Development of a Recombinant Protein-Based Immunoassay for Detection of Antibodies Against Karolinska Institute and Washington University Polyomaviruses.

To develop polyomavirus VP1 recombinant protein-based immunoassay, the expression of two polyomavirus (Karolinska Institute Polyomavirus; KIPyV, and Washington University Polyomavirus; WUPyV) VP1s in insect cells was investigated using an improved baculovirus system (BacMagic). The reliability of the purified VP1 to serve as antigens in serological tests was confirmed by the establishment of an enzyme-linked immunosorbent assay (ELISA). Two panels of serum samples were used, with Panel I comprising 60 sera (20 KIPyV-positive, 20 WUPyV-positive, and 20 negative) and Panel II consisting of 134 sera with unknown status. The seroprevalence of KIPyV and WUPyV in the study population was determined to be 62% and 50%, respectively. Antibody-negative sera exhibited low reactivities in both ELISAs, whereas antibody-positive sera displayed high reactivity with median optical density values of 1.37 and 1.47 in the KIPyV and WUPyV ELISAs, respectively. The differences in seroreactivities between antibody positive and negative for each virus were statistically significant (p < 0.0001; with 95% confidence interval). The study suggests that seroconversion for KIPyV and WUPyV occurs in childhood, with KIPyV seropositivity reaching 70% and WUPyV seropositivity reaching 60% after the age of 5 years. Adult seroprevalence for polyomaviruses was high, with more than 64% and 51% of the adult population being seropositive for KIPyV and WUPyV, respectively. The constant prevalence of KIPyV and WUPyV antibody in the age groups suggested that this antibody persists for life. The fact that antibody titers were generally stable over time revealed a persistent infection of polyomaviruses in the human population. The insect cell-derived recombinant VP1-based ELISA has been demonstrated to be valuable as a serological assay, offering a valid, reliable, fast, nonlaborious, and economical procedure.

The frequency of IgG anti-varicella and anti-rubella antibodies in female students of Shiraz University of Medical Sciences, Iran.

Background: As infection with rubella and varicella-zoster viruses (VZV) can lead to congenital syndrome and its dangerous complications, assessing immunity to these congenital infections can represent the biological risk assessment related to their exposure in high-risk groups. Therefore, we aimed to evaluate the frequency of IgG anti-varicella/rubella antibodies (Abs) in female students at Shiraz University of Medical Sciences (SUMS), Iran. Patients and Methods: In this study, a total of 434 female students were included. Sera were isolated from blood samples and stored at -20°C for later analysis. A questionnaire form was documented and contained demographic data as well as the history of vaccinations. Enrolled students were divided into recipients of either one or two doses of the measles/rubella (MR) vaccine. Serum samples were analyzed for rubella and VZV IgG Abs using commercial IgG immunoassays. Results: The students were 21.6±4.25 years old on average. Out of the 434 enrolled students, 292 (67.3%) and 287 (66.1%) students were positive for anti-varicella and anti-rubella IgG-Abs, respectively. The frequency of anti-rubella IgG Ab was significantly higher in those who received one dose of MR vaccine (P<0.001). In addition, 205 (47.2%) and 59 (13.6%) students were double-positive (anti-varicella/rubella IgG Abs) and double-negative, respectively. Conclusion: Our results indicated that an additional dose of rubella vaccine may be required for those who received two doses of the vaccine. In addition, we recommend the inclusion of the VZV vaccine in Iran's routine vaccination program. Further studies are recommended to verify these results.

Salivary Vascular Endothelial Growth Factor and Epidermal Growth Factor Levels in Patients with Recurrent Aphthous Stomatitis: a Meta-Analysis.

Recurrent aphthous stomatitis (RAS) has been identified as a common oral lesion with an unknown pathogenesis. Various studies have been conducted to show the important role of two factors named epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) in RAS, but certain results have not been achieved. The present meta-analysis was conducted to evaluate the salivary levels of EGF and VEGF in patients with RAS. For this purpose, the related articles in the Web of Science, PubMed, Embase, ProQuest and Scopus databases until January 2022 were searched and their abstracts were studied. Google scholar and scientific information database were also searched for articles in Persian. The searches were completed by the medical subject heading terms considering "recurrent aphthous stomatitis" and "saliva" in combination with "EGF" or "VEGF" keywords. All case control studies that evaluated the salivary levels of EGF and VEGF in patients with RAS were included in this study. To evaluate statistical heterogeneity between the studies, Cochrane Q and I2 tests were adopted. The extracted data then were used in the analysis process based on comprehensive meta-analysis software. Originally, 619 articles were considered, of which 7 articles were selected. According to this meta-analysis, salivary EGF and VEGF levels were significantly lower in the active and remission period of RAS than in healthy individuals (pValue< 0.05). In addition, salivary levels of these factors were significantly lower in the active stage of RAS than in the healing phase. This review study suggests that decreasing of salivary EGF and VEGF levels have significant role in the development of RAS.

Investigation of the Seroprevalence of Antimeasles Immunoglobulin G Antibody in Students at Shiraz University of Medical Sciences.

Measles is an acute, highly contagious disease with a high mortality rate in children. Although vaccination has reduced measles incidence, outbreaks still occur. Therefore, in this study, we aimed to investigate the frequency of antimeasles immunoglobulin G (IgG) antibody (Ab) among students at Shiraz University of Medical Sciences (SUMS). Four hundred fifty SUMS students were enrolled in this cross-sectional study. Information on demographics and measles vaccination history was collected using a questionnaire. Participants were divided into two groups, including A and B, according to routine doses of measles vaccine and the national measles/rubella immunization program. The antimeasles IgG Abs were tested using a commercial Enzyme-Linked Immunosorbent Assay Kit. Participants ranged in age from 18 to 48 years, with a mean age of 22.2 (±4.3). Fifty percent of the subjects were male. Our results showed that 63.6% of the cases were positive for antimeasles IgG Abs. The seroprevalence of IgG Abs between groups A and B did not differ significantly (p = 0.612). There was also no significant correlation between the seroprevalence of antimeasles IgG Abs and the age (p = 0.43) or sex (p = 0.24) of the subjects. The results showed that the frequency of antimeasles IgG Abs is lower than required to prevent the measles virus from circulating. Therefore, a booster vaccination may be necessary.

A comprehensive narrative review of the cutaneous manifestations associated with COVID-19.

The systemic and respiratory clinical manifestations of coronavirus disease 2019 (COVID-19) include fever, coughing, sneezing, sore throat, rhinitis, dyspnea, chest pain, malaise, fatigue, anorexia and headache. Moreover, cutaneous manifestations have been reported in 0.2% to 20.4% of cases. Early diagnosis of COVID-19 leads to a better prognosis; knowledge of its cutaneous manifestations is one way that may help fulfil this goal. In this review, PubMed and Medline were searched with the terms "dermatology", "skin" and "cutaneous", each in combination with "SARS-CoV-2" or "COVID-19". All articles, including original articles, case reports, case series and review articles published from the emergence of the disease to the time of submission, were included. In this comprehensive narrative review, we tried to provide an analysis of the cutaneous manifestations associated with COVID-19, including maculopapular rash, urticaria, Chilblain-like, vesicular lesions, livedo reticularis and petechiae in asymptomatic/symptomatic COVID-19 patients that might be the first complication of infection after respiratory symptoms. Immune dysregulation, cytokine storms, side effects of antiviral drugs, environmental conditions and high-dose intravenous immunoglobulin (IVIG) therapy might be involved in the pathogenesis of the cutaneous manifestations in COVID-19 patients. Therefore, knowledge of cutaneous COVID-19 manifestations might be vital in achieving a quick diagnosis in some COVID-19 patients, which would help control the pandemic. Further research is very much warranted to clarify this issue.

Epstein-Barr Virus Nuclear Antigen 1 Increases the Expression of Viral Oncogenes and Cellular Genes in the HeLa Cell Line.

Epstein-Barr virus (EBV) represents one of the most important viral carcinogens. EBV nuclear antigen-1 (EBNA1) can induce the expression of different cellular and viral genes. In this study, we evaluated the EBNA1 effects on the expression patterns of human papillomavirus type 18 (HPV-18) E6 and E7 oncogenes and three cellular genes, including BIRC5, c-MYC, and STMN1, in a cervical adenocarcinoma cell line. HeLa cells were divided into three groups: one transfected with a plasmid containing the EBNA1 gene, one transfected with a control plasmid, and one without transfection. In all three groups, the expression levels of E6, E7, BIRC5, c-MYC, and STMN1 genes were checked using real-time PCR. Pathological staining was used to examine changes in cell morphology. Real-time PCR results showed that the expression level of HPV-18 E6 (P=0.02) and E7 (P=0.02) oncogenes significantly increased in HeLa cells transfected with the EBNA1 plasmid compared to cells transfected with control plasmid. Also, the presence of EBNA1 induced the expression of BIRC5 and c-MYC, which increased tenfold (P=0.03) and threefold (P=0.02), respectively. Regarding the STMN1 cellular gene, although the expression level in HeLa cells transfected with EBNA1 plasmid showed a twofold increase, this change was insignificant (P=0.11). Also, EBNA1 expression caused the creation of large HeLa cells with abundant cytoplasm and numerous nuclei. The EBV-EBNA1 could increase the expression levels of HPV-18 E6 and E7 viral oncogenes as well as c-MYC and BIRC5 cellular genes in the HeLa cell line. These findings indicate that the simultaneous infection of cervical cells with HPV-18 and EBV might accelerate the progression of cervical cancer.