The role of Herpesviridae infection in the progression of comorbid somatic pathology

Background. The objective was to study the peculiarities of cellular and humoral immunity in patients with chronic obstructive pulmonary disease (COPD) group B and C, GOLD 2–3, who have signs of metabolic syndrome (MS) and Herpesviridae infection. Materials and methods. Forty-two patients with B and C groups of COPD, GOLD 2–3, associated with MS were examined. Eighteen individuals had COPD combined with MS, and Herpesviridae infection. The average age of patients was 51.3 ± 4.2 years. The comparison group included 24 people with COPD and MS without signs of Herpesviridae infection. All patients underwent determination of herpes simplex virus (HSV) type 1 and cytome­galovirus (CMV) antigens in the blood and saliva, titer of specific immunoglobulin (Ig) G and IgM antibodies to HSV type 1 and CMV, a comprehensive immunological examination with a study of cellular and humoral immunity indexes, cytokine status. Results. The study of the activity of herpes viruses in the blood of patients of the main group did not reveal active replication of HSV type 1 and CMV; in the saliva of 15 individuals (83.3 %), an active replication of HSV type 1 was detected, and in 12 people (66.7%) — of CMV. All patients in the main group had a severe course of Herpesviridae infection with an exacerbation rate of more than 6 times a year. The immunological status of patients of the main group showed immunodeficiency mainly of T-cell immune system and NK-cells, a significant increase in the relative number of T- and B-lymphocytes with early and late markers of activation on the background of autoimmune manifestations and inflammatory changes in peripheral blood. Conclusions. The presence of chronic persistent infection of HSV type 1 and CMV causes a severe course of COPD associated with MS, induces the development of infectious exacerbations of COPD and more significant manifestations of systemic inflammation in atherosclerosis as a morphological substrate of MS.

Characterization of Episomal Replication of Bovine Papillomavirus Type 1 DNA in Long-Term Virion-Infected Saccharomyces Cerevisiae Culture

We have previously reported that bovine papillomavirus type 1 (BPV-1) DNA can replicate its genome and produce infectious virus-like particles in short term virion-infected S. cerevisiae (budding yeast) cultures (Zhao and Frazer 2002, Journal of Virology, 76:3359–64 and 76:12265–73). Here, we report the episomal replications of BPV-1 DNA in long term virion-infected S. cerevisiae culture up to 108 days. Episomal replications of the BPV-1 DNA could be divided into three patterns at three stages, early active replication (day 3–16), middle weak replication (day 23–34/45) and late stable replication (day 45–82). Two-dimensional gel electrophoresis analysis and Southern blot hybridization have revealed further that multiple replication intermediates of BPV-1 DNA including linear form, stranded DNA, monomers and higher oligomers were detected in the virion-infected yeast cells over the time course. Higher oligomers shown as covalently closed circular DNAs (cccDNAs) are the most important replication intermediates that serve as the main nuclear transcription template for producing all viral RNAs in the viral life cycle. In this study, the cccDNAs were generated at the early active replication stage with the highest frequencies and then at late stable replication, but they appeared to be suppressed at the middle weak replication. Our data provided a novel insight that BPV-1 genomic DNA could replicate episomally for the long period and produce the key replication intermediates cccDNAs in S. cerevisiae system.

Application of Ex-Vivo/3D Organoid Models in COVID-19 Research

COVID-19 treatment methods based on 3D organoids and ex-vivo platforms are analyzed in this chapter. Initially, the platforms available for cell culture and its working characteristics are explained. Subsequently discusses the organoids with their definition and included their uses in various applications. Further, the chapter extends to describe the uses of different organoids with their use in different stages. Most of these methods utilized the 3D ex-vivo cell culture method to develop organoids and test them over infected tissues. Based on the study in this chapter, it is found that the demonstration of active replication of the human organoids culture system of lungs is found to be more helpful for COVID-19 treatment.

The Effect of a Unique Region of Parvovirus B19 Capsid Protein VP1 on Endothelial Cells

Parvovirus B19 (B19V) is a widespread human pathogen possessing a high tropism for erythroid precursor cells. However, the persistence or active replication of B19V in endothelial cells (EC) has been detected in diverse human pathologies. The VP1 unique region (VP1u) of the viral capsid has been reported to act as a major determinant of viral tropism for erythroid precursor cells. Nevertheless, the interaction of VP1u with EC has not been studied. We demonstrate that recombinant VP1u is efficiently internalized by rats’ pulmonary trunk blood vessel-derived EC in vitro compared to the human umbilical vein EC line. The exposure to VP1u was not acutely cytotoxic to either human- or rat-derived ECs, but led to the upregulation of cellular stress signaling-related pathways. Our data suggest that high levels of circulating B19V during acute infection can cause endothelial damage, even without active replication or direct internalization into the cells.

SMARCAD1 Mediated Active Replication Fork Stability Maintains Genome Integrity

ABSTRACTStalled fork protection pathway mediated by BRCA1/2 proteins is critical for replication fork stability that has implications in tumorigenesis. However, it is unclear if additional mechanisms are required to maintain replication fork stability. We describe a novel mechanism by which the chromatin remodeler SMARCAD1 stabilizes active replication forks that is essential for resistance towards replication poisons. We find that loss of SMARCAD1 results in toxic enrichment of 53BP1 at replication forks which mediates untimely dissociation of PCNA via the PCNA-unloader, ATAD5. Faster dissociation of PCNA causes frequent fork stalling, inefficient fork restart and accumulation of single-stranded DNA resulting in genome instability. Although, loss of 53BP1 in SMARCAD1 mutants restore PCNA levels, fork restart efficiency, genome stability and tolerance to replication poisons; this requires BRCA1 mediated fork protection. Interestingly, fork protection challenged BRCA1-deficient naïve- or PARPi-resistant tumors require SMARCAD1 mediated active fork stabilization to maintain unperturbed fork progression and cellular proliferation.