Association of GST null genotypes with anti-tuberculosis drug induced hepatotoxicity in Western Indian population.

BACKGROUND: The first line anti-tubercular (anti-TB) treatment normally involves isoniazid, rifampicin, pyrazinamide, and ethambutol. Clearance of these drugs depends on the activity of several enzymes such as N-acetyl transferase 2, cytochrome P450 oxidase and glutathione S-transferase (GST). Some of these enzymes are highly polymorphic leading to significant inter-individual variation in their activity thereby increasing the risk of drug induced hepatotoxicity (DIH). AIM: To investigate the possible association of anti-TB DIH with genetic polymorphism of GST genes in Western Indian population. MATERIAL AND METHODS: A prospective case-control study was undertaken on patients who received anti-TB treatment. Cases (n = 50) were distinguished from controls (n = 246) based on occurrence of DIH during anti-tubercular treatment. A multiplex polymerase chain reaction was employed to identify homozygous null mutation at GSTM1 and GSTT1 loci. Results. Homozygous null mutation in GSTM1 gene alone or in both GSTM1 and T1 genes was found to be significantly associated with anti-TB DIH at p < 0.02 and p < 0.007, respectively, in our study population. CONCLUSIONS: This is the first study to report GSTM1 null and combined GSTM1 and T1 null genotypes to be risk factors of anti-TB DIH in Western Indian population. Screening of patients for these genotypes prior to anti-TB regimen would provide better control of hepatotoxicity.

Impact of diabetes mellitus on outcome of HCC.

BACKGROUND: Diabetes mellitus (DM) is recently identified risk factor for development and progression of chronic liver disease as well as hepatocellular carcinoma (HCC). We planned a prospective analysis to identify impact of DM in Indian patients with HCC. METHODS: During last 10 years, 160 consecutive patients of HCC were evaluated. Demographic profile like age of presentation, clinical features, etiology of HCC, tumor size at presentation, management and ultimate outcome was compared diabetic with non-diabetic HCC patients. RESULTS: During last 10 years, 160 consecutive patients of HCC were evaluated (Mean age = 59.6 +/- 12.9 years, sex ratio (M: F) = 5.4: 1). Etiology for HCC were hepatitis B in 45 (28.2%), hepatitis C in 18 (11.3%), alcohol in 27 (16.8%), alcohol with hepatitis B in 12 (7.5%), alcohol with hepatitis C in 1 (0.6%), non-alcoholic steatohepatitis in 4 (2.5%) and cryptogenic in 53 (33.2%) patients. Patients of HCC with DM (group-A, n =46, age = 62.6 +/- 9.5 years, sex (M: F) = 6.6:1) were compared with patient of HCC without DM (group-B, n =114, age = 66.7 +/- 13.7 years, sex (M: F) = 5.4:1). Duration of diabetes in group-A was 7.6 +/- 3.2 years. Patients in group-A had more advanced HCC (size of lesion > 5 cm and >3 lesions of 3 cm or more diameter, portal vein thrombosis or intra-hepatic bile duct involvement) than group-B [34 (73.9%) vs 72 (54.3%)]. Mortality with in one year was significantly more in group-A compared to group-B [36 (78.2%) vs 56 (49.1%)]. CONCLUSION: DM is associated with more advanced lesion and poor outcome in patient with HCC.

Prevalence of non-alcoholic fatty liver disease: population based study.

BACKGROUND AND AIM: Non-alcoholic fatty liver disease (NAFLD) is a common cause of chronic liver disease and liver transplantation in western countries. Increasing incidence of NAFLD has been well documented from Asian countries like Japan and China. Diabetes mellitus (DM), obesity, hyperinsulinemia are predisposing factors for NAFLD. There is increase in incidence of DM, obesity and insulin resistance in India in last two decades. Hence it is logical to expect increase in incidence of NAFLD in India. There is limited data on the prevalence of NAFLD from India. Majority of data comes from hospital based studies including small number of patients. Therefore this study was planned to estimate the prevalence of NAFLD in general population. MATERIAL AND METHODS: Residents of two Railway colonies were evaluated on history, clinical examination, anthropometric measurements, biochemical tests and abdominal ultrasound. RESULTS: 1,168 participants were evaluated. Persons with any amount of alcohol consumption, HBs Ag positive, Anti HCV positive, persons with other known liver diseases and taking medications causing liver disease were excluded. Prevalence of NAFLD on ultrasound was 16.6%. Out of 730 subjects above the age of 20 years (341 male 384 female 389) mean age 39.08 +/- 12.3 years, 4% had diabetes, 57% had central obesity. Prevalence of NAFLD based on the ultrasound above 20 years of age was 18.9%. NAFLD was more prevalent in male than female (24.6% vs 13.6%, p < 0.001). Risk factors associated with NAFLD were age more than 40 years, male gender, central obesity, high BMR > 25, elevated fasting blood sugar, raised AST and ALT. CONCLUSION: Prevalence of NAFLD in Indian population is comparable to the west.

Angiogenesis in chronic liver disease.

BACKGROUND: Chronic liver disease is characterized by inflammation and fibrosis. As a consequence angiogenesis leading to new vasculature may have prognostic value in disease progression. Interfering with angiogenesis may be a potential target to avoid progression of liver disease. Hence we planned to evaluate the CD34 and vascular endothelial growth factor (VEGF), the markers for angiogenesis in chronic liver disease. METHOD: Liver biopsies from 79 patients of chronic liver disease and 21 cases of HCC (M: F = 4:1, age range 22 to 80) were stained for routine HE, CD 34 and VEGF immunostaining (Dako Corp & Santa Cruz respectively). Etiologies of chronic liver disease were alcoholic liver disease, HBV, HCV infection, NAFLD, autoimmune liver disease, and cryptogenic liver disease. Thirty biopsies from normal liver obtained at autopsy were taken as controls. Expressions of CD 34 and VEGF were compared with the stage of fibrosis. RESULTS: Out of 79 patients, angiogenesis was seen in 45.5% cases of chronic liver disease. None of the case with normal liver histology was CD 34 or VEGF positive. No significant correlation of angiogenesis was found between any etiologies of chronic liver disease. CD 34 was positive in 18/21 (85.7%) cases of hepatocellular carcinoma. CD 34 and VEGF positivity was 20.9% and 46.5% in stage 1 and 2 fibrosis while it was 75% and 80% in stage 3 and 4 fibrosis respectively. VEGF appeared more common as compared to CD 34 in early fibrosis. CONCLUSION: Angiogenesis was present in 45.5% cases of chronic liver disease. It was proportional to the increase in stage of fibrosis. Expression of VEGF was commonly found in early stages of fibrosis. Hence, therapeutic strategies of inhibiting VEGF expression may be of importance in preventing the progression of chronic liver disease in its early stage.

An experience with covered transjugular intrahepatic portosystemic shunt for refractory ascites from western India.

BACKGROUND: In refractory ascites/hydrothorax (RA), uncovered transjugular intrahepatic portosystemic shunt (TIPS) is shown to be superior to large-volume therapeutic paracentesis (LVP) for long-term control of ascites, but at a cost of increased risk of hepatic encephalopathy (HE). Use of covered TIPS has shown to improve shunt patency rate over uncovered TIPS. This retrospective analysis was performed on patients with RA to assess efficacy of TIPS, both covered and uncovered. METHODS: Over 10-year period, patients with RA, patients either required LVP at least 2 times in a month, or were intolerant to LVP, or were unwilling to undergo further LVP, were treated with TIPS (Group-A = 12 patients with uncovered TIPS {Wallstent = 10, Memotherm = 1, SMART = 1}, age = 56.1 +/- 4.5 years, male: female = 5:1; Group-B = 11 patients with e-PTFE-covered TIPS {Viatorr = 11}, age = 55.8 +/- 5.2 years, male: female = 8:3). They were followed-up with clinical and ultrasonography/ Doppler examination every monthly for 3 months and every 3 monthly thereafter (mean = 9.6 +/- 4.2 months). Clinical success (disappearance of ascites at 1-month), technical success (post-TIPS reduction of portosystemic pressure gradient {PPG} < 12 mmHg), appearance of encephalopathy, TIPS-dysfunction (> 50% reduction in flow-velocity, > 50% shunt stenosis or increase in PPG > 12 mmHg in presence of symptoms) and mortality were noted. Data were analyzed using chi-square test and t test. RESULTS: Baseline clinical and biochemical characteristics were similar in both groups. TIPS placement was possible in 11/12 group-A and 11/ 11 group-B patients. Fall in PPG after TIPS was similar in both groups. One patient in group-A was lost followup after the procedure. On comparison of group-A and group-B, clinical success (63.3% and 81.8%), technical success (90.9% and 100%), occurrence of HE (60% and 54.4%) and mortality at 1-year (70% and 63.3%) were not significantly different. TIPS-dysfunction requiring re-intervention was significantly more common in group-A (50%) than group-B (0%). CONCLUSIONS: Covered TIPS was superior to uncovered TIPS, because of less TIPS-dysfunction without increasing chances of HE; but failed to offer any survival advantage.

Combination of Peginterferon alpha-2b (12 kDa) and Lamivudine in difficult-to-treat chronic hepatitis B- an Indian experience.

INTRODUCTION: Treatment strategies for chronic hepatitis B (CHB) using either interferon or Lamivudine can achieve sustained response in 30-40 % in HBeAg positive CHB. Retreatment of treatment failure in CHB and treatment of HBeAg negative patients pose major therapeutic challenge. Because of success story of Peginterferon in hepatitis C and some preliminary data in CHB, we undertook this open-labeled prospective study to study the response of Peginterferon alpha-2b (PegIFN) and Lamivudine combination in patients of CHB. MATERIALS AND METHODS: Following 4 groups of patients of CHB with persistently elevated transaminases were treated with Lamivudine 100 mg PO daily and PegIFN 1.5 microg/kg SC once a week- 1) HBeAg negative treatment naive patients- for 12 months 2) HBeAg negative patients who were nonresponsive to at least two treatment regimens for 12 months, 3) HBeAg positive treatment naive patients- for 6 months 4) HBeAg positive patients who were treatment failure at least with two regimens- for 6 months. Patients were tested for LFT, HBeAg, antiHBe, quantitative HBVDNA (only in HBeAg negative CHB) and liver biopsy when possible at inclusion. During treatment period, LFT was tested at monthly interval and HBeAg, antiHBe and HBVDNA (in HBeAg negative CHB) at 3 monthly intervals. End of treatment response (EOR) was assessed at end of treatment period and sustained response (SR) at 6 months post treatment. Treatment response was defined in HBeAg positive patients with normalization of enzymes and disappearances of HBeAg & in antiHBe positive patients with normalization of enzymes and loss of detectable HBVDNA. RESULTS: Total 25 patients with CHB were included in this study with mean age of 37.9 +/- 4.6 years (range 20-56) and male: female= 11.5:1. 1) In HBeAg negative treatment naive patients, 4/6 patients achieved EOR (66.6%), whereas in 3/6 patients (50%) had SR. 2) In HBeAg negative treatment failure group, 4/5 patients had EOR (80%) and 3/5 patients achieved SR (60%), of which 1 patient lost HBsAg. 3) In HBeAg positive treatment naive patients, 4/5 patients achieved EOR (80%) and 3/5 patients obtained SR (60%). One patient in this group lost HBsAg. 4) In HBeAg positive treatment failure group, 5/9 patients achieved EOR (55.5%), whereas 3/9 patients (33.3%) obtained SR. CONCLUSION: Overall, in difficult-to-treat patients (considering groups 1, 2 & 4- total 20 patients) with combination of PegIFN and Lamivudine, EOR was seen in 13 patients (65%), SR was obtained in 9 patients (45%) and loss of HBsAg was seen in 1 patient (5%).

Extrahepatic manifestations of viral hepatitis.

UNLABELLED: Viral hepatitis has been shown to be associated with various extrahepatic manifestations. These can be seen in both acute and chronic liver disease, may precede or follow overt liver disease. AIMS AND OBJECTS: To study the prevalence of extrahepatic manifestations of viral hepatitis and follow the course of the disease in response to antiviral therapy whenever indicated. METHODS: Prospectively 448 patients of viral hepatitis were evaluated for extrahepatic manifestations and patients of glomerulonephritis (GN), polyarteritis nodosa (PAN) and cryoglobulinemia were tested for viral markers. All patients were investigated for liver and kidney function tests, hematological workup and viral markers such as HBsAg, HBeAg, Anti HBeAg, HCV RNA, IgM anti HAV and IgM anti HEV. Serum electrophoresis and kidney biopsies were done whenever indicated. In 10 cases of hepatitis B glomerulonephritis immunohistochemistry was done on kidney biopsies for demonstration of hepatitis B surface and core antigen. RESULTS: Of total 448 cases 181 (40.4%) had hepatitis B infection, 142 (31.6%) had hepatitis C infection, 86 (19.1%) hepatitis E and 39 (8.7%) had hepatitis A infection. Extrahepatic manifestations were seen in 29 (6.4%) cases and these were cases of GN, PAN, cryoglobulinemia, thrombocytopenia, agranulocytosis, aplastic anemia and pancreatitis. Patients with hepatitis A with extrahepatic manifestations showed complete recovery in both hepatitis and extrahepatic manifestations. Six patients with PAN were treated with interferon of which 4 showed excellent response. Three patients of hepatitis B and hepatitis C related GN were given interferon and 4/6 responded well to treatment. CONCLUSION: Prevalence of extrahepatic manifestations with viral hepatitis was found to be 6.4%. These manifestations recover completely with recovery from viral hepatitis.