Virology Research Research Articles

The incidence and structure of dental pathology in children with chronic viral hepatitis C

To develop a complex of therapeutic measures for various lesions of the oral mucosa in children with chronic viral hepatitis C. To solve the tasks and achieve the goals of the study, general clinical, biochemical, immunological, microbiological, virological, dental and statistical research methods were used in the work. The study was conducted in the period from 2018 to 2021. Data collection was carried out on the basis of the Republican Specialized Scientific and Practical Medical Center of Pediatrics of the Ministry of Health of the Republic of Uzbekistan (Department of Chronic Hepatitis. The head of the department is Academician of the Academy of Sciences of the Republic of Uzbekistan Inoyatova F.I.), as well as on the basis of the pediatric dental clinic of the Tashkent State Dental Institute. The main group of patients was formed, including 112 children with HCV and pathology of COPD. Results: The analysis showed that the proposed method of treatment had a significantly positive effect on the condition of the oral mucosa in children with hepatit C

Plasma-Like Culture Medium for the Study of Viruses.

Viral infections attract more and more attention, especially after the emergence of novel zoonotic coronaviruses and the monkeypox virus over the last 2 decades. Research on viruses is based to a great extent on mammalian cell lines that are permissive to the respective viruses. These cell lines are usually cultivated according to the protocols established in the 1950s to 1970s, although it is clear that classical media have a significant imprint on cell growth, phenotype, and especially metabolism. So, recently in the field of biochemistry and metabolomics novel culture media have been developed that resemble human blood plasma. As perturbations in metabolic and redox pathways during infection are considered significant factors of viral pathogenesis, these novel medium formulations should be adapted by the virology field. So far, there are only scarce data available on viral propagation efficiencies in cells cultivated in plasma-like media. But several groups have presented convincing data on the use of such media for cultivation of uninfected cells. The aim of the present review is to summarize the current state of research in the field of plasma-resembling culture media and to point out the influence of media on various cellular processes in uninfected cells that may play important roles in viral replication and pathogenesis in order to sensitize virology research to the use of such media.

Multiplication of defective Ebola virus in a complementary permissive cell line

In an effort to develop safe and innovative in vitro models for Ebola virus (EBOV) research, we generated a recombinant Ebola virus where the glycoprotein (GP) gene was substituted with the Cre recombinase (Cre) gene by reverse genetics. This defective virus could multiply itself in a complementary permissive cell line, which could express GP and reporter protein upon exogenous Cre existence. The main features of this novel model for Ebola virus are intact viral life cycle, robust virus multiplication and normal virions morphology. The design of this model ensures its safety, excellent stability and maneuverability as a tool for virology research as well as for antiviral agent screening and drug discovery, and such a design could be further adapted to other viruses.

Biological properties of domestic strain vRub-Ant of rubella virus

Introduction. Rubella is a mild infectious disease affecting mainly children and is caused by the rubella virus, part of the Matonoviridae family, genus Rubivirus. Rubella causes congenital rubella syndrome (CRS) and is the main cause of developmental abnormalities, especially blindness and deafness.
 There is no specific treatment for rubella and CRS. In order to avoid possible complications from rubella infection, a live attenuated rubella vaccine based on the foreign strain of Wistar RA 27/3 rubella virus is used. However, the actual, more effective and preferred vaccine strain the rubella virus for the Russian Federation is considered to be a viral strain of rubella circulating on its territory.
 The aim of the study was to study the biological properties of the developed domestic cold-adapted strain vRub-Ant circulating in the territory of the Russian Federation.
 Materials and methods. Following cell cultures were used in the study human embryo lung diploid cell strain LECH-3, transferable cell line from embryonic kidney cells of green monkeys Vero CCL-81 and Vero ECC, human mesenchymal stem cells, human peripheral blood mononuclear cells (PBMC). Cell cultures were grown on a DMEM/F12 nutrient medium with the addition of 5% fetal bovine serum. Swabs from the pharynx and nasal passages from a child with rubella were used as clinical virus-containing material. Monoclonal anti-idiotypic antibodies m(anti-ID)Ab were used to assess the expression level of alpha/beta and gamma interferon receptors (/ and IFN-R)Ab, imitating the biological effects of alpha/beta and gamma interferons (/ and IFN) of humans. The cultural, virological, immunochemical and serological research methods were applied in the study.
 Results. Attenuation of the vRub-Ant clinical isolate of rubella virus was carried out for 20 consecutive passages on LECH-3 diploid cells at a reduced temperature of 30C. The main biological markers of attenuation were determined to be ts and ca phenotypes. The avirulence of the attenuated viral strain (att-phenotype) was assessed by the level of expression of / and IFN-R. A lower level of / and IFN-R expression was found on the membranes of human PBMC induced by the vaccine strain vRub-Ant in comparison with the parent wild variant of the rubella virus. This trait,the att phenotype, is characteristic of attenuated viral strains. It has been shown that the vaccine strain vRub-Ant has lost neurotropism and was unable to bind to the membrane receptors of the brain (MRB) of guinea pig embryos, unlike its parent rubella virus strain. The high immunogenicity of the domestic cold-adapted strain vRub-Ant was confirmed by high titers of neutralizing rubella antibodies observed in guinea pigs immunized subcutaneously with one vaccination dose of the virus.
 Conclusion. A domestic attenuated vaccine strain vRub-Ant of the rubella virus that has the main biological markers of attenuation (ts-ca and att phenotypes) has been developed. The vaccine strain vRub-Ant induces a high levels of neutralizing antibodies in guinea pigs following the immunization with a single vaccination dose of the vaccine. The viral strain vRub-Ant has lost its tropism to the MRB of guinea pig embryos, unlike its parent variant.

The Use of Synchrotron Radiation in Virology

Introduction: The demand for new rapid methods for development of medicines for health care is becoming more relevant. Detailed knowledge of the spatial structure of viral proteins and their complexes formed when the organism is infected with viruses is important for effective development of vaccines and antiviral drugs. Timely selection of sensitive animal models and the study of the pathogenesis of an infectious disease on them are important for quality testing of biological preparations. The synchrotron radiation source is a new powerful biological research tool. Objective: To analyze the potential of existing sources of synchrotron radiation for conducting virology research, from macroorganisms to individual viral proteins. Materials and methods: We searched for literary sources published in 1996–2022 and devoted to the use of synchrotron radiation in virology and its importance for public health in the future using relevant keywords in the PubMed and PDB databases. Fifty-one full-text publications were found eligible for inclusion in the review. Results: Currently, there are over 70 different synchrotron radiation sources worldwide and many of them are used for diverse biological studies of living systems. Phase-contrast X-ray imaging makes it possible to visualize soft tissues in vivo with resolution up to 1 µm in the absence of contrast agents. Synchrotron radiation allows real-time 3D-histology without the necessity to prepare ultra-thin slices. Obtaining the structure of viral proteins in solution and protein crystallography realized with synchrotron radiation sources has been actively used in antiviral drug development and the study of fundamental properties of viruses since 2000. Conclusion: The X-ray techniques realized in synchrotron radiation sources discussed in this review constitute the fundamental basis of many virology studies and have a promising future for ensuring biological safety of Russia. Keywords: synchrotron radiation, virology, animal microtomography, histology, protein crystallography.

APPROBATION TRIALS OF A VACCINE AGAINST NEWCASTLE DISEASE FROM THE LA SOTA STRAIN

the paper presents the results of an approbation test of the physical and biological properties of an experimental series of a vaccine against Newcastle disease from the La-Sota strain in the conditions of the National Reference Center for Veterinary Medicine of the Committee for Veterinary Control and Supervision of the Ministry of Agriculture of the Republic of Kazakhstan (CVC and SMA RK) and the Research Institute of Biological Problems security (RIBSP). The aim of the study was to test a vaccine against Newcastle disease from the La-Sota strain. When performing the work, general virological and immunological research methods were used. The results of approbation studies showed that the vaccine was free of contaminants and its biological activity was 9,03 ± 0,14 lg EID50/cm3. The vaccine is harmless to chickens of 40 days of age. The vaccine is immunogenic for chickens. The average titers of antibodies to the Newcastle disease virus on the 16th day after vaccination is 1:127,2 in the hemagglutination inhibition test (HITA). The vaccine meets all the required requirements in terms of its physico-biological properties and is suitable for specific prophylaxis of Newcastle disease. Approbation tests of the physical, biological properties of the vaccine were completed with positive results and protocols, research acts were signed by members of the Commission of the CVC and SMA RK. It is recommended to use the vaccine on a large scale in the conditions of poultry farms, private farmsteads for the specific prevention of Newcastle disease.

Determination of the optimal immunizing dose of heterologous goat pox virus vaccine (Poxviridae: Chordopoxvirinae: Capripoxvirus) against lumpy skin disease

Lumpy skin disease (LSD), sheep pox and goat pox are dangerous diseases of domestic ruminants. Representatives of the genus of capripoxviruses are antigenically similar and can be used as a vaccine for three infections, as in the case of representatives of the genus of orthopoxviruses, which includes viruses of smallpox, monkeypox, and cowpox, that all belong to a single family Poxviridae. In this study, the vaccine strain G20-LKV of the goat pox virus and the virulent strain RIBSP2019/K of the LSD virus were used. The experiments were carried out on clinically healthy cattle of the Kazakh White-headed breed, aged six to eight months. Virological and serological research methods were used in the work. All immunized animals that received different doses of the vaccine showed resistance to the infection challenge, without showing any clinical signs of the disease. In animals that received the lowest doses of the vaccine 15,000, 30,000 and 40,000 TCID50, no adverse events, skin and temperature reactions were observed at the injection site. Those vaccinated with high doses of the vaccine had a local reaction in the form of swelling at the site of vaccine administration. Control animals infected with a virulent virus showed clinical signs of the cattle lumpy skin disease . The vaccine, prepared based on the "G20-LKV" strain of the goat virus, is protective for cattle against infection with a virulent LSD virus at immunizing doses from 15,000 to 80,000 TCID50, which are dependent on the LSD epizootic situation in particular region.

Generation and characterization of SARS-CoV-2 pseudoviruses

BACKGROUND: Pseudovirus technology is a versatile and valuable tool for both fundamental and applied virological research. Pseudotyped viruses provide the same cell entry mechanism as SARS-CoV-2 and are widely used to investigate the virus entry mechanism, cell tropism, and virus neutralization assays.
 AIM: The aim of the work is to obtain pseudotyped SARS-CoV-2 viruses and evaluate their transducing activity.
 MATERIALS AND METHODS: Using genetic engineering methods, a genetic construct carrying the SARS-CoV-2 glycoprotein S gene was obtained, as well as the pLenti-Luc-GFP reporter plasmid encoding the green fluorescent protein (GFP) and firefly luciferase genes. Pseudovirus particles were generated by transfection of eukaryotic cells. The transducing activity of pseudoviral particles displaying SARS-CoV-2 glycoprotein S on their surface was studied using HEK293, HEK293-hACE2, and HEK293-hACE2-TMPRSS2 (t) cell cultures.
 RESULTS: Based on the second-generation lentiviral platform, pseudoviruses were obtained that exhibit SARS-CoV-2 S glycoprotein on their surface. It was found that the pseudoviruses penetrate more efficiently into HEK293-hACE2-TMPRSS2 cells than into HEK293-hACE2. Pseudoviruses have been shown to be sensitive to neutralization by recombinant monoclonal antibodies that interact with the receptor-binding domain (RBD) of the SARS-CoV-2 S glycoprotein.
 CONCLUSIONS: The pseudoviruses can be used both to search for antiviral drugs that would be able to block the penetration of SARS-CoV-2 into the target cell, and to evaluate the effectiveness of the developed monoclonal antibodies and vaccines against SARS-CoV-2.

Global Scientific Productivity of Top Four Journals of Virology Over Three Decades: A Scientometrics Study

Trends in Virology research have been inclined after outbreak of COVID 19 pandemic. The main objective of this study is to identifying and visualizing the scientometric indicators of top four highly productive journals publishing papers on area related to Virology research. The data collected on 5 January 2022 from Web of Science all relevant global publications of Virology research. Then, publications were restricted to top 4 highly productive journals in this field. An Exploratory and descriptive analysis of bibliographic data (number of publication/citations yearly output, growth rate, authorship and collaborative pattern, productive countries) by using Bibexcel, MS Excel and VOS viewer software packages were used. The top ranked journals of virology ranked as Journal of Virology (N= 37205, 51.50%), Virology Journal (N= 15052, 20.83%), Journal of General Virology (N= 10745, 14.87%) and Archives of Virology (N= 9238, 12.79%) respectively. Cross-country comparison reveals that the USA is currently taking the leading role, followed by the UK, Germany & Japan and China is a newcomer with potential contributions to make in this research area. The present study offers important quantitative information on top journals of Virology. In addition, researchers are increasingly investigating the intellectual structures of disciplines by using various bibliometric techniques.

Counteracting Immunosuppression in the Tumor Microenvironment by Oncolytic Newcastle Disease Virus and Cellular Immunotherapy.

An apparent paradox exists between the evidence for spontaneous systemic T cell- mediated anti-tumor immune responses in cancer patients, observed particularly in their bone marrow, and local tumor growth in the periphery. This phenomenon, known as "concomitant immunity" suggests that the local tumor and its tumor microenvironment (TME) prevent systemic antitumor immunity to become effective. Oncolytic Newcastle disease virus (NDV), an agent with inherent anti-neoplastic and immune stimulatory properties, is capable of breaking therapy resistance and immunosuppression. This review updates latest information about immunosuppression by the TME and discusses mechanisms of how oncolytic viruses, in particular NDV, and cellular immunotherapy can counteract the immunosuppressive effect of the TME. With regard to cellular immunotherapy, the review presents pre-clinical studies of post-operative active-specific immunotherapy and of adoptive T cell-mediated therapy in immunocompetent mice. Memory T cell (MTC) transfer in tumor challenged T cell-deficient nu/nu mice demonstrates longevity and functionality of these cells. Graft-versus-leukemia (GvL) studies in mice demonstrate complete remission of late-stage disease including metastases and cachexia. T cell based immunotherapy studies with human cells in human tumor xenotransplanted NOD/SCID mice demonstrate superiority of bone marrow-derived as compared to blood-derived MTCs. Results from clinical studies presented include vaccination studies using two different types of NDV-modified cancer vaccine and a pilot adoptive T-cell mediated therapy study using re-activated bone marrow-derived cancer-reactive MTCs. As an example for what can be expected from clinical immunotherapy against tumors with an immunosuppressive TME, results from vaccination studies are presented from the aggressive brain tumor glioblastoma multiforme. The last decades of basic research in virology, oncology and immunology can be considered as a success story. Based on discoveries of these research areas, translational research and clinical studies have changed the way of treatment of cancer by introducing and including immunotherapy.

Generation of a VeroE6 Pgp gene knock out cell line and its use in SARS-CoV-2 antiviral study

Vero cells are widely used for antiviral tests and virology research for SARS-CoV-2 as well as viruses from various other families. However, Vero cells generally express high levels of multi-drug resistance 1 (MDR1) or Pgp protein, the efflux transporter of foreign substances including many antiviral compounds, affecting the antiviral activity as well as interpretation of data. To address this, a Pgp gene knockout VeroE6 cell line (VeroE6-Pgp-KO) was generated using CRISPR-CAS9 technology. These cells no longer expressed the Pgp protein as indicated by flow cytometry analysis following staining with a Pgp-specific monoclonal antibody. They also showed significantly reduced efflux transporter activity in the calcein acetoxymethyl ester (calcein AM) assay. The VeroE6-Pgp-KO cells and the parental VeroE6 cells were each infected with SARS-CoV-2 to test antiviral activities of remdesivir and nirmatrelvir, two known Pgp substrates, in the presence or absence of a Pgp inhibitor. The compounds showed antiviral activities in VeroE6-Pgp-KO cells similar to that observed in the presence of the Pgp inhibitor. Thus, the newly established VeroE6-Pgp-KO cell line adds a new in vitro virus infection system for SARS-CoV-2 and possibly other viruses to test antiviral therapies without a need to control the Pgp activity. Removal of the Pgp inhibitor for antiviral assays will lead to less data variation and prevent failed assays.

APOBEC3B: Future direction of liver cancer research.

Liver cancer is one of the most common cancers in the world, and the rate of liver cancer is high due to the of its illness. The main risk factor for liver cancer is infection with the hepatitis B virus (HBV), but a considerable number of genetic and epigenetic factors are also directly or indirectly involved in the underlying pathogenesis of liver cancer. In particular, the apolipoprotein B mRNA editing enzyme, catalytic peptide-like protein (APOBEC) family (DNA or mRNA editor family), which has been the focus of virology research for more than a decade, has been found to play a significant role in the occurrence and development of various cancers, providing a new direction for the research of liver cancer. APOBEC3B is a cytosine deaminase that controls a variety of biological processes, such as protein expression, innate immunity, and embryonic development, by participating in the process of cytidine deamination to uridine in DNA and RNA. In humans, APOBEC3B has long been known as a DNA editor for limiting viral replication and transcription. APOBEC3B is widely expressed at low levels in a variety of normal tissues and organs, but it is significantly upregulated in different types of tumor tissues and tumor lines. Thus, APOBEC3B has received increasing attention in various cancers, but the role of APOBEC3B in the occurrence and development of liver cancer due to infection with HBV remains unclear. This review provides a brief introduction to the pathogenesis of hepatocellular carcinoma induced by HBV, and it further explores the latest results of APOBEC3B research in the development of HBV and liver cancer, thereby providing new directions and strategies for the treatment and prevention of liver cancer.

A New Strategy for Efficient Screening and Identification of Monoclonal Antibodies against Oncogenic Avian Herpesvirus Utilizing CRISPR/Cas9-Based Gene-Editing Technology.

Marek’s disease virus (MDV) is an important oncogenic α-herpesvirus that induces Marek’s disease (MD), characterized by severe immunosuppression and rapid-onset T-cell lymphomas in its natural chicken hosts. Historically, MD is regarded as an ideal biomedical model for studying virally induced cancers. Monoclonal antibodies (mAbs) against viral or host antigenic epitopes are crucial for virology research, especially in the exploration of gene functions, clinical therapy, and the development of diagnostic reagents. Utilizing the CRISPR/Cas9-based gene-editing technology, we produced a pp38-deleted MDV-1 mutant—GX0101Δpp38—and used it for the rapid screening and identification of pp38-specific mAbs from a pool of MDV-specific antibodies from 34 hybridomas. The cross-staining of parental and mutated MDV plaques with hybridoma supernatants was first performed by immunofluorescence assay (IFA). Four monoclonal hybridomas—namely, 4F9, 31G7, 34F2, and 35G9—were demonstrated to secrete specific antibodies against MDV-1’s pp38 protein, which was further confirmed by IFA staining and confocal analysis. Further experiments using Western blotting, immunoprecipitation (IP), liquid chromatography–tandem mass spectrometry (LC–MS/MS), and immunohistochemistry (IHC) analysis demonstrated that the pp38-specific mAb 31G7 has high specificity and wide application potential for further research in MD biology. To the best of our knowledge, this is the first demonstration of the use of CRISPR/Cas9-based gene-editing technology for efficient screening and identification of mAbs against a specific viral protein, and provides a meaningful reference for the future production of antibodies against other viruses—especially for large DNA viruses such as herpesviruses.

Lung Organoids as Model to Study SARS-CoV-2 Infection.

Coronavirus disease-2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global pandemic and has severely affected socio-economic conditions and people’s life. The lung is the major target organ infected and (seriously) damaged by SARS-CoV-2, so a comprehensive understanding of the virus and the mechanism of infection are the first choices to overcome COVID-19. Recent studies have demonstrated the enormous value of human organoids as platforms for virological research, making them an ideal tool for researching host–pathogen interactions. In this study, the various existing lung organoids and their identification biomarkers and applications are summarized. At the same time, the seven coronaviruses currently capable of infecting humans are outlined. Finally, a detailed summary of existing studies on SARS-CoV-2 using lung organoids is provided and includes pathogenesis, drug development, and precision treatment. This review highlights the value of lung organoids in studying SARS-CoV-2 infection, bringing hope that research will alleviate COVID-19-associated lung infections.

STUDY OF HEPAITIS A AND HEPATITIS E VIRUS INFECTION IN ACUTE HEPATITIS PATIENTS

Background: Acute viral hepatitis is a major public health problem worldwide. Hepatitis-A virus (HAV) and Hepatitis E virus (HEV) are two major hepatotropic viruses responsible for enterically transmitted acute viral hepatitis. To know seroprevalence , clinico- Objectives: epidemiological factors and co-infection of hepatitis A & hepatitis E virus infection. The study was carried out in the Virology Research Methods: & Diagnostic Laboratory (VRDL) under Department of Microbiology IGGMC, Nagpur from January to October 2019. Cases included were children & young adults of both sexes having clinical hepatitis attending OPD and IPD. Blood sample was collected aseptically from each suspected patient. Serum was separated and tested for presence of IgM antibody against HAV and HEV by using Enzyme Linked Immunosorbant assay. Out of 184 samples, HAV seropositivity was seen in 13.04 %, HEV seropos Results: itivity in 35.32 %. Maximum cases were seen in age group of 0-10 years for HAV and 21-30 years for HEV. Male to female ratio was almost equal for both HAV and HEV. Urban population was most affected Source of water in maximum cases was tap water in urban and well water in rural area. Prevalence of HEV infection among Conclusion: the acute viral hepatitis cases is much more common than HAV. HEV being caused by feco-oral route, it is very important to have supply of safe drinking water. Awareness regarding sanitation, hand and food hygiene is the key factor to prevent hepatitis A & E infection.

Human Platelet Lysate Induces Antiviral Responses against Parechovirus A3.

Human platelet lysate (hPL) contains abundant growth factors for inducing human cell proliferation and may be a suitable alternative to fetal bovine serum (FBS) as a culture medium supplement. However, the application of hPL in virological research remains blank. Parechovirus type-A3 (PeV-A3) belongs to Picornaviridae, which causes meningoencephalitis in infants and young children. To understand the suitability of hPL-cultured cells for PeV-A3 infection, the infection of PeV-A3 in both FBS- and hPL-cultured glioblastoma (GBM) cells were compared. Results showed reduced PeV-A3 infection in hPL-cultured cells compared with FBS-maintained cells. Mechanistic analysis revealed hPL stimulating type I interferon (IFN) antiviral pathway, through which phospho-signal transducer and activator of transcription 1 (STAT1), STAT2, interferon regulatory factor 3 (IRF3) were activated and antiviral genes, such as IFN-α, IFN-β, and Myxovirus resistance protein 1 (MxA), were also detected. In addition, an enhanced PeV-A3 replication was detected in the hPL-cultured GBM cells treated with STAT-1 inhibitor (fludarabine) and STAT1 shRNA. These results in vitro suggested an unexpected effect of hPL-activated type I IFN pathway response to restrict virus replication and that hPL may be a potential antiviral bioreagent.

Expert symposium on severe acute hepatitis of unknown cause in children, a paradigm of exchanges and sharing between Chinese and Western medicine professionals.

Expert symposium on severe acute hepatitis of unknown cause in children, a paradigm of exchanges and sharing between Chinese and Western medicine professionals.

Designing a Science Based Strategy to Prepare for the Next Pandemic

Since late 2019, the COVID-19 pandemic has ravaged countries and peoples across the globe. As of August 27, 2021, 215 million people have been infected. Most countries have utilized different responses, which significantly influenced the impact of the disease on their respective populations. In this paper, we used an investigative approach to identify the proper measures needed to prepare for the next pandemic. Various National COVID-19 response strategies were identified and compared. In addition, scientific findings of the characteristics of the COVID-19 virus were utilized to identify appropriate responses. It was found that measures including swift lockdowns, border closings, and travel restrictions, accompanied by mask-wearing, mass testing, contact tracing, and healthcare system adjustments were key pieces of pandemic response. In addition, virology research, equipment stockpiling, manufacturing plans, healthcare worker benefits, and healthcare re-education are key to be prepared in advance. These findings are crucial in ensuring that the world is adequately prepared for the next pandemic.

Phylogenetic analysis of BKV genetic variations, based on the whole sequence of the genome and different genomic sections.

BK polyomavirus (BKV) primarily infects humans in their early life stages, and in later life stages, immunosuppressed patients may develop asymptomatic infections. The nucleotides 1744-1812 in the VP1 gene are traditionally used to determine this virus's genotype. The complete genome of the BKVsamples from patients referred to Masih Daneshvari Hospital's Virology Research Center was amplified by previously known primer sets. The phylogenetic diversity of the whole genome, different genomic sections, and the noncoding control region of BKVsamples were investigated. Using software Mega X and references, the samples' genotype was determined in separate genomic fragments and the whole genome. The samples were classified into two genotypes (I and IV) and five subtypes (Ia, Ib-1, Ib-2, IVc-1, and IVc-2), but none of the isolates belonged to genotypes II, III, V, or VI. The large T antigen-based phylogenetic tree provided 100% bootstrap values for these divisions, which were superior to those (96%-100%) used in the VP1 sequence. Among the genomic segments, large tumor antigen and VP1 had the most mutations. The noncoding control area contained mutations at the O41 position in the granulocyte/macrophage stimulus gene and the P31 position in the NF-1 gene. The validity of the phylogenetic analysis was supported by sequence analysis, which found single-nucleotide polymorphisms(SNPs) that could be useful for subclassifying isolates. More research with a large number of samples and in the wider geographical areas is needed to understand the genetic diversity of the BKV in Iran and also to determine these SNPs' clinical significance in terms of patient outcome and viral load dynamics.

Efficacy decametoxin in vitro for quick inactivation of respiratory coronavirus

Despite the fact that specific prophylaxis agents have already been widely introduced into medical practice in all countries of the world, and antiviral drugs are being developed and are undergoing the first stages of clinical trials, SARS-CoV-2 continues to spread in the human population. In this regard, an urgent medical problem today is the expansion of the arsenal of effective disinfectants and antiseptics, the action of which would be aimed at the rapid and complete inactivation of extracellular coronavirus, which is a very important element in controlling the spread of COVID-19.
 The aim of the study was to evaluate the ability of decamethoxin to have a virucidal effect against SARS-COV-2 and other human coronaviruses on the model of respiratory coronavirus IBV (infectious bronchitis virus) with an exposure time of 30, 60 and 120 seconds.
 Classical and modern virological research methods were used in the work: determination of the cytotoxic effect of decamethoxin in cell culture by the effect on their viability, cultivation, accumulation and determination of the infectious titer of IBV by cytopathic action in cell culture; assessment of the virucidal effect of decamethoxin by the suspension method to determine the residual infectious titer of the virus in cell culture by the method of limiting dilutions.
 The effectiveness of the antiseptic decamethoxin from the group of quaternary ammonium compounds was studied in relation to the prototype strain of the IBV (infection bronchitis virus) coronavirus family in vitro. It has been established that an isotonic solution of decamethoxin at a concentration of 100 μg/ml completely inactivates 3.0 lg(TCD50/0.1 ml) of the prototype respiratory coronavirus strain with a clinically significant contact time of 30–120 seconds at room temperature(18–24 оС). Decamethoxine has been shown to be an effective, fast-acting antiseptic capable of completely inactivating a prototype coronavirus strain. The revealed virucidal properties of decamethoxine in pharmacopoeially significant concentrations in relation to coronavirus allow to recommend it as an antiseptic in the development of methods for non-specific prevention of coronavirus infection in humans.