TRAIL receptor I (DR4) polymorphisms C626G and A683C are associated with an increased risk for hepatocellular carcinoma (HCC) in HCV-infected patients

<p>Abstract</p> <p>Background</p> <p>Tumour surveillance via induction of TRAIL-mediated apoptosis is a key mechanism, how the immune system prevents malignancy. To determine if gene variants in the TRAIL receptor I (<it>DR4</it>) gene affect the risk of hepatitis C virus (HCV)-induced liver cancer (HCC), we analysed <it>DR4 </it>mutations C626G (rs20575) and A683C (rs20576) in HCV-infected patients with and without HCC.</p> <p>Methods</p> <p>Frequencies of <it>DR4 </it>gene polymorphisms were determined by LightSNiP assays in 159 and 234 HCV-infected patients with HCC and without HCC, respectively. 359 healthy controls served as reference population.</p> <p>Results</p> <p>Distribution of C626G and A683C genotypes were not significantly different between healthy controls and HCV-positive patients without HCC. <it>DR4 </it>variants 626C and 683A occurred at increased frequencies in patients with HCC. The risk of HCC was linked to carriage of the 626C allele and the homozygous 683AA genotype, and the simultaneous presence of the two risk variants was confirmed as independent HCC risk factor by Cox regression analysis (Odds ratio 1.975, 95% CI 1.205-3.236; p = 0.007). Furthermore HCV viral loads were significantly increased in patients who simultaneously carried both genetic risk factors (2.69 ± 0.36 × 10⁶ IU/ml vs. 1.81 ± 0.23 × 10⁶ IU/ml, p = 0.049).</p> <p>Conclusions</p> <p>The increased prevalence of patients with a 626C allele and the homozygous 683AA genotype in HCV-infected patients with HCC suggests that these genetic variants are a risk factor for HCC in chronic hepatitis C.</p

Alignment and regional community in Southeast Asia: ASEAN diplomacy from 1967-1999

This study examines the alignment behaviour of ASEAN towards external parties between 1967 and 1999 with the goal of establishing which factors have been most important for determining patterns of cooperation and conflict in Southeast Asia. For this purpose, the study uses an integrative perspective that primarily builds upon the insights from realism and social constructivism but also incorporates some ideas borrowed from selective cognitive theories. The study pursues three goals. Firstly, on the theoretical level, it aims at making a contribution to debates in security studies about the causes of alignment. In particular, this study assesses the relevance of three different explanations of alignment in the ASEAN context: balance-of-threat theory, identity-based accounts, and balance-of-interest theory. Secondly, it evaluates to what extent ASEAN has evolved into a regional community, based on the existence of a collective identity. The notion of regional community is subjected to critical analysis by gauging ASEAN diplomacy against three indicators that are used to operationalize the concept of regional identity: shared problem representations, mutual identifications and norm compliance with the 'ASEAN way'. Thirdly, with regard to the study of international relations in Southeast Asia, this study intends to explore the nature of state interaction in the region. What factors have guided alignment of ASEAN members between 1967 and 1999? The findings of this study are threefold. Firstly, only limited support is found for both threat-based and identity-based explanations of ASEAN alignment, whereas balance-of-interest theory has high plausibility for most cases of ASEAN alignment. Secondly, with regard to the community idea, this study concludes that while ASEAN's diplomacy has partly been based on the idea of community, ASEAN has constituted a rule-based community, not one based on a shared regional identity. Thirdly, as to the nature of state interaction in the region, this study finds that, on a fundamental level, ASEAN behaviour has been characterized by continuity since 1967: alignment has been in support of the status quo. Moreover, security relations in Southeast Asia have become characterized by a high degree of ambiguity, yet this ambiguity has not been adequately accounted for by Western security theory

Theories of international cooperation and the GATT/WTO regime: beyond the dichotomy of rational and cognitive approaches

This thesis aspires to assess the explanatory value of different theories of international cooperation for the case of the world trade regime of GATT/WTO and subsequently strives to reach a satisfactory interpretation of the instance of cooperation. The world trade regime embarked on a process of transformation with the signing of the Marrakech Agreements of 15th April 1994. The event marked the conclusion of the Uruguay Round and, with the establishment of the WTO, the beginning of a new era for the world trade regime. The thesis endeavours to establish the substance of the regime change from GATT to the WTO. It outlines the most significant provisions of the agreement of the Uruguay Round and, subsequently, analyses the change on the level of regime norms underlying the world trade regime. The analysis of regime norms yields insights about the essence of the regime transformation and as to what factors proved to be conducive to cooperation in the sphere of the world trade. The GATT/WTO regime with its extended scope and more sophisticated institutional structures can be regarded as a prime example of successful cooperation. However, the prospects for cooperation between states in an anarchic environment without central authority for enforcement are the subject of a remarkably intense scholarly debate. Therefore it is worthwhile to examine which theoretical framework proves to be most adept at elucidating the circumstances of this instance of cooperation. This thesis applies different theories of international cooperation to the case of the GATT/WTO regime. While a large array of rational theories attempts to explain cooperation from a perspective which focuses on interests and capabilities, a different strand of theories, that of cognitive approaches, emphasizes the paramountcy of ideas and beliefs as variables which explain cooperation. They endogenize the process of interest formation. This thesis seeks to synthesise the strong points of rational and cognitive approaches and thus to reconcile the divergent schools of thought. Its further purpose is to set out factors which are conducive to cooperation

The PNPLA3 rs738409 148M/M genotype is a risk factor for liver cancer in alcoholic cirrhosis but shows no or weak association in hepatitis C cirrhosis.

BACKGROUND: An isoleucine>methionine mutation at position 148 in the PNPLA3 gene (p.I148M, rs738409) has recently been identified as a susceptibility factor for liver damage in steatohepatitis. Here, we studied whether the PNPLA3 rs738409 polymorphism also affects predisposition to hepatocellular carcinoma (HCC). METHODS: We compared distributions of PNPLA3 genotypes in 80 and 81 Caucasian patients with alcoholic and hepatitis C virus (HCV)-associated HCC to 80 and 81 age- and sex-matched patients with alcohol-related and HCV-related cirrhosis without HCC, respectively. PNPLA3 genotypes in 190 healthy individuals from the same population served as reference. Potential confounders obesity, diabetes, HCV genotype and HBV co-infection were controlled by univariate and multivariate logistic regression with forward variable selection. RESULTS: PNPLA3 genotypes were in Hardy-Weinberg equilibrium for all study groups. The frequency of the 148M allele was significantly (p<0.001) increased in alcoholic cirrhosis with (53.7%) and without HCC (36.2%) but was not different between healthy controls (22.9%) and patients with cirrhosis (25.3%; p = 0.545) and HCC (30.2%; p = 0.071) due to hepatitis C. HCC risk was highest in 148M/M homozygous patients with alcoholic liver disease (odds ratio (OR) 16.8 versus healthy controls; 95% confidence interval (CI) 6.68-42.43, p<0.001). Finally, multivariate regression confirmed 148M/M homozygosity (OR 2.8; 95%-CI: 1.24-6.42; p = 0.013) as HCC risk factor in alcoholic cirrhosis. In HCV-related cirrhosis only HCV genotype 1 was confirmed as a HCC risk factor (OR 4.2; 95%-CI: 1.50-11.52; p = 0.006). CONCLUSION: The PNPLA3 148M variant is a prominent risk factor for HCC in patients with alcoholic cirrhosis, while its effects are negligible in patients with cirrhosis due to HCV. This polymorphism provides an useful tool to identify individuals with particularly high HCC risk in patients with alcoholic liver disease that should be taken into account in future HCC prevention studies

Genetic Analyses Reveal a Role for Vitamin D Insufficiency in HCV-Associated Hepatocellular Carcinoma Development.

BACKGROUND: Vitamin D insufficiency has been associated with the occurrence of various types of cancer, but causal relationships remain elusive. We therefore aimed to determine the relationship between genetic determinants of vitamin D serum levels and the risk of developing hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). METHODOLOGYPRINCIPAL FINDINGS: Associations between CYP2R1, GC, and DHCR7 genotypes that are determinants of reduced 25-hydroxyvitamin D (25[OH]D3) serum levels and the risk of HCV-related HCC development were investigated for 1279 chronic hepatitis C patients with HCC and 4325 without HCC, respectively. The well-known associations between CYP2R1 (rs1993116, rs10741657), GC (rs2282679), and DHCR7 (rs7944926, rs12785878) genotypes and 25(OH)D3 serum levels were also apparent in patients with chronic hepatitis C. The same genotypes of these single nucleotide polymorphisms (SNPs) that are associated with reduced 25(OH)D3 serum levels were found to be associated with HCV-related HCC (P = 0.07 [OR = 1.13, 95% CI = 0.99-1.28] for CYP2R1, P = 0.007 [OR = 1.56, 95% CI = 1.12-2.15] for GC, P = 0.003 [OR = 1.42, 95% CI = 1.13-1.78] for DHCR7; ORs for risk genotypes). In contrast, no association between these genetic variations and liver fibrosis progression rate (P&gt;0.2 for each SNP) or outcome of standard therapy with pegylated interferon-α and ribavirin (P&gt;0.2 for each SNP) was observed, suggesting a specific influence of the genetic determinants of 25(OH)D3 serum levels on hepatocarcinogenesis. CONCLUSIONSSIGNIFICANCE: Our data suggest a relatively weak but functionally relevant role for vitamin D in the prevention of HCV-related hepatocarcinogenesis

A common polymorphism in the NCAN gene is associated with hepatocellular carcinoma in alcoholic liver disease

BACKGROUND & AIMS: The genetic background of alcoholic liver diseases and their complications are increasingly recognized. A common polymorphism in the neurocan (NCAN) gene, which is known to be expressed in neuronal tissue, has been identified as a risk factor for non-alcoholic fatty liver disease (NAFLD). We investigated if this polymorphism may also be related to alcoholic liver disease (ALD) and hepatocellular carcinoma (HCC). METHODS: We analysed the distribution of the NCAN rs2228603 genotypes in 356 patients with alcoholic liver cirrhosis, 126 patients with alcoholic HCC, 382 persons with alcohol abuse without liver damage, 362 healthy controls and in 171 patients with hepatitis C virus (HCV) associated HCC. Furthermore, a validation cohort of 229 patients with alcoholic cirrhosis (83 with HCC) was analysed. The genotypes were determined by LightSNiP assays. The expression of NCAN was studied by RT-PCR and immunofluorescence microscopy. RESULTS: The frequency of the NCAN rs2228603 T allele was significantly increased in patients with HCC due to ALD (15.1%) compared to alcoholic cirrhosis without HCC (9.3%), alcoholic controls (7.2%), healthy controls (7.9%), and HCV associated HCC (9.1%). This finding was confirmed in the validation cohort (15.7% vs. 6.8%, OR=2.53; 95%CI: 1.36-4.68; p=0.0025) and by multivariate analysis (OR=1.840; 95%CI: 1.22-2.78; p=0.004 for carriage of the rs2228603 T allele). In addition, we identified and localised NCAN expression in human liver. CONCLUSIONS: NCAN is not only expressed in neuronal tissue, but also in the liver. Its rs2228603 polymorphism is a risk factor for HCC in ALD, but not in HCV infection

Influence of the CXCL1 rs4074 A allele on alcohol induced cirrhosis and HCC in patients of European descent

BACKGROUND AND AIMS: CXCL1 (CXC chemokine-ligand-1) is a ligand for CXC chemokine receptor 2 expressed on hepatic stellate cells (HSC). Thus, CXCL1 might contribute to HSC activation and fibrogenesis. In the present study, we investigated the influence of the CXCL1 rs4074 polymorphism on the occurrence of alcohol induced liver cirrhosis and hepatocellular carcinoma (HCC). METHODS: The study involved 458 patients with alcoholic cirrhosis (170 with HCC), 115 alcoholics without liver disease and 342 healthy controls. All subjects were genotyped for the CXCL1 rs4074 polymorphism and CXCL1 serum levels of 132 patients were measured. In vitro CXCL1 secretion in TLR-transfected cell lines were studied by ELISA. RESULTS: Distribution of the CXCL1 genotypes (GG/GA/AA) was 159/219/80 in patients with alcoholic cirrhosis, 52/44/19 in alcoholic controls and 158/140/44 in healthy controls. Patients with alcohol-induced cirrhosis were significantly more often carriers of the CXCL1 rs4074 A allele (65.3%) than alcoholics without liver disease (54.8%, OR=1.55; 95%CI=1.025-2.350; p=0.04) and healthy controls (53.8%, OR=1.62; 95%CI=1.212-2.151; p=0.001). Accordingly, the frequency of the CXCL1 rs4074 A allele was significantly higher in the cirrhotic patients than in the subjects without cirrhosis (41.4% vs. 33.9%, OR=1.38, 95% CI:1.14-1.66, p=0.001). Furthermore cirrhotic carriers of the CXCL1 rs4074 A allele had significantly higher CXCL1 serum levels than carriers of the GG genotype. In contrast to sera from healthy controls, sera from patients with alcoholic cirrhosis induced CXCL1 secretion in TLR2- (p=0.016) and TLR4- (p=0.008) transfected HEK293 cells. This finding indicates that sera from patients with alcoholic cirrhosis contain soluble ligands that can induce CXCL1 production via stimulation of TLRs. CONCLUSION: The enhanced CXCL1 serum levels in carriers of the rs4074 A allele together with their increased frequency in patients with alcohol induced cirrhosis suggest the CXCL1 rs4074 A allele as a genetic risk factor for alcoholic cirrhosis

Compartment-specific distribution of human intestinal innate lymphoid cells is altered in HIV patients under effective therapy

<div><p>Innate lymphocyte cells (ILCs), a novel family of innate immune cells are considered to function as key orchestrators of immune defences at mucosal surfaces and to be crucial for maintaining an intact intestinal barrier. Accordingly, first data suggest depletion of ILCs to be involved in human immunodeficiency virus (HIV)-associated damage of the intestinal mucosa and subsequent microbial translocation. However, although ILCs are preferentially localized at mucosal surfaces, only little is known regarding distribution and function of ILCs in the human gastrointestinal tract. Here, we show that in HIV(-) individuals composition and functional capacity of intestinal ILCs is compartment-specific with group 1 ILCs representing the major fraction in the upper gastrointestinal (GI) tract, whereas ILC3 are the predominant population in ileum and colon, respectively. In addition, we present first data indicating that local cytokine concentrations, especially that of IL-7, might modulate composition of gut ILCs. Distribution of intestinal ILCs was significantly altered in HIV patients, who displayed decreased frequency of total ILCs in ileum and colon owing to reduced numbers of both CD127(+)ILC1 and ILC3. Of note, frequency of colonic ILC3 was inversely correlated with serum levels of I-FABP and sCD14, surrogate markers for loss of gut barrier integrity and microbial translocation, respectively. Both expression of the IL-7 receptor CD127 on ILCs as well as mucosal IL-7 mRNA levels were decreased in HIV(+) patients, especially in those parts of the GI tract with reduced ILC frequencies, suggesting that impaired IL-7 responses of ILCs might contribute to incomplete reconstitution of ILCs under effective anti-retroviral therapy. This is the first report comparing distribution and function of ILCs along the intestinal mucosa of the entire human gastrointestinal tract in HIV(+) and HIV(-) individuals.</p></div

Association between plasma markers of gut epithelial damage/ microbial translocation and colon NCR(+)ILC3 frequency in HIV(+) individuals.

<p><b>(A)</b> Plasma serum levels of intestinal fatty acid binding protein (I-FABP; ng/ml), a marker for gut barrier defects, sCD14 (μg/ml), sCD163 (μg/ml), and LPS binding protein (LBP, μg/ml) were quantified by ELISA in HIV patients (red) and HIV(-) controls (blue) (* p≤0.05). Next, association between plasma levels of I-FABP (ng/ml) <b>(B)</b> and sCD14 (μg/ml) <b>(C)</b>, respectively, and frequency of colon NCR(+)ILC3 (left panel) or colon NCR(-)ILC3 (right panel) were examined. Pearson correlation coefficient was computed for analysing correlations.</p

Compartment-specific distribution and function of intestinal ILCs in HIV(-) individuals.

<p><b>(A)</b> Frequency of ILCs (black dots, left panel) and cNK cells (white dots; right panel) expressed as percentage of CD45(+) lymphocytes in the different parts of the human GI tract (ILCs: oesophagus, n = 10; stomach n = 15; duodenum, n = 21; ileum, n = 13; colon: n = 13; cNK: oesophagus, n = 12; stomach n = 15; duodenum, n = 21; ileum, n = 14; colon: n = 13; grey columns) in comparison to peripheral blood (n = 16; white columns). <b>(B)</b> Distribution of CD103(+)ILC1 (upper left panel), CD127(+)ILC1 (upper right panel), ILC2 (lower left panel), and ILC3 (lower right panel) in the human intestinal tract (black columns) and the circulating blood (white columns). <b>(C)</b> Composition of CD127(+)ILC1 (blue), intraepithelial ILC1 (red), ILC2 (green) and ILC3 (violet) in the peripheral blood and the different segments of the gastrointestinal tract. <b>(D)</b> Distribution of NCR(-) (left panel) and NCR(+) ILC3 (right panel) in the peripheral blood and the different segments of the gastrointestinal tract. <b>(E)</b> Percentage comparison between NCR(-) and NCR(+) ILC3 subsets in those compartments. For functional characterisation ILCs derived from PBMC or biopsy samples (oesophagus, stomach, duodenum, Ileum and colon) obtained from HIV(-) individuals, were stimulated with PMA/ionomycin and then tested for IFN-γ production in CD103(+)ILC1 <b>(F)</b>, CD127(+)ILC1 <b>(G)</b>,or IL-22 production in ILC3 <b>(H)</b>, respectively. * p≤0.05; ** p≤0.01; *** p≤0.001 (FDR-adjusted P values, parametric ANOVA).</p