almlysis, age >45 years (Odd Ratio (OR) = 4.1; confidence intervals (el) (1.7-10.33) and BMI>30 (OR = 3 .9 CI (16-9.433 were the only independent risk tactors of cirrhosis. Conclusion: Our msnhs suggest that overweight is a risk factor for ALD by inducing nmre associated diseases and cirrhosis. It could be explained by a po~ible potentiation of metabolic ettects of ethanol ingestion through excess weight in alcoholic patients.

M1376

Non-invasive Markers of Fibrotic Nonalcoholic Steatohepatitis Sangik O h Aaron Beusnl}, Joseph Grossman, Imad Nasser, Michael P Curry, Nezam H. Afdhal

BACKGROUND: The challenge fi.~r the clinician in assessing the patients with nonalcoholic laity liver disease (NAFLD) is to diflerentiate those with steatosis alone from those with progressive liver disease. At the present time, liver biopsy remains as the gold standard in staging hepatic fibrosis OBJECTIVES: To assess the role of serum YKL-40 and HA in differentiating patients with fibrotie nonalcohohc steatohepatitis(NASH) from those with simple steatosis. METHODS We perfbrnmd a cross sectional analysis of 60 consecutive patients unth NAFLD Vanons clinical and biochemical data were obtained including estima- tion of insulin resistance by" using homeostasis model assessment (HOMA), Serum levels of YKL-40 and F24 were measured by ELISA and compared to histological staging of biopsies by Brunt Score and Computerized in,age Analysis. RESULTS: Thirty-one patients had simple steatosls and 29 patients were found to have steatosls plus fibrosis. Our univariate analysis showed that the fibrosis group consisted of more &abetics(21.7 % vs. 0%, p = 0.02), higher insulin resistance based on HOMA index (5.99 vs. 3.47, p = 001) and higher AST/ALT ratio (0.73 vs 0 .55 p = 001). "Fnere were no significant difli'rences in mean age, number of females, body mass index (BMI) and totaI cholesterol levels between the two groups. The mean of serum YKL-40 w~ls significantly higher (142 vs 72 ng/ml, p < 0.05) in patients with fibrosis (stage 1-4) than snnple steatosis. Tbere was a trend towards higher serum levels of hyalurunic acid in the fibrosis group (61 vs. 36.3 U/ml, p=0.06) but this did not reach statistical significarme with currant sample size. However, when we compared simple steatosts to stage 2 to 4 librosis, both serum markers were sigulficandy higher in the fibrosis group. A mcewer operator cupee (ROC) curve for serum YKL-40 revealed a sensitivity of 75% and a specihcity of 90% in detecnng fibrosis stage greater than or equal to Stage 2 when cutofl concentration was 116 ng/mh Hyaluronic acid had sensitivity of 50% and specificity ot 95% in detecting patients with fibrosis stage greater than or equal to Stage 2 when cntoft concentration was 84.6 U/mL When two serum fibrosis markers are combined to detect hbrosLs stage greater than or equal to 2, the sensitivity was 100% and specificity was 50%. CONCLUSIONS: Serum YKL-40 and HA are useful markers m differentiating patients with simple steatosis ti'mn NASH patients with fibrosis stage greater than or equal to 2. in asmciation with clinical parameters, they may identify suitable patients fur liver biopsy.

M1377

Undetected Flepatomegaly in Obese Children by Primary Care Physicians: A Pitfall in the Diagnosis of Pediatric NAFLD Mark Fishbein, Chris Mogren, Jennifer Mogren, Ryan ,Jennings

Intruducuon: Non-alcoholic tally liver disease (NAFLD) is the most common occurring liver disease in American adults, in obese children, NAFLD is also likely to be prominent. There ate no current recommendations tbr the prunary care physician regarding the evaluation of NAFLD in children. The diagnosis usually relies on the detection of hepatomegaly or the identification of abnormal serum liver chemistries. In this investigation, we assess the primary care physician's aptitude for detecting hepatomegaly by abdominal examination in obese children and requesting the appropriate tests to pursue an evaluation for NAFLD. Methods: Eighteen pnmar?" care physicians (6 pediatricians, 6 Ihmily practitioners, 6 internists/pediatri- cians) were enlisted as 'examiners" thr the study. In order to blind these indiwduals, they were instructed that the purpose of the study was to "compare physical examination findings m obese children among pnrnary care physicians" Eleven (9M, 2F) obese children (BMI > 95th %) ages 8 to 17 yv, were selected as study participants. Two children, evaluated previously by a pediatric gastroenterologist, possessing hepatmnegaly (liver edge > 2 cm below right costal margin), elevated serum arranotransferases and t~atty liver (established by MR1) were inchided in this group Each child was situated in a private examination room equipped with stethoscope and table Examination findings were recorded on a standard school physical torm that contained patient identifier and vital signs. A questionnaire regard- ing luether evaluation and consultation was also included Examiners were allotted five to ten minutes per child and then rotated to the next station Tire presence or absence of bepatomegaly was then confirmed on all children by tire pediatric gastroenterologist. Results: Four children displayed hepatomegaly (two identified previously + two identified presently). Hepatomegaly was identified by 1 of 18 (55%) exarmners in a single child. Serum liver chemistries ~ r e requested by 5 of 18 (27.8%) examiners tbr at least one child with hepatomegaly None ot the examiners requested imaging studies of the liver. Conclusion: Approximately 75% of primary care' physmians were unable to detect hepatomegaly or request the appropriate diagnostic studies necessary to pursue an evaluation of NAFLD in the pediatric population. Therdhre, the diagnosis of NAFLD is likely to be delayed, perhaps until adulthood, in most att~,cted children

M1378

TNFa Inhibits Expression of Betaine-homocysteine Methyl Transferase, Cystathionine ~-Synthase and Methionine Synthase in Murine Primary Hepatoc~tes Cheng Ji, Nell Kaplowitz

BACKGROUND/,a, IM: We pmvmusly examined the expression of 1200 toxicology-related genes over time in the routine model of intragasmc ethanol teeding and demonstrated that ttm npregulation of gera's was associated with endoplasmic teticulum (ER) stress response and that there was a link between ethanol-induced elevation of homocysteine, ER stress,

and alcoholic liver injury (Alcoholism 26(5): 138A, 2002). Since increased TNF~ production by Kupffer cells has been reported b':," others to play important role in alcohol-induced liver injury in mice, we are interested in studying the relationship between TNFor and homocysteine metabolisnr. METHODS: Primary" hepatocytes fi'om C57 BU6 mice were used for the experi- ment. The hepatocytes were treated with TNF,~ (1,000 U/mL or 5,000 U/mL), homocysteine (5 mM), and TNFa plus homocysteine in the presence or absence of antioxidams (vitamin E and bntylated hydroxytohiene) for 20 hours, mRNA for betaine-homocysteine methyl transferase (BHMT), cystationine [3-synthase (CBS) and methioinne symhase (MS) was ana- lyzed by RT-PCR and quantified with the Phosphorlmage Intracellular levels of homocysteine were determined using the IMx System RESULTS: TNF treatment at lower dose and high dose respectively reduced BHMT expression by 34% and 57%, CBS expression by 25% and 41%, and MS expression by 21% and 33%. Homocysteine treatment had little dfect on the expression of BHMT, CBS or MS Intraceflular homocysteine levels *','ere greater in TNF treated cells (2.9 _+ 1.2 nmol/mg protein at low dose; 4.1 _+ 1.4 nmol/mg protein at high dose) than in control (1.5 _+ 1.1 nmol/mg protein) Similar results could be obtained in the presence or absence of antioxidant treatment indicating that TNF-induced oxidative stress does not cause this effect. CONCLUSIONS: These results suggest that disturbance of homocysteine metabolism by TNFe~ might lead to the devation of homocysteine in the bepatocytes.

M1379

The Impact of Anti-Diabetic Medications on the Gene Expression of Diabetics with Non-Alcoholic Fatty Liver Disease (NAFL) Karen Schlauch, Janus P. Ong, Luca Del Gaicco, Francesco Goretta, Gen'i Grant, Hazem Elariny, Abraham Younoszai, Zachary Goodman, Alan Christensen, Vikas Chandhoke, Zobair M. Younossi

Treatment of type-2 diabetes (DM) may' aftect the pattern of gene expression and natural history of patients with NAFL. Aim: To determine the rmpact of anti-diabetm nmdications on the pattern of differential gene expression in NAFL Methods: Our NAFL database was used to identify" patients with biopsy-proven NAFL tor whom a liver specimen was snap- frozen at the time of biopsy' From each sample, total RNA was exu~cted, amplified and reverse transcribed into cDNA probes with aminoallyl nucleotides and coupled with fluores- cent cyanine 3 (Cy3)- or cyanine 5 (Cy5) dyes. Customized microarray gene chips were developed containing 5297 relevant genes, cDNA probes were hybridized with the chip at 42 C and scanned using GSI Lumonics Scan Array 5000. Each sample was run in triplicate and genes with statistically significant up- or down-regulation of more than 2-tbld were confirmed using real-time RT-PCR. Additionally', for each patient, extensive chmcal data, history of DM, currem medications dncluding insulin, mettormin and thiazolidmediones (Rosiglitazone and Piogfitazone)~ were available. All other forms of liver disease, had been excluded and a single pathologist had reviewed all the slides. Results: Gene expression data was available for 16 diabetic patients with NAFL Of these, 13 were requinng medical treatment and 3 *vere not taking any medication. In comparison to diabetic NAFL patients who were not taking a W medications, diabetic NAFL patients on thiazofidinediones displayed 113 genes which were significantly- up-regulated by more than 2-fold (p<0.05) (ghitathione S-transferases, apolipoprotein, ubNuitin-conjugating enzymes, fatty-acid binding protein, akohol dehydrogenase) and 26 genes that were significantly down-regulated by" more than 2-fold (p<O.05) (collagens, cTtochrome P450). The same comparison for those taking metformin indicated 261 genes that were significantly up-regulated (apoltpoproteins, alcohol dehydrogenases, insulin receptor substrate 1, glutathione S-transferases, ubiquinn-conjugat- ing enzymes, fatty acid binding protein 4, leptin receptor), and 70 genes that were significantly down-regulated (cytochrome P450, collagen type VII, alpha 1, ATP synthase). Conclusions: Anti-diabetic medication may cause change the hepatic gene expressions, potentially" affecting the progressive course of NAFL

M1380

Prevalence and Predictive Factors of Non Alcoholic Steatohepatitis (NASH) in Obese Patients Undergoing Bariatrie Surgery: a Prospective Study in 92 Patients Florence Harnois, Simon Msika, Jeanine Barge, Jean Marc Sabate, Eric BIezel, Jean Pierre Farmachidi, Benoit Coffin

Obesity has been identified as a risk factor of NASH, its prevalence in obese patients varies from 0 to 25% in previous studies Aims: to determine the prevalence of NASH and extensive fibrosis in a prospecnve cohort of panents with morbid obesity (BMI>40 kg/m2 or >35 kg/m2 in association with comorbidity) requiring bariatric surgery (ring gastmplasty or gastric bypass) and to identi~y predictive tactors of NASH. Methods: From 07/01 to 09/02, 96 consecutive patients underwent fiver biopsy during hariatric surgery, 4 were excluded. The diagnosis of NASH was established on steatosis associated to at least 2 of the following criteria: lobular necrotic and inflammatory tbci, ballooning degeneration with or without Mallory bodies and periceUular fibrosis (Lee R, Hepatology 1995; 21:1742-3). Climcal and biochemical data were analysed by univariate and multivariate analysis. Results (mean+/- 5EM): 92 patients (85 women, age :38 +/- II years) were analDed. Mean BMI was 45.7 +/- 5.1 kg/m2. Viral C and B serologies were negative Alcohol intake was <20 g/d. Arterial hypertension was observed in 20 patients (21.7%) and a melfitus diabetes in 11 (11.9%). Steatosis was observed in 90 patients (97.8%), lobular inflammation and necrosis in 17 (18.5%). Steatosts associated with lobular necrotic and inflammatory loci and ballooning degeneration or pericellular fibrosis was noticed in 9 patients (9.8%). Portal fibrosis was observed in 9 patients: 2 had NASH, I a periceflular fibrosis w4thout inflammatory loci and 6 inflammatory or necrotic lesions. No cirrhosis or extensive fibrosis was found. The overall prevalence of NASH in this population was 9.8% Univariate analysis identified 3 predictive tactors of NASH: total cholesterol (p = 0.046), triglycerides (p = 0.03) and BMI (p = 0.003). Only BMI was an independent factor in multivariate analysis. Conclusion: In this prospective cohort of obese patients requirirtg bariatric surgery, the prevalence of NASH was 98%, no patient had cirrhosis or extensive fibrosis but patients were relatively young. Only BMI was a predictive factor of NASH

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