Background: The disease caused by hepatitis C virus (HCV) is asymptomatic, silent, and progressive liver disease. In HCV-infected patients the increase in serum HA is associated with the development of hepatic fibrosis and disease progression. Methods: HCV-RNA detection was performed in all serological samples of blood donors that tested positive using HCV Ultra ELISA. Determination of hyaluronan (HA) was performed in positive HCV samples using ELISA-like fluorometric method. The HA content was compared to HCV viral load, genotype of the virus, liver fibrosis as well as ALT and GGT liver biomarkers. Results: Persistently normal ALT (<40 U/L) and GGT (<50 U/L) serum levels were detected in 75% and 69% of the HCV-Infected blood donors, respectively. Based on ROC analysis, the HA value < 34.2 ng/mL is an optimal cut-off point to exclude HCV viremia (specificity = 91%, NPV = 99%). Applying HA value ≥34.2 ng/mL significant liver fibrosis (≥F2) can be estimated in 46% of the HCV-infected blood donors. HA serum level (≥34.2 ng/mL) associated with a high ALT level (>40 U/mL) can correctly identify HCV infection and probable liver fibrosis (sensitivity = 96% and specificity = 90%) in asymptomatic blood donors. Conclusions: A high level of HA (≥34.2 ng/mL) in association with ALT (≥40 U/L) in serum can provide a good clinical opportunity to detect HCV-infected asymptomatic persons that potentially require a liver biopsy confirmation and antiviral treatment to prevent the development of advanced liver fibrosis or cirrhosis.
CONTEXT AND OBJECTIVE: Heparanase-1 degrades heparan sulfate and has been correlated with tumor progression. Although the isoform heparanase-2 has no catalytic activity, it seems to be important for modulating heparanase-1 activity. Cathepsin B is a proteinase involved in tumor metastasis. The aim of this study was to analyze heparanase isoform expression and cathepsin B activity in plasma samples from patients with gastrointestinal carcinomas, compared with healthy individuals (control group). DESIGN AND SETTING: This was an analytical cross-sectional study. Peripheral blood samples were collected at a Brazilian public hospital, from 21 patients with histopathological diagnoses of gastrointestinal carcinomas and from 43 healthy individuals. The analyses were performed in two Brazilian medical schools. METHODS: Heparanase isoforms were identified and quantified in plasma samples by means of Western blot. The enzymatic activities of heparanase-1 and cathepsin B were also measured. RESULTS:The results demonstrated that the expression of both heparanase isoforms was significantly greater in plasma samples from gastrointestinal carcinoma patients, compared with the control group. Logistic regression analysis showed that increased heparanase-1 and heparanase-2 expression was exclusively dependent on the tumor. There was a significant increase in heparanase-1 and cathepsin B activity in the patients' plasma. CONCLUSION: Overexpression of heparanase-1 and heparanase-2, along with increased heparanase-1 and cathepsin B activity in plasma, is associated with the diagnosis of gastrointestinal carcinoma. These findings provide support for using non-invasive assays (plasma samples) as an auxiliary method for diagnosing gastrointestinal tumors. RESUMO CONTEXTO E OBJETIVO:A heparanase-1 degrada heparam sulfato e está relacionada à progressão de tumor. Apesar de a isoforma heparanase-2 não possuir atividade catalítica, parece ser importante para modular a atividade da heparanase-1. A catepsina B é uma proteinase envolvida na metástase de tumores. O objetivo deste estudo foi analisar a expressão das isoformas de heparanase e atividade da catepsina B em amostras de plasma de pacientes com carcinomas gastrointestinais, comparando-se com indivíduos saudáveis (grupo controle). TIPO DE ESTUDO E LOCAL: Este é um estudo transversal analítico. Foram coletadas amostras de sangue periférico, em hospital público brasileiro, de 21 pacientes com diagnóstico histopatológico de carcinoma gastrointestinal e 43 indivíduos saudáveis. As análises foram realizadas em duas faculdades de medicina brasileiras. MÉTODOS: As isoformas da heparanase foram identificadas e quantificadas em amostras de plasma por Western blot. As atividades enzimáticas de heparanase-1 e catepsina B foram também mensuradas. RESULTADOS: Os resultados demonstraram que as expressões das isoformas de heparanase foram significativamente maiores nas amostras de plasma de pacientes com carcinoma gastrointestinal em comparação com o grupo controle. A análise feita por regres...
Background: Mutations critical for ABO group phenotypes which encode the catalytic domain of ABO glycosyltransferases have been studied in healthy blood donors. Weakening of A and B transferases have been reported in patients with leukemia, particularly those in which the myeloid lineage is involved. The purpose of this study was to use sequence specific PCR (PCR-SSP) to perform ABO genotyping of leukemic patients. Material and Methods: A total of 108 unrelated leukemic patients were initially classified serologically using tube test technique and gel test for ABO typing and subgroup detection. Genomic DNA was prepared from peripheral blood cells. All samples were then screened by 32 different PCR-SSP, each specific for a single nucleotide variation to screen 16 known polymorfic sites of exons 6 and 7 (modified from Seltsam, Transfusion 2003,43(4):428–39. Results: The results of PCR screening were conclusive and consistent with the ABO phenotypes in 71 patients (65.07%). Common ABO alleles were found in only 15 (13.8%) of patients studied. Unusual ABO alleles were found in 26 of 28 blood group O patients (ABO*O05, ABO*O21, ABO*O30, ABO*O33, ABO*O35, ABO*O36, ABO*O43 and ABO*O45), in 20 of 43 blood group A patients (ABO*A103, ABO*A104, ABO*A106, ABO*A202, ABO*A203), in 8 of 10 blood group B patients (ABO*B104, ABO*B105) and in 2 of 4 AB blood group patients (ABO*Bx01, ABO*B103). In 37 patients (34.2%), PCR screening revealed allele combinations that were incompatible with known ABO allele combinations or subgroups predicted by serologic analysis. Conclusion: This is the first report of using PCR-SSP for ABO genotyping of leukemic patients. The pathophysiologic role of unusual ABO allele combinations in patients with leukemia warrants further investigation.
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