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 cutoff 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).

Zinc, an important micronutrient for the storage, structural stabilization, secretion and action of insulin, is present in highest concentration in pancreas. The transport of zinc occurs through the zinc transporter-8 (ZnT8) to the insulin secretory vesicles. Zinc Transporter-8 Autoantibodies (ZnT8A) has been found to be associated with Type 2 Diabetes Mellitus. Recently it is recognized as a new autoantigen in Type 1 Diabetes Mellitus (T1DM) and its autoantibodies have been found in 50-60% of individuals with T1DM. Moreover, ZnT8A exhibit humoral auto reactivity which is not displayed by any of the other islet autoantigen like glutamine decarboxylase (GAD), insulin or tyrosine phosphate-related molecules (IA-2). Immunity against ZnT8 is dependent on clinical characteristics, which may provide evidence for early recognition highlighting the importance of this transporter in the pathogenesis of T1DM. Information regarding this article was retrieved through PubMed, Google Scholar and other search engines available in the University by using the keywords zinc, ZnT, ZnT8, SLC30A8 (Solute carrier 30 member 8) and Type 1 Diabetes Mellitus. Information was gathered through original researches, reviews and epidemiological studies published up to August 2019.The aim of this review is to summarize the emerging role of ZnT8A in diagnosis and understanding the genetic basis of Type 1 Diabetes Mellitus.

Background: Coronavirus disease 2019 (COVID-19) pandemic emerged in Karachi and rapidly spread throughout Pakistan Since February 26, 2020.  Objectives: Vaccination is currently one of the most effective COVID-19 eradication approach. The purpose of this study was to gather data on the adverse effects of the COVID-19 vaccine. Methodology: It was an observational study that was carried out between the 11th and 23rd of April 2021, and the participants were Karachi residents. We looked at the proportion of self-reported local and systemic adverse effects within seven days of immunization in people who filled out Google forms and received one or two doses of the vaccine. Results: The vaccination ratio for male was slight higher than females. Participants aged between 51 to 60 years and 41 to 50 years had higher number of vaccinations. Sinopharm is by far the most widely used vaccine. After the first dose of vaccination, the majority of participants complained of fever, chills, muscle pain, and arm pain, whereas after the second dose, the majority of participants had no symptoms, with a few participants complaining of fever, chills, diarrhea, and muscle pain. Conclusion: The first and second doses' post-vaccination adverse effects were mild and predictable, and there were no hospitalizations; this data can help lessen vaccine hesitancy.

Type 1 Diabetes Mellitus (T1DM), is a genetic disease, the prevalence of which is increasing due to delays in the diagnosis because of the absence of standard antibodies. Therefore, there is a need for a molecular variant that can help in early diagnosis in these patients. The polymorphism in the SLC30A8 gene can identify the people at risk of T1DM. However, there is a controversy in the association of the SLC30A8 gene in the pathogenesis of T1DM and a dearth of data in our part of the world. Therefore, we aimed to determine the most frequent autoantigen SLC30A8 genotype and its association with T1DM in the Pakistani population. This case-control study was carried out at Ziauddin Medical University (ZMU) jointly with Baqai Institute of Diabetology and Endocrinology (Baqai Medical University) from June to October 2019. A total of 50 subjects were enrolled (25 cases and 25 controls) in the study. Cases included 25 diagnosed patients of T1DM meeting American Diabetes Association (ADA) new criteria and controls were their first-degree relatives. Blood was drawn, DNA was extracted and amplified through PCR. The RFLP of PCR product was done using a restriction enzyme, Alu I for genotyping. The most frequently observed genotype was CC among cases as well as controls. The CC genotype of rs13266634 was not found significantly associated with T1DM (p=0.08) but the OR was 2.7; CI=0.86 -9.00. Similarly other genotypes were also not found statistically significant. However, HbA1C and Fasting Blood Sugar (FBS) were found statistically significant (p=0.001, p=0.000) in T1DM patients compared to controls. The autoantigenic variants of SLC30A8 of rs13266634 were not found statistically significant with T1DM. The role of this variant as a susceptibility gene in T1DM development should be further confirmed by carrying out studies with a larger sample size.

Background: To determine KRAS gene in circulating tumor DNA in comparison with histological grading through liquid biopsy in colorectal cancer patients. Methods: This dual-centered cross-sectional study included 73 diagnosed patients of colorectal cancer at different grading levels [Grade I, well differentiated (n = 7, 9.5%); Grade II, moderately differentiated (n = 14,18.9%); and Grade III, poorly differentiated (n = 52, 70%)]. Blood was collected, and plasma was separated. ctDNA was extracted, using magnetic bead-based technique (MagMAX Cell-Free DNA kit). KRAS gene was quantified through qPCR. STRING database was used to find KRAS interactomes. Results: Mean threshold cycle (CT value) of KRAS gene in Grade III samples showed significantly higher (P = 0.001) levels of ctDNA (2.7 ± 1.14) compared with Grade II and Grade I (3.1 ± 0.68, 2.3 ± 0.60), respectively. Grading characterization showed that rectal cancer (n = 22, 42.3%) with Grade III (68.8%) was more prevalent than colon and sigmoid cancer (n = 19, 36.5%, n = 11, 21%, respectively). STRING database showed 10 functional genes interacting with KRAS expressed as gene/proteins. Conclusion: Liquid biopsy can be used to detect ctDNA in plasma of CRC patients and enabled to detect the KRAS gene by qPCR. The technique being less invasive and cost-effective is convenient for multiple biopsies in different cancers.