Manish Kumar Tripathi

Role of Proinflammatory Cytokines (Interferon Gamma) and Anti-Inflammatory Cytokine (Interleukin-10) Gene Polymorphisms in Chronic Hepatitis B Infection: An Indian Scenario

Immune-mediated mechanisms have been found to play an important role in the progression of hepatitis B virus (HBV) infection. The outcomes of infection do not appear to be determined by viral strains. Instead, allelic variants in human genome are likely to affect the disease progression. Allelic variation of proinflammatory cytokines such as interferon gamma (IFN-γ) participates in the elimination of HBV, and interleukin-10 (IL-10) helps in inhibition of Th1 effector mechanisms for host defense. The aim of this study was to determine the influence of host genetic factors in chronic HBV infection and gene promoter polymorphism or single-nucleotide polymorphism analysis of IFN-γ+874 and IL-10 (-1082, -592, and -819) on disease progression and persistence. A total of 232 patients along with 76 healthy controls were included. Allele-specific primers for IFN-γ and restriction fragment length polymorphism for IL-10 were used. The study indicated that low IFN-γ expression probably impairs host immune response to HBV, rendering these subjects more prone to HBV infection. No significant differences were detected between the 2 groups in the distributions of IL-10 genotype at the -1082, -819, and -592 positions. Odds ratio indicated that heterozygosity of genotypes -819 CT and -592 AC was more strongly associated with liver chronicity. Significantly, AA homozygous genotype was dominant in chronic hepatitis B cases in IFN-γ+874 and IL-10 (-1082 and -592) and is associated with increased risk of persistent infection.

Ulcerative Colitis and Its Association with Salmonella Species

Ulcerative colitis (UC) is characterized by presence of ulcer in colon and bloody diarrhea. The present study explores the possibility of association between Salmonella and ulcerative colitis. The present study comprised 59 cases of UC, 28 of colon cancer (CC), 127 of irritable bowel syndrome (IBS), and 190 of healthy control. The serological study was done by Widal and Indirect Haemagglutination Assay (IHA) for ViAb. Nested PCR was performed targeting fliC, staA, and stkG gene for Typhi and Paratyphi A, respectively. A total of 15.3% patients were positive for Salmonella “O” antigen among them 18.6% UC, 35.5% CC, 12.6% IBS, and 15.3% healthy control. A total of 36.9% patients were positive for “H” antigen including 39.0%, 57.1%, and 67.7% UC, CC, and IBS, respectively. About 1.73% show positive agglutination for AH antigen including 3.4%, 3.6%, and 1.6%, UC, CC, and IBS. A total of 10.89% were positive for ViAb. While 6.8% of UC, 10.7% of CC, 11.0% of IBS, and 12.1% of healthy subjects were positive for the antibody, the PCR positivity rates for Salmonella specific sequences were 79.7% in UC, 53.6% in CC, 66.1% in IBS, and 16.3% in healthy controls. The present study suggested that higher prevalence of Salmonella might play important role in etiopathogenesis of UC, IBS, and CC.

P-096 YI Comparative Response of Combination Therapy vs. Mono Therapy in Steroid Dependent Ulcerative Colitis Patients

Background:Ulcerative colitis (UC) is an idiopathic inflammatory bowel disease (IBD) characterized by a chronic relapsing/intermittent clinical course. Steroids offer efficient and prompt relief of symptoms in a high proportion of ulcerative colitis (UC). Although most patients initially respond to corticosteroids, after 1 year, approximately 25% become steroid-dependent. The aim of this prospective study was to assess whether the administration the efficacy of azathioprine (AZA) and mesalamine combination to AZA monotherapy in maintaining remission of steroid-dependent ulcerative colitis (UC). Methods:The present study was carried out in the Department of Gastroenterology, IMS, BHU, Varanasi, India. A total of 136 patients age sex matched were included in the study after written informed consent fulfilling inclusion and exclusion criterias. Patients with steroid-dependent UC in remission were randomized to receive AZA alone (1.5 mg/kg) (group 1) or in combination with mesalamine (800 mg tid) (group 2). Each group had 59 patients. Patients were considered in remission when steroid was withdrawn, an Ulcerative Colitis Clinical Activity Index (UCCAI) score of <2, an Ulcerative Colitis Disease Activity Index (UCDAI) score of 0, and a negative colonoscopy and histology. Followup was carried out every monthly for 2 year. Clinical examination, hematological and biochemical test were carried out routinely. Sigmoidoscopy and completion of inflammatory bowel disease quality-of-life questionnaire (IBD-Q) and UCDAI every 3 months; and 3) total colonoscopy with biopsies at the end of the study. Results:Out of 136 patients 110 patients were analysed and rest 18 (13.23%) were lost in followup and 4 patients in each group developed side effects or poor compliance with treatment and were withdrawn from the study. Five patients receiving monotherapy (group 1) relapsed after the first year of the study and 3 after the second year of the study (14.54%). In the combination group (group 2) 2 patients relapsed after the first year of study and 2 after the second year of the study (7.27%) having statistically significant when compared with group 1 (p<0.05). Relapse rates were significant whether analyzed by intention-to-treat or per protocol. There were no significant differences between groups in time to relapse or discontinuation of treatment, UCCAI, UCDAI, or IBD-Q scores. Adverse events were similar both the groups and statistically non significant. Conclusions:Patients with steroid-dependent UC successfully maintained in remission on AZA and mesalamine and however relapse rate were more common in patients on azathioprine alone.

P-157 YI Elevated Macrophage Migration Inhibitory Factor Sera Level in Ulcerative Colitis Patients

Background:Ulcerative colitis (UC) is a form of inflammatory bowel disease (IBD) and is often confused with Irritable bowel Syndrome (IBS). Ulcerative colitis is treated as an autoimmune disease. A number of cytokines have been implicated in chronic inflammatory lesions of UC. It has been reported that inflammatory cytokines play important role in the development of the disease. Macrophage migration inhibitory factor (MIF) is important pro inflammatory cytokine and plays a critical role in regulating broad range of immune and inflammatory activities. Higher levels may be related to the pathogenesis and induction of inflammation in UC. Aim of the present study was to evaluate the serum concentration of MIF in patients with ulcerative colitis, Irritable Bowel Syndrome (IBS), colon cancer and healthy control. Methods:Study was carried out in the Department of Gastroenterology, IMS, BHU, Varanasi, India. A total of 275 subjects age sex matched (59 UC, 127 IBS, 28 colon cancer and 61 Healthy controls) were enrolled for the study fulfilling inclusion and exclusion criteria. Blood samples were obtained after informed consent for serum separation. Enzyme linked immune sorbent assay (ELISA) was performed using Bioassay Technology Laboratory kit for human macrophage migration inhibitory factor following the manufacturers protocol. A p value of less than 0.05 was considered significant. Statistical analysis was conducted by using SPSS 16.0 (SPSS, Chicago, IL). Results:MIF level in UC was found to be significantly higher (P < 0.0003) having mean SD 3.55 ± 1.3 (Z score = 3.36) when compared with healthy control having mean SD 0.89 ± 0.48 whereas in IBS patients MIF level was found to be significantly elevated (p < 0.0006) having mean SD 3.96 ± 1.39 (Z score = 3.19) when compared to healthy control and that of colon cancer was also found to be high (P < 0.00471) having mean SD 3.02 ± 1.76 (Z score = 2.59) when compared to healthy control. Conclusions:MIF concentration significantly elevated in UC, colon cancer and IBS in respect to control subject which may immobilize macrophages and can lead to production of several inflammatory cytokine which supports the claim that inflammation could be an effective factor in the pathogenesis of disease.

Clinical Profile and Outcome in Acute on Chronic Liver Failure (ACLF)

Background and Aims: ACLF is defined as an acute deterioration of liver function in a patient with previously fairly well-compensated chronic liver disease. The aims of this study were to understand the natural history, etiology of acute insult and underlying chronic liver disease, prognostic outcome, and whether there was any subgroup of chronic liver disease which had a higher rate of decompensation. Materials and Methods: This was a prospective study conducted in the Gastroenterology department, Institute of Medical Sciences, BHU, Varanasi from December 2013 to November 2014. Data including demographics, clinical presentation, hospital course and outcome of 86 patients were recorded. The etiologies of superimposed acute event and chronic liver disease were investigated on the basis of routine investigations, viral markers, autoimmune markers, Wilson disease panel, serum ferritin and liver biopsy (if feasible). Study variables like coagulopathy, hepatic encephalopathy, sepsis, hyponatremia, renal failure, hepatorenal syndrome and various prognostic scores were considered. Results:Most of the patients were young adults (55%) and males were more common than females (M: F = 2.7:1). The most common presenting features were coagulopathy and ascites (95%) whereas encephalopathy (28%) and GI bleed were less common. The most common cause of acute insult leading to Fig. 1 Etiology of acute events.

Inflammatory cytokine IL-10 polymorphism in patients of ulcerative colitis and irritable bowel syndrome

S 21 ANNUAL CONFERENCE—2013 M isella n eo u s Corresponding author: Jitendra Km Choudhary. E-mail: jitendra709@gmail.com INFLAMMATORY CYTOKINE IL-10 POLYMORPHISM IN PATIENTS OF ULCERATIVE COLITIS AND IRRITABLE BOWEL SYNDROME Manish Kumar Tripathi, Ashok Kumar Jain, Vinod Kumar Dixit, Pankaj Kaushik, Jitendra Kumar Choudhary, Manjita Srivastava, Neha Singh, Arttrika Ranjan, Neha Gupta, Sangey Chopel Lamtha, Smita Verma Department of Gastroenterology, Institute of Medical Sciences, BHU, Varanasi Background: Ulcerative colitis (U.C.) is a complex multifactorial trait involving both environmental and genetic factors. UC is characterized by activation of macrophages and T lymphocyte; pro-inflammatory cytokine, chemokines and adhesion molecule expression and an inability to adequately down-regulate immune activation. Aim: The aim of the present study is to examine Interleukin 10 gene polymorphism in patients with UC and IBS. Methods: Age and sex matched 56 patients of IBS and 36 patients of ulcerative colitis were enrolled in the study. Blood samples were obtained after informed consent for DNA isolation by phenol-chloroform method. PCR for IL 10 -1082, -819, -592 was done followed by restriction digestion by site specific enzyme Mnl1, RsaI and MaeIII respectively, the results was analyzed on 2% agarose gel. Results: The percentage distribution of genotypic frequencies of IL-10 -1082 homozygotes and heterozygotes are given in the table 1. Table 1 Genotypic frequencies of IL10 -1082, -819, -592 between IBS and UC patients Cytokines Genotype Frequency IL-10 (1082) AA AG GG IBS 32(57.1) 16(28.6) 8(14.3) Ulcerative Colitis 22(61.1) 8(22.2) 6(16.7) c2=0.479, p=0.787 IL-10 (819) TT TC CC IBS 13(23.2) 21(37.5) 22(39.3) Ulcerative Colitis 4(11.1) 20(55.6) 12(33.3) c2=3.550, p=0.169 IL-10 (592) AA AC CC IBS 36(64.3) 10(17.9) 10(17.9) Ulcerative Colitis 16(44.4) 12(33.3) 8(22.2) c2=3.934, p=0.140 S124 Conclusion: Genotypic frequencies of IL-10 between UC and IBS patients showed that there is no statistically significant difference in IL-10 -1082, -819, -592 cytokine genes. Genotype ATA/ATA correlate with low level of IL-10 production Hence the results of this study indicate low production of anti-inflammatory cytokines IL-10 in patients with UC and IBS. Corresponding author: V.K.Dixit. E-mail: manish.imsbhu@gmail.com