MECHANISM OF SARS-COV-2 INVASION INTO THE LIVER AND HEPATIC INJURY IN PATIENTS WITH COVID-19

A large number of studies have shown that patients with Coronavirus disease 2019 (COVID-19) have different degrees of liver injury. However, the mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) invasion into the liver are still not fully understood. This review mainly summarizes the recently published works on the abnormal liver biochemical indicators and the mechanism of viral invasion with liver injury in COVID-19 patients. Generally, SARS-CoV-2 infection of the liver was caused by blood circulation or retrograde infection of digestive tract, which led to the liver injury through direct cytopathic effect induced by virus or immunopathological effect caused by excessive inflammation. Besides these, hypoxia, endothelial injury and drug-induced jury were also the main reasons of liver injury in COVID-19 patients. In the liver function indicators, elevated alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transpeptidase, and lactate dehydrogenase levels with reduced albumin levels were observed in COVID-19 patients.

Identification and Semisynthesis of (–)-Anisomelic Acid as an Oral Antiviral Agent against SARS-CoV-2 in Mice

(–)-Anisomelic acid (AA), isolated from Anisomeles indica (L.) Kuntze (Labiatae) leaves, is a macrocyclic cembranolide with a trans-fused α-methylene-β-lactone motif. Cytopathic effect assays showed that the anti-SARS-CoV-2 effect of AA (IC50 = 4.3 μM) is comparable to that of remdesivir (IC50 = 2.1 μM), and more potent than that of molnupiravir (IC50 = 27.8 μM). Challenge studies in SARS-CoV-2-infected K18-hACE2 mice showed that oral administration of AA and remdesivir can both reduce the viral titers in the lung tissue at the same level. To facilitate drug discovery, we used a semisynthetic approach to shorten the project timelines. The enantioselective semisynthesis of AA from the naturally enriched and commercially available starting material (+)-costunolide was achieved in five steps with a 27% overall yield. The developed chemistry provides opportunities for developing AA-based novel ligands for selectively targeting proteins involved in viral infection.

A Study of the Impact of Graphene Oxide on Viral Infection Related to A549 and TC28a2 Human Cell Lines

Graphene has been one of the most tested materials since its discovery in 2004. It is known for its special properties, such as electrical conductivity, elasticity and flexibility, antimicrobial effect, and high biocompatibility with many mammal cells. In medicine, the antibacterial, antiviral, and antitumor properties of graphene have been tested as intensively as its drug carrying ability. In this study, the protective effect of graphene oxide against Rubella virus infection of human lung epithelial carcinoma cells and human chondrocyte cells was examined. Cells were incubated with graphene oxide alone and in combination with the Rubella virus. The cytopathic effect in two incubation time periods was measured using DAPI dye as a percentage value of the changed cells. It was shown that the graphene oxide alone has no cytopathic effect on any of tested cell lines, while the Rubella virus alone is highly cytopathic to the cells. However, in combination with the graphene oxide percentage of the changed cells, its cytotopathicity is significantly lower. Moreover, it can be concluded that graphene oxide has protective properties against the Rubella virus infection to cells, lowering its cytopathic changes to the human cells.

REACTIVATION OF HEPATITIS B IN PATIENTS RECOVERING FROM COVID-19

Пандемия новой короновирусной инфекции, вызванной вирусом SARS-CoV-2, явилась вызовом системе здравоохранения во всем мире. На данный момент мы обладаем большей информацией об этом заболевании, проявляющемся в основном симптомами респираторной инфекции, от легких проявлении ОРВИ до тяжелого поражения легких. Также коронавирусная инфекция проявляется симптомами поражения ЖКТ чаще всего в виде рвоты, диареи и боли в животе. Кроме того, за год наблюдении, начиная с самого начала пандемии в Китае, описаны нарушения функции печени различного генеза у пациентов с коронавирусной инфекцией. Среди возможных причин называются прямое цитопатическое действие вируса, способного связываться с AПФ2β рецепторам гепато-билиарной системы, иммуноопосредованное повреждение гепатоцитов, в том числе при «цитокиновом шторме» и гепатотоксичности препаратов, применяемых при коронавирусной инфекции. Кроме этих механизмов повреждения печени есть еще реактивация хронических персистирующих инфекции. В частности, речь идет о реактивации хронического гепатита В. Статей, описывающих такие случаи, значительно меньше, чем тех, которые практически детально описывают различные изменения ферментов печени у пациентов, наблюдавшихся с коронавирусной инфекцией. Кроме бремени инфекции в манифестный период, есть не менее тяжелые последствия у реконвалесцентов или у тех, кто перенес заболевание в легкой форме, о чем мы и должны помнить, чтобы принимать необходимые меры в ближайшем и отдаленном периоде выздоровления после COVID-19. В данной статье мы приводим собственные наблюдения реактивации хронического гепатита В у 4 пациентов, перенесших COVID-19 в манифестной форме. The pandemic of the new coronavirus infection caused by the SARS-CoV-2 virus is a challenge to the health system around the world. At the moment, we have more information about this disease, which is manifested mainly by symptoms of a respiratory infection, from mild manifestations of ARVI to severe lung damage. Also, coronavirus infection often manifests itself as symptoms of a gastrointestinal disease, in the form of vomiting, diarrhea and abdominal pain. In addition, over a year of observation, starting from the very beginning of the pandemic in China, liver dysfunctions of various origins have been described in patients with coronavirus infection. Possible reasons include the direct cytopathic effect of the virus capable of binding to ACE2β receptors of the hepato-biliary system, immune-mediated damage to hepatocytes, including during a "cytokine storm" and hepatotoxicity of drugs used in coronavirus infection. In addition to these mechanisms of liver damage, there is also a reactivation of chronic persistent infections. In particular, we are talking about the reactivation of chronic hepatitis B. In addition to the burden of infection in the manifest period, there are no less severe consequences for convalescents or those who have suffered a mild illness, which we must remember in order to take the necessary measures in the near and distant period of recovery after COVID -19. In this article, we present our own observations of the reactivation of chronic hepatitis in 4 patients who underwent manifest COVID-19.

Antiviral Activity of Ribavirin against Tilapia tilapinevirus in Fish Cells

The outbreak of the novel Tilapia tilapinevirus or Tilapia lake virus (TiLV) is having a severe economic impact on global tilapia aquaculture. Effective treatments and vaccines for TiLV are lacking. In this study, we demonstrated the antiviral activity of ribavirin against TiLV in E-11 cells. Our findings revealed that at concentrations above 100 μg/mL, ribavirin efficiently attenuates the cytopathic effect of the TiLV infection in fish cells. When administered in a dose-dependent manner, ribavirin significantly improved cell survival compared to the untreated control cells. Further investigation revealed that the cells exposed to ribavirin and TiLV had a lower viral load (p < 0.05) than the untreated cells. However, at concentrations above 1000 μg/mL, ribavirin led to cell toxicity. Taken together, our results demonstrate the efficacy of this antiviral drug against TiLV and could be a useful tool for future research on the pathogenesis and replication mechanism of TiLV as well as other piscine viruses.

Isolation and Molecular Characteristics of a Novel Recombinant Avian Orthoreovirus From Chickens in China

In recent years, the emergence of avian orthoreovirus (ARV) has caused significant losses to the poultry industry worldwide. In this study, a novel ARV isolate, designated as AHZJ19, was isolated and identified from domestic chicken with viral arthritis syndrome in China. AHZJ19 can cause typical syncytial cytopathic effect in the chicken hepatocellular carcinoma cell line, LMH. High-throughput sequencing using Illumina technology revealed that the genome size of AHZJ19 is about 23,230 bp, which codes 12 major proteins. Phylogenetic tree analysis found that AHZJ19 was possibly originated from a recombination among Hungarian strains, North American strains, and Chinese strains based on the sequences of the 12 proteins. Notably, the σC protein of AHZJ19 shared only about 50% homology with that of the vaccine strains S1133 and 1733, which also significantly differed from other reported Chinese ARV strains. The isolation and molecular characteristics of AHZJ19 provided novel insights into the molecular epidemiology of ARV and laid the foundation for developing efficient strategies for control of ARV in China.

Microfluidic characterisation reveals broad range of SARS-CoV-2 antibody affinity in human plasma

The clinical outcome of SARS-CoV-2 infections, which can range from asymptomatic to lethal, is crucially shaped by the concentration of antiviral antibodies and by their affinity to their targets. However, the affinity of polyclonal antibody responses in plasma is difficult to measure. Here we used microfluidic antibody affinity profiling (MAAP) to determine the aggregate affinities and concentrations of anti–SARS-CoV-2 antibodies in plasma samples of 42 seropositive individuals, 19 of which were healthy donors, 20 displayed mild symptoms, and 3 were critically ill. We found that dissociation constants, Kd, of anti–receptor-binding domain antibodies spanned 2.5 orders of magnitude from sub-nanomolar to 43 nM. Using MAAP we found that antibodies of seropositive individuals induced the dissociation of pre-formed spike-ACE2 receptor complexes, which indicates that MAAP can be adapted as a complementary receptor competition assay. By comparison with cytopathic effect–based neutralisation assays, we show that MAAP can reliably predict the cellular neutralisation ability of sera, which may be an important consideration when selecting the most effective samples for therapeutic plasmapheresis and tracking the success of vaccinations.

Etiological significance of Corynebacterium spp. in the development of diseases of the respiratory tract

Corynebacterium spp. It is associated with inflammatory diseases of the respiratory tract (tracheitis, pharyngitis, rhinosinusitis, bronchitis, pneumonia, etc.). C. pseudodiphtheriticum can be the causative agent of bacterial coinfection in patients with a new coronavirus infection (COVID-19). The aim is to determine the pathogenic properties and resistance to antimicrobial drugs of Corynebacterium spp. strains to establish their etiological significance in the development of inflammatory diseases of the respiratory tract. Strains of Corynebacterium spp. isolated from patients with inflammatory diseases of the respiratory tract (43 pcs.) and practically healthy individuals (29 pcs.). Isolates were identified by mass spectrometric method (MALDI-TOF MS), their cytopathic effect in CHO-K1 cell culture, hemolytic, urease activity, antimicrobial drug resistance were determined. Strains of Corynebacterium spp. isolated from patients in the amount of 105 CFU/ml or more, practically healthy - 104 CFU/ml or less. Isolates of Corynebacterium spp. patients had a more pronounced cytopathic effect (83.7±11.1%) and were more often resistant to antimicrobial drugs than those isolated from practically healthy. To establish the etiological significance of Corynebacterium spp. isolates. in the development of inflammatory diseases of the respiratory tract, it is advisable to determine their amount in biological material (105 CFU/ml or more), the cytopathic effect on CHO-K1 cell culture, as well as the presence of multiple resistance to antimicrobial drugs. Differences in the characteristics of Corynebacterium spp. isolates. from patients with respiratory tract pathology and practically healthy individuals are associated with the strain, not the species, of corynebacteria.

Studies on Growth Characteristics and Cross-Neutralization of Wild-Type and Delta SARS-CoV-2 From Hisar (India)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly evolved to generate several antigenic variants. These variants have raised concerns whether pre-existing immunity to vaccination or prior infection would be able to protect against the newly emerging SARS-CoV-2 variants or not. We isolated SARS-CoV-2 from the coronavirus disease 2019 (COVID-19)-confirmed patients in the beginning of the first (April/May 2020) and second (April/May 2021) waves of COVID-19 in India (Hisar, Haryana). Upon complete nucleotide sequencing, the viruses were found to be genetically related with wild-type (WT) and Delta variants of SARS-CoV-2, respectively. The Delta variant of SARS-CoV-2 produced a rapid cytopathic effect (24–36 h as compared to 48–72 h in WT) and had bigger plaque size but a shorter life cycle (~6 h as compared to the ~8 h in WT). Furthermore, the Delta variant achieved peak viral titers within 24 h as compared to the 48 h in WT. These evidence suggested that the Delta variant replicates significantly faster than the WT SARS-CoV-2. The virus neutralization experiments indicated that antibodies elicited by vaccination are more efficacious in neutralizing the WT virus but significantly less potent against the Delta variant. Our findings have implications in devising suitable vaccination, diagnostic and therapeutic strategies, besides providing insights into understanding virus replication and transmission.

Persistence of SARS-CoV-2 infection on personal protective equipment (PPE)

Background SARS-CoV-2 stability and infection persistence has been studied on different surfaces, but scarce data exist related to personal protective equipment (PPE), moreover using realist viral loads for infection. Due to the importance for adequate PPE management to avoid risk of virus infection, RNA stability was evaluated on PPE. Methods Persistence of SARS-CoV-2 infection and detection of genomic RNA in PPE (gowns and face masks) were determined by in-vitro assays and RT-qPCR, respectively. Samples were infected with a clinical sample positive for SARS-CoV-2 (Clin-Inf), and with a heat-inactivated SARS-CoV-2 strain sample (Str-Inf) as a control. Results PPE samples infected with Clin-Inf were positive for the 3 viral genes on gowns up to 5 days post-infection, whereas these overall genes were detected up to 30 days in the case of face masks. However, gowns and FFP2 masks samples contaminated with Clin-Inf showed a cytopathic effect over VERO cells up to 5–7 days post-infection. Conclusions SARS-CoV-2 RNA was detected on different PPE materials for 5 to 30 days, but PPE contaminated with the virus was infectious up to 5–7 days. These findings demonstrate the need to improve PPE management and to formulate strategies to introduce viricidal compounds in PPE fabrics.