Small nucleolar RNAs (snoRNAs) are appreciable players in gene expression regulation in human cells. The canonical function of box C/D and box H/ACA snoRNAs is posttranscriptional modification of ribosomal RNAs (rRNAs), namely, 2′-O-methylation and pseudouridylation, respectively. A series of independent studies demonstrated that snoRNAs, as well as other noncoding RNAs, serve as the source of various short regulatory RNAs. Some snoRNAs and their fragments can also participate in the regulation of alternative splicing and posttranscriptional modification of mRNA. Alterations in snoRNA expression in human cells can affect numerous vital cellular processes. SnoRNA level in human cells, blood serum, and plasma presents a promising target for diagnostics and treatment of human pathologies. Here we discuss the relation between snoRNAs and oncological, neurodegenerative, and viral diseases and also describe changes in snoRNA level in response to artificial stress and some drugs.
Under the COVID-19 pandemic, healthcare workers were at the highest risk of getting infected with the disease; this necessitates specialized studies in this occupational group. The aim of the study was to identify non-occupational risk factors and laboratory markers indicating that severe clinical forms of new coronavirus infection would probably develop in healthcare workers in the initial period of the pandemic. The study included 366 workers who suffered COVID-19 in 2020–2021. The disease was confirmed by examining smears from the pharynx and nose with PCR. Some of the samples were examined using the SARS-CoV-2 whole genome sequencing technology. To determine laboratory prognostic indicators evidencing the development of more severe forms of the disease (pneumonia), a number of healthcare workers underwent laboratory examination during the acute period of the disease, namely: general clinical and biochemical blood tests, immunophenotyping of lymphocytes, analysis of the hemostasis system and cytokine levels. To study non-occupational risk factors of pneumonia, all healthcare workers after recovery were asked to fill in a Google form developed by the authors. The most severe clinical forms of COVID-19 were registered in healthcare workers who were older than 40 years, with low physical activity and a body mass index higher than 25.0, had diabetes mellitus and chronic diseases of the genitourinary system. When analyzing the results of laboratory tests, markers indicating development of pneumonia were identified and their critical values (cut-off points) were determined: the level of lymphocytes (below 1.955•109/l), T-cytotoxic lymphocytes (below 0.455•109/l), T-helpers (below 0.855•109/L), natural killers (below 0.205•109/l), platelets (below 239•109/L), erythrocyte sedimentation rate (above 11.5 mm/h), D-dimer (above 0.325 mcg/ml), total protein (below 71.55 g/L), lactate dehydrogenase (above 196 U/L), C-reactive protein (above 4.17 mg/l), and interleukin-6 (above 3.63 pg/l). The study identified non-occupational risk factors causing development of severe COVID-19 and established laboratory prognostic indicators.
Система генетического контроля реакции врожденного иммунитета на гриппозную инфекцию и функции генов позволяет вести разработку системного лечения гриппа с ориентацией на фенотипические проявления мутаций с учетом наследственной предрасположенности индивида к тяжелому течению заболевания и/или развитию осложнений.
Abstract The Omicron variant of SARS-CoV-2 has rapidly spread globally in late 2021 - early 2022, displacing the previously prevalent Delta variant. Before December 16, 2021, community transmission had already been observed in tens of countries globally. However, in Russia, the majority of reported cases at that time had been sporadic and associated with travel. Here, we report an Omicron outbreak at a student dormitory in Saint Petersburg between December 16 - 29, 2021, which was the earliest known instance of large-scale community transmission in Russia. Out of the 465 sampled residents of the dormitory, 180 (38.7%) tested PCR positive. Among the 118 residents for whom the variant has been tested by whole-genome sequencing, 111 (94.1%) carried the Omicron variant. Among these 111 residents, 60 (54.1%) were vaccinated or had reported previous COVID-19. Phylogenetic analysis confirmed that the outbreak was caused by a single introduction of the BA.1.1 sublineage of Omicron. The dormitory-derived clade constituted a significant proportion of BA.1.1 samples in Saint-Petersburg and has spread to other regions of Russia and other countries. The rapid spread of Omicron in a population with preexisting immunity to previous variants underlines its propensity for immune evasion.
BACKGROUND: The application of CRISPR/Cas9 is one of the most rapidly developing areas in biotechnology. This method was used to obtain clones of а human origin cell line with knockout of one or more genes of the IFITM family, representing host restriction factors for influenza infection. Amphotericin B has previously been shown to promote influenza infection by blocking IFITM3 function. AIM: The aim of this study was to evaluate the effect of amphotericin B on the sensitivity of IFITM knockout cells to influenza A virus infection. MATERIALS AND METHODS: WI-38 VA-13 cells and mutant clones with IFITM3 knockout (F3 clone) or IFITM1, IFITM3 knockout (clone E12) were infected with influenza virus A/PR/8/34 (H1N1) in the presence or absence of amphotericin B. Forty-four hours after infection, the culture medium was taken to determine the infectious activity of the virus by titration in the MDCK cell culture, as well as the hemagglutinating activity of the virus. The infected cells were stained with fluorescently labeled antibodies against the viral NP protein, and the number of NP-positive cells was determined by flow cytometry. RESULTS: The addition of amphotericin B increased the hemagglutinating and infectious activity of the virus in WI-38 VA-13cells, while the difference was insignificant for clones with IFITM gene knockout. A similar dependency was obtained for the percent of infected cells. CONCLUSIONS: Mutant cells with a knockout of one or several genes of the IFITM family were equally susceptible to influenza infection regardless of the addition of amphotericin B, which confirms the crucial importance of a defect in the IFITM3 protein in increasing the permissiveness of cells to influenza A virus.
The emergence of novel highly pathogenic avian influenza viruses (HPAIVs) in migratory birds raises serious concerns as these viruses have the potential to spread during fall migration. We report the identification of novel HPAIV A(H5N8) clade 2.3.4.4 virus that was isolated from sick domestic duck at commercial farm during the second wave of spread that began in October and affected poultry (ducks; chiсkens) in several European regions of Russia and Western Siberia in 2016. The strain was highly lethal in experimental infection of chickens and mice with IVPI = 2.34 and MLD50 = 1.3log10 EID50, accordingly. Inoculation of chickens with the HPAIV A/H5N8 demonstrated neuroinvasiveness, multiorgan failure, and death of chickens on the 3rd day post inoculation. Virus replicated in all collected organ samples in high viral titers with the highest titer in the brain (6.75±0.1 log10TCID50/ml). Effective virus replication was found in the following cells: neurons and glial cells of a brain; alveolar cells and macrophages of lungs; epithelial cells of a small intestine; hepatocytes and Kupffer cells of a liver; macrophages and endothelial cells of a spleen; and the tubular epithelial cells of kidneys. These findings advance our understanding of histopathological effect of A(H5N8) HPAIV infection.
ABSTRACT In this paper, we analyze the etiology of the diseases occurring during two consecutive influenza epidemic seasons in St. Petersburg, Russian Federation. The analysis is based on the results of the PCR diagnostics of the clinical samples collected from patients hospitalized in three St. Petersburg hospitals with influenza like illnesses (ILI). It was shown that the influenza virus A(H1N1)pdm09 was the dominant causative agent during the 2012-2013 epidemic season while, in the 2013-2014 season, A(H3N2) virus was predominant among adults and children. The influenza B virus activity was high in the 2012-2013 season and low in the 2013-2014 season. During both seasons, the main causative agent for the hospitalization of young children was respiratory syncytial virus (RSV), followed by rhinovirus and influenza virus. The rate of involvement of parainfluenza, adenovirus, metapneumovirus and coronavirus was low and was negligible for bocavirus. Children 0-2 and 3-6 years old formed the group of patients that was affected by acute respiratory infection agents the most. Children younger than 3 months old were the major group of the intensive care unit (ICUs) patients and only 27.5% of them were adults. RSV and rhinovirus were the leading cause of ILI among the children admitted to ICU. Among the adult patients admitted to the ICU, only influenza A(H1N1)pdm09, A(H3N2) and B viruses were detected during both influenza seasons. According to the results of the antigenic and genetic analysis, most influenza A(H1N1)pdm09 and A(H3N2) viruses circulating in St. Petersburg matched the vaccine strains recommended by the WHO for vaccine composition in the 2012-2013 and 2013-2014 seasons.
Human seasonal coronaviruses (hCoVs) are a group of viruses that affect the upper respiratory tract. While seasonal patterns and the annual variability of predominant hCoV species are well-documented, their genetic and species diversity in St. Petersburg and across Russia remains largely unexplored. In this study, we developed a two-pool, long-amplicon (900-1100 bp) PCR primer panel for the whole-genome sequencing of four seasonal hCoV species. The panel was validated using nasopharyngeal swab samples collected within the Global Influenza Hospital Surveillance Network (GIHSN) project. Over a period of six epidemiological seasons from 2017 to 2023, we retrospectively analyzed 14,704 nasopharyngeal swabs collected from patients hospitalized in St. Petersburg clinics. Of these samples, 5010 (34.07%) tested positive for respiratory viruses, with 424 (2.88% of all samples) identified as seasonal human coronaviruses. The assessment of species diversity showed that predominant hCoV species alternate between seasons. Whole-genome sequences for 85 seasonal human coronaviruses (hCoVs) with >70% genome coverage were obtained, including 23 hCoV-OC43, 6 hCoV-HKU1, 39 hCoV-229E, and 17 hCoV-NL63. These represent the first near-complete genomes of seasonal hCoVs from the Russian Federation, addressing a significant gap in the genomic epidemiology of these viruses. A detailed phylogenetic analysis of the sequenced genomes was conducted, highlighting the emergence of hCoV-229E subclades 7b.1 and 7b.2, which carry numerous substitutions in the Spike protein. Additionally, we sequenced a historical hCoV-229E isolate collected in the USSR in 1979, the oldest sequenced 229E virus from Eurasia, and demonstrated that it belongs to Genotype 2. The newly developed PCR-based sequencing protocol for seasonal hCoVs is straightforward and well-suited for genomic surveillance, providing a valuable tool to enhance our understanding of the genetic diversity of human seasonal coronaviruses.
Whole genome sequencing (WGS) is considered the best instrument to track both virus evolution and the spread of new, emerging variants. However, WGS still does not allow the analysis of as many samples as qPCR does. Epidemiological and clinical research needs to develop advanced qPCR methods to identify emerging variants of SARS-CoV-2 while collecting data on their spreading in a faster and cheaper way, which is critical for introducing public health measures. This study aimed at designing a one-step RT-qPCR assay for multiplex detection of the Omicron lineage and providing additional data on its subvariants in clinical samples. The RT-qPCR assay demonstrated high sensitivity and specificity on multiple SARS-CoV-2 variants and was cross-validated by WGS.
A panel of five monoclonal antibodies (MAbs) to the HA1 molecule of the influenza B virus of the Victorian lineage with high virus-neutralizing activity was developed. For identification of the virus neutralizing epitopes in HA1 escape mutants (EM) of the influenza B/Shandong/07/97 and B/Malaysia/2506/04 virus were selected using virus-neutralizing antibodies (MAbs). Three EMs had single, two double and one triple amino acid substitutions (AAS) in HA1 (H122N, A202E, K203T, K203i, K203N or A317V). In addition, AAS N197S was detected in three EMs. A correlation of AAS identified with peculiarities of interaction of EMs with Mabs was discussed.
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