Introduction PIIINP has recently been shown to discriminate between simple steatosis (SS) and NASH both in patients without advanced fibrosis and in patients with all degrees of fibrosis1. In this study we validated PIIINP as a biomarker of NASH in a cohort of patients with biopsy proven NAFLD and evaluated its performance at the proposed diagnostic thresholds. Methods 71 patients with NAFLD and no evidence of other liver disease were included in this study. Liver biopsies were performed on all patients and analysed by a expert liver histopathologist. All liver biopsies were of suitable size for analysis (> 12mm and > 5 portal tracts) and classified in a dichotomous manner into those with SS or histological NASH. Fibrosis was assessed using the Scheuer classification. Serum samples were taken at the time of liver biopsy. Results 14 of the 60 patients with non-advanced fibrosis (4-F0, 18-F1.1-F2) and all 11 patients with advanced fibrosis (9-F3, 2-F4) had NASH respectively. The AUROC of PIIINP in discriminating between SS and NASH in patients with non-advanced fibrosis and all degrees of fibrosis was 0.81 (CI 0.69–0.94) and 0.87 (CI 0.79–0.96) respectively. In comparison, the ability of ALT to discriminate between SS and NASH ranged between 0.43–0.45. The performance of the recently proposed thresholds PIIINP in their ability to diagnose NASH in our population is displayed in the table. Abstract PWE-139 Table PIIINP Thresholdng/ml F0–2:NPV/PPV % F0–4:NPV/PPV % 5.2 92/42 88/56 7.2 85/68 83/85 11 77/100 72/100 Conclusion PIIINP discriminates between SS and NASH. The performance of the proposed diagnostic thresholds is comparable to that reported in the original publication of this biomarker. Our results suggest that PIIINP can be used to detect the minority of patients with NAFLD who have NASH and are at risk of developing progressive fibrosis. Disclosure of Interest None Declared. Reference Tanwar S et al. Validation of Terminal Peptide of Procollagen III for the Detection and Assessment of Nonalcoholic Steatohepatitis in Patients with Nonalcoholic Fatty Liver Disease. Hepatology. 2013 Jan; 57(1):103–11. doi: 10.1002/hep.26030. PMID: 22930399

Background: Hepatic stellate cells (HSCs) are a major fibrogenic cell type that contributes to collagen accumulation during chronic liver disease. With increasing interest in developing antifibrotic therapies, there is a need for cell lines that preserve the in vivo phenotype of human HSCs to elucidate pathways of human hepatic fibrosis. We established and characterised two human HSC cell lines termed LX-1 and LX-2, and compared their features with those of primary human stellate cells.Methods and results: LX-1 and LX-2 were generated by either SV40 T antigen immortalisation (LX-1) or spontaneous immortalisation in low serum conditions (LX-2). Both lines express a smooth muscle actin, vimentin, and glial fibrillary acid protein, as visualised by immunocytochemistry. Similar to primary HSCs, both lines express key receptors regulating hepatic fibrosis, including platelet derived growth factor receptor beta (betaPDGF-R), obese receptor long form (Ob-R-L), and discoidin domain receptor 2 (DDR2), and also proteins involved in matrix remodelling; matrix metalloproteinase (MMP)-2, tissue inhibitor of matrix metalloproteinase (TIMP)-2, and MT1-MMP, as determined by western analyses. LX-2 have reduced expression of TIMP-1. LX-2, but not LX-1, proliferate in response to PDGF. Both lines express mRNAs for alpha1(I) procollagen and HSP47. Transforming growth factor beta1 stimulation increased their alpha1(I) procollagen mRNA expression, as determined by quantitative reverse transcription-polymerase chain reaction. LX-2, but not LX-1, cells are highly transfectable. Both lines had a retinoid phenotype typical of stellate cells. Microarray analyses showed strong similarity in gene expression between primary HSCs and either LX-1 (98.4%) or LX-2 (98.7%), with expression of multiple neuronal genes.Conclusions: LX-1 and LX-2 human HSC lines provide valuable new tools in the study of liver disease. Both lines retain key features of HSCs. Two unique advantages of LX-2 are their viability in serum free media and high transfectability.

Background & Aims: Liver fibrosis and cirrhosis result from the excessive secretion of matrix proteins by hepatic stellate cells (HSCs). Previously considered irreversible, we have studied a model of cirrhosis to determine the mechanisms mediating and limiting spontaneous recovery. Methods: A micronodular cirrhosis was induced in rats after 12 weeks of CCl4 intoxication. Livers were analyzed for evidence of matrix degradation, matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) expression, stellate cell apoptosis, tissue transglutaminase (tTg) expression, and matrix cross-linking during spontaneous recovery of up to 366 days. Results: Over 366 days of recovery, micronodular cirrhosis underwent significant remodeling to a macronodular cirrhosis. Expression of collagen-1 and TIMP messenger RNA (mRNA) decreased significantly and active MMPs were shown in livers during remodeling of fibrosis. Resolution also was characterized by apoptosis of HSCs, predominantly at the margins of fibrotic septa. Residual septa, not remodeled at 366 days, were characterized by tTg-mediated cross-linking and relative hypocellularity. Conclusion: Recovery from comparatively advanced cirrhosis is possible and results in remodeling from a micronodular cirrhosis to a macronodular cirrhosis. We suggest resolution is limited by tTg-mediated matrix cross-linking and a failure of HSC apoptosis.

Apoptosis of hepatic stellate cells (HSC) has previously been shown to occur during spontaneous resolution of experimental liver fibrosis. TIMP-1 has also been shown to have a key role because of its ability to inhibit apoptosis of HSC via matrix metalloproteinase (MMP) inhibition. This has led to further study of novel substrates for MMPs that might impact on HSC survival. N-Cadherin is known to mediate cell-cell contacts in fibroblasts. In this study we demonstrate that N-Cadherin is expressed by activated rat HSC. Furthermore, during apoptosis of HSC, the N-Cadherin is cleaved into smaller fragments. Apoptosis of HSC may be inhibited by TIMP-1. This is associated with reduced fragmentation of N-Cadherin. N-Cadherin may have an important role in supporting HSC survival while N-Cadherin cleavage may play a part in promoting HSC apoptosis in recovery from liver fibrosis.

Liver fibrosis is characterized by the activation of hepatic stellate cells (HSC) with an increased synthesis of matrix proteins and decreased extracellular matrix degradation. Matrix metalloproteinases (MMPs) are the principal family of enzymes involved in the cleavage and degradation of matrix proteins. Because of their ability to destroy normal matrices and cause tissue injury, the extracellular activity of MMPs is carefully regulated. They are secreted initially as proenzymes, which are converted by a number of mechanisms to active forms following cleavage of a C-terminal propeptide. Active MMPs can be inhibited by the action of tissue inhibitors of metalloproteinases (TIMPs), which bind to their catalytic site. Studies of culture-activated HSC demonstrate these cells to be a major source of TIMP-1 and TIMP-2 and their secretion leads to net inhibition of MMP activity. Studies in human liver disease and in animal models of liver fibrosis have indicated that TIMPs play a major role in regulating matrix degradation in the liver. Alteration of the balance between TIMPs and MMPs contributes significantly to the progression of liver fibrosis. In animal models of regression of liver fibrosis, improvement occurs because of the combination of apoptosis of activated hepatic stellate cells and by the increased degradation of the fibrillar liver matrix.

Participants will be able to: 9 Summarize recent knowledge about the mechanisms that lead to non-ulcer dyspepsia and HP/NSAID-negative ulcers and rank the relative utilities of different medical and surgical treatments for GERD and gastroparesis; 9 Identify the factors that control nutrient assimilation and metabolism, design a multidisciplinary approach to obesity management, better understand the effectiveness and side effects of herbal and complementary medications, and review the rationale for using parenteral and parenteral nutrition; 9 Apply recent knowledge about the management and treatment of Crohn's disease, ulcerative colitis, sprue and irritable bowel syndrome; 9 Characterize effective strategies to screen for colon cancer and provide surveillance for patients with a history of polyps.Discuss the evidence to guide screening for hepatocellular carcinoma, gastric cancer and Barrett's esophagus; 9 Examine new treatment approaches that are being developed to manage Hepatitis C treatment failures, NASH, and autoimmune hepatitis;

The activated hepatic stellate cell (HSC) is central to liver fibrosis as the major source of collagens I and III and the tissue inhibitors of metalloproteinase-1 (TIMP-1). During spontaneous recovery from liver fibrosis, there is a decrease of TIMP expression, an increase in collagenase activity, and increased apoptosis of HSC, highlighting a potential role for TIMP-1 in HSC survival. In this report, we use tissue culture and in vivo models to demonstrate that TIMP-1 directly inhibits HSC apoptosis. TIMP-1 demonstrated a consistent, significant, and dose-dependent antiapoptotic effect for HSC activated in tissue culture and stimulated to undergo apoptosis by serum deprivation, cycloheximide exposure, and nerve growth factor stimulation. A nonfunctional mutated TIMP-1 (T2G mutant) in which all other domains are conserved did not inhibit apoptosis, indicating that inhibition of apoptosis was mediated through MMP inhibition. Synthetic MMP inhibitors also inhibited HSC apoptosis. Studies of experimental liver cirrhosis demonstrated that persistent expression of TIMP-1 mRNA determined by PCR correlated with persistence of activated HSC quantified by alpha smooth muscle actin staining, while in fibrosis, loss of activated HSC correlated with a reduction in TIMP-1 mRNA. We conclude that TIMP-1 inhibits apoptosis of activated HSC via MMP inhibition.

Autoimmune hepatitis is a liver disease associated with:•hyperglobulinaemia•autoantibodies in the serum•a history of other autoimmune disorders.It was previously known as ‘chronic active hepatitis’, but this term is now outmoded, in recognition of the acute presentation in some cases. It typically occurs in young women (age at onset 20–40 years), but it may develop at any age in either sex.

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Comment on: Impact of pegylated interferon alfa-2b and ribavirin on liver fibrosis in patients with chronic hepatitis C.