Alteration in serum miR126 expression in healthy adults observing Navratri fast.

Background: Fasting is practiced by various religions in the world. The previous studies show the effect of fasting on biochemical markers in healthy subjects; however, no study is available on its effect on gene expression or epigenetic markers. In the present study, miR126, a microRNA, was measured in serum samples of healthy adult subjects, and their correlation with biochemical profile was carried out during the short-term fasting of the Navratri festival. Methods: A total of 30 subjects who underwent fasting for 07 days during the Navratri festival were recruited for the study. The fasting blood samples were obtained at three different time points; day 1 of fasting, day 7 of fasting, and day 7 after completion of fasting period. The miR126 expression, fasting plasma glucose, and lipid profile were measured in all the three samples. Results: The miR126 levels showed a decreasing trend with a significant difference across the three time points (p-value = 0.006). Fasting plasma glucose increased continuously across three time points without showing any statistical significance. Serum total cholesterol (p = 0.001) and triglycerides (p = 0.001) levels were decreased initially and then increased after resuming normal diet. There was a medium-level negative correlation (-0.332) between baseline fasting glucose level and miR126 level (p = 0.068). Conclusion: The study revealed that serum levels of total cholesterol and triglyceride were more dynamic than the miR126 levels. A significant decrease in the miR126 expression across three time points is a promising outcome of this pilot study and indicates its role in short-term fasting. However, the fasting plasma glucose showed heterogeneous values without significant correlation with miR126 levels.

A pilot study on DNA hypermethylation status in promoter region of P16 gene in patients with sporadic breast cancer.

Background: Epigenetic modification of cancer-related genes plays a role over and above their genetic alterations and contributes to the tumor initiation and progression of breast cancer. Promoter methylation of tumor suppressor genes is one such epigenetic modification, which can be potential biomarker. In this study, promoter methylation status of p16 gene was studied in blood samples of patients with breast carcinoma. Methods: Seventy-five patients, freshly diagnosed with carcinoma of breast and 20 age and sex matched healthy control subjects were recruited for the study. DNA extracted from EDTA blood sample was bisulfite converted and subjected to methylation-specific PCR to amplify the p16 promoter region. Results: Out of 75 patients, 25 (33%) patients showed hypermethylation in promoter region of p16 gene, which was statistically significant in comparison with the control group (p < 0.05). In subgroup analysis, lymph node involvement, cancer grade, and histopathological finding did not show any difference with methylation status of p16 promoter. Conclusion: Significant hypermethylation of p16 promoter region in the blood of histopathologically proven cases of breast cancer was observed suggesting promoter hypermethylation of p16 may be a possible mechanism accounting for sporadic carcinoma of breast.

Association of promoter methylation status of NRF2 and PNPLA3 genes in alcoholic liver disease.

BACKGROUND: Alcoholic liver disease (ALD) is the leading cause of chronic liver disease. In the liver, metabolism of alcohol occurs through multiple mechanisms and it results in the generation of various toxic products. Multiple genetic causes have been identified that are associated with the development and progression of ALD. The present study assessed the promoter site methylation status of nuclear factor erythroid 2-related factor 2 (NRF2) and patatin-like phospholipase domain-containing protein-3 (PNPLA3) genes in different subgroups of ALD. METHODS: The patients recruited were cases of alcohol dependence syndrome with hepatic dysfunction, compensated cirrhosis, decompensated cirrhosis, and acute-on-chronic liver failure due to alcohol as an etiology along with healthy control subjects. Routine biochemical investigations were performed along with methylation-specific polymerase chain reaction (MS-PCR) to qualitatively assess the promoter methylation status of NRF2 and PNPLA3 in all these cases. RESULTS: There was significant difference in methylation status of NRF2 gene in ALD when compared to healthy controls but there was no such difference in PNPLA3. All biochemical and clinical parameters studied were significantly different in subgroups of ALD except the serum aspartate aminotransferase (AST) level. Subgroups of ALD did not show any significant association with NRF2 or PNPLA3 methylation status. Gamma-glutamyl transferase (GGT) and creatinine levels in serum were significantly associated with the methylation status of NRF2 gene while no such association was seen with PNPLA3 gene. Model for end-stage liver disease (MELD) score varied differentially with NRF2 methylation and PNPLA3 methylation but there was no statistical significance. CONCLUSIONS: The present study showed that methylation status of NRF2 and PNPLA3 genes could not differentiate between subgroups of alcoholic liver diseases. However, the unmethylation of NRF2 promoter is associated with higher serum levels of GGT.

Immunohistochemical analysis and correlation of cyclooxygenase-2 expression status with clinicopathological parameters in head and neck squamous cell carcinomas: An Indian perspective.

CONTEXT: Head and neck squamous cell carcinoma (HNSCC) poses a major health problem and despite the advancements in its diagnosis and management the overall survival has not improved significantly. A search for newer diagnostic and prognostic markers along with fresh molecular targets is required for its prevention and cure. AIMS: The study aims to study the expression of cyclooxygenase-2 (COX-2) in HNSCCs and investigate its correlation with the clinicopathological profile of these cases. This study was performed to determine the significance of COX-2 expression in the Indian context. SETTINGS AND DESIGN: This study incorporated 90 cases of HNSCCs; both prospectively and retrospectively in a tertiary care center. MATERIALS AND METHODS: Expression of COX-2 on immunohistochemistry (IHC) was evaluated in correlation with the histological grade, maximum tumor size, tumor depth, nodal status and lymphovascular/perineural invasion (lvi/pni). The study received a waiver from the institutional ethics committee. STATISTICAL ANALYSIS USED: Statistical analysis of the data was done using SPSS software. RESULTS: COX-2 expression was found in 97.8% of the cases. A statistically significant correlation of COX-2 immunopositivity was found with the histological grade, clinical staging (tumor size and nodal status), maximum tumor depth and lvi/pni in our study (P < 0.05). CONCLUSIONS: COX-2 is expressed by most of the cases in this study. Its expression is related to tumor growth, differentiation and aggressiveness and therefore can be used as a good independent prognostic marker in HNSCCs. There is also possible scope of using it for targeted therapy in HNSCCs.

Prevalence of unsuspected glucose intolerance in coronary artery disease (CAD) patients: Importance of HbA1c.

BACKGROUND: The mortality and morbidity rates are two to fourfold higher among Coronary Artery Disease (CAD) patients with type 2 diabetes mellitus (DM). American Diabetes Association (ADA) and World Health Organization (WHO) define different criteria for the diagnosis of glucose intolerance. This study compares the available diagnostic criteria for DM in Indian men and their importance in CAD patients. METHODS: This cross-sectional study was done on 794 male volunteers; 483 individuals from general population and 311 patients undergoing angiography for evaluation of CAD. Individuals with previous clinical history of diabetes mellitus were excluded. RESULTS: More than 90% of diabetics by ADA criteria could be diagnosed by Fasting plasma glucose (FPG) and HbA1c criteria while FPG and pg2h plasma glucose (WHO criteria) could detect only 74%. Impaired Fasting Glucose (IFG) or Impaired Glucose Tolerance (IGT) was present in 36.7% of individuals diagnosed to be diabetic based on HbA1c; more in CAD +ve group (53.8%) than in general population (23.6%). ROC analysis suggests >121 mg/dl of FPG or >6.2% of HbA1c as optimum cut-off for the diagnosis of DM. FPG and HbA1c criteria have higher Relative Risk for presence of coronary artery occlusion and HOMA-IR. CONCLUSION: Inclusion of HbA1c in the criteria for diagnosis of DM (ADA criteria) can detect large number of cases with persistent hyperglycemia in the non-diagnostic range of DM (IFG or IGT) among general population and CAD patients. This has special relevance to epidemiological studies as the diagnosis of DM can be made on single fasting blood sample.

Visceral leishmaniasis mimicking as second line anti retroviral therapy failure.

Visceral leishmaniasis (VL) has increased as a complicating infection in subjects with human immunodeficiency virus (HIV) in developing countries. Both infections tend to lower the cell-mediated immunity resulting in poor drug response. In HIV-positive subjects the clinical course as well as organ involvement of VL simulates tuberculosis, another very common tropical infection. We present a case of VL/HIV co-infection where the individual failed to respond to first and second line antiretroviral therapy with persistently low CD4 counts. This patient was also subjected empirically to antitubercular therapy with no clinical improvement; he was finally diagnosed as a case of VL in HIV upon revelation of amastigotes in bone marrow despite the initial negative serology on two occasions. He showed dramatic improvement in CD4 counts and clinical status on Amphotericin B therapy. In endemic areas and in HIV positive subjects a systemic and careful parasitology follow-up is necessary to ensure that no clinical form of leishmaniasis is overlooked.