Molecular Biomarkers & Diagnosis

Purpose: To determine the Hepatitis C virus (HCV) load in peripheral blood specimens of patients with renal abnormality reporting to the nephrology unit and to correlate the viral load with different biomarkers in serum.

Materials and Methods: Fifty peripheral blood specimens were obtained from patients reporting to the nephrology unit and these patients were categorized into three groups as Group I: Renal Transplant patients, Group II: Dialysis patients, Group III: Other patients. with elevated liver enzymes and renal pathology. Peripheral blood specimens collected from kidney transplant recipients (n = 11), dialysis (n=17) and others (n =22) were subjected to detection of antibodies to HCV, determination of viral load by Real Time PCR and biochemical profiling consisting of estimation of bilirubin, total protein, alanine aminotransferase, and alkaline phosphatase. Antibodies to HCV were detected by ELISCAN™ HCV and the viral load by using HCV RG RTPCR kit (QIAGEN, Hilden). Statistical analysis - T test and the logistic regression analysis assessing the correlation between viral load and serum bilirubin, Serum glutamate pyruvate transaminase (SGPT), Alkaline phosphatase, total protein were performed using SPSS software version 14.0

Results: Antibodies to HCV were detected by ELISA in 39 (78%) peripheral blood samples and genomic HCV was detected in 31 (62%) by RTPCR. In 8 (16%) patients, HCV antibodies only were detected and RTPCR did not reveal the presence of HCV in these specimens. Logistic regression analysis performed on biochemical parameters and viral load revealed correlation between alkaline phosphatase enzyme levels and viral load (Hosmer and Lemehow test P value< 0.05 statistically significant).

Conclusion: Determination of viral load is a reliable diagnostic test in detection of HCV infection. Elevated levels of alkaline phosphatase enzyme could be associated with increased viral load. To the best of our knowledge, this finding is the first being reported in Indian literature.

Metabolic profiles of hydrophilic and lipophilic cell extracts from three cancer cell lines, Miapaca-2, Panc-1 and AsPC-1, and a non-cancerous pancreatic ductal epithelial cell line, H6C7, were examined by proton nuclear magnetic resonance spectroscopy. Over twenty five hydrophilic metabolites were identified by principal component and statistical significance analyses as distinguishing the four cell types. Fifteen metabolites were identified with significantly altered concentrations in all cancer cells, e.g. absence of phosphatidylgrycerol and phosphatidylcholine, and increased phosphatidylethanolamine and cholesterols. Altered concentrations of metabolites involved in glycerophospholipid metabolism, lipopolysaccharide and fatty acid biosynthesis indicated differences in cellular membrane composition between non-cancerous and cancer cells. In addition to cancer specific metabolites, several metabolite changes were unique to each cancer cell line. Increased N-acetyl groups in AsPC-1, octanoic acids in Panc-1, and UDP species in Miapaca-2 indicated differences in cellular membrane composition between the cancer cell lines. Induced glutamine metabolism and protein synthesis in cancer cells were indicated by absence of glutamine other metabolites such as acetate, lactate, serine, branched amino acids, and succinate. Knowledge of the specifically altered metabolic pathways identified in these pancreatic cancer cell lines may be useful for identifying new therapeutic targets and studying the effects of potential new therapeutic drugs.