IgG antibodies were detected in 44 (33,1%) samples from a population of 9 federal subjects. Intensive circulation of WNV lineage 2 in the epizootic cycle in the southern part of European Russia was confirmed. 12 WNV isolates were obtained, including those isolated for the first time in the Republic of Crimea, the Karachay-Cherkess Republic and the Stavropol Territory. The heterogeneity of circulating WNF causative agent's strains related to the two clades of the WNV lineage 2 formed in 2021 and 2022 was established. \u0000Conclusion. In the course of comprehensive monitoring studies, a new northern border of the WNF range was established, which as of 2022 passes through the territory of the Tver region. Data from seroepidemiological studies, including the detection of low-avidity IgG antibodies, confirm fairly intense, but undiagnosed contact of the population of the European part of Russia with the WNF pathogen. In the southern region of the European part of Russia, WNV genotype 2 predominantly circulates, belonging to at least two clades formed in 2021 and 2022.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"13 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140247953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. Faizuloev, A. Gracheva, E. Korchevaya, Yulia I. Ammour, D. Smirnova, Darya M. Khokhlova, Andrey O. Drokov, A. A. Pankratov, G. Trunova, V. Khokhlova, M. S. Vorontsova, I A Leneva, O. A. Svitich, V. V. Zverev
Introduction. Despite the end of the COVID-19 pandemic, the problem of vaccine prevention of this disease appears highly relevant. The emergence and widespread distribution of the Omicron SARS-CoV-2 variant of concern (VOC) and its sublineages has dramatically reduced the efficacy of vaccination. The possible approach to solving this problem is to develop a nasal live attenuated vaccine capable of activating humoral, mucosal, and cell-mediated immunity, providing a prolonged immune response and cross-protection against different VOCs. The aim of the study was to determine the immunization efficacy with attenuated cold-adapted Wuhan-like SARS-CoV-2 D-D2 strain against homologous and heterologous challenges. Materials and methods. The study was conducted on an animal model of coronavirus pneumonia in golden Syrian hamsters. The efficacy of immunization was assessed by comparing the dynamics of weight, viral load in organs and histopathological changes in the lungs in immunized and unimmunized animals. Results. Single intranasal immunization of golden Syrian hamsters with D-D2 strain showed its high immunogenicity: seroconversion was evident in all immunized animals. Wuhan-like D-D2 strain provides highly effective protection of hamsters against the development of productive infection and pneumonia when challenged both with ancestral virus and heterologous strains related to Delta (AY.122) and Omicron (sublineages BA.1.1 and BA.5.2) variants. Conclusion. SARS-CoV-2 attenuation is a promising strategy for the development of a highly effective nasal live COVID-19 vaccine.
{"title":"Single intranasal immunization with attenuated Wuhan-like SARS-CoV-2 provides highly effective cross-protection against Delta and Omicron variants of concern","authors":"E. Faizuloev, A. Gracheva, E. Korchevaya, Yulia I. Ammour, D. Smirnova, Darya M. Khokhlova, Andrey O. Drokov, A. A. Pankratov, G. Trunova, V. Khokhlova, M. S. Vorontsova, I A Leneva, O. A. Svitich, V. V. Zverev","doi":"10.36233/0372-9311-496","DOIUrl":"https://doi.org/10.36233/0372-9311-496","url":null,"abstract":"Introduction. Despite the end of the COVID-19 pandemic, the problem of vaccine prevention of this disease appears highly relevant. The emergence and widespread distribution of the Omicron SARS-CoV-2 variant of concern (VOC) and its sublineages has dramatically reduced the efficacy of vaccination. The possible approach to solving this problem is to develop a nasal live attenuated vaccine capable of activating humoral, mucosal, and cell-mediated immunity, providing a prolonged immune response and cross-protection against different VOCs. \u0000The aim of the study was to determine the immunization efficacy with attenuated cold-adapted Wuhan-like SARS-CoV-2 D-D2 strain against homologous and heterologous challenges. \u0000Materials and methods. The study was conducted on an animal model of coronavirus pneumonia in golden Syrian hamsters. The efficacy of immunization was assessed by comparing the dynamics of weight, viral load in organs and histopathological changes in the lungs in immunized and unimmunized animals. \u0000Results. Single intranasal immunization of golden Syrian hamsters with D-D2 strain showed its high immunogenicity: seroconversion was evident in all immunized animals. Wuhan-like D-D2 strain provides highly effective protection of hamsters against the development of productive infection and pneumonia when challenged both with ancestral virus and heterologous strains related to Delta (AY.122) and Omicron (sublineages BA.1.1 and BA.5.2) variants. \u0000Conclusion. SARS-CoV-2 attenuation is a promising strategy for the development of a highly effective nasal live COVID-19 vaccine.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"567 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140246776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guzel S. Isaeva, A. Z. Zaripova, L. Bayazitova, Ralina M. Khusainova, T. A. Chazova, O. F. Tyupkina, E. Nikitina, I. Tsvetkova
Objective: to investigate the regional peculiarities of Streptococcus pneumoniae carriage in the pediatric population and characterize the dominant serotypes of the pathogen. Materials and methods. The clinical study group consisted of 509 healthy children attending preschool institutions. Examination of nasopharyngeal samples for the detection of S. pneumonae was carried out by classical bacteriological and molecular biological methods. The serotype was determined by real-time PCR. Genome-wide sequencing of the serogroups 15 and 11 isolates and bioinformatic analysis were performed. Results. The S. pneumoniae bacterial carriers in the group of healthy children was detected in 207 children (40.7%), while the frequency of detection of S. pneumoniae in urban children living in Kazan was significantly higher than in children living in rural area and amounted to 53.4 and 31.1%, respectively (p 0.05). Among children vaccinated with the 13-valent pneumococcal conjugate vaccine (PCV-13), S. pneumoniae carriers were not detected in 57.5% of cases. There were no significant differences in the degree of nasopharyngeal contamination depending on the vaccination status. Analysis of the serotype composition indicates the predominance of vaccine serotypes (57.7%), while the share of serotypes included in the PСV-13 vaccine accounts for only 24.7%, the share of non-vaccine serotypes was 32.1%, untyped — 10.2%. In unvaccinated children, vaccine serotypes that are part of the PCV-13 and 23-valent polysaccharide pneumococcal vaccine prevailed (PPSV-23): 6ABCD (21%), 11 AD (15%), 14 (13%). In vaccinated children, serotypes not included in the active vaccines dominated: 15AF (17.4%), 23A (19.2%), as well as 11AD (19.6%) (11А is included in PPSV-23). The 27 Kz isolate (serotype 15C) belonged to one of the most common sequence types ST1025. The 105_Kz isolate (serotype 11D) belonged to another common sequence type ST 62. Conclusion. In order to improve epidemiological surveillance of pneumococcal infection, it is necessary to introduce the monitoring of circulating clonal complexes of dominant S. pneumoniae serogroups and analyze the genetic determinants of antibiotic resistance and virulence depending on the sequence type.
{"title":"Characteristics of Streptococcus pneumoniae carriage in the pediatric population","authors":"Guzel S. Isaeva, A. Z. Zaripova, L. Bayazitova, Ralina M. Khusainova, T. A. Chazova, O. F. Tyupkina, E. Nikitina, I. Tsvetkova","doi":"10.36233/0372-9311-445","DOIUrl":"https://doi.org/10.36233/0372-9311-445","url":null,"abstract":"Objective: to investigate the regional peculiarities of Streptococcus pneumoniae carriage in the pediatric population and characterize the dominant serotypes of the pathogen. \u0000Materials and methods. The clinical study group consisted of 509 healthy children attending preschool institutions. Examination of nasopharyngeal samples for the detection of S. pneumonae was carried out by classical bacteriological and molecular biological methods. The serotype was determined by real-time PCR. Genome-wide sequencing of the serogroups 15 and 11 isolates and bioinformatic analysis were performed. \u0000Results. The S. pneumoniae bacterial carriers in the group of healthy children was detected in 207 children (40.7%), while the frequency of detection of S. pneumoniae in urban children living in Kazan was significantly higher than in children living in rural area and amounted to 53.4 and 31.1%, respectively (p 0.05). Among children vaccinated with the 13-valent pneumococcal conjugate vaccine (PCV-13), S. pneumoniae carriers were not detected in 57.5% of cases. There were no significant differences in the degree of nasopharyngeal contamination depending on the vaccination status. Analysis of the serotype composition indicates the predominance of vaccine serotypes (57.7%), while the share of serotypes included in the PСV-13 vaccine accounts for only 24.7%, the share of non-vaccine serotypes was 32.1%, untyped — 10.2%. In unvaccinated children, vaccine serotypes that are part of the PCV-13 and 23-valent polysaccharide pneumococcal vaccine prevailed (PPSV-23): 6ABCD (21%), 11 AD (15%), 14 (13%). In vaccinated children, serotypes not included in the active vaccines dominated: 15AF (17.4%), 23A (19.2%), as well as 11AD (19.6%) (11А is included in PPSV-23). The 27 Kz isolate (serotype 15C) belonged to one of the most common sequence types ST1025. The 105_Kz isolate (serotype 11D) belonged to another common sequence type ST 62. \u0000Conclusion. In order to improve epidemiological surveillance of pneumococcal infection, it is necessary to introduce the monitoring of circulating clonal complexes of dominant S. pneumoniae serogroups and analyze the genetic determinants of antibiotic resistance and virulence depending on the sequence type.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"2020 43","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140246072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Y. Popova, M. Y. Shchelkanov, N. V. Krylova, Alexey A. Belik, Lyubov M. Semeikina, Tatyana S. Zaporozhets, Vyacheslav Y. Smolenskiy, Elena V. Persianova, M. N. Prosyannikova, Yuriy A. Belov, O. V. Iunikhina, Anastasiya B. Pott, Tatyana F. Khomichuk, A. Simakova, S.A. Abramova, O. B. Romanova, T. N. Detkovskaya, S. P. Kryzhanovskiy, N. Besednova
Introduction. The COVID-19 pandemic, etiologically related to SARS-CoV-2, was the longest-lasting pandemic for an acute respiratory disease and had a significant impact on demography, economics and politics globally. Experiences with this pandemic are significant for the sustainable development of human society. A detailed analysis of these experiences in epidemic control should include details of the pathogen evolution down to the regional level. The aim of the study was to establish the regularities of the COVID-19 epidemic process in connection with the change of the pathogen genetic variants on the territory of Primorsky Krai. Materials and methods. A retrospective epidemiological analysis of COVID-19 incidence and dynamics of circulation of SARS-CoV-2 genetic variants during the pandemic of this disease (11.03.2020–05.05.2023) in Primorsky Krai was carried out. Data was gathered from the Department of Rospotrebnadzor in Primorsky Krai, the Centre of Hygiene and Epidemiology in Primorsky Krai, as well as the G.P. Somov Research Institute of Epidemiology and Microbiology of the Russian Federation and Russian Platform for Aggregation of Information on Virus Genomes (VGARus). In particular, 1055 nucleotide sequences of full-length SARS-CoV-2 genomes from Primorsky Krai, among which 553 were sequenced at the G.P. Somov Research Institute of Epidemiology and Microbiology were included in analysis. Results. When analyzing the epidemic dynamics of COVID-19 in Primorsky Krai (2020–2023), 7 rises in incidence with different clinical and epidemiological symptoms depending on the genetic variants of the pathogen were identified. At the beginning of the pandemic in Primorsky Krai, as well as throughout Russia, Wuhan-like variants of SARS-CoV-2 were predominant, though later, Delta and Omicron genetic variants were in the majority. By the end of April — beginning of May 2023, the proportion of Omicron sub-variants (XBB.1.9.2 and XBB.1.16) in Primorsky Krai was higher than the Russian average and comparable to that in neighboring countries (Republic of Korea and Japan). Conclusion. Due to the ongoing evolution of SARS-CoV-2, the possibility of the emergence of new pathogens, the peculiarities of the geographical location as well as political and economic importance of Primorsky Krai, it is necessary to consistently improve regional capabilities for operational molecular virological monitoring.
{"title":"Genotypic portrait of SARS-CoV-2 in Primorsky Krai during the COVID-19 pandemic","authors":"A. Y. Popova, M. Y. Shchelkanov, N. V. Krylova, Alexey A. Belik, Lyubov M. Semeikina, Tatyana S. Zaporozhets, Vyacheslav Y. Smolenskiy, Elena V. Persianova, M. N. Prosyannikova, Yuriy A. Belov, O. V. Iunikhina, Anastasiya B. Pott, Tatyana F. Khomichuk, A. Simakova, S.A. Abramova, O. B. Romanova, T. N. Detkovskaya, S. P. Kryzhanovskiy, N. Besednova","doi":"10.36233/0372-9311-497","DOIUrl":"https://doi.org/10.36233/0372-9311-497","url":null,"abstract":"Introduction. The COVID-19 pandemic, etiologically related to SARS-CoV-2, was the longest-lasting pandemic for an acute respiratory disease and had a significant impact on demography, economics and politics globally. Experiences with this pandemic are significant for the sustainable development of human society. A detailed analysis of these experiences in epidemic control should include details of the pathogen evolution down to the regional level. \u0000The aim of the study was to establish the regularities of the COVID-19 epidemic process in connection with the change of the pathogen genetic variants on the territory of Primorsky Krai. \u0000Materials and methods. A retrospective epidemiological analysis of COVID-19 incidence and dynamics of circulation of SARS-CoV-2 genetic variants during the pandemic of this disease (11.03.2020–05.05.2023) in Primorsky Krai was carried out. Data was gathered from the Department of Rospotrebnadzor in Primorsky Krai, the Centre of Hygiene and Epidemiology in Primorsky Krai, as well as the G.P. Somov Research Institute of Epidemiology and Microbiology of the Russian Federation and Russian Platform for Aggregation of Information on Virus Genomes (VGARus). In particular, 1055 nucleotide sequences of full-length SARS-CoV-2 genomes from Primorsky Krai, among which 553 were sequenced at the G.P. Somov Research Institute of Epidemiology and Microbiology were included in analysis. \u0000Results. When analyzing the epidemic dynamics of COVID-19 in Primorsky Krai (2020–2023), 7 rises in incidence with different clinical and epidemiological symptoms depending on the genetic variants of the pathogen were identified. At the beginning of the pandemic in Primorsky Krai, as well as throughout Russia, Wuhan-like variants of SARS-CoV-2 were predominant, though later, Delta and Omicron genetic variants were in the majority. By the end of April — beginning of May 2023, the proportion of Omicron sub-variants (XBB.1.9.2 and XBB.1.16) in Primorsky Krai was higher than the Russian average and comparable to that in neighboring countries (Republic of Korea and Japan). \u0000Conclusion. Due to the ongoing evolution of SARS-CoV-2, the possibility of the emergence of new pathogens, the peculiarities of the geographical location as well as political and economic importance of Primorsky Krai, it is necessary to consistently improve regional capabilities for operational molecular virological monitoring.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"530 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140247100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tatyana A. Kuznetsova, N. Besednova, M. R. Aliev, M. Y. Shchelkanov
The aim of the review is to give a brief characteristic of cell cultures obtained from mammalian tissues and to consider the current possibilities and prospects for their use in virology. The analysis of the literature data presented in the main databases, such as Web of Science, PubMed, Scopus, Elsevier, Google Scholar and RSCI (as of July 2023), indicates that various types of cell cultures are currently used in virological studies. The use of cell culture has a number of advantages over other in vitro and in vivo methods of virological research. The review provides numerous examples on the development of new methods of obtaining cell cultures for the cultivation of viruses. Among them are sensitive and reporter cell systems, the design of which can be a promising tool for diagnostics of existing and new unknown viral infections. Cell cultures are characterized as potential in vitro models in virology for developing new diagnostic test-systems and antiviral drugs. An important area of cell cultures application is their use as a substrate for the production of culture-derived vaccines. Another aspect of the cell cultures application is also highlighted, such as the study of the effect of the viruses on the host immune system or the mechanisms of immunopathogenesis of viral infections. It is concluded that the use of cell cultures remains currently and in the near future one of the most important methods in practical virology and in scientific research.
本综述旨在简要介绍从哺乳动物组织中获得的细胞培养物的特点，并探讨其目前在病毒学中应用的可能性和前景。对主要数据库（如 Web of Science、PubMed、Scopus、Elsevier、Google Scholar 和 RSCI，截至 2023 年 7 月）中的文献数据进行的分析表明，目前有各种类型的细胞培养物被用于病毒学研究。与其他体外和体内病毒学研究方法相比，细胞培养法具有许多优势。这篇综述提供了大量实例，说明如何开发新的细胞培养方法来培养病毒。其中包括敏感细胞和报告细胞系统，这些系统的设计可以成为诊断现有和新的未知病毒感染的有前途的工具。细胞培养物是病毒学中潜在的体外模型，可用于开发新的诊断测试系统和抗病毒药物。细胞培养物的一个重要应用领域是用作生产培养物衍生疫苗的基质。此外，还强调了细胞培养物应用的另一个方面，如研究病毒对宿主免疫系统的影响或病毒感染的免疫发病机制。总之，细胞培养物的使用在当前和不久的将来仍然是实用病毒学和科学研究中最重要的方法之一。
{"title":"The cell cultures in virology: from the past to the future","authors":"Tatyana A. Kuznetsova, N. Besednova, M. R. Aliev, M. Y. Shchelkanov","doi":"10.36233/0372-9311-421","DOIUrl":"https://doi.org/10.36233/0372-9311-421","url":null,"abstract":"The aim of the review is to give a brief characteristic of cell cultures obtained from mammalian tissues and to consider the current possibilities and prospects for their use in virology. \u0000The analysis of the literature data presented in the main databases, such as Web of Science, PubMed, Scopus, Elsevier, Google Scholar and RSCI (as of July 2023), indicates that various types of cell cultures are currently used in virological studies. The use of cell culture has a number of advantages over other in vitro and in vivo methods of virological research. The review provides numerous examples on the development of new methods of obtaining cell cultures for the cultivation of viruses. Among them are sensitive and reporter cell systems, the design of which can be a promising tool for diagnostics of existing and new unknown viral infections. Cell cultures are characterized as potential in vitro models in virology for developing new diagnostic test-systems and antiviral drugs. An important area of cell cultures application is their use as a substrate for the production of culture-derived vaccines. Another aspect of the cell cultures application is also highlighted, such as the study of the effect of the viruses on the host immune system or the mechanisms of immunopathogenesis of viral infections. It is concluded that the use of cell cultures remains currently and in the near future one of the most important methods in practical virology and in scientific research.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"30 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140245360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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