Association of interleukin-10 with hepatitis B virus (HBV) mediated disease progression in Indian population.

Background & objectives: Interleukin (IL)-10, an anti-inflammatory Th2 cytokine, is one of the key coordinators of the inflammatory responses involved. The present study was designed to evaluate the impact of IL-10 (-819/-592) genotypes, haplotypes, mRNA and the protein levels with risk for hepatitis B virus (HBV) related hepatocellular carcinoma (HCC) development in India. Methods: A total of 390 subjects (145 controls, 62 inactive HBV-carriers, 64 chronic-active HBV patients, 60 HBV related cirrhotics and 59 HBV- HCC subjects) were enrolled in the study. Allele specific (AS)-PCR, ELISA and RT-PCR methods were used for assessing polymorphism, spontaneous blood levels and the mRNA expression, respectively of IL-10. Results: The study revealed that the CC/TA genotype acted as a risk factor for cirrhosis (ORa=2.02; P<0.05) and the subsequent HCC development (ORa=2.20; P<0.05), with controls as reference. However, no significant association was found between the two haplotypes (CC and TA) observed and HCC risk. Moreover, the IL-10 protein and mRNA levels in peripheral blood mono nuclear cells (PBMCs) showed a significant elevation as the disease progressed to cirrhosis. But, no variation was observed in the IL-10 levels in subjects with different IL-10 genotypes. Interpretation & conclusions: These preliminary results suggest a strong association of IL10 (-819/-592) with the HBV infection mediated disease progression, from inactive carrier state to malignancy, in Indian population.

Mechanism of intestinal folate transport during folate deficiency in rodent model.

Background & objectives: Folate deficiency is a public health problem and is the most notable for its association with neural tube defect in developing embryo, megaloblastic anaemia, cancers and cardiovascular diseases. The mechanisms of the intestinal folate uptake process have been earlier characterized. However, much less is known about regulation. In this study we evaluated the mechanistic insights of folate absorption in an in vivo model of folate deficiency. Methods: Male Wistar rats were fed folate-containing diet (2 mg/kg folic acid) or a folic acid-free diet over 3 months and folate transport was studied in intestinal brush border membrane vesicles (BBMV). Results: The characterization of the folate transport system in intestinal brush border membrane (BBM) suggested it to be a carrier mediated, acidic pH stimulated, and Na+ independent. Folate deficiency increased the folate transport by altering the Vmax without changing the Km of folate transport process. The increased transport efficiency of the BBM was associated with upregulation of folate transporters at both mRNA and protein level. Interpretation & conclusions: Folate deficiency resulted in significant upregulation of intestinal folate uptake, by increasing number of transporters without any change in specificity of transporters towards its substrate. The observed upregulation was associated with significant increase in reduced folate carrier (RFC) and proton coupled folate transporter (PCFT) expressions, suggesting the transcriptional and translational regulation of folate uptake during folate deficiency.

Indocyanine green clearance test (using spectrophotometry) and its correlation with Model for End Stage Liver Disease (MELD) score in Indian patients with cirrhosis of liver.

Background: Child Turcotte Pugh (CTP) score and Model for End Stage Liver Disease (MELD) are used commonly to assess the prognosis of liver disease but the disadvantage of these static tests is their inability to identify the functional reserve of the liver. Among all quantitative liver function tests indocyanine green (ICG) clearance test is most widely used and has been used to determine operative risk before hepatectomy and to assess prognosis of patients with cirrhosis. Aim: To correlate indocyanine green (ICG) clearance test with MELD score in patients with cirrhosis of liver. Methods: Forty patients with cirrhosis of liver were included and divided into two groups according to their CTP scores. Group A had 20 patients with CTP class A and group B had 20 patients with CTP class B. After ICG injection, ICG retention at 15 minutes (ICGR15) and ICG clearance rate were calculated. Results: In group A, the mean ICGR15 was 32.86% + 6.4% while in group B it was 51.08% + 12.8% (p <0.001). ICG clearance rates were 4.3% + 2.8% and 3.5% + 3.8% per minute in group A and B respectively. MELD score had a strong positive correlation with ICGR15 but a negative correlation with ICG clearance rate. On ROC curve analysis, AUC for MELD was 0.805 vs. 0.88 for ICGR15 in assessing prognosis of patients with cirrhosis. The sensitivity and specificity of MELD score was 60% and 80% respectively while that of ICGR15 was 85% and 90% respectively. Conclusion: ICGR15 has a higher sensitivity and specificity than MELD score in assessing the prognosis of patients with cirrhosis of liver.

Distribution of XRCC1 genotypes in north Indian population.

Background & objectives: XRCC1, a major DNA repair gene, acts as a scaffold of different activities involved in repair by interacting with components of base excision repair (BER) at the site of damage. Polymorphisms in this gene are associated with variations in the repair efficiency which might predispose an individual to cancer risk. To associate a gene polymorphism with disease risk, it is imperative to have the data for its genotype distribution in normal population. The present study was therefore carried out to find distribution of XRCC1 polymorphisms (codons 194, 280 and 399) in normal north Indian population. Methods: Healthy volunteers hailing from north India (150) were enrolled in the study. DNA was isolated from blood samples and genotyping of codons 194, 280 and 399 of XRCC1 gene was done by PCRrestriction fragment length polymorphism (RFLP), using specific primers. Results: The frequencies obtained for heterozygous genotype of codons 194 and 399 were 45 and 49 per cent respectively and were higher than wild and variant genotypes. For codon 280, the highest frequency (59%) was obtained for the wild genotype. Frequencies of the variant genotypes of codons 194 and 399 were higher in males and females respectively. The allele frequencies also followed the similar trends. Interpretation & conclusions: A significant distribution of variant and heterozygous XRCC1 genotypes was noticed that warrants further studies on the association between these genotypes and disease risk in our study population.