Sabina Mahmood

Immunohistochemical Evaluation of Oxidative Stress Markers in Chronic Hepatitis C

Oxidative stress (OS) plays a major role in chronic hepatitis C. Various OS markers have been found to be elevated in hepatitis C virus (HCV)-related liver disease. This study detected the presence of OS in serum and liver biopsy specimens of HCV patients. Reactive oxygen molecules (ROM) in sera of 54 HCV patients were compared with 23 controls. OS markers 8-hydroxydeoxyguanosine (8-OHdG), 4-hydroxy-2-nonenal, malondialdehyde, and thioredoxin were measured in liver biopsy specimens of 18 HCV patients with fibrosis staging F1 (six); F2 (two), F3 (four), and F4 (six). The interferon (IFN) response and hepatocellular carcinoma (HCC) occurrence in the presence of OS markers were also evaluated. The level of ROM in HCV patients was 318 +/- 56.7 Carr compared with 248 +/- 40.8 Carr in controls (p=0.032). Multivariate analysis found age (p=0.0236) to be the only independent variable associated with increase in ROM in sera. In liver biopsy specimens, OS markers were found mainly around the area of piecemeal necrosis or the periportal area. The presence of OS markers seemed to increase with fibrosis staging, although not significantly. The OS DNA damage marker 8-OHdG was detected in the nucleus of hepatocytes. Thirteen patients received IFN therapy. During the 4-year follow-up period, HCC developed in four nonresponders to IFN and in one untreated patient. OS markers were stained in both HCC cells and non-HCC cells in HCC patients. OS markers were found in serum and liver specimens of HCV-associated liver disease and in HCC tissue. Detection of OS markers may be important for monitoring disease progression in HCV patients. Antioxidant therapy in combination with antiviral therapy may minimize liver damage and aid in the prevention and subsequent development of HCC.

Influence of viral load and genotype in the progression of Hepatitis B-associated liver cirrhosis to hepatocellular carcinoma.

Abstract: Aim/Background: Hepatitis B virus (HBV) is an important factor in the development of hepatocellular carcinoma (HCC). We studied the influence of HBV viral load on HCC occurrence in HBV related liver cirrhosis (LC).

Effect of Vitamin E on Serum Aminotransferase and Thioredoxin Levels in Patients with Viral Hepatitis C

Objectives: Oxidative stress induces cellular responses such as cell death, gene activation and cell proliferation, in the liver. Vitamin E (Vit. E) has been found to protect the liver against oxidative stress in animal experiments. Thioredoxin (TRX) is a stress inducible, multifunctional protein, secreted during oxidative stress. This study evaluated effects of Vit. E on serum TRX and aminotransferase levels in hepatitis C virus (HCV) patients, partly non-responsive to initial interferon (IFN), with higher than average level of serum alanine aminotransferase (ALT) after receiving anti-inflammatory drug treatment. Methods: Seventeen HCV patients (male=3; female=14) of age 62±7.65 years receiving anti-inflammatory drug therapy, at least 6 months prior to Vit. E administration, were given d-α´-tocopherol 500 mg/day, orally, for a period of 3 months. ALT, aspartate aminotransferase (AST), TRX and Vit. E were measured at 0, 1, 2 and 3 months and 1 month after end of treatment. As controls, the same patients biochemical data, 3 months from the start of therapy were used. Patients were divided into three categories: total patients “T”, low ALT group “L” (ALT<70 IU/l) and high ALT group “H”(ALT>70 IU/l), respectively.Results: The ALT level was lowered, significantly in group H, in the 1st, 2nd, 3rd and 1-month post therapy, compared to the initial value. But group L showed little or no change in ALT. Post Vit. E therapy, in groups T and H, the TRX level was elevated but remained below initial levels, whereas in group L, TRX level remained significantly lower than the pretreatment value. Groups T and L, showed significant reduction (p<0.05) in serum TRX levels in the 2nd and 3rd month. Group H showed a tendency towards TRX reduction, but not significantly. Serum Vit. E levels increased significantly (p<0.0001) from the 1st to 3rd month in all three T, H and L groups. Conclusion: Oxidative stress induced liver damage is reduced by Vit. E in patients with viral hepatitis C, particularly those with initial ALT levels >70 IU/l. Vit. E treatment causes reduction of oxidative stress markers as TRX and ALT in sera. Therefore, Vit. E can act as a supportive therapy to combat liver damage caused by oxidative stress, in such patients with continuously high levels of ALT even after anti-viral and anti-inflammatory drug therapy.

Clinical significance of intrahepatic interleukin-8 in chronic hepatitis C patients

Background/AIM: Interleukin-8 (IL-8) is known as a chemotactic and angiogenetic cytokine and is a potential mediator of host response to injury or inflammation. In order to identify the role of IL-8 in the pathogenesis of chronic hepatitis C (CHC), we assessed semiquantitatively the messenger RNA (mRNA) expression of IL-8 and other cytokines in liver biopsy specimens of CHC patients. METHOD: Liver biopsy specimens were obtained under peritoneoscopy from 35 patients with CHC. The mRNA expression of IL-8 and other cytokines in the liver were determined by real-time PCR and the correlation between the mRNA expression and histological classification of liver were studied. Liver histology was classified by both staging of fibrosis (F0-F4) and grading of activity (A1, mild; A2, moderate and A3, severe). RESULTS: Patients were classified into F1, 8; F2, 9; F3, 9 and F4, 9 and A1, 6; A2, 14 and A3, 15, by staging of fibrosis and grading of activity, respectively. Expression of IL-8 mRNA increased with staging of fibrosis (F1, 0.402+/-0.65; F2, 0.413+/-0.246; F3, 1.388+/-2.166; F4, 1.991+/-1.879) and grading of activity (A1, 0.560+/-0.808; A2, 0.780+/-1.268; A3, 1.548+/-1.957). The mRNA expressions of IL-2, IL-1alpha, IL-1beta, IL-15 and TNF-alpha were found to be closely correlated with IL-8 mRNA (R=0.638; 0.522; 0.487; 0.465 and 0.495, respectively, in all P<0.05). CONCLUSION: In CHC, intra-hepatic expression of both IL-8 and IL-2 increased with fibrosis and inflammatory activity. Positive correlations were found between IL-8 and other cytokines and between cytokines themselves. These findings suggest that these interacting cytokines play an active role in the pathogenesis of CHC, and maybe involved in the upregulation or induction of one and other.

Long term follow-up of a group of chronic hepatitis C patients treated with anti-inflammatory drugs following initial interferon therapy

BACKGROUND: A relationship between hepatocellular carcinoma (HCC) recurrence and serum alanine aminotransferase (ALT) in a group of hepatectomized patients has been reported. Another study suggested the development of HCC is more rapid in a high ALT group of hepatitis C virus (HCV)-associated cirrhotic patients. To find a relationship between ALT and HCC occurrence, we observed changes in ALT over a period of 6 years, in a group of non-cirrhotic, chronic hepatitis C (CHC) patients treated with anti-inflammatory drugs post interferon (IFN) therapy. METHOD: Eighty three CHC patients, with fibrosis stage 1, 2, 3 (F1, F2, F3) who had a partial (PR) or non-response (NR) to initial IFN therapy, were treated with anti-inflammatory drugs for 6 years. Over a period of 6 years HCC developed in nine patients. Of them, one belonged to F2 and eight to F3. Within the first 2 years HCC developed among two patients in F3. Multivariate analysis revealed that in F3, the 6 year average ALT activity (odds ratio 5.59; P<0.05) was the only significant variable associated with HCC occurrence. All other variables remained insignificant. Among the six F3 patients in whom HCC developed, the likelihood of HCC occurrence was found to be significantly higher (odds ratio 1.89; P<0.001) in patients who showed elevated ALT activity (>80 IU) two or more times during the 6 year period, compared to those with ALT (>80 IU) for less than 2 years. CONCLUSION: These findings suggest that continuous elevation of ALT seems to be important for HCC diagnosis. Patients with ALT >==80 IU for 2 years or more are at a greater risk of HCC development. It is necessary to continue treatment with anti-inflammatory drugs, following initial IFN therapy to suppress ALT below 80 IU, to prevent HCC occurrence or delay the time of HCC occurrence in order to prolong life.

Assessment of Health Related Quality of Life in Chronic Liver Disease Patients Using the Japanese Versions of CLDQ and SF-36

Liver disease affects health-related quality of life. The CLDQ is a liver disease specific questionnaire. This study attempted to translate the original CLDQ into Japanese and compare it with SF-36 in chronic liver disease patients, mainly chronic hepatitis C. The Japanese versions of CLDQ and SF-36 were administered to 120 CHC; 45 CHB; 29 NAFLD; 21 HCC post treatment and 50 healthy controls, between February and March, 2008. CLDQ scores of CHC pa- tients and controls were unaffected by sex and age. SF-36 scores of female CHC patients were significantly lower (P = 0.0081) compared to male in the domain of physical function. CHC patients over 70 years had significantly lower SF-36 scores in multiple domains compared to CHC patients below 70. CLDQ scores of CHC patients were lower than controls in all 6 domains. CHB & NAFLD patients had significantly low scores in 3 domains, compared to controls. CHC patients scored significantly lower than CHB & NAFLD patients in 2 domains. Significant differences in SF-36 scores between controls, CHC, CHB and NAFLD patients were not observed. CLDQ scores of treatment naive CHC patients, having ALT levels (�40 or� 40) IU/l; liver cirrhosis patients (child Pugh A) and HCC patients post treatment, revealed that HCC and cirrhosis patients had similar CLDQ scores and significantly lower scores compared to CHC patients with ALT �40 IU/l in 5 out of 6 domains. CHC patients with ALT �40 IU/l had significantly lower CLDQ scores than CHC pa- tients with ALT �40 IU/l in 3 domains. Similar differences was not observed using the SF-36. CLDQ gave a better un- derstanding of HRQL in patients with different forms of chronic liver disease and also disease progression. Age and sex did not affect CLDQ scores. CLDQ appears to be a more convenient tool to study the HRQL in chronic liver disease pa- tients.

An Analysis of Risk Factors for Developing Hepatocellular Carcinoma in a Group of Hepatitis C Patients with Stage 3 Fibrosis following Interferon Therapy

The risk of Hepatocellular carcinoma (HCC) is high in HCV-infected patients who have biochemically and histologically active chronic hepatitis. To observe the long prognosis of Chronic Hepatitis C (CHC) patients with stage 3 fibrosis (F3), 55 CHC patients after initial Interferon (IFN) therapy were followed up for up to 12 years (average 9.8 ± 2.3 years). According to the annual average alanine aminotransferase (ALT) levels, patients were grouped into, low (ALT ≤g 30 IU/l); moderate (ALT >30 <80 IU/l) and high (ALT ≥ 80 IU/l) ALT groups. Eleven patients were re-treated with IFN. During the follow-up period of 12 years, HCC developed in 26 patients with an average annual incidence of 3.9%. Biochemical responders to initial IFN therapy (n = 8) and those re-treated with IFN (n = 10), except 1, did not develop HCC. Cox regression analysis to evaluate risk factors for HCC occurrence, found development of Liver Cirrhosis within 3 years of initial IFN therapy(P = 0.05) and the 3 year annual average ALT post initial IFN therapy (P = 0.033) to be significant. The 12 year annual average ALT was also found to be significantly related to HCC occurrence (P = 0.016), on univariate analysis. Patients belonging to the continuously low ALT group (ALT ≤ 30 IU/l for ≥3 years), did not develop HCC or receive IFN re-treatment. In CHC patients with F3, after initial IFN therapy, keeping ALT continuously low, below 30 IU/l for 3 years or more seems important. Continuing treatment with anti-inflammatory drugs along with subsequent IFN re-treatment may prevent or delay HCC even in elderly patients.

Clinical usefulness of the branched DNA assay version 2 in predicting the efficacy of Interferon treatment in a group of chronic HCV patients based on serotype.

Interferon (IFN) response depends upon pretreatment viral loads and viral genotype/serotype. This study investigated the difference in response to IFN in different viral load groups of 96 chronic hepatitis C patients and compared version 1 (bDNA1.0) and version 2 (bDNA2.0) of the branched DNA assay, according to serotype. On univariate analysis, viral load (P=0.0001, by bDNA 1.0; P=0.0020, by bDNA 2.0,), serotype (P=0.0053) and age (P=0.0073) were significant predictors of IFN response. On multivariate analysis, serotype (odds ratio, 5.44; 95% confidence interval, 1.94-15.24; P<0.01) was a stronger predictor of IFN response than age or viral load (by bDNA2.0). In very high (>6.7 log eq/ml), high (6.0 approximately 6.69 log eq/ml) and low (<6 log eq/ml) viral load groups (by bDNA2.0), complete response was 25, 55 and 92.6% in serotype 2 (sero-2), and 10, 20 and 71.4% in serotype 1 (sero-1), respectively. In sero-2, bDNA2.0 can detect high viral loads underestimated by bDNA1.0. In a low viral load group (by bDNA2.0), IFN response is dependent upon serotype; sero-2 responded better than sero-1. However, in high and very high viral load groups, sensitivities of bDNA1.0 and bDNA2.0 are not effective in clinically distinguishing CR from non-response, and aid in patient selection for IFN therapy.