A school-based intervention of screening a movie to increase hepatitis B vaccination levels among students in Uttar Pradesh, India: impact on knowledge, awareness, attitudes and vaccination levels.

BACKGROUND: India is home to one in 14 of all chronic hepatitis B virus (HBV) cases, meaning that it is important to develop HBV interventions that are applicable in the Indian context. Vaccination is the foremost tool for interrupting the HBV infection cycle. HBV vaccination was not included in India's government-sponsored expanded immunisation program until 2011, and many children born earlier remain unvaccinated. This study sought to observe the impact of the HOPE Initiative's school-based intervention to increase vaccination coverage by increasing HBV awareness among students in Lucknow, Uttar Pradesh. METHODS: At 430 schools in the administrative areas within and surrounding Lucknow, students viewed an educational documentary film on HBV and completed two questionnaires, one immediately before the screening and the other six weeks later. Both questionnaires asked the same 14 questions, which were organized into five domains: knowledge of the magnitude of the problem of HBV; knowledge of modes of HBV transmission; knowledge of consequences of HBV infection; awareness of HBV; and attitudes regarding HBV. The baseline questionnaire also asked students whether they had been vaccinated against HBV. At two-year follow-up, researchers measured vaccination levels at a subset of 30 intervention schools and six non-intervention schools to further assess the impact of the intervention. RESULTS: Baseline questionnaires were completed by 11,250 students, and post-intervention questionnaires, by 9698 students. Scores for knowledge about the magnitude of the HBV problem improved from 41% at baseline to 74% at follow-up, and scores for knowledge about modes of transmission, from 38% to 75% (p < 0.05 for both). The baseline HBV vaccination level among students receiving the intervention was 21%. Two years after the intervention, 45% of students (N = 4284) reported being vaccinated at intervention schools compared to 22% (N = 1264) at non-intervention schools. CONCLUSIONS: The observed increases in HBV awareness, knowledge and vaccination levels in this study indicate that school-based interventions can be used to achieve higher vaccination coverage among Indian children. The documentary film was found to be an affordable tool for reaching large audiences. More studies are needed to validate the impact of this intervention and to explore its applicability to other social causes.

Association of interleukin-10 polymorphisms with chronic hepatitis C virus infection in a case-control study and its effect on the response to combined pegylated interferon/ribavirin therapy.

We conducted a case-control study involving 150 genotype 3 chronic hepatitis C virus (HCV) patients and 150 healthy controls to investigate the association of polymorphisms in the interleukin-10 (IL-10) gene with chronic HCV infection and the association of these polymorphic variants with the combination of pegylated interferon (Peg-IFN) and ribavirin therapy response. Our data revealed that the GG genotype of IL-10 -1082A/G exhibited significant association with genotype 3 chronic HCV infection compared to controls. Treatment response data also showed a significant increase in risk for the GG genotype of IL-10 -1082A/G in response-relapse patients or non-responder patients compared to sustained virological response patients. Further, a significant increase in risk was also revealed for the CC genotype of IL-10 -592A/C in response-relapse patients or non-responder patients compared to sustained virological response patients, suggesting a role of the GG genotype of IL-10 -1082A/G and CC genotype of IL-10 -592A/C in the treatment outcome of combined Peg-IFN/ribavirin therapy.

Alterations of p53 gene in gallbladder cancer patients of North India.

BACKGROUND: Mutations in p53 gene are found in a majority of human malignancies and usually occur in the exons 5, 6, 7 and 8. Mutated p53 protein is more stable and gets accumulated in the cells that induce the host to develop anti-p53 antibodies in sera of cancer patients. AIM: This study is aimed to observe the frequency and nature of mutations in exons 5-8 of p53 gene and to evaluate its correlation with prevalence of serum p53 antibodies in Indian patients with gallbladder cancer (GBC). METHODS: Mutation studies were done in cancer tissues obtained from 62 patients with proven GBC (40 cytologically proven cases and 22 resected gallbladder cancer tissues) by polymerase chain reaction (PCR), restriction fragment length analysis (RFLP) and single strand conformation polymorphism (SSCP). Presence of serum p53 antibodies was determined using highly specific enzyme linked immunosorbent assay (ELISA) kit in 50 patients with GBC and 30 patients of cholelithiasis. Clinicopathologic characteristics of these patients were given attention. RESULTS: Antibodies to p53 protein was present in the serum in 34% (17/50) of GBC patients and in 3.3% (1/30) patients with cholelithiasis (p < 0.018). RFLP failed to detect common mutations in the exons 5- 8 of the p53 gene in 62 samples. Using SSCP analysis we could detect frameshift mutation in p53 gene in 2 of 22 (9.1%) GBC cases. Mutated samples were sequenced and found to have insertion of adenine at codon 271 (GAG) in exon 8 region. CONCLUSION: Our results show that 1//3rd of the north Indian patients with GBC have antibodies to p53 protein. The commonest identifiable alteration in the p53 gene was a frameshift mutation at codon 271.

High rates of early HBeAg seroconversion and relapse in Indian patients of chronic hepatitis B treated with Lamivudine: results of an open labeled trial.

BACKGROUND: The use of Lamivudine in chronic hepatitis B (CHB) is well known, however the reported rate of HBeAg sero-conversion and its durability post-treatment have varied considerably. We undertook the present study to study the effect of Lamivudine on HBeAg loss and seroconversion rates in Indian patients of CHB in relation to frequency, predictors and durability. METHODS: We treated 60 patients of e antigen positive CHB (with active viral replication and ongoing necro-inflammatory activity) with Lamivudine. They were followed up by monthly aminotransferases, and 3 monthly HBeAg and anti-HBe. Those who attained HBeAg sero-conversion were advised to discontinue Lamivudine after 6 months and followed up every 3 months thereafter, to see for relapse. Treatment was given for maximum of 3 years if not sero-converted. RESULTS: The annual incremental loss of HBeAg in patients receiving Lamivudine was 25 (41.6%) at end of 1st year, 33 (55%) at 2nd year and 35 (58.3%) at 3rd year. The corresponding rates for full sero-conversion were 17/60 (28.6%), 22/60 (36.6%) and 24/60 (40%) in the 3 years. HBeAg loss correlated with increased pre-therapy ALT levels (p = 0.002) and decreased pretreatment HBV-DNA levels (p = 0.004). The presence of cirrhosis had no influence on the rate of HBeAg loss. Relapse occurred in 35% (7/20) post-treatment at median time of 6 months. CONCLUSION: Indian patients showed a higher rate of HBeAg sero-conversion in the first year of Lamivudine treatment. This correlated with baseline ALT and inversely with HBV-DNA levels. Relapse rate after treatment was high and occurred soon after stopping treatment.

Effect of endoscopic sphincterotomy on gall bladder bile lithogenicity and motility.

BACKGROUND: Endoscopic sphincterotomy has been shown to inhibit stone formation in the gall bladder of experimental animals. AIMS: To investigate the alterations in bile composition and gall bladder motility after endoscopic sphincterotomy. PATIENTS: A study was performed of gall bladder bile composition and gall bladder motility in patients with gallstone disease ((n = 20; age 40-60 years, median age 55 years: seven men), with gall bladder calculi (n = 12) and with diseased gall bladder (chronic inflammation) without gall bladder calculi (n = 8)), who had received endoscopic sphincterotomy for common bile duct stones. Age and sex matched disease controls comprised 20 patients with gallstone disease but without stones and an intact sphincter of Oddi (with gall bladder calculi (n = 10) and diseased gall bladder without gall bladder calculi (n = 10)). METHODS: Gall bladder motility was assessed by ultrasound. Duodenal bile collected by nasoduodenal tube after stimulation of gall bladder by intravenous ceruletid infusion was analysed for cholesterol, phospholipid, and bile acid concentrations, cholesterol saturation index, and nucleation time. RESULTS: There was a significant reduction in mean (SEM) fasting volume (12.5 (1.7) ml v 26.4 (2.5) ml; p < 0.001) and mean (SEM) residual volume (4.34 (0.9) ml v 14.7 (0.98) ml; p < 0.001), and increase in mean (SEM) ejection fraction (65.7 (4.2)% v 43.6 (5.52)%; p < 0.001) and mean (SEM) rate constant of gall bladder emptying (-0.031/min v -0.020/min; p < 0.01) in patients who had been subjected to endoscopic sphincterotomy. Median nucleation time was significantly longer (17 days v 6 days; p < 0.006) in treated patients. There was a reduction in total mean (SEM) lipid concentrations (6.73 (0.32) g/dl v 7.72 (0.84) g/dl; p < 0.05), cholesterol (5.6 (1.5) mmol/l v 10.3 (2.23) mmol/l; p < 0.001) and CSI (0.72 (0.15) v 1.32 (0.31); p < 0.001). There was no significant change in mean (SEM) phospholipid (25.6 (3.5) mmol/l v 23.4 (6.28) mmol/l) and bile acid (93.7 (7.31) mmol/l v 105.07 (16.6) mmol/l) concentrations. CONCLUSIONS: After endoscopic sphincterotomy there was enhanced contractility of the gall bladder, accompanied by a prolongation of nucleation time and reduction in cholesterol saturation index.

Polarizing microscopy of partially dissolved gallstone powder: a simple technique for studying gallstone composition.

A quick and reliable method for estimating the proportion of constituents of a stone may be useful in determining the prevalence of chemical type of gallstones in different geographic areas or ethnic groups. Chemical and infrared spectroscopy estimation facilities are not commonly available in many parts of the world and visual inspection may not be reliable for scientific purposes. To investigate whether identification of microcrystalline solids in partially dissolved stone powder could accurately predict the chemical nature of the gallstone we undertook a blind study on 40 samples of gallstones and compared our results with those of visual inspection and quantitative infrared spectroscopy. There were 29 cholesterol stones (mean cholesterol 83.4 +/- 7.5%), six intermediate cholesterol stones (mean cholesterol 48.0 +/- 21.2%) and five pigment stones (mean cholesterol 14.3 +/- 4.3%) as determined by quantitative infrared spectroscopy. Microscopic examination of partially dissolved gallstone powder in ethanol correctly identified all 29 cholesterol gallstones (sensitivity 100%, specificity 92%), four of six intermediate cholesterol stones (sensitivity 67%, specificity 100%) and all five pigment stones (sensitivity 100%, specificity 97%). It also detected microspheroliths of calcium carbonate in 14 of 16 calcium carbonate containing gallstones (sensitivity 88%, specificity 100%). The chemical grouping of stones on the basis of microscopic examination was correct in 37 (93%) of 40 samples and was especially useful in identifying 'intermediate' cholesterol stones which cannot be recognized by visual inspection. Thus microscopic examination of powdered gallstones appears to be a simple and reliable method of determining gallstone composition.

Geographic variations in structure and composition of gallstones and their correlation with brittleness.

Cholesterol gallstones obtained from different geographic regions have been reported to show significant differences in their minor chemical constituents. We undertook the present study with two objectives: (i) to investigate the possible physiochemical and radiological differences between cholesterol gallstones obtained from Indian and German patients; and (ii) to compare the brittleness of the two stone groups. Forty Indian and 36 German gallstones (matched in size and shape) were subjected to assessment of physical characteristics, in vitro computed tomography (CT) and chemical analysis. German stones more often had a stone density distribution index of > or = 50 Hounsfield units (HU) (26 vs 14; P < 0.01), peripheral calcification (18 vs 9; P < 0.02), maximum CT density > or = 90 HU (17 vs 9; P < 0.05) and significant calcium carbonate (9 vs 5; P < 0.05), compared to Indian stones. The in vitro lithotripsy performed with the Siemens Lithostar Plus machine at a constant energy level showed the German stones to be more brittle (easy to fragment), more often requiring < 1000 shocks for fragmentation (25/36; 69%) compared to Indian stones (18/40; 45%, P < 0.05). Gallstones from different geographic regions may show significant variations in their physicochemical characteristics that may explain the differences in their brittleness to lithotripsy.

Utility of biliary microcrystal analysis in predicting composition of common bile duct stones.

The high failure rate of contact litholytic therapy for common bile duct stones with currently available cholesterol solvents has been attributed to the inclusion of patients with pigment stones, as no pretreatment investigation is undertaken to distinguish the two stone types. In 36 patients with common bile duct stones we prospectively evaluated the utility of microscopic examination of bile collected from the biliary tree in predicting stone composition. The bile, obtained by means of either an endoscopically placed nasobiliary catheter (n = 27) or a surgically placed T-tube (n = 9), was subjected to microscopic examination, and findings were compared with the composition of stones retrieved subsequently. On the basis of quantitative infrared spectroscopy, stones were classified as cholesterol (n = 28) or pigment (n = 8) stones. The presence of cholesterol crystals in bile correctly identified 24 of 28 patients with cholesterol stones (sensitivity, 86%; specificity, 100%; positive predictive value, 100%). Calcium bilirubinate granules, when present alone (without cholesterol crystals), correctly identified all eight patients with pigment stones (sensitivity, 100%; specificity, 89%; positive predictive value, 73%). Thus, microscopic examination of bile from the biliary tree, if used as a pretreatment screening test, may help distinguish between patients with cholesterol and pigment stones and thereby considerably improve the results of litholytic therapy of common bile duct stones with currently available solvents.

Duodenal bile examination in identifying potential non-responders to bile salt treatment and its comparison with gall bladder bile examination.

The results of bile salt treatment in patients with radiolucent stones and a functioning gall bladder have been poor. In 42 of these patients awaiting cholecystectomy we determined the value of duodenal bile examination in predicting gall stone composition, and thus identifying those less likely to respond to bile salt therapy. Based on chemical analysis and scanning electron microscopy, 28 of 42 (67%) gall stones retrieved at surgery were potentially insoluble. Microscopic examination of duodenal bile correctly identified 21 (75%) of them: it predicted all four (100%) pigment stones, three of six (50%) calcium carbonate containing cholesterol stones, and 14 of 18 (78%) cholesterol stones with pigment shells. It was nearly as reliable as microscopic examination of bile aspirated directly from the gall bladder during surgery (21 (75%) v 23 (82%); p = NS). Furthermore, the presence of cholesterol crystals in duodenal bile was a more sensitive indicator than chemical detection of supersaturation (34 of 38 (89%) v 25 of 35 (71%); p < 0.05) for prediction of cholesterol gall stones. Microscopic examination of duodenal bile, if used as a screening test, could help to exclude potential non-responders and thereby improve considerably the results of oral bile salt treatment for gall stone dissolution.

Brittleness of gallstones to lithotripsy: effect of physicochemical and ultrastructural characteristics.

Why some gallstones do not fragment easily to lithotripsy is unclear. The aim of the present study was to determine gallstone factors associated with resistance to fragmentation. Seventy-six cholesterol rich gallstones were subjected to physical evaluation, in vitro computed tomography, chemical analysis by infra red spectroscopy, and ultrastructural studies (of 26 matched stones) by scanning electron microscopy and energy dispersive X-ray microanalysis. In vitro lithotripsy was performed with the Siemens Lithostar Plus machine at a constant energy level. Of 76 stones, 33 required > 1000 shocks for effective fragmentation (largest diameter < or = 5 mm); they had significantly higher maximum diameter (P < 0.001), volume (P < 0.001), weight (P < 0.001) and lower stone density distribution (SDD) index (P < 0.05) and lower calcium bilirubinate content (P < 0.001) than those requiring < or = 1000 shocks. Large (> 15 mm; P < 0.05), round stones (P < 0.02) and gallstones with SDD index of < 50 HU (P < 0.02) and gallstones with SDD index of < 50 HU (P < 0.05) were more difficult to fragment. Clinical application of these parameters may help in better selection of patients for this non-surgical mode of therapy for gallstones.

Is duodenal bile representative of gallbladder bile? A comparative study.

Thirty-nine patients with cholelithiasis were prospectively studied to evaluate the qualitative and quantitative differences between duodenal bile and gallbladder bile. Duodenal bile obtained before cholecystectomy by nasoduodenal intubation and ceruletide injection was qualitatively similar to gallbladder bile obtained during surgery. Microscopic cholesterol crystals as an indicator of cholesterol gallstones (n = 35) could be detected in 31 (89%) and 35 (100%; p = NS), respectively. Moreover, there was no difference in the molar percentage of three biliary lipids and the mean cholesterol saturation index (1.54 +/- 0.72 and 1.74 +/- 0.42; p = NS) of the two sources of bile. Duodenal bile was, however, dilute as compared with gallbladder bile, as evidenced by lower cholesterol crystal counts (167 +/- 247 versus 705 +/- 978; p < 0.01), lower total lipid concentration (5.8 +/- 2.7 versus 11.1 +/- 5.6 g/dl; p < 0.001), and lower concentrations (in mmol/l) of the three bile lipids--that is, total bile acids, phospholipids and cholesterol (p < 0.001). Good concentrated bile (total lipid concentration > or = 5 g/dl) could be obtained in 74% of duodenal bile samples, compared with 90% of gallbladder bile (p = NS). Our study shows that, although duodenal bile is dilute as compared with gallbladder bile, it is qualitatively similar to gallbladder bile and, because of the ease and safety of its collection, can be used to study serial alterations in biliary composition in individual subjects.