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.

A study to assess the relationship between donor uric acid levels and supernatant hemolysis in stored packed red blood cell units.

BACKGROUND: Most of the red blood cell (RBC) storage lesions can be attributed to oxidative stress encountered by the RBCs throughout the duration of their storage. Various donor variables at the time of donation may be responsible for the total antioxidant capacity of the supernatant and thus, the "storability" and the magnitude of development of these RBC storage lesions. It is known that uric acid (UA) is responsible for more than 60% of the TAC of the blood. This study aims to explore the relationship between donor UA levels and the difference in percentage hemolysis, an important RBC storage lesion, on day 1 and day 21, in stored packed RBCs (PRBCs) units. MATERIALS AND METHODS: The serum UA of 100 healthy voluntary male blood donors was estimated at the time of blood donation. The percentage hemolysis in the supernatant of the leukoreduced citrate phosphate dextrose/saline-adenine-glucose-mannitol RBC units (n = 100) prepared from these donors was calculated on day 1 and day 21. The difference in percentage hemolysis between donors with high normal serum UA levels (>7 mg/dL) was compared to that of the donors with low normal serum UA levels (<5 mg/dL) to observe the effect of donor UA levels on the difference in percentage hemolysis. RESULTS: The mean of the differences in percentage hemolysis in the supernatant in low UA group (<5 mg/dL) was higher than the mean of the differences in percentage hemolysis in the supernatant in high UA group (>7 mg/dL) and this was statistically significant (P < 0.001). The donor serum UA level and difference in percentage hemolysis on day 21 and day 1 were found to be negatively co-related. CONCLUSION: Higher levels of serum UA of blood donors seem to have a protective effect on the stored PRBC units as shown in this study. Hence, the potential of UA as one of the constituents of RBC additive solutions might lead to the enhancement of the quality of stored PRBC units by decreasing the RBC storage lesions.

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.

Analysis of short-term variation and long-term drift during reagent kit lot change in an NABL accredited clinical biochemistry laboratory.

BACKGROUND: Kit lot change in clinical biochemistry labs leads to variations in patient results. This study planned to identify variations during 60 reagent lot changes in our laboratory during the period from June 2018 to May 2019. METHODS: A statistical analysis was performed to identify the difference between patient samples results variations and QC results. The long term drift was analyzed using a regression test. RESULTS: There was a significant difference between the patient and QC results in 16.7% of reagent lot changes. Moreover, the extent of variation in QC results was 3.3%. No long-term drift was seen in three analytes which were studied using regression analysis. CONCLUSIONS: Our results showed that, during reagent kit lot change, along with QC material, the patient samples should also be run in order to identify the variation. However, this practice is presently ignored by most of the laboratories. There was no accumulated effect in our laboratory due to reagent kit lot change.

Association of Glycated Haemoglobin and Serum Apolipoproteins with Diabetic Retinopathy: An Indian Overview.

INTRODUCTION: India is presently facing an epidemic of diabetes mellitus and the risks of chronic complications from the disease are associated with the duration of the disease as well as the degree of hyperglycaemia. Diabetic retinopathy is a known microvascular complication of diabetes mellitus and is the most common cause of blindness in the western countries.Apolipoproteins are the protein component of lipoproteins. Apart from acting as structural proteins, they also act as cofactors to various enzymes. AIM: To measure the levels of serum apolipoproteins and glycated haemoglobin in cases of diabetic retinopathy and to assess their association with the stages of diabetic retinopathy. MATERIAL AND METHODS: The 135 diabetic cases [with (110) and without (125) retinopathy] attending the Ophthalmology OPD of this tertiary care hospital were included in the present study. Following retinoscopy, the patients were classified as Non-Proliferative Diabetic Retinopathy (NPDR) (n=75) and Proliferative Diabetic Retinopathy (PDR) (n=35). The controls (n=100) were age and sex matched patients who did not have diabetes. The cases and controls were assessed for HbA1c, total cholesterol, triglycerides, HDL cholesterol, Apo A-I and Apo B-100. RESULTS: The HbA1c was found to be higher in diabetics without retinopathy (7.02%) as compared to controls (5.58%) (p<0.05) and the highest value was seen in the mild NPDR group (8.82%). The mean value of Apo A-I was found to be lowest in the diabetics without retinopathy at 88 mg/dl and the highest in severe NPDR at 167 mg/dL. The mean value of Apo B-100 was found to be highest in severe NPDR at 114 mg/dL. The mean value of HDL cholesterol was lowest in moderate NPDR at 36.6 mg/dl. Total cholesterol was highest in severe NPDR at 280.88mg/dl while triglyceride was highest in severe NPDR at 286.4mg/dl. CONCLUSION: In our study, the level of HbA1c was found to range from 5.58% in non-diabetic to 8.82% in mild NPDR. There was a clear association between Apo B-100 and total cholesterol, triglycerides with the highest value of each parameter seen in the severe NPDR group. There was a discordance noted in the levels of HDL and Apo A-I in various groups. Apo B-100 values may be of value in prognosis of diabetic retinopathy as higher values may result in progression of the disease. Further studies involving Lp(a) and homocysteine may be required in cases of diabetic retinopathy.