Identification of a Single Nucleotide Polymorphism of Vitamin D Receptor (VDR) and Vitamin D Binding Protein (VDBP) Gene and Its Dysregulated Pathway Through VDR-VDBP Interaction Network Analysis in Vitamin D-Deficient Infertile Females.

INTRODUCTION: The prevalence of female infertility in Pakistan is currently estimated at 22%, and emerging research suggests that vitamin D (VD) deficiency (VDD) may play a significant role in influencing female fertility. The focus of this study was to investigate the single nucleotide polymorphism (SNP) patterns within the VD binding protein (VDBP). The study aimed to explore dysregulated pathways and gene enrichment through an interaction network analysis, specifically focusing on the interplay between the VD receptor (VDR) and VDBP in females experiencing unexplained infertility (UI) coupled with VDD. METHODS: A cross-sectional study was conducted on VD-deficient, fertile, and UI female subjects. VDBP and VDR were assessed by enzyme-linked immunoassay and genotyping performed. FunRich (version 3.1.3; http://funrich.org/index.html) was employed for analysis of the identified proteins: VDR and VDBP and with their mapped gene datasets, gene enrichment, and protein-protein interaction (PPI) network. RESULTS: The mean VD and VDR values of infertile females were significantly lower than those of fertile females. VDBP in infertile females (median (IQR)): 296.05 (232.58-420.23)) was lower than that of fertile females (469.9 (269.57-875.55), (p=0.01)). On sequence analysis, a mutation rs 4588 SNP (Thr 436 Lys) was found in exon 11 of the VDBP gene of UI females, but no mutation in exons 8 and 9 of the VDR gene, with some insignificant intronic variants, was observed. The proteins such as plasma membrane estrogen receptor signaling pathway (p < 0.001), VDR, SMAD3, NCOR1, CREBBP, NCOA1, STAT1, GRB2, PPP2CA, TP53, and NCOA2 were enriched after biological pathway grouping when VDR was made the focused gene and directly interacting with VDBP. CONCLUSION: The females with UI exhibited significantly low VD, VDBP, and VDR. The plasma membrane estrogen receptor signaling pathway was enriched in VDD infertile females.

Serum Levels of OX40 in Early and Late-Stage Oral Squamous Cell Carcinoma.

Background The tumor necrosis factor receptor superfamily, member 4 (OX40) and its ligand (OX40L) are members of the tumor necrosis factor superfamily and play roles as costimulatory immunomodulators to combat infectious diseases as well as cancers. Presently, many therapeutic agents focused on OX40 and OX40L are in trials for antitumor efficacy. In Pakistan, oral squamous cell carcinoma (OSCC) is the second most prevalent cancer with a mortality of 50% despite the availability of various therapeutic modalities. Data regarding serum levels of OX40 in patients with OSCC is lacking. Therefore, the study aimed to assess the OX40 levels in serum and their association with the clinicopathological features of the tumor. Methodology A cross-sectional study was conducted and serum samples of 78 biopsy-confirmed OSCC patients were collected prior to any treatment along with 10 healthy persons after informed consent. Serum levels of OX40 were measured via sandwich enzyme-linked immunosorbent assay (ELISA). Results The mean serum levels of OX40 were 1.65 ± 0.64 ng/ml and 2.39 ± 0.58 ng/ml in early and late-stage disease patients of OSCC, respectively (p =<0.005). However, based on gender and tumor site, male gender and buccal mucosa tumors in late-stage OSCC patients showed higher mean levels of OX40, 2.42± 0.58 ng/ml and 2.41 ± 0.58 ng/ml (p =<0.05), respectively. Patients with well-differentiated tumors demonstrated mean serum levels of 2.28 ng/ml, and in moderately differentiated tumors, the mean levels were 2.19 ng/ml (p =0.47). Conclusions A high OX40 level is associated with advanced-stage disease and a poor prognosis, possibly reflecting the immune-exhausted status against OSCC.

Knowledge, attitudes, and practices against the growing threat of COVID-19 among medical students of Pakistan.

BACKGROUND: On account of the COVID-19 pandemic, many changes have been implicated in university medical students. We are cognizant that pandemic can be controlled with dedicated contributions from all involved in the healthcare profession. Therefore, it is important to know the pandemic and application of knowledge by the medical students to formulate a further line of management in Pakistan. OBJECTIVE: We aimed toinvestigate the knowledge, attitudes, and practices (KAP) regarding COVID-19 and its impact on medical students of Pakistan. METHODS: A cross-sectional survey was conducted in June 2020 by a validated self-administered questionnaire. The survey instrument was tailored from a published questionnaire comprised of questions on demographics (6), knowledge (14), attitudes (4), and practices (6). RESULTS: Of the 1474 medical students in the study, 576(39.1%) were males, and 898(60.1%) were females. Two-thirds of the participants 1057(71.7%) had adequate knowledge, and almost all the students had positive attitudes (1363(92.5%), and good practices 1406(95.4%) to COVID-19. Two-thirds of the medical students 1023(69%) believed that the COVID-19 outbreak had affected their social, mental, and psychological well-being. One-quarter of the medical students 391(26%) become more religious, 597(40%) realized the importance of life, and 1140(77%) became careless because of the pandemic. The female medical students were 2.545 times (p < .001) and 4.414 times (p < .001) more likely to have positive attitudes and good practices toward COVID-19 as compared to males. CONCLUSION: Medical students, especially females and senior year scholars, were well-versed with desired levels of knowledge, attitudes, and preventive measures toward COVID-19. Most of them recognized COVID 19, is shaping their social, mental, and psychological well-being and encroaching on the healthcare system and economy. The information acquired by the KAP study may help to devise effective preventive strategies for future events.

Association of the JAZF1 Variant in Adults With a Parental History of Type 2 Diabetes Mellitus In Pakistan.

Background Type 2 diabetes mellitus (T2DM) is a chronic multifactorial condition and quickly growing disease in Pakistan. Many genes together with Zinc finger protein 1 (JAZF1) have already been described earlier in the literature but the role of JAZF1 in this subset of the population is yet to define. This study was aimed at identifying JAZF1 polymorphism and the risk of developing T2DM in persons with a parental history of T2DM in the Pakistani population. Methods DNA samples from 75 non-diabetic Pakistani participants with a family history of T2DM and 75 controls were evaluated by using a polymerase chain reaction (PCR) and the restriction fragment length polymorphism method. Results The alleles AA and AG and the GG genotype of JAZF1 (rs864745) varied considerably in frequency distribution between cases and control (p<0.05). The GG was independently and significantly associated with cases who had a family history of T2DM [odds ratio (OR) 2.6 (95% confidence interval (Cl) 1.3-5.1); p=0.005] while the AA allele was significantly associated with controls without a family history of T2DM [odds ratio (OR) 0.39 (95% confidence interval (Cl) 0.2-0.7); p=0.0059] and the allele AG has no significance and was equally distributed among control and cases with p-value=1.000. Conclusion Genotype GG of the JAZF1 variant was found significantly associated with the risk of developing type 2 diabetes mellitus in the Pakistani subset of the population.

Association of Zinc Transporter-8 Autoantibody (ZnT8A) with Type 1 Diabetes Mellitus.

BACKGROUND: Zinc transporter 8 autoantibody (ZnT8A), discovered through bioinformatics, is identified as another major biomarker for type 1 diabetes mellitus (T1DM), expanding the panel of diagnostic autoantibodies. The absence of standard autoantibodies in T1DM patients and the presence of ZnT8A in individuals before disease development has led the researchers to evaluate ZnT8A to gather information about the frequency and its association. Therefore, we aim to find out the concentration of ZnT8A and its association with T1DM. METHODS: A case-control study with 25 type 1 diabetes mellitus patients and 25 first-degree relatives of cases as controls was conducted at Ziauddin University in collaboration with the Baqai Institute of Diabetology and Endocrinology (BIDE), Karachi. Demographic data were collected from patients on a standard questionnaire. Blood samples were collected, after approval from Ziauddin Ethics Review Committee, from subjects and serum was separated to estimate ZnT8A by using sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean age at diagnosis of T1DM patients was 13.40±5.05 years, and the duration of diabetes was 7.74±5.85 years. The frequency of ZnT8A was found higher in cases (19 (76%)) compared to controls (6 (24%)). ZnT8A concentrations were significantly higher in cases (13.82 ng/ml) compared to the controls (8.78 ng/ml; p= 0.024). The cut-off value of 9 ng/ml was selected for measuring sensitivity, specificity, and accuracy, which were determined as 76%, 76%, and 76%, respectively. CONCLUSIONS: ZnT8A was found significantly associated with T1DM. Subjects with ZnT8A values ≥ 9 ng/ml are 10 times more at risk to develop T1DM (p = 0.000).