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Objective: The aim of the present study was to characterize the predominant hepatitis B virus (HBV) strains and their molecular variants present in the HBV isolates of the different genotypes found among the chronic carriers of the virus in our community. Methods: Precore/core and core promoter regions of HBV DNA were amplified by polymerase chain reaction and then subjected to direct sequencing. Of the 64 hepatitis B surface antigen (HBsAg)-positive chronic HBV carriers investigated, 44 were HBeAg negative and 20 were HBeAg positive. Results: In addition to genotype D, which was the predominant genotype, 12 genotype C (18.7%) and 6 genotype A (9.4%) were also detected. Presence of T at nt 1858 has often been related to the development of precore stop mutation at nt 1896, while that of C has been related to the development of 1762–1764 double mutation. In our study group, 39 of the 44 HBeAg-negative samples have T1858. The precore stop codon mutation was found in only 8 (18%) of the HBeAg-negative samples. More than half of the HBeAg-negative samples had wild-type sequence in the precore region. The core promoter region could be sequenced from 40 samples, and 1762–1764 double mutation was detected in 13 (32.5%) of them. No significant changes could be detected in the core amino acid sequence of these isolates. Conclusion: The pattern of core promoter and precore mutation of HBV isolates in the present study is atypical and not in accordance with reports from other parts of the world, where genotype D and genotype C with T at codon 1858 are common.

Metformin is a popular oral antidiabetic medicine of the Biguanide class. It is widely chosen as the first line therapy for treating Type 2 Diabetes Mellitus. The drug is believed to be a comparatively safer agent for controlling blood sugar. This case report aims at describing an unusual phenomenon of Metformin hypersensitivity in a female patient who presented with perioral lesions. JCMCTA 2016 ; 27 (1) : 75 - 76

Virulence-associated genes and neutral DNA markers of Helicobacter pylori strains from the Santhal and Oroan ethnic minorities of West Bengal, India, were studied. These people have traditionally been quite separate from other Indians and differ culturally, genetically, and linguistically from mainstream Bengalis, whose H. pylori strains have been characterized previously. H. pylori was found in each of 49 study participants, although none had peptic ulcer disease, and was cultured from 31 of them. All strains carried the cag pathogenicity island and potentially toxigenic s1 alleles of vacuolating cytotoxin gene ( vacA ) and were resistant to at least 8 μg of metronidazole per ml. DNA sequence motifs in vacA mid-region m1 alleles, cagA , and an informative insertion or deletion motif next to cagA from these strains were similar to those of strains from ethnic Bengalis. Three mobile elements, IS 605 , IS 607 , and IS Hp608 , were present in 29, 19, and 10%, respectively, of Santhal and Oroan strains, which is similar to their prevalence in Bengali H. pylori. Thus, there is no evidence that the gene pools of H. pylori of these ethnic minorities differ from those of Bengalis from the same region. This relatedness of strains from persons of different ethnicities bears on our understanding of H. pylori transmission between communities and genome evolution.

Limited information is available about the prevalence and genotype distribution of hepatitis C virus (HCV) in the general population of India. A community-based epidemiologic study was carried out in a district in West Bengal, India. By a 1:3 sampling method, 3,579 individuals were preselected from 10,737 inhabitants of 9 villages of the district, of whom 2,973 (83.1%) agreed to participate. Twenty-six subjects (0.87%) were HCV antibody positive. The prevalence increased from 0.31% in subjects

We compared putative molecular markers of virulence ( vacA , cagA , and iceA ) of Helicobacter pylori strains isolated from 52 adult duodenal ulcer patients from West Bengal, India, with those of H . pylori strains isolated from 48 adult healthy volunteers from the same region. On the basis of genotyping by PCR, we conclude that the H. pylori strains isolated from the two study groups were indistinguishable and that there are geographic variations in the association of certain putative H. pylori virulence genes with clinical status.

Increasing significance of tumor–stromal interaction in development and progression of cancer implies that signaling molecules in the tumor microenvironment (TME) might be the effective therapeutic targets for hepatocellular carcinoma (HCC). Here, the role of microRNA miR-199a-3p in the regulation of TME and development of HCC has been investigated by several in vitro and in vivo assays. Expression of miR-199a-3p was observed significantly low in HCC tissues and its overexpression remarkably inhibited in vivo tumor growth and metastasis to lung in NOD-SCID mice. In vitro restoration of miR-199a-3p expression either in endothelial cells (ECs) or in cancer cells (CACs) significantly diminished migration of ECs in co-culture assay. Again incubation of miR-199a-3p transfected ECs with either conditioned media (CM) of CACs or recombinant VEGF has reduced tube formation, in ECs and it was also dropped upon growth in CM of either anti-VEGF antibody-treated or miR-199a-3p-transfected CACs. In addition, bioinformatics and luciferase-reporter assays revealed that miR-199a-3p inhibited VEGF secretion from CACs and VEGFR1 and VEGFR2 expression on ECs and thus restricted cross talk between CACs and ECs. Again, restoration of miR-199a-3p in hepatic stellate cells (HSCs) reduced migration and invasion of CACs in co-culture assay, while it was enhanced by the overexpression of HGF suggesting miR-199a-3p has hindered HSC-CACs cross talk probably by inhibiting HGF and regulating matrix metalloproteinase MMP2, which were found as targets of miR-199a-3p subsequently by luciferase-reporter assay and gelatin zymography, respectively. Thus, these findings collectively highlight that miR-199a-3p restricts metastasis, invasion and angiogenesis in HCC and hence it may be considered as one of the powerful effective therapeutics for management of HCC patients.

Hepatitis B is major health concern globally, specially in the Asia-Pacific region. The virus results in countless premature mortalities andgreater number of morbidities. While the scientific community is more focused toward HBV-related liver disease burden, the toll of this viruson our economies remain largely undermine. Proper planning and active participation of the government machinery and decision makersare vital to the best utilization of resources of our resource-constrained countries for the implementing best possible measures againsthepatitis B.

The coastal aquaculture mainly shrimps constitute major export sector in Bangladesh and is increasingly shaped by international trade conditions and by national responses to those stringent quality and safety standards. PCR based validated methods for detection of major bacterial pathogens in shrimp might be very useful tool for ensuring quality and safety standards of exportable shrimps. The objective of this study was to evaluate overall performance (sensitivity and specificity) of the multiplex PCR assay for detection of Vibrio cholerae, Vibrio parahaemolyticus, Salmonella sp. and Escherichia coli O157:H7 from spiked shrimp samples. The targeted genes were ompW for V. cholerae, tdh for V. parahaemolyticus, sefA for Salmonella spp. and hlyEHEC for E. coli O157:H7. The genomic DNA was extracted by using standard method and amplified accordingly. Sensitivity of the assay was tested by inoculating the shrimp homogenate with viable cells of laboratory references strains (target pathogens). The genes were amplified individually both from culture homogenate and spiked samples. Twenty different uniplex and multiplex PCR assay were performed; the results showed that the sensitivity and specificity of multiplex PCR are comparable to that of the results of uniplex PCR for the samples. DNA extracted from shrimp samples spiked with non-target pathogen (Bacillus cereus, Shigella flexneri and Staphylococcus aureus) yielded negative results.

Background Malnutrition is a global health problem affecting more than 300 million pre-school children worldwide. It is one of the major health concerns in India since around 50% of children below the age of two suffer from various forms of malnutrition. The gut microbiome plays an important role in nutrient pre-processing, assimilation and energy harvest from food. Consequently, dysbiosis of the gut microbiota has been implicated in malnutrition. Methodology/Principal Findings Metagenomics approach was adopted to investigate the gut microbiome sampled from 20 rural Indian children with varying nutritional status. The changes in the abundances of various taxonomic and functional groups were investigated across these gut microbiomes. A core set of 23 genera were observed across samples, with some showing differential abundances with varying nutritional status. One of the findings of the current study is the positive/negative associations of specific taxonomic and functional groups with the nutritional status of the children. Notable alterations in the architecture of the inter-microbial co-occurrence networks were also observed with changes in nutritional status. A key example is the clustering of potentially pathogenic groups into a distinct hub in severely malnourished gut. Our data does not demonstrate causality with the microbiome patterns that we observed, rather a description of some interesting patterns, whose underlying mechanism remains to be uncovered. Conclusions The present study envisioned interrelationships between the pattern of gut microbiome and the nutritional status of children. The cause of this pattern needs to be explored. However, insights obtained from the present study form the basis for further metagenomic investigations on larger population of children. Results of such studies will be useful in identifying the key microbial groups that can be utilized for targeted therapeutic interventions for managing severe acute malnutrition.

Diffie-Hellman (D-H) key exchange protocol, first published in 1976 is one of the most distinguished and widely used secured key exchange protocols. The protocol makes it possible to exchange a symmetric key between two parties over an unsecured communication channel without pre-agreement of any shared secret. The D-H key exchange protocol however requires a pre-set public parameters P (a large prime) and g a generator of ℤp to be public i.e. known to all users of the system. This paper proposes a key exchange mechanism without any public parameter. Unlike D-H key exchange mechanism our scheme uses encryption to transmit intermediate data during key exchange stage, thus it is non-vulnerable to man-in-the-middle attach.

Gut hypersensitivity has been shown to be present in irritable bowel syndrome. The current study sought to determine the involvement or hypersensitivity of the gut afferents, objectively, by recording cerebral evoked potential after rectal stimulation.In 13 patients with irritable bowel syndrome and nine healthy controls, rectal perception thresholds to electrical stimulation were measured, and cerebral evoked potentials were recorded from 2 cm behind vertex (Cz') after rectal stimulation electrically (frequency 1 Hz, duration 0.5 ms) at an intensity 50% above perception threshold and with filter setting 1-250 Hz.Perception thresholds to rectal electrical stimuli in patients with irritable bowel syndrome were lower than controls (p0.05). Rectal stimulation led to recognizable and reproducible cerebral evoked potentials. P1, N1, P2 latencies in patients with irritable bowel syndrome were shorter than that in controls (p0.05). P1/N1 amplitude was greater in patients with irritable bowel syndrome than in controls (p0.05).The shorter latency and increased amplitude of cerebral evoked potential after rectal stimulation in patients with irritable bowel syndrome compared to controls provide objective evidence supporting visceral afferent hypersensitivity as the underlying mechanism in irritable bowel syndrome.

Helicobacter pylori is a gastric pathogen that chronically infects more than half of all people worldwide (for reviews see references 48 and 64) and constitutes a major cause of peptic ulcer disease and an early risk factor for gastric cancer. It may also contribute to childhood malnutrition and increase the risk or severity of infection by other gastrointestinal pathogens such as Vibrio cholerae, especially in developing countries (18, 19). It appears to be one of the most genetically diverse of bacterial species, because DNA fingerprinting can distinguish any given isolate from most others (5, 56) and because of the ∼3 to 5% DNA sequence divergence typically found in essential genes from unrelated strains (3, 29). This mutational diversity is enhanced by a rich history of interstrain recombination (29, 40, 54). In contrast, most other well-studied bacterial species characterized to date are much more strongly clonal (see, e.g., references 28, 31, and 53). The observed genetic diversity implies a lack of population-wide selection for just one or a few universally most fit H. pylori genotypes. Some of this may reflect preferential transmission within families and among people in close contact, not in large epidemics (11, 24, 50). Such a pattern means that no individual strain would compete simultaneously against many others (12, 38, 55). H. pylori diversity would also be enhanced if humans differ in traits that are important to individual strains (e.g., specificity or strength of immune and inflammatory responses or availability of receptors used for H. pylori adherence [26, 33]). There are also indications of significant geographic differences among strains. For example, only one-half to two-thirds of U.S. and European strains carry the cag pathogenicity island (PAI), a 40-kb DNA segment many of whose genes seem to help induce interleukin 8 and thereby a strong and potentially damaging inflammatory response; such strains are recovered preferentially from persons with overt disease (4, 8, 9, 17). In contrast, nearly all east Asian strains carry the cag PAI independent of disease status (34, 47). Similarly, somewhat more than half of U.S. and European strains carry toxigenic (vacAs1) alleles of the vacuolating cytoxin gene, with other strains carrying nontoxigenic (vacAs2) alleles (in general, vacAs1 strains carry the cag PAI (65); nearly all east Asian strains carry vacAs1 alleles. Potentially more significant in terms of host interaction and evolution were findings that east Asian and Western strains differ markedly in DNA sequence motifs in the vacA and cagA genes (3, 34, 35, 46, 58, 62), since the proteins these genes encode probably each interact directly with host factors (CagA protein is translocated to host cells and is tyrosine phosphorylated in them but is not needed for interleukin 8 induction) (7, 45, 51). Even though less geographic partitioning was found in a sampling of housekeeping genes (3, 58), it is not clear whether the regional differences in cagA and vacA alleles reflect natural selection, random genetic drift including founder effects, or both during H. pylori evolution. It is noteworthy in this context that the two strains whose genome sequences have been determined (6, 57) and compared to better understand H. pylori genetic diversity (23) are from ethnic European patients (26695 from the United Kingdom [6]; J99 from a Caucasian in Pulaski, Tenn. [57; T. L. Cover, personal communication]) and thus may not be fully representative of H. pylori worldwide. We began studies of genotypes of H. pylori strains of India, motivated by the differences between strains of east Asia and the West found to date and a sense that the peoples of the vast Indian subcontinent (some one-fifth of all humanity) may have been sufficiently isolated during much of human history to have allowed the emergence of a distinct H. pylori gene pool (37, 42, 63). It is well established that H. pylori infection and peptic (especially duodenal) ulcer disease are very common in India (1, 36, 44, 52) and that a large fraction of strains may be resistant to metronidazole (Mtzr) (2), but to our knowledge there has been very little analysis to date of the genotypes of the underlying H. pylori strains (20). Here we identify several markers that help distinguish H. pylori strains from Calcutta from those of east Asia and the West.

DNA motifs at several informative loci in more than 500 strains of Helicobacter pylori from five continents were studied by PCR and sequencing to gain insights into the evolution of this gastric pathogen. Five types of deletion, insertion, and substitution motifs were found at the right end of the H. pylori cag pathogenicity island. Of the three most common motifs, type I predominated in Spaniards, native Peruvians, and Guatemalan Ladinos (mixed Amerindian-European ancestry) and also in native Africans and U.S. residents; type II predominated among Japanese and Chinese; and type III predominated in Indians from Calcutta. Sequences in the cagA gene and in vacAm1 type alleles of the vacuolating cytotoxin gene (vacA) of strains from native Peruvians were also more like those from Spaniards than those from Asians. These indications of relatedness of Latin American and Spanish strains, despite the closer genetic relatedness of Amerindian and Asian people themselves, lead us to suggest that H. pylori may have been brought to the New World by European conquerors and colonists about 500 years ago. This thinking, in turn, suggests that H. pylori infection might have become widespread in people quite recently in human evolution., published_or_final_version

Background The dynamics of physiological and behavioral risk factors of diabetes in rural India is poorly understood. Using data from a health and demographic surveillance site of Birbhum district in West Bengal, India, this study aims to assess the risk factors associated with type 2 diabetes mellitus. Research design and methods A total of 7674 individuals aged ≥18 years participated in a cross-sectional study. Venous plasma glucose method was used for measuring and reporting glucose concentrations in blood, categorized as individuals with diabetes, pre-diabetes or impaired, and normoglycemic. Aside from a set of physiological and behavioral risk factors, a range of socioeconomic confounders of diabetes was computed. Bivariate analysis with χ 2 test, and multivariate ordered logit regression methods were deployed to attain the study9s objective. Results Overall 2.95% and 3.34% of study participants were diagnosed as individuals with diabetes and pre-diabetes or impaired, respectively. Compared to the poorest, the richest have higher probability (β: 0.730; 95% CI 0.378 to 1.083) of being diagnosed with diabetes. As compared to people with normal body mass index, overweight/obese people are more prone to being diagnosed with diabetes (β: 0.388; 95% CI 0.147 to 0.628). With a decreasing level of physical activity, people are more likely to be diagnosed with diabetes. Conclusions To curb the level of diabetes, this study recommends a culturally sensitive, focused intervention for the adoption of physical activity with more traditional dietary practices, to control the level of overweight/obesity. Attention should be paid to relatively older patients with diabetes or adults with pre-diabetes.

Genetic susceptibility is an important modifier of clinical outcome and natural history of progression in Alcoholic liver disease (ALD). While the significance of ethnicity in this evolution is very clear, subtle inter-individual genetic variant(s) might be important and thus we investigated those in an Indian population. Fourteen markers were genotyped within two alcohol metabolism genes [Alcohol dehydrogenase (ADH) gene clusters (ADH1B and ADH1C) and Aldehyde dehydrogenase (ALDH2)], one microsomal ethanol oxidizing enzyme cytochrome p450 (CYP2E1) and three oxidative stress response (OSR) genes (MnSOD, GSTT1 and GSTM1) among 490 Bengali individuals (322 ALD and 168 control) from Eastern and North-Eastern India and validation was performed in a new cohort of 150 Bengali patients including 100 ALD and 50 advanced non-alcoholic steatohepatitis (NASH). Out of 14 genetic variants, carriage of 5 genotypes (rs2066701CC in ADH1B, rs1693425TT in ADH1C, rs4880TT in MnSOD and GSTT1/GSTM1 null, p-value

Summary Genome diversity is a hallmark of hepatitis B virus (HBV), which allowed its classification into 10 genotypes (A–J) and numerous subgenotypes. Among them, Genotype D is currently segregated into eight subgenotypes (D1–D8). Here, we report the identification and characterization of a novel subgenotype within genotype D of HBV from chronic hepatitis B e antigen (HBeAg)-negative patients of Eastern India. Phylogenetic tree analysis based on complete genome sequences revealed that six of 39 HBV/D isolates formed a distinct cluster supported by high bootstrap value and had nucleotide divergence >4% relative to the known D subgenotypes (D1–D8), justifying their assignment into a new subgenotype (D9). By comparing the amino acid sequences of the four ORFs of HBV/D9 with D1–D8, 36 specific residues, including a unique one (E112 in the core region), were identified that could be considered as a signature of D9. Further analysis by Simplot, BootScan and jpHMM demonstrated that D9 resulted from a discrete recombination with genotype C over the precore–core region. This type of recombination has not been described previously as all C/D recombinants reported so far possessed genotype C backbones with mosaic fragments derived from HBV/D. Interestingly, compared to other subgenotypes of HBV/D, D9 isolates had a higher frequency of mutations (A1762T and G1764A) in the basal core promoter region that had been implicated in the development of hepatocellular carcinoma. Further investigations are needed to determine the overall prevalence and clinical significance of these newly characterized D9 strains and to assess the impact of inter-genotypic recombination on viral properties.

Background Infection with Helicobacter pylori strains that express CagA is associated with gastritis, peptic ulcer disease, and gastric adenocarcinoma. The biological function of CagA depends on tyrosine phosphorylation by a cellular kinase. The phosphate acceptor tyrosine moiety is present within the EPIYA motif at the C-terminal region of the protein. This region is highly polymorphic due to variations in the number of EPIYA motifs and the polymorphism found in spacer regions among EPIYA motifs. The aim of this study was to analyze the polymorphism at the C-terminal end of CagA and to evaluate its association with the clinical status of the host in West Bengal, India. Results Seventy-seven H. pylori strains isolated from patients with various clinical statuses were used to characterize the C-ternimal polymorphic region of CagA. Our analysis showed that there is no correlation between the previously described CagA types and various disease outcomes in Indian context. Further analyses of different CagA structures revealed that the repeat units in the spacer sequences within the EPIYA motifs are actually more discrete than the previously proposed models of CagA variants. Conclusion Our analyses suggest that EPIYA motifs as well as the spacer sequence units are present as distinct insertions and deletions, which possibly have arisen from extensive recombination events. Moreover, we have identified several new CagA types, which could not be typed by the existing systems and therefore, we have proposed a new typing system. We hypothesize that a cagA gene encoding higher number EPIYA motifs may perhaps have arisen from cagA genes that encode lesser EPIYA motifs by acquisition of DNA segments through recombination events.

Since the beneficial effects of viable probiotic bacteria as dietary supplements have gained huge research interest, Lactobacillus spp. with probiotic characteristics are widely used to prepare fermented dairy products such as yoghurts, milk-shakes etc. In this study, eight (08) homemade yoghurt samples were collected from different regions in the country for isolation of probiotic Lactobacillus spp. Among the samples, four (04) isolates were identified as Lactobacillus plantarum based on their growth and biochemical characteristics. The Isolates were resistant to NaCl (1-9%) and bilesalt (0.05-0.3%) and showed good growth in the acidic condition, while maximum growth was observed at pH around 6.0. The isolates were examined for their antibacterial activity against nine (09) different test pathogens and found all pathogens are inhibited their growth to some extent but maximum zone of inhibition was observed against Bacillus cereus (53.20 mm) and minimum was against Staphylococcus aureus (19 mm) after 72 hour incubation. The results of the present study indicate that, homemade yoghurts in Bangladesh are potential source of probiotic Lactobacillus spp. Further extensive research on isolation and characterization of probiotic organisms from local fermented foods and their growth optimization might be required for development of probiotic enriched food supplements in our country.

This paper proposes to put forward an innovative algorithm for symmetric key block cipher named as “Triple Prime Symmetric Key Block Cipher with Variable Key-Spaces (TPSKBCVK)” that employs triple prime integers as private key-spaces of varying lengths to encrypt data files. Principles of modular arithmetic have been elegantly used in the proposed idea of the cipher. Depending on observations of the results of implementation of the proposed cipher on a set of real data files of several types, all results are registered and analyzed. The strength of the underlying design of the cipher and the liberty of using a long key-space expectedly makes it reasonably non-susceptible against possible cryptanalytic intrusions. As a future scope of the work, it is intended to formulate and employ an improved scheme that will use a carrier media (image or multimedia data file) for a secure transmission of the private keys. General Terms Computer Security, Symmetric Key, Cryptography, Algorithm

Introduction: Transient Elastography (TE) of liver is a well established tool to measure liver stiffness, mainly used for assessment of hepatic fibrosis due to chronic hepatitis. Liver biopsy is the gold standard test for measurement of liver iron concentration (LIC) whereas T2* MRI is the best available non-invasive method for the same in thalassemia. We intended to use hepatic TE as an alternative cheaper tool to assess hepatic iron overload so that it can be applied to larger number of patients. Objective: To assess degree of liver stiffness by TE in patients with HbE beta thalassemia and correlate the findings with LIC calculation by T2* MRI of liver. Materials and Method: 53 patients with HbE beta thalassemia from the thalassemia clinic of Institute of Haematology and Transfusion Medicine, Medical College, Kolkata were enrolled for the study. Patients with known liver disease were excluded. Baseline data like HbE%, mutations, transfusion requirement, growth status, serum ferritin level etc were collected. All of them underwent TE of liver in the School of Digestive and Liver Diseases, IPGMER using the FibroScan Touch 502 machine (Di Marco et al, British Journal of Haematology, Volume 148,3, 476-479, February 2010). 20 randomly selected patients were also assessed by T2*MRI of liver for hepatic iron assessment at the same time. LIC calculation was done from T2* value (J S Hankins et al, Blood, 14 May 2009, Volume 113:20). Data were analyzed by SPSS software-19, IBM. Results: The patients with HbE beta thalassemia had a mean HbE level of 53.66 (±18.45) %. Common beta mutations [mostly IVS-1-5(G-C)] usually found in this part of India, were detected. Mean and median age of the study population was 24.11±13.11 years and 20 years, respectively. Median age of 1st transfusion was 11 years. 35.84% patients were non-transfusion dependent. 39/53 patients had facial deformity and growth retardation. Mean baseline hemoglobin was 7.10±0.76 gm/ dl. Mean serum ferritin level was 3183.66±338.45 ng/ml. TE showed 30.18 % patients had severe liver stiffness (Liver stiffness measurement, LSM >15 kPa) whereas 43.34% had minimum stiffness (LSM≤7 kPa). No significant statistical correlation was found between serum ferritin and LSM. 12/20 patients showed very high calculated LIC (>15 mg/g) and lower T2* value ( Discussion: There is lack of data regarding hepatic iron overload in HbE beta thalassemia and so also from this part of India. There was a trend that higher the age, higher was the LSM irrespective of the serum ferritin level though not found statistically significant (Figure 1). Serum ferritin level was also not significantly correlated with the calculated LIC in those 20 patients assessed with T2* MRI. 2 patients with mildly elevated LIC had a high ferritin level. Preliminary report indicates that with increase in LSM there was increase in calculated LIC also. Statistical analysis revealed patients with LSM≥7.2 kPa had moderate or severe hepatic iron overload and thus undermine the need for routine T2*MRI. The cut off value signifies that patients with LSM Conclusion: In resource-poor countries like India, TE may be a relatively cheap tool to be used as a marker of hepatic iron overload in future. Table 1. Finding Cut off: ROC (TE-value and LIC categories), n=20 Positive if Greater Than or Equal Toa Sensitivity 1 - Specificity 2.3 1.00 1.00 3.4 1.00 .50 4.4 .94 .50 5.7 .88 .50 6.2 .83 .50 6.5 .77 .50 7.2 .77 .00 8.2 .72 .00 8.85 .66 .00 9.45 .61 .00 10.2 .55 .00 11.85 .50 .00 13.85 .44 .00 15.75 .38 .00 18.3 .33 .00 22.9 .27 .00 27.9 .22 .00 35.9 .16 .00 44.7 .11 .00 48.0 .05 .00 49.8 .00 .00 Table 2. The smallest cutoff value is the minimum observed test value minus 1, and the largest cutoff value is the maximum observed test value plus 1. LSM more than 7.2 had a sensitivity of 77.2 % and specificity of 100%. Figure 2. Figure 2. Disclosures No relevant conflicts of interest to declare.

Variants in the UGT1A1 gene and its promoter are known to determine levels of unconjugated bilirubin (UCB), but do not explain all cases of unconjugated hyperbilirubinemia. To discover associations with variants in genes other than UGT1A1, we undertook a genome-wide association study. We recruited 200 participants to cover the entire range of quantitative variation in UCB level. The data set -- after data curation, including analyses for population stratification and cryptic relatedness -- comprised genotypes at 512,349 SNP loci on 182 individuals. Quantitative trait locus (QTL) association analyses were performed, after adjusting the UCB level for effects of age, gender, and genotype at the dinucleotide (TA) insertion locus in UGT1A1 that is known to significantly modulate UCB level. A significant association of a polymorphic marker (rs2328136) near the NUP153 gene (which produces a 153 kDa nucleoporin) was obtained (p = 0.002, after multiple-testing correction). The frequency of the variant allele (A) at the rs2328136 locus in our study population is 40%, higher than most global populations. NUP153, whose product is a major regulatory factor in bidirectional transport of biomolecules across nucleus to cytosol, is associated with the transport of biliverdin reductase, which is important for bilirubin conjugation.

Hepatitis B virus (HBV) strains isolated from members of the primitive Paharia ethnic community of Eastern India were studied to gain insight into the genetic diversity and evolution of the virus. The Paharia tribe has remained quite separate from the rest of the Indians and differs culturally, genetically, and linguistically from the mainstream East Indian population, whose HBV strains were previously characterized. Full-length HBV DNA was PCR amplified, cloned, and sequenced. Phylogenetic relationships between the tribal sequences and reference sequences from the mainstream population were assessed, and divergence times of subgenotypes of HBV genotype D were estimated. HBV was found in 2% of the Paharias participating in the study. A predominance of hepatitis B e antigen-negative infection (73%) was observed among the Paharias, and the genome sequences of the HBV strains exhibited relative homogeneity, with a very low prevalence of mutations. The novel feature of Paharia HBV was the exclusive presence of the D5 subgenotype, which was recently identified in Eastern India. Analysis of the four open reading frames (ORFs) of these tribal HBV D5 sequences and comparison with previously reported D1 to D7 sequences enabled the identification of 27 specific amino acid residues, including 6 unique ones, that could be considered D5 signatures. The estimated divergence times among subgenotypes D1 to D5 suggest that D5 was the first to diverge and hence is the most ancient of the D subgenotypes. The presence of a specific, ancient subgenotype of HBV within an ethnically primitive, endogamous population highlights the importance of studies of HBV genetics in well-separated human populations to understand viral transmission between communities and genome evolution.

Data regarding the outcome of hepatitis B virus (HBV)-related cirrhosis after the onset of decompensation is scanty.From January 1998 to December 2008, a retrospective-prospective inception cohort study involving HBV-related decompensated cirrhotics was performed. Predictors of death and clinical events after the onset of decompensation were evaluated. Patients with co-infection with hepatitis C virus and/or human immunodeficiency virus, alcohol consumption to any degree and diabetes diagnosed before the detection of liver disease were excluded.Two hundred and fifty-three patients (231 males, 139 e-negative), including 102 untreated patients, were analysed. The mean (+/-SD) age was 43.0 (+/-12.0) years. The mean (+/-SD) follow-up period was 47 (+/-47) months. Decompensation was the first presentation of liver disease in 210 (83%) patients. Ascites (70%) and variceal bleed (28%) were predominant modes of decompensation. Forty-three (17%) patients died (22 vs 14% in untreated and treated cohort, respectively; P=0.002). Type 2 hepato-renal syndrome was the commonest cause of death (32%). Survival was independent of e-antigen status. In the total cohorts, predictors of death were occurrence of sepsis with systemic inflammatory response (SIRS), ascites as the initial mode of decompensation, absence of antiviral therapy and events of high-grade hepatic encephalopathy [hazards ratios (HR) of 4.4, 3.6, 2.2 and 1.7 respectively]. In the untreated cohort, initial decompensation with ascites and development of sepsis with SIRS were independent predictors of death (HR 8.5 and 2.3 respectively), while 5-year survival was higher in patients having initial decompensation with variceal bleed vs ascites (29 vs 16%, respectively, P=0.002).Decompensation with ascites and sepsis with SIRS predict reduced survival. Antiviral therapy beyond 6 months improves outcome.

There is a paucity of community-based epidemiological data on nonalcoholic fatty liver (NAFL) among nonaffluent populations in developing countries. Available studies are radiological and/or biochemical and lack histological assessment, limiting their strength. We conducted a prospective epidemiological study comprising a 1:3 subsample of all adult (18 years) inhabitants of a rural administrative unit of West Bengal, India. Subjects positive for hepatitis B virus and/or hepatitis C virus infection and consuming any amount of alcohol were excluded. Diagnosis of NAFL was by dual radiological screening protocol consisting of ultrasonographic and computed tomographic examination of the liver. Transient elastographic examination and liver biopsy were performed in a subset to identify significant liver disease. The risk factors of having NAFL were analyzed. A total of 1,911 individuals were analyzed, 7% of whom were overweight and 11% of whom had abdominal obesity. The prevalence of NAFL, NAFL with elevated alanine aminotransferase, and cryptogenic cirrhosis was 8.7%, 2.3%, and 0.2%, respectively. Seventy-five percent of NAFL subjects had a body mass index (BMI)25 kg/m(2), and 54% were neither overweight nor had abdominal obesity. The subjects with the highest risk of having NAFL were those with a BMI25 kg/m(2) (odds ratio 4.3, 95% confidence interval 1.6-11.5). Abdominal obesity, dysglycemia (fasting plasma glucose100 mg/dL or elevated homeostatic model assessment of insulin resistance), and higher income were the other risk factors. Even having a normal BMI (18.5-24.9 kg/m(2)) was associated with a 2-fold increased risk of NAFL versus those with a BMI18.5 kg/m(2).There is a significant prevalence of NAFL and potentially significant liver disease, including cryptogenic cirrhosis, in this predominantly nonobese, nonaffluent population in a developing country. NAFL will be a major determinant of future liver disease burden in countries of the developing world.

Treatment failure is a major cause of concern for the Helicobacter pylori -related gastroduodenal diseases like gastritis, peptic ulcer, and gastric cancer. Curcumin, diferuloylmethane from turmeric, has recently been shown to arrest H. pylori growth. The antibacterial activity of curcumin against 65 clinical isolates of H. pylori in vitro and during protection against H. pylori infection in vivo was examined. The MIC of curcumin ranges from 5 μg/ml to 50 μg/ml, showing its effectiveness in inhibiting H. pylori growth in vitro irrespective of the genetic makeup of the strains. The nucleotide sequences of the aroE genes, encoding shikimate dehydrogenase, against which curcumin seems to act as a noncompetitive inhibitor, from H. pylori strains presenting differential curcumin MICs showed that curcumin-mediated growth inhibition of Indian H. pylori strains may not be always dependent on the shikimate pathway. The antimicrobial effect of curcumin in H. pylori -infected C57BL/6 mice and its efficacy in reducing the gastric damage due to infection were examined histologically. Curcumin showed immense therapeutic potential against H. pylori infection as it was highly effective in eradication of H. pylori from infected mice as well as in restoration of H. pylori -induced gastric damage. This study provides novel insights into the therapeutic effect of curcumin against H. pylori infection, suggesting its potential as an alternative therapy, and opens the way for further studies on identification of novel antimicrobial targets of curcumin.

Hepatic fibrosis, a reparative response to different types of liver injury, has emerged as the primary determinant of outcome in advancing chronic liver disease, including cirrhosis. Once considered irreversible, today there is enough clinical as well as laboratory data available for us to be optimistic and expect regression of liver fibrosis in clinical situations, with resultant improvement in outcome. The primary premise of this approach to modify liver fibrosis has been its success in treating the basic pathology underlying persistent liver inflammation and injury, often with the reversal of cirrhosis. However, more focussed anti-fibrotics altering the dynamics of collagen deposition and resorption are undergoing evaluation and will be available shortly. In this changing scenario, there is a need for precise, easy to use endpoints of success/failure of anti-fibrotic therapy. A future scenario may be envisaged as one of a more positive, aggressive approach to treatment of chronic liver disease- treating the cause as well as using anti-fibrotics. This "hit the enemy and repair the hut" approach ushers in a new era from the hitherto barren pessimism in the treatment of chronic liver disease.

a) Absence of pathognomonic symptoms in patients with early phase of hepatocellular carcinoma (HCC) often leads to untreatable disease when diagnosed. Alpha-fetoprotein (AFP) with radiological images is the only potential HCC diagnosis option while disease prognosis remains dismal mandating necessity of biomarker identification with diagnostic, prognostic potentials as well as ability to assist in therapy. Thus this study aimed to identify non-invasive biomarker correlated with HCC pathogenesis by analyzing deregulated miRNAs in premalignant liver cirrhosis (LC) and in HCC. b) Among 148 study subjects, 117 were chronic Hepatitis B virus (HBV) (NAsymptomatic- control = 28, NLC = 30, NHCC = 59), 14 chronic Hepatitis C virus (HCV) infected (NLC = 7, NHCC = 7) and 17 were uninfected control. Differential expression profiling of miRNAs in each of 4 HBV infected LC and HCC tissues were performed by comparing with 8 asymptomatic controls using microarray and validated by qRT-PCR. R packages were used to determine area under receiver operating characteristics (AUROC) curve and other statistical analysis. Different softwares were used for prediction of target pathways associated with HCC development. c) Microarray analysis leads to identification of significantly altered [> or < 1.5 fold, p value d) In conclusion, combination of miR-126 with AFP may be a better predictor of HCC in high-risk chronic hepatitis patients and miR-126 alone can show better performance in detection of HCC with low AFP. Citation Format: Amit Ghosh, Alip Ghosh, Somenath Datta, Debanjali Dasgupta, Soumyajit Das, Sukanta Ray, Subash Gupta, Simanti Datta, Abhijit Chowdhury, Saroj Kant Mohapatra, Soma Banerjee. Hepatic microRNA as biomarker for detection of hepatocellular carcinoma in high-risk chronic Hepatitis B patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3969. doi:10.1158/1538-7445.AM2015-3969

Background and Aims: Chronic hepatitis has emerged as a distinct clinical outcome in large DILI registries. AT drugs being the commonest DILI agents in India, we investigated the pathways that mediate the development of hepatic fibrosis in long-term treatment of AT drugs in mice. Methods: In a combined in vivo and in vitro experiment-based study, the extent of liver injury and development of hepatic fibrosis were evaluated in wild type BALB/c mice at different time points of AT drugs (cotreatment of INH 50 mg/kg and RMP 100 mg/kg body weight) treatment for 24 weeks. Markers of oxidative stress, hepatic inflammation, apoptosis and fibrosis were evaluated. In vitro studies with mediators released from INH treated E47 cells on LX2 cells were carried out to establish the pathways in greater detail. Results: Development of hepatic oxidative stress associated with increased expression of NADPH oxidase was observed in mice during long-term treatment of AT drugs in mice. Immunohistochemistry and quantitative polymerase chain reaction demonstrated a gradual increase of HSC activation during AT drugs treatment. Histological evidence of liver fibrosis was first observed at 3 months of these drugs treatment. In addition, increasing apoptosis of hepatocytes associated with duration of AT drugs treatment accelerated hepatic fibrosis, suggesting a causative contribution of apoptosis in this process. In vitro study on stable human cell lines also confirmed the involvement of mediators (products of oxidative stress) released from INH-treated hepatocytes in the activation of hepatic stellate cell and increased collagen synthesis. Conclusions: In mice, long-term treatment of AT drugs contributes HSC activation leading to development of hepatic fibrosis. These findings will help in understanding of the pathobiology of AT druginduced liver fibrosis.

Background and Aims: Clinical outcomes of liver abscess are changing, even in South Asia, where parasitic infections are common. We wanted to capture this emerging spectrum in ‘‘real life’’ through assessment of clinical features, management strategies, and outcomes of a cohort of treated liver abscess patients at a single center. Methods: In this retrospective study of patients admitted to a referral institution between February 2010 and June 2014, patients with liver abscess, who had received prior treatment in another hospital and then been referred for complicated illness, were studied; their demographics, presentations, evolution, and outcomes were analyzed. Results: A total of 154 patients [males 109, 70.78%), mean age 43.71 16.49 years] were included in this study. Alcohol was the commonest (n = 36, 23.38%) predisposing factor, followed by biliary obstruction (n = 32, 20.78%) and diabetes (n = 17, 11.04%). The most common presenting features were fever, pain abdomen, and jaundice [143 (92.86%), 133 (86.36%) and 32 (20.78%) respectively]. Majority of patients (90, 58.44%) had solitary liver abscess, right lobe being commonest location (94, 61.04%). Median abscess volume was 180 mL (7–1524 mL); abscess rupture occurred in 21 (13.64%) cases, most commonly to subdiaphragmatic location (12/21). Overall, 97 (63%) patients required abscess drainage [single time aspiration 15 (15.46%), percutaneous indwelling catheter in 80 (82.47%), and surgical drainage in 2 (2.06%)]. Patients who required percutaneous drainage were mostly males, had higher abscess volumes, and were more frequently alcohol users and diabetics (data provided in table). Six (3.9%) of the patients died during study period. Higher bilirubin level, shortness of breath, higher volume of abscess, and requirement of surgery were more frequent in non-survivors. Conclusion: While large abscess, alcoholism, and diabetes were associated with increased need for catheter drainage; 37% can be treated by drug therapy alone. Careful selection of subjects for drainagemight avoid premature mortality in this benign condition.

To identify the variants in UDP-glucuronosyltransferase 1 (UGT1A1) gene in Gilbert's syndrome (GS) and to estimate the association between homozygosity for TA insertion and GS in India, as well as the frequency of TA insertion and its impact among normal controls in India.Ninety-five GS cases and 95 normal controls were selected. Liver function and other tests were done. The promoter and all 5 exons of UGT1A1 gene were resequenced. Functional assessment of a novel trinucleotide insertion was done by in silico analysis and by estimating UGT1A1 promoter activity carried out by luciferase reporter assay of appropriate constructs in Hep G2 cell line.Among the GS patients, 80% were homozygous for the TA insertion, which was several-fold higher than reports from other ethnic groups. The mean UCB level was elevated among individuals with only one copy of this insertion, which was not significantly different from those with two copies. Many new DNA variants in UGT1A1 gene were discovered, including a trinucleotide (CAT) insertion in the promoter found in a subset (10%) of GS patients, but not among normal controls. In-silico analysis showed marked changes in the DNA-folding of the promoter and functional analysis showed a 20-fold reduction in transcription efficiency of UGT1A1 gene resulting from this insertion, thereby significantly elevating the UCB level.The genetic epidemiology of GS is variable across ethnic groups and the epistatic interactions among UGT1A1 promoter variants modulate bilirubin glucuronidation.

Interferon treatment is the established option for the treatment of patients with chronic hepatitis B without decompensated liver disease. However, such treatment is expensive. We report here our data of a multi-center, open-label trial of the use of an indigenously produced interferon in the treatment of chronic HBeAg-positive chronic hepatitis B. Adult patients with chronic HBeAg-positive hepatitis B with elevated serum transaminase activity and positive serum HBV DNA test were treated with 5 MU/day of an indigenously produced interferon (Shanferon; Shantha Biotechnics, Hyderabad, India) for 4 months, and were then followed up for 6 months. Of the 39 patients enrolled, 36 completed the treatment and 33 completed the post-treatment follow-up. Of the 33 patients who completed the study, end-of-treatment biochemical and virological responses were observed in 10 (30%) and 5 (15%) respectively. Sustained biochemical and virological responses were observed in 15 (45%) and 7 (21%), patients respectively. Adverse effects led to the discontinuation of treatment in only one patient. Our data suggest that safety and efficacy of the indigenously produced interferon were similar to those previously reported results with interferon from other sources.

Summary Background : Helicobacter pylori infection is very common in India, as in other developing countries, but few data exist on the susceptibility of H. pylori to antimicrobial agents commonly used for eradication here. Aim : To determine the antimicrobial susceptibility of H. pylori strains from Kolkata, in eastern India. Methods : A total of 67 H. pylori strains isolated from gastritis and peptic ulcer patients of Kolkata were examined in the study. Minimum inhibitory concentration to the antibiotics was determined by the agar dilution method. Results : Most of the strains (85%) were resistant to at least 8 μg/mL of metronidazole and 7.5% strains were resistant to tetracycline, which was high when compared with other reports in India. All Kolkata strains were highly sensitive to clarithromycin, furazolidone and amoxicillin. Conclusions : Our results differed significantly from the few available reports on drug sensitivity profile of H. pylori from other parts of India, namely, Hyderabad, Mumbai and Lucknow. This finding supports the need for rigorous susceptibility testing as a guide to empirical treatment and more generally, to define the resistance patterns of H. pylori in particular geographical areas.

Background/Aims To evaluate the role of mitochondrial oxidative stress and permeability transition (MPT) in isoniazid (INH) and rifampicin (RMP) induced hepatotoxicity in mice. Methods Liver damage was induced by co-treatment of INH (50mg/kg) and RMP (100mg/kg). Pre-treatment with either methionine or phorone was done to modulate hepatic GSH level. Liver cell injury was assessed biochemically and histologically.Evidence of apoptosis was sought by TUNEL test, caspase assay and histology. Results INH and RMP co-treatment caused steatosis and increased apoptosis of the hepatocytes, hepatic oxidative stress, particularly in the mitochondrial fraction with increased mitochondrial permeability transition (MPT).Mitochondrial oxidative stress as well as liver cell injury was increased by prior treatment with phorone. This was attenuated by pretreatment with methionine suggesting a glutathione (GSH) dependent phenomenon. Conclusions Oxidative stress in the mitochondria and inappropriate MPT are important in the pathogenesis of apoptotic liver cell injury in INH-RMP hepatotoxicity. The phenomenon is GSH dependent and methionine supplementation might have a protective role.

We developed and evaluated a simple, novel multiplex PCR assay for rapid detection of Helicobacter pylori infection and for the determination of vacA and cagA genotypes directly from gastric biopsy specimens. This assay did not require culturing of strains or extraction of DNA from biopsy samples. This multiplex PCR assay would be of particularly great value for laboratories in developing countries.