cases vs 30% (39/130) in group 1 (p< 0.01).Conclusions: of all the NAFLD 6% presentedacute hepatitis like features: younger pts, intense symptoms and high level of ALT wereclinical characteristics. Further and prolonged studies are necessary to confirm this data andto investigate the prognostic significance.

M1994

Molecular Factors Involved in Determining Genetic Susceptibility to Mallory-Denk Body FormationNatasha T. Snider, Shinichiro Hanada, Sujith Weerasinghe, Philip C. Andrews, BishrOmary

Background: Mallory-Denk bodies (MDBs) are hepatocyte inclusions associated with alcoholicand nonalcoholic steatohepatitis, and some forms of drug-induced liver injury. Understandingthe biological significance of MDBs and devising means to alter their formation has clinicalrelevance to protein aggregation diseases beyond the liver. Feeding mice 3,5-diethoxycar-bonyl-1,4-dihydrocollidine (DDC) produces, in a strain-selective manner, inclusions resem-bling human MDBs. Aims: We hypothesized that proteomic comparison of livers from MDB-prone C57BL/6 mice and MDB-resistant C3H mice, before and after DDC treatment, willilluminate the molecular mechanisms regulating susceptibility of C57BL/6 mice to MDBformation. Methods: 2-dimensional (2D) differential-in-gel electrophoresis and mass spectro-metry of liver protein homogenates resulted in the identification of >200 charged isoformswith >2 fold differences between the two strains. The results for some of the proteins werevalidated in control and DDC-treated livers, and isolated hepatocytes using 1D/2D gelanalysis followed by immunoblotting. Enzymatic activities were also measured. Results:Proteins involved in cellular homeostasis and survival, such as carbonic anhydrase 3 (CA3)and GAPDH are expressed at lower levels in C57BL/6 livers. Whereas DDC treatment stronglyinduces CA3 and GAPDH expression in isolated C3H hepatocytes, it has no effect in C57BL/6 hepatocytes. There is significantly higher expression of reactive oxygen species-generatingcytochrome P450 and detoxifying glutathione-S-transferase enzymes, and lower expressionof the antioxidant enzyme peroxiredoxin 6 in the C57BL/6 livers, indicating a state ofelevated oxidative stress. Furthermore, the livers, but not other organs, of untreated andDDC-fed C3H mice have a 2-fold higher expression of nucleoside diphosphate kinase B(NDPK-B), a multifunctional protein involved in regulating intracellular energy pools andprotection from oxidative stress. Additionally, isolated C3H hepatocytes have 6-fold higherNDPK expression and activity relative to C57BL/6 hepatocytes, indicating that differencesin total liver NDPK levels are accounted for by differences in hepatocytes. Conclusions:Genetic predisposition to MDB formation is likely related, at least in part, to deficiencies insystems involved in regulating intracellular homeostasis, energy generation and protectionfrom oxidative stress. NDPK may be an upstream effector functioning in a strain-dependentand tissue-selective manner to limit MDB formation. Similar molecular determinants mightcontribute to the genetic differences in human liver diseases that are associated withMDB formation.

M1995

Spontaneously Hypertensive Rat Develops Pronounced Steatohepatitis Inducedby Choline Deficient Diet: An Evidence for Hypertension as the CriticalEnhancer for Nonalcoholic Fatty Liver DiseaseTakuya Ikuta, Keishi Kanno, Sosuke Matsuda, Nobusuke Kishikawa, Keiko Fujita,Susumu Tazuma

Background & Aim: Hypertension is frequently accompanied by a variety of metabolicdisturbances including hepatic steatosis. Hypertension has also been reported to promotethe decline of mitochondrial function as well as to alter the redox system of reactive oxygenspecies (ROS) in the liver. However, the causal link between hypertension and steatohepatitisremains to be established. The aim of this study was to test the hypothesis that the existenceof high blood pressure modulates the development of steatohepatitis. Method: We utilizedspontaneously hypertensive rat (SHR) which develops essential hypertension from the ageof 5w. Six to 8w old female SHR and Wister Kyoto rat, a normotensive control (n=6/group)were fed choline deficient (CD) diet for 5 weeks. Hepatic steatosis was evaluated histologicallyand by hepatic triglyceride (TG) content. The hepatic expressions of genes involving lipidmetabolism and antioxidant status were quantified by real-time PCR. Hepatic lipid peroxida-tion was evaluated by measuring thiobarbituric acid reactive substances (TBARS) levels.Plasma levels of insulin, glucose, and alanine aminotransferase (ALT) were measured enzym-atically. Results: CD diet resulted in significant body weight loss in both groups withsubstantial reduction in SHR. By contrast to WKY rat, the liver from SHR fed CD dietexhibited pronounced hepatic steatosis as depicted histologically. This was further confirmedby hepatic TG content, which revealed significant 8-fold increase in SHR with no significant1.2-fold increase in WKY rat. Consistent with marked decrease in mRNA levels of hepaticantioxidant enzymes,hepatic TBARS levels as well as plasma ALT levels were increased onlyin SHR following CD diet. Substantial steatosis in SHR might be explained by 87% reductionin mRNA levels of peroxisome proliferator activated receptor (PPAR) α with 75% reductionof acyl-Coenzyme A oxidase (ACOX) 1, a PPARα target gene. In addition, mRNA expressionsof microsomal triglyceride transfer protein (MTP) and apolipoprotein B (APOB), both ofwhich are requisite for TG secretion form liver, were also down-regulated in SHR by 70%.Consistently, plasma TG level was significantly lower in SHR (-65%) compared to WKY raton CD diet. Conclusion: Our data indicates SHR exhibited reduced fatty acid oxidation aswell as TG secretion from the liver when fed CD diet, resulting in severe hepatic steatosis.Taken together with the finding that hepatic TBARS levels and plasma ALT levels wereincreased only in SHR on CD diet, hypertension is suggested to be one of the criticalenhancers in the development of steatohepatitis.

S-455 AGA Abstracts

M1996

UGT1A1 Promoter Gene Polymorphism Increases Risk of GallstonesFormation in Patients With Wilson's DiseaseMirjana Kalauz, Dora Grgic, Davor Radic, Irena Hrstic, Milorad Opacic, Rajko Ostojic,Tomislav Brki, Boris Vucelic

Introduction: Wilson's disease is an autosomal recessive inherited disorder of hepatic coppermetabolism characterized by copper overload and toxicity in the liver and several othertissues such as brain and kidney. The hepatic course is variable and includes asymptomaticelevation of liver enzymes without morphological changes, chronic hepatitis, cirrhosis andacute liver failure. Patients with Wilson's disease may be more prone to gallstone formationowing to increased intravascular hemolysis and coexistence of liver cirrhosis. We have nowtested the hypothesis that the coinheritance of the common UGT1A1 A(TA)7TAA alleleassociated with Gilbert syndrome increases the risk for gallstones formation in patients withWilson's disease. Patients and methods: Thirty-one patients with Wilson's disease (18 males,14 females) were included in the study. Wilson's disease was diagnosed according to typicalclinical and laboratory features (Leipzig score of≥4) and most patient were tested for ATP7Bgene mutations. The UGT1A1 promoter [(A(TA)7TAA] was genotyped by direct sequencingof PCR products. All patients (23 with hepatic course, 5 with neurological course, 2 withmixed form, 1 asymptomatic) were examined with transabdominal US for the presence ofgallstones. Results: In our cohort of Wilson's disease patients , 8 patient were heterozygousfor the A(TA)7TAA allele, while 7 patients were homozygous for the A(TA)7TAA allele. Incomparison to stone-free patients with Wilson's disease, significantly more Wilson's diseasepatients with gallstones either carried the heterozygous A(TA)7TAA allele (p=0,037) or thehomozygous A(TA)7TAA allele (p=0,002). Conclusion: Our results suggest that the A(TA)7-

TAA allele of the UGT1A1 promoter is additional risk factor for developing gallstones inpatients with WD. This finding suggest that decreased bilirubin conjugation due to diminishedactivity of UDP glucuronosyltransferase (UGT) and increased unconjugated bilirubin depos-ition in billiary tree may be additional lithogenic factors in patients with Wilson's disease.

M1997

The Impact of Transferrin Gene Polymorphism to Phenotypic Expression inPatients With Iron OverloadSandra Milic, Smiljana Ristic, Nada Starcevic Cizmarevic, Davor Stimac

Introduction: Products of the transferrin (TF) and hemochromatosis (HFE) genes interactin iron metabolism by competing for binding to transferrin receptor. The combination ofthe TF polymorphism (C1/C2) and HFE (C282Y and H63D) mutations may result in anexcess of free iron and consequently iron overload. We must emphasize that the TF C2allele has been reported to be associated with various conditions related to free radicals,suggesting differences in iron metabolism of TF C2 carriers. Aim: To determine the impactof TF polymorphism (C1/C2) to phenotypic expression in patients with iron overload inwhich genotyping for HFE mutations (C282Y and H63D) was performed earlier. Patientsand Methods: The study included 160 patients with previously defined HFE (C282Y andH63D) and they had high iron parameters (iron or transferrin saturation or ferritin): 13(8.1%) C282Y homozygotes, 9 (5.6%) compound heterozygotes C282Y/H63D, 8 (5%) H63Dhomozygotes, 14 (8.7%) C282Y heterozygotes, 48 (30%) H63D heterozygotes, while 66(41.2%) had no HFE mutations. Transferrin genotypes were analysed by PCR-RFLP method.Results:The TF C2 allele was determined in 44 observed patients (27.50%). We did notshow that interactions between HFE and TF gene polymorphism, primarily TF C2 allele,markedly raised any iron parameters (iron or transferrin saturation or ferritin), we onlyobserved the existence of higher values of iron, transferrin saturation and ferritin in carierrsof C282Y and TF C2 allele and H63D and TF C2 allele but these tendecies failed to reachstatistical significance (p>0.05). Conclusion: Our study indicated that iron overload dependsof HFE genotypes and supports the functional role of the HFE protein in iron metabolismwhereas the interaction with TF C2 allele polymorphism seems to have no effect on higheriron parameters but warrants further studies in larger series of iron overload patients.

M1998

Frequency and Factors Associated With Elevated ALT Levels in MorbidlyObese Young Adults Awaiting Bariatric SurgeryHarmony Allison, Daria Homenko, Kathleen Viveiros

Introduction: Obesity and other factors have been associated with fatty liver disease inchildren and adults. Few studies have addressed liver disease in morbidly obese youngadults. We aim to identify the prevalence and factors associated with elevated alanineaminotransferase (ALT) levels in morbidly obese young adults. Methods: This is a retrospect-ive, cross-sectional, study. Subjects were included in the study if they were aged 18 - 25,had pre-operative laboratory data and underwent bariatric surgery from January 1, 2002 -December 31, 2008. Subjects with known liver disease (other than fatty liver disease) wereexcluded. All subjects completed an application, which included past medical history, familyhistory, medications, review of systems, and habits as part of their evaluation for bariatricsurgery. Subjects also had a complete medical evaluation by a staff physician. Our databasewas created from these medical records. Elevated ALT was defined as an ALT greater than19 for women and greater than 30 for men. Very elevated ALT was defined as an ALT 1.5times normal (29 for women, 45 for men). A univariate and multivariate logistic regressionwas used to evaluate factors associated with elevated and very elevated ALT. Results: Onehundred and eleven patients met inclusion criteria: 84% were female, 77% white, with anaverage age of 22.6 years. The mean body mass index (BMI) was 48.2. Sixty-six subjectshad elevated ALT (60%) and of these thirty-three had very elevated ALT (50%). Comorbiditesincluded hypertension (21.7%), sleep apnea (35.1%), dyslipidemia (16.2%) and diabetes(3.6%). In the univariate analysis, elevated ALT was not associated with alcohol, diabetes,dyslipidemia, or BMI. Fasting blood glucose levels were not available for this study; however,neither hemoglobin A1C (HA1C) as a continuous variable nor HA1C divided into tertileswas associated with elevations of liver enzymes. There was a trend towards significance withhigh blood pressure (p=.06) and lower high density lipoprotein (HDL) (p=.07) and elevated

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