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smanner with significant inhibition noted at 5.0 and 10.0 μM. 18β-GA washout restored anormal rate of cell migration. No inhibition of migration was noted between 0.625 to 2.5μM despite reaching near complete inhibition (90-95%) of GJIC at these concentrations.The Cx-43 polypeptide exists in three isoforms under control conditions: two predominantphosphorylated forms (P2 and P1) and one minor non-phosphorylated form (P0). 18β-GAshifts the phosphorylation state of Cx-43 from nearly equal amounts of P2 and P1 to apredominantly P0 isoform at concentrations between 0.625 to 2.5 μM. Confocal microscopyrevealed that Cx-43 is not present at the leading-edge lamellipodia extensions of migratingcells, but localizes primarily to the retracting edge at points of contact between cells andalong lines of cortical actin filaments. Conclusions: Although a correlation exists betweenGJIC and Cx-43 phosphorylation state, GJIC and maintenance of the P2 isoform are notrequired for IEC-6 migration. Based on the specific subcellular localization of Cx-43 inmigrating cells, we speculate that regulation of actin cytoskeleton dynamics may be a novelfunction for gap junction proteins and might regulate epithelial cell migration In Vivo alongthe intestinal villous. (Supported by an Intramural Research Grant for New Faculty fromMichigan State University).

M1703

Intestinal Permeability and Bacterial Translocation Are Modulated By theGenetic Background of the HostSamuel M. Alaish, Jose Greenspon, Daniel Eyvazzadeh, Terez Shea-Donohue, Alan Cross,Alessio Fasano, James P. Nataro

Failure of the intestinal barrier is a characteristic feature of cholestasis, as bacteria from theintestinal lumen are believed to enter the circulation and initiate a systemic inflammatoryresponse, which can lead to impairment of multiple organs. A person's genetic backgroundcan affect the immune and inflammatory regulation of their intestinal barrier to commensalorganisms. Indeed, this has been shown in some familial cases of Crohn's disease. We havepreviously observed disparate mortalities in two different strains of inbred mice followingcommon bile duct ligation. We hypothesized that the genetic background of the hostmodulates the intestinal barrier following cholestasis. Two different strains of male inbredmice, A/J and C57BL/6J, were subjected to common bile duct ligation. Sham-operatedanimals served as controls. Following one week of ligation, jejunal intestinal permeabilitywas measured by trans-epithelial electrical resistance (TEER). Bacterial translocation tomesenteric lymph nodes, liver, spleen and blood was also studied in these same strainsfollowing three weeks of common bile duct ligation. Jejunal TEER was significantly reducedin each ligated strain compared to their sham counterpart (p<0.05, Student's t-test); jejunalTEER was also significantly lower in both sham and ligated C57BL/6J compared to shamand ligated A/J mice, respectively. In agreement with the TEER data, bacterial translocationto mesenteric lymph nodes was significantly increased in C57BL/6J mice (67% of mice)versus A/J mice (24%), (p=0.03, Fisher's exact test). There was also a marked disparity inpositive blood cultures. Whereas 4 (27%) of 15 C57BL/6J mice were bacteremic, none ofthe 17 A/J mice were (p<0.05, Fisher's exact test). No significant differences were found inliver or spleen cultures. Our study demonstrates that mice of different genetic backgroundsexhibit different intestinal barrier resistance. Following common bile duct ligation, differencesin both TEER and bacterial translocation were found between two strains of inbred mice.Expanding on our previous observation of higher mortality in ligated C57BL/6J mice com-pared to A/J mice, these differences support the notion that failure of the intestinal barriermay be genetically predisposed in some cholestatic patients, resulting in increased episodesof sepsis and higher mortality. Further experiments are warranted to clarify the underlyingmechanisms and identify the genes responsible.

M1704

Changes in the Expression of Claudins in Active Ulcerative ColitisTadayuki Oshima, Hiroto Miwa, Takayuki Matsumoto, Takashi Joh

Background and Aims: Epithelial barrier function is impaired in ulcerative colitis (UC),but the pathophysiological mechanisms leading to this barrier defect are still far from clear.Because epithelial barrier function is primarily regulated by the most apical intercellularjunction, referred to as the tight junction (TJ), we investigated the expression of TJ proteinson rectal epithelial mucosa in UC. Methods: Biopsies from the rectum of patients withactive UC and of controls were studied. Tight-junction proteins were dual stained usingspecific antibodies (claudin-1, -2, -3, -4, and -7) as primary antibodies, and Cy-3 conjugatedanti-rabbit IgG and Alexa488-conjugated anti-mouse IgG as secondary antibodies. Sampleswere analyzed by immunofluorescence microscopy. Immunoblotting and real-time poly-merase chain reaction were performed to quantify TJ proteins and mRNA, respectively. Thestudy design was approved by the local ethics committee. The subject gave informed consentand patient anonymity was preserved. Results: At tight junctions claudin-4 and -7 stainingwas down-regulated in active UC, whereas claudin-2 staining was up-regulated. Claudin-4and -7 proteins were significantly down-regulated on immunoblotting (32 ± 19 and 51 ±10 % of control respectively, p < 0.001), whereas claudin-2 protein was up-regulated inactive UC (11491 ± 327 % of control, p < 0.001). Claudin-1 and -3 expression levels wereunchanged in controls and active UC. In active UC claudin-2 mRNA was significantlyincreased (6.2 ± 1.2 times of control, p < 0.001), whereas claudin-4 and -7 mRNA weresignificantly decreased (0.45 ± 0.13 and 0.3 ± 0.08 times of control respectively, p < 0.005).Conclusions: Down-regulation of claudin-4 and claudin-7, and up-regulation of claudin-2, might lead to altered TJ structure and be related to the impaired epithelial function inactive UC.

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M1705

Bifidobacterium Treatment Improves Intestinal Adherens Junction Structure inExperimental Necrotizing EnterocolitisKaterina Dvorak, Ludmila Khailova, Kelly M. Arganbright, Toshi Kinouchi, BohuslavDvorak

Background: Neonatal necrotizing enterocolitis (NEC) is a major cause of morbidity andmortality in premature infants. Adherens junctions (AJs) are cellular connections anchoringcells one to another and attaching to components of the intracellular matrix. AJs are composedof transmembrane and cytosolic components - cadherins and catenins. Previously, we haveshown that oral administration of probiotics normalizes intestinal tight junction structurein an experimental model of NEC. Objective: The aim of this study was to evaluate theeffect of bifidobacterium treatment on formation of intestinal adherens junctions in a ratmodel of NEC. Methods: Premature rats were divided into three groups: dam fed (DF),hand-fed with formula (NEC), or hand-fed with formula supplemented with 5 x 106 CFUBifidobacterium bifidum OLB6378 per day (BB). All groups were exposed to asphyxia/coldstress to develop NEC. Incidence of NEC, quantification and localization of E-cadherin, α-and β-catenin were evaluated in the site of injury - the terminal ileum. Results: Treatmentwith Bifidobacterium bifidum significantly lowered the incidence of NEC in neonatal ratmodel. In the ileum of DF and BB rats, E-cadherin staining was localized mainly in thecrypt enterocytes. Staining for α-catenin exhibited diffused signal in interocytes of the NECgroup, whereas localization of this protein in the DF and BB group was clearly associatedwith the membrane. β-catenin signal was significantly reduced in the NEC group, whereasthe DF and BB groups exhibited strong membrane-associated signal. Conclusions: AJs playan important role in organogenesis of epithelial tissues. Abnormal histological localizationof AJs proteins in the ileum of NEC animals suggests irregularity in the formation of epithelialintracellular structures during NEC pathogenesis. Treatment with B. bifidum OLB6378normalizes these changes. Supported by the NIH Grant HD-39657 (to B.D.) and a gift fromMeiji Dairies Corporation.

M1706

Basolateral Potassium Channels Play a Key Role in the Forskolin-Induced ButNot in the Carbachol-Induced Reduction in Colonic Epithelial ResistanceJenny Gustafsson, Sara K. Lindén, Henrik Sjovall

Background: Ulcerative colitis is a common inflammatory bowel disorder that may be dueto a defective colonic barrier function. Tight junctions between cells play a key role inbarrier maintenance. In human colonic biopsies, there is a drop in paracellular electricalresistance during stimulation of secretion with carbachol, a response that was enhanced inpatients with ulcerative colitis in remission (1). We therefore proceeded to determine themechanism behind the secretagogue-induced resistance drop, in mouse colon In Vitro.Materials and methods: Segments of mouse distal colon were mounted in miniaturizedUssing chambers (exposed area 0.45cm2). Tight junction resistance (Rp) and membranecurrent (Im)were recorded by square wave current analysis. Chloride secretionwas stimulatedby serosal addition of either forskolin (10μM) which activates the cAMP-CFTR system,or with carbachol (1mM) which acts mainly by opening Ca2+ gated potassium channels.Mechanisms behind Rp-effects were studied by treating the tissue with bumetanide (0.1mM,blocks basolateral Na-K-Cl co-transport) or BaCl2 (5mM, blocks several types of basolateralpotassium channels), prior to addition of secretagogues. Results: Forskolin and carbacholboth induced a significant decrease in Rp (forskolin: ΔRp -19.3±2.6Ω*cm2 p<0.001; carba-chol: ΔRp -9.9±0.8Ω*cm2 p<0.001) and stimulated chloride secretion (Im: forskolinΔ307±38μA/cm2 p<0.001; carbachol Δ222±21μA/cm2 p<0.001). As a control experiment,the response to forskolin was also reproduced in cultured colonocytes (CaCo2-cells, n=4experiments). In mouse colon, pretreatment with bumetanide had no significant effect onthe Rp response (forskolin: Δ-19.3±2.6 to Δ-21.9±2.4Ω*cm2 n.s; carbachol: -9.9±0.8 to-6.4±1.5 Ω*cm2 n.s) but reduced both the forskolin and carbachol effects on Im (forskolin:Δ307±38 to Δ106±20μA/cm2 p<0.001; carbachol: Δ222±21 to Δ50±14μA/cm2 p<0.001).In contrast, pretreatment with Ba2+ reduced the forskolin induced effects on Rp (Δ-19.3±2.6to Δ-5.4±2.0Ω*cm2 p<0.001) and Im (Δ307±38 to 63±8μA/cm2 p<0.001) but lacked effecton the carbachol response (Rp and Im). Conclusions: Activation of both cAMP-mediatedand Ca2+-mediated secretion is accompanied by a reduction in epithelial resistance. Ba2+

sensitive K+ channels play a central role in the cAMP-mediated but not in the Ca2+-mediatedresistance response. The response can occur independently of effects on membrane current.1. Gustafsson J, Eklund L, Stage L and Sjövall H, Cholinergic activation alter epithelialresistance and the response is increased in sigmoid colon of patients with Ulcerative colitisin remission, Gastroenterology (2007) vol. 132, no. 4, suppl 1

M1707

Infections At First Decompensation of Liver Cirrhosis Predict HigherMortality At Hospital Admission and At Long TermCarlos Noronha Ferreira, Teresa Rodrigues, Helena Cortez-Pinto, Fatima Serejo, FernandoRamalho, Jose Velosa

Background: Patients with liver dysfunction are prone to infection which is frequently aprecipitant of upper gastrointestinal bleeding, encephalopathy and renal failure. Bacterialinfections in patients with cirrhosis are poorly tolerated and responsible for 30-50% ofdeaths. Aims and methods: To determine the significance of community acquired andnosocomial infections and the effect on mortality at hospital admission, at 1 and 3 years inpatients hospitalized with first episode of decompensation of liver cirrhosis. The records of297 consecutive patients were analyzed. Patients were followed up till death or study closuredate on 31st August 2007. Exploratory analysis, Fishers Tests, Logistic regression, KaplanMeier Survival Analysis with Log Rank Test, Time-dependent Cox model and ProportionalHazards Cox regression analysis were used. (SPSS 16). Results: Mean age - 57 ± 13.7 years,male sex - 78.8% (234/301). Child Pugh (CP) score - 9.5 ± 2.4, MELD score - 16.2 ± 7.Median follow up- 12.7 months (IQR - 29.3) Community infection (≤ 48h admission) -24.6% (69/280) and nosocomial infection (> 48h admission) - 15.4% (43/236). Community

and / or nosocomial infection were more frequent in Child Pugh score ≥ 10 (OR 5.72, p=0.002, 95% CI 1.9 - 17), MELD score ≥ 15 (OR 2.1, p = 0.006, 95% CI 1.24 - 3.58), andHDL cholesterol < 30mg/dL (OR 2.98, p = 0.001, 95% CI 1.52 - 5.84). The age, sex, etiologyof liver cirrhosis, type of decompensation, serum sodium < 130mEq/L and total cholesterol< 100mg/dL were poor predictors of infection. Mortality rates: First admission - 14.3% (43/301), at 1 year - 39.8% (100/251) and at 3 years - 58.1% (91/217). Kaplan Meier survivalanalysis showed decreased long term survival in patients with infections at first admissionfor decompensated liver cirrhosis (p <0.001). Conclusions:Community and nosocomialinfections have a high incidence at first episode of decompensation of liver cirrhosis andare independent predictors of mortality at hospital admission, at 1 and 3 years. These patientsmay benefit from intensive treatment and early referral for liver transplantation.

OR = Odds ratio, HR = Adjusted Hazards Ratio, p = p value, CI = Confidence interval

M1708

Liver Transplantation for Hepatic Sarcoidosis: Long Term Follow-Up andRecurrence After Liver Transplantation, a Single Center ExperienceMuhammad Bilal, Mankanwal S. Sachdev, Mohammad K. Ismail

AIM:Hepatic sarcoidosis as a pure indication for orthotopic liver transplantation (OLT)remains a rare entity. Consequently, the studies evaluating this subset of patients and theirlong term course are scarce.The aim of our study was to report our single center experiencewith hepatic sarcoidosis patients undergoing OLT while evaluating the pre-operative charac-teristics, post-op long term follow up and complications in such patients. METHODS:Aretrospective chart review of 760 patients who underwent liver transplant (TX) betweenJanuary 1993 and October 2008 at the University of Tennessee, Memphis, was undertaken.10 patients (1.3%) were identified who received OLT purely for hepatic sarcoidosis. Thediagnosis of sarcoidosis was established by the presence of characteristic clinical findingsand presence of non-caseating granulomas in the native liver explants. Graft and patientsurvival rates were compared using Kaplan-Meier Analysis.RESULTS: The median age was46.5 yrs (29 - 61 yrs), and male to female ratio was 3:7. 8 pts were African-American and2 were Caucasian. All pts presented with liver as a primary site of manifestation of sarcoidosis.Extrahepatic manifestations were present in 6 pts (5 with pulmonary disease, 2 with hypercal-cemia, 1 with cutaneous, 1 with ophthalmologic and 1 with hepatic artery arteritis). Liverenzymes typically showed a cholestatic pattern with median total bilirubin of 10.7 mg/dland median alkaline phosphatase of 375 IU/L. All 10 were transplanted, and at present 8pts remain alive with a median post-TX follow-up of 3.2 yrs (0.1-15 yrs). One patientexpired after 6 yrs post transplant, due to multi-organ failure. The second patient died 1month post-op from immediate post-OLT complications. Of the 8 who are still alive, post-transplant complications have been noted in 2 pts. Both of them developed end stage renaldisease (ESRD) due to chronic tacrolimus toxicity requiring hemodialysis. One pt developedhypercalcemia-related pancreatitis. Recurrence of hepatic sarcoidosis was found in 1 pt at4 months of transplant. Recurrence was characterized by an increase in alkaline phosphatase>400 IU/L and GGT >1000 IU/L. Liver biopsy showed multiple non-caseating granulomasin portal tract and hepatic parenchyma. Staining was negative for fungal and acid fastorganism. Despite recurrence, graft function remained stable. Of note, allograft did not haveevidence of sarcoidosis on biopsy at time of donation. CONCLUSIONS: Advanced liverdisease secondary to sarcoidosis is rare. OLT is an effective treatment for hepatic sarcoidosiswith excellent long-term graft and pt survival.

M1709

Low Total Cholesterol Levels At First Decompensation of Liver Cirrhosis AreAssociated with Advanced Liver Disease and Predict Higher Mortality AtHospital Admission and At 1 YearCarlos Noronha Ferreira, Teresa Rodrigues, Helena Cortez-Pinto, Fatima Serejo, FernandoRamalho, Jose Velosa

Background: The liver has a central role in the metabolism of lipoproteins. Recent studiesindicate that low total and HDL cholesterol levels may reflect deteriorating liver functionand thus be used as indicators of liver function and prognosis. Aims and methods: Todetermine the significance of low total and HDL cholesterol with regard to etiology, severityand type of decompensation of liver cirrhosis and influence on in-hospital mortality andmortality at 1 and 3 years. The records of 297 consecutive patients admitted with the firstdecompensation of liver cirrhosis were analyzed. Statistical analysis with Kruskal Wallis test,Mann Whitney U and Logistic regression Kaplan Meier Survival Analysis with Log RankTest, Time-dependent Cox model and Proportional Hazards Cox regression analysis (SPSS16).was performed. Results: Average age - 57 ± 13.7 years, male sex - 79% (239). Medianfollow - up - 12.7 months (IQR - 29.3). Mean total cholesterol - 129.6 ± 54.4 mg/dL, andHDL cholesterol 30.5 ± 18.9 mg /dL. Total and HDL cholesterol were significantly lowerin Child Pugh class C (p< 0.001; p< 0.001), MELD score ≥15 (p < 0.04; p < 0.0005),creatinine ≥ 1.3 mg/dL (p < 0.012: p < 0.0045). There was no association between lowertotal and HDL cholesterol and age ≥ 50 years, etiology of cirrhosis, type of decompensationand serum sodium < 130meq/L. Mortality rates: At admission - 13.5% (40/297), at 1 year- 39.8% (100/251) and at 3 years - 58.1% (91/217). Mortality at first admission wassignificantly higher in patients with total cholesterol < 100mg/dL (adjusted OR 2.8, p =

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0.005, 95% C.I. 1.37 - 5.73) and HDL cholesterol < 30mg/dL (adjusted OR 2.74, p = 0.037,95% C.I. 1.06 - 7.06). At 1 year, only total cholesterol < 100 mg/dL (adjusted HR 2.74,p = 0.005, 95% C.I. 1.37 - 5.73) predicted higher mortality. Total cholesterol < 100mg/dLand HDL cholesterol < 30mg /dL were poor predictors of mortality at 3 years. Kaplan Meieranalysis revealed that total cholesterol < 100mg /dL (p = 0.048) was associated with poorlong-term survival. Conclusions: Total and HDL cholesterol levels are significantly lower inadvanced liver cirrhosis and renal insufficiency. At first hospital admission for decompensatedliver cirrhosis, total cholesterol < 100mg/dL is an independent predictor of in-hospitalmortality and mortality at 1 year, while HDL cholesterol < 30mg/dL only predicts in- hospitalmortality. Prognostic models including these two factors may identify high risk patientswho could benefit from intensive care and early referral for liver transplantation.

M1710

Predictors of Short and Long Term Mortality At First Decompensation of LiverCirrhosis. Total Cholesterol Levels and Infections Influence MortalityCarlos Noronha Ferreira, Teresa Rodrigues, Helena Cortez-Pinto, Fatima Serejo, FernandoRamalho, Jose Velosa

Introduction: Survival in patients with decompensated liver cirrhosis depends on the severityof liver insufficiency and complications of portal hypertension. Aims and Methods: Toidentify factors influencing mortality at 1 and 3 years after first decompensation of livercirrhosis.The case records of 297 consecutive patients admitted with first decompensationof liver cirrhosis were analyzed. Exploratory analysis, Kaplan Meier survival analysis withLog Rank Test, Time-dependent Coxmodel and Proportional Hazards Cox regression analysiswere used.(SPSS 16). Results: Average age - 57± 13.7 years, male sex - 78.8% (234). ChildPugh (CP) score - 9.5 ± 2.4 and MELD score - 16.2 ± 7. Median follow up- 12.7 months(IQR - 29.3). Mortality rates: In-hospital mortality - 13.5% (40/297), at 1 year - 39.8%(100/251) and at 3 years - 58.1% (91/217). By proportional hazard Cox regression analysis,CP score ≥ 10, MELD score ≥ 15, hepatocellular carcinoma, serum creatinine > 1.3mg/dL, serum sodium < 130mEq/L, Total cholesterol < 100mg/dL, community and noscomialinfections predicted higher mortality. The age, sex, aetiology and type of decompensationof liver cirrhosis, HDL cholesterol < 30mg/dL and alcoholic abstinence were poor predictorsof mortality. Kaplan Meier survival analysis identified age < 50 years (p = 0.001), CP classA (p < 0.001), MELD score < 15 (p < 0.001), upper GI hemorrhage (p = 0.002), totalcholesterol > 100 mg/dL (p = 0.048), serum creatinine < 1.3mg/dL (p < 0.001), < 2 hospitalre-admissions (p < 0.001) as being associated with better long term survival. A survivalbenefit of alcoholic abstinence (p = 0.04) was observed only in patients in CP class C (p =0.014). Conclusions: Community and nosocomial infections are independent predictors ofmortality at 1 and 3 years. Total cholesterol < 100mg/dL predicts higher mortality at 1 year.The combination of these two factors with commonly used prognostic models could identifya sub-group of patients at higher risk for mortality thus justifying early referral to a trans-plant centre.Predictive factors of mortality at 1 and 3 years.

HR -Adjusted Hazards Ratio, p = p value, CI = Confidence interval

M1711

Are the Characteristics of Patients with Cirrhosis and DecompensationChanging? a Study Comparing Two Time PeriodsRania N. Rabie, Eberhard Renner, Florence Wong

Introduction: It appears that the characteristics of patients with decompensated cirrhosisare changing. Aim: To assess whether there has been a change in the demographics, etiologicfactors, and clinical features of patients with decompensated cirrhosis over the span of adecade. Methods: Consecutive patients with decompensated cirrhosis enrolled from thestart of the decade (Jan 1st - Dec 31st, 2000; n=50) were compared to consecutive patientsenrolled from this year (Jan 1st 2008 - present; n=50). Baseline demographics, etiologicfactors, mode(s) of decompensation, severity of liver disease (MELD, Child-Pugh scores)and clinical and biochemical parameters were analyzed. Patients with a history of livertransplantation or an active malignancy, including hepatocellular carcinoma, were excluded.Results: Significant changes were observed in the etiology of liver disease with more patientshaving alcoholic cirrhosis and less having viral related cirrhosis in the present cohort. Patientsin the previous cohort had more severe liver disease at enrollment as reflected by significantlyhigher Child-Pugh and MELD scores with a significantly lower albumin and a trend towardsmore hepatic encephalopathy. All other components of the Child-Pugh and MELD scoreswere similar between both groups. No significant differences were observed in gender, ageor mode of first decompensation, or the presence of specific complications of cirrhosis(baseline grade of ascites or coagulopathy, history of variceal bleeding or spontaneousbacterial peritonitis) (Table 1). Conclusions: Over the course of 8 years, there has been ashift in the etiology of liver cirrhosis with alcohol now surpassing viral etiologies. This mayreflect more widespread and improved treatment of viral hepatitis. Patients in the present

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