Aberrant expression of miRNAs predicts recurrence and survival in stage-II colorectal cancer patients from Egypt

Background: Patients with stage II CRC have a varying survival outcome. Therefore, it is critical to identify prognostic biomarkers that can define more aggressive forms of the disease. We assessed the expression levels of five miRNAs that have been previously addressed in relation to the development and progression of solid and hematological tumors. Methods: We measured the expression levels of miR-21, miR-137, miR-145, miR-320 and miR-498in stage II CRC patients from Egypt (124 tissues and 41 blood samples) by quantitative real time PCR (qPCR). The results were correlated with relevant clinicopathological factors, response to treatment and survival rates of the patients. Results: miR-137, miR-145 and miR-320 were significantly reduced in 39.5%, 48.4% and 52.4%; respectively whereas miR-21 and miR-498 were significantly overexpressed in 48.4% and 40.3% of the CRC tissues compared to the control group. In patients' blood, miR-137, miR-145 and miR-320 were significantly reduced in 46.3%, 46.3% and 51. 2%; respectively whereas mir-21 and miR-498 were significantly overexpressed in 46.3% and 43.9% of the cases, respectively. The concordance between tissue and blood was weak for miR-320 and miR-145 (kappa 40-65%), intermediate for miR-498 and miR-137 (kappa 65-75%) and strong for miR-21 (kappa 75-85%). In univariate analysis performance status, over-expression of miR-21 and miR-498 and reduced miR-137, miR-145, and miR-320 associated significantly with reduced DFS and OS. However, in multivariate analysis, miR-498 and miR-320 were independent prognostic factors for DFS whereas miR-21 was independent prognostic factors for OS. Conclusions: miRNAs play an important role in the development and progression of stage II CRC. A five markers panel (miR-21, miR-498, miR-137, miR-145 and miR-320) can predict recurrence and survival in stage II CRC patients from Egypt (AU)

Anti-leishmanial polyclonal antibodies to assess the performance characteristics of leishmanial antigen detection ELISA

The polyclonal anti bodies raised in rabbits against amastigote antigen extract were purified and fractionated, and IgG class antibodies and from the same antibodies, a peroxidase conjugate [labeled antibodies] reagent were prepared. The antibodies and the labeled antibodies were analyzed for efficacy of the homologous extracted antigens by capture ELISA. The titration curves of the anti-amastigote IgG antibody against extracted antigens showed that both free antibody and corresponding labeled antibody reacted with the original amastigote antigens. Further analysis involved the interaction between the antibody and two leishmanial stages; mammalian amastigote and infective promasitgote by immunoflourescene technique. The strong interaction was not only with surface antigenic components of the stages but also with their internal components. Capture-ELISA system was done to detect specific leishmanial antigens in urine and sera from visceral leishmaniasis patients [VL]. Most of the urine samples were positive [90% sensitivity] for leishmanial antigens without cross-reactivity [100% specificity] with any other tested samples from heterologous parasitic infections. But, only 61% sensitivity and 53% specificity were obtained when the capture ELISA was done to detect the specific leishmanail antigens in sera from VL