Search

BACKGROUND/AIMS: Intercross studies in inbred mice susceptible or resistant to liver fibrosis revealed complement factor 5 as a quantitative trait gene associated with the development of fibrosis. In 277 patients with hepatitis C, two C5 SNPs, rs17611 and rs2300929, have been associated with advanced fibrosis. METHODS: We investigated the association of these C5 SNPs with advanced fibrosis in 1435 HCV infected patients and in 1003 patients with other liver diseases. We performed genotyping with melting curve analysis using fluorescence resonance energy transfer probes in the LightCycler. RESULTS: The defined high-risk genotypes (AA and TT) and alleles (A and T) were not associated with advanced fibrosis in HCV patients when Chi square testing and logistic regression analysis were applied (rs17611A 0.45 in F0-1 vs. 0.43 in F2-4, P=0.31; rs2300929T 0.91 F0-1 and 0.91 in F2-4, P=0.82). In the group of patients with liver diseases other than HCV we neither found an association of the C5 SNPs with advanced fibrosis nor an overrepresentation of the SNPs in patients with cirrhosis. CONCLUSIONS: We found no evidence that these C5 SNPs are genetic risk factors for the development of advanced fibrosis in chronic HCV infection or other chronic liver diseases.

Background/Aims Aim of our study was to analyze fibrosis progression after liver transplantation (OLT) in hepatitis C virus (HCV)-infected patients based on protocol liver biopsies and to identify risk factors, which may play a role in the development of severe fibrosis stages. Methods One hundred and eighty-three liver graft recipients who had a histological follow-up evaluation of 1 year after OLT were analyzed. Overall 1039 protocol liver biopsies were performed after 1-, 3-, 5-, 7- and 10 years and staged according to the Scheuer score. Results The fibrosis progression rate was not linear. The fibrosis scores were 1.2 after one, 1.7 after three, 1.9 after five, 2.1 after 7 and 2.2 after 10 years. The 39 recipients with fibrosis stages 3 or 4 in the 1-year biopsy had a significantly reduced survival rate, while fibrosis stage 0–2 indicated excellent survival. Independent risk factors for progression of fibrosis at 1 year were HCV genotype 1 and 4 ( P =0.01) and donor age>33 years ( P =0.01), whereas risk factors for development of cirrhosis (30/183 recipients (16%)) were donor age ( P =0.002) and multiple steroid pulses ( P =0.05). Conclusions These data provide information on the course of recurrent hepatitis C and may be helpful to individualize the treatment of transplanted patients.

Background/Aims Hepatic injury induced by ischemia/reperfusion following surgery, transplantation, or circulatory shock combined with resuscitation is a major clinical problem. Methods In this study, hypoxic and inflammatory conditions were mimicked by exposing human hepatocytes to N 2 (at 4 and 37°C) or to cytokines/endotoxin to investigate the potential protective effects of heme oxygenase-1 (HO-1). Incubation of human hepatocytes with single cytokines (IFN-γ, IL-1β, TNF-α) or LPS, as well as a combination of all four stimuli (CM, cytomix) caused a time-dependent HO-1 mRNA expression over 12h and a decline by 24h. In parallel, we observed a time-dependent membrane leakage for LDH and AST and a maximum HO-1 protein expression between 3–24h. Results Warm and cold hypoxia showed similar results in HO-1 mRNA and protein expression and the release of LDH and AST. CoPP, a potent HO-1 inducer, and bilirubin, a co-product of the HO-pathway, protected human hepatocytes from warm and cold hypoxia. HO-1 enzyme activity was highest during warm hypoxia, followed by cold hypoxia and CM which was confirmed by intracellular Fe 2+ formation. Conclusions Taken together, we demonstrated, that HO-1 induction protected human hepatocytes against warm and cold hypoxia. Our results also suggest that HO-1 induction may have therapeutic potential against inflammatory insults.

Background/Aims Graft reperfusion in liver transplantation is usually performed by initial portal reperfusion (IPR) and delayed rearterialization. Its influence on graft microcirculation is unknown. This study aimed to assess sinusoidal perfusion in dependence to this reperfusion technique during human living-donor liver transplantation. Methods Hepatic microcirculation was measured both in the donor and the recipient ( n =14) by using the orthogonal polarization spectral imaging technique. By using initial portal reperfusion, the mean time interval between portal venous and hepatic arterial reperfusion was 27.7±13.3 min. Results Hepatic nutritive perfusion, as given by the functional sinusoidal density and the volumetric blood flow, was found significantly decreased during portal reperfusion when compared to baseline. Rearterialization resulted in hyperperfusion of individual sinusoids at a decreased density of the sinusoidal network. Interestingly, the time interval between portal venous and hepatic arterial reperfusion significantly correlated with the changes of the liver grafts' microcirculation. Conclusions The study indicates graft microcirculatory dysfunction as a major determinant of postischemic liver injury. Moreover, microvascular impairment was significantly influenced by the interval between portal venous and hepatic arterial reperfusion, which suggests the reinforcement of the pathomechanism of injury involving hypoxia and rapid graft rewarming due to initial portal reperfusion.

Methods : Following liver transplantation, a 26-year old female suffered from primary non-function of the transplant. The patient was subsequently treated with a modular extracorporeal liver support concept until a suitable organ became available. A bioreactor was charged with human liver cells, obtained from a discarded cadaveric graft (470 g, viability: 60%). The bioreactor was integrated into an extracorporeal circuit with continuous single pass albumin dialysis and continuous veno-venuous hemodiafiltration for detoxification and fluid reduction. Results : Over the total system application time of 79 h, a significant reduction of the plasma levels of total bilirubin (21.1 mg/dl at start, 10.1 mg/dl at end of therapy) and ammonia (100 versus 22.7 μmol/l) was achieved. During treatment the patient's neurological status significantly improved from coma stage IV to I permitting extubation. Recovery of kidney function with a urine output of 1325 ml/24 h compared to 45 ml/24 h prior to system application, was noted. Over the treatment period, an improvement of coagulation status was observed. Adverse events were absent. Conclusions : This first successful clinical treatment of a patient with liver failure suggests that a modular approach combining both primary human liver cell bioreactor technology and detoxification methods is promising.

Background/Aims The results of liver transplantation for Budd–Chiari syndrome (BCS) are poorly known and the role and timing of the procedure are still controversial. The aim of this study was to investigate the results of transplantation for BCS, focusing on overall outcome, on prognostic factors and on the impact of the underlying disease. Methods An enquiry on 248 patients representing 84% of the patients transplanted for BCS in the European Liver Transplantation Registry between 1988 and 1999. Results Of the 248 patients, 70.4% were female and 29.6% male. The mean age was 35.7 years. The overall actuarial survival was 76% at 1 year, 71% at 5 years and 68% at 10 years. 77% of deaths occurred in the first 3 months: 47% were due to infection and multiple organ failure, and 18% to graft failure or hepatic artery thrombosis. Late mortality (>1 year) occurred in nine patients, due to BCS recurrence in four of them. The only pre-transplant predictors of mortality on multivariate analysis (Cox) were impaired renal function and a history of a shunt. Conclusions Liver transplantation for BCS is an effective treatment, irrespective of the underlying cause, and should be considered before renal failure occurs.

Ischemic preconditioning (IP) is known to protect hepatic tissue from ischemia-reperfusion injury. However, the mechanisms involved are not fully understood yet.Using intravital multifluorescence microscopy in the rat liver, we studied whether IP exerts its beneficial effect by modulating postischemic Kupffer cell activation, leukocyte-endothelial cell interaction, microvascular no-reflow, mitochondrial redox state, and, thus, tissue oxygenation.Portal triad cross-clamping (45 min) followed by reperfusion induced Kupffer cell activation, microvascular leukocyte adherence, sinusoidal perfusion failure (no-reflow) and alteration of mitochondrial redox state (tissue hypoxia) (P0.05). This resulted in liver dysfunction and parenchymal injury, as indicated by decreased bile flow and increased serum glutamate dehydrogenase (GLDH) levels (P0.05). IP (5 min ischemia and 30 min intermittent reperfusion) was capable to significantly reduce Kupffer cell activation (P0.05), which was associated with a slight attenuation of leukocyte adherence. Further, IP markedly ameliorated sinusoidal perfusion failure (P0.05), and, thereby, preserved adequate mitochondrial redox state (P0.05). As a consequence, IP prevented the decrease of bile flow (P0.05) and the increase in serum GLDH levels (P0.05).IP may exert its beneficial effects on hepatic ischemia-reperfusion injury by preserving mitochondrial redox state, which is guaranteed by the prevention of reperfusion-associated Kupffer cell activation and sinusoidal perfusion failure.

To the Editor:Therapeutic measurements to control hepatitis C reinfection afterorthotopic liver transplantation (OLT) still pose many problemsand remain unsatisfactory [1]. While hepatitis C viremiadecreases strongly during the anhepatic phase of OLT, initiationof viral replication does occur within hours or days [2,3]. At thisearly stage after OLT, i.e. once chronic HCV reinfection is estab-lished, the virologic response is often markedly reduced.Although specifically targeted antiviral therapeutic regimes forhepatitis C (STAT-C) are currently extensively studied, none ofthem have been tried in the post-OLT period treatment.However, as recently described by Ferenci et al. [4], high doseintravenous silibinin (Legalon SIL ) can express potent antiviralactivity. This effect was shown to be driven by direct inhibitionof the viral RNA polymerase NS5B [5,6].In this letter, we can report the first successful prevention ofHCV reinfection after OLT by the administration of silibinin(1400 mg/d). This drug was applied immediately after OLT bydaily infusions for 14 days.At the time of OLT, the 57-year-old male patient exhibited aMELD Score of 23, Child-Pugh stage C liver cirrhosis, and a25 mm hepatocellular carcinoma in the left lobe. HCV infection(genotype 3a) was first diagnosed in 1997 and three interferon-based treatment regimens – at last PegInterferon alpha and Riba-virin were given for 48 weeks 6 years before OLT – failed toinduce a sustained virologic response (SVR). The anhepatic phaseduring OLT surgical procedures lasted 61 min and postoperativecare transpired without complication after a short phase of renalinsufficiency and haemodialysis treatment on day 1 and 2.Immunosuppressive therapy included methyl-prednisolone,mycophenolatmofetil, and belatacept. Aminotransferase levelsreached normal values within 12 postoperative days. Bilirubinlevels rose to a maximum of 9.5 mg/dl 7 days after OLT andshowed a protracted mild elevation until 4 weeks later. Thepatient was discharged from the hospital 21 days after OLT.The levels of HCV RNA 3 months prior to OLT were rather low(17.800 IU/ml), and further declined significantly during theanhepatic phase. Silibinin infusions were started 8 h after OLT.At this time HCV RNA levels measured 182 IU/ml, and droppedagain to 127 IU/ml after 48 h (RNA levels measured directly afterhaemodialysis are not available). Already from day 3 onwards,HCV RNA levels were below

185 THE PROTECT STUDY: EVEROLIMUS MONOTHERAPY vs. CALCINEURIN INHIBITOR-BASED THERAPY RESULTS IN PROGRESSIVE RENAL FUNCTION BENEFIT OVER 35 MONTHS IN LIVER TRANSPLANT RECIPIENTS M. Sterneck, G. Kaiser, N. Richter, N. Heyne, F. Rauchfuss, P. Neuhaus, P. Schemmer, L. Fischer, E.M. Paulus, M. Mertens, H.J. Schlitt. University Medical Center Hamburg Eppendorf, Department of Hepatobiliary and Transplant Surgery, Hamburg, Essen University Hospital, Department of General, Visceral and Transplantation Surgery, Essen, Hannover Medical School (MHH), Clinic for General, Abdominal and Transplant Surgery, Hannover, University Hospital Tuebingen, Department of General, Visceral and Transplant Surgery, Tubingen, Jena University Hospital, Department of General, Abdominal and Vascular Surgery, Jena, Charite University Medical Center Berlin, Clinic for General, Adbominal and Transplant Surgery, Berlin, University Hospital Heidelberg, Department of General Visceral and Transplant Surgery, Heidelberg, Novartis Pharma, Nurnberg, University Hospital Regensburg, Department of Surgery, Regensburg, Germany E-mail: sterneck@uke.de

1304 TH2 CYTOKINE-MEDIATED BILIARY EPITHELIAL CELL (BEC) BARRIER DYSFUNCTION MAY CONTRIBUTE TO IMMUNOGLOBULIN G4 (IGG4) ASSOCIATED SCLEROSING CHOLANGITIS (IAC) T. Muller, C. Beutler, A. Hurtado Pico, E. Schott, A. Fischer, A. Pascher, P. Neuhaus, B. Wiedenmann, T. Berg. Medizinische Klinik m. S. Hepatologie und Gastroenterologie, Klinik fur Allgemein-, Visceralund Transplantationschirurgie, Charite – Universitatsmedizin Berlin, CVK, Berlin, Klinik und Poliklinik fur Gastroenterologie und Rheumatologie, Sektion Hepatologie, Universitatsklinikum Leipzig, Leipzig, Germany E-mail: tobiashp.mueller@charite.de

Reinfection of the graft with hepatitis B virus (HBV) and hepatitis delta virus (HDV) is a potential complication in patients undergoing orthotopic liver transplantation (OLT). Therefore, we added recombinant interferon-alpha (rIFNa) to the standard immunosuppressive regimen in 11 patients who received transplants following liver failure attributed to cirrhosis B (n = 10, with HDV co-infection in four cases) or fulminant hepatitis B (n = 1). Patients were treated with rIFNa for periods ranging from 2 to 3 months between the first and the 13th month after OLT. All patients received immunosuppressive treatment with low-dose corticosteroids, azathioprine and cyclosporine. Anti-HBs hyperimmune globulin was also administered. None of the patients showed evidence of severe allograft rejection. Seven patients suffered HBV reinfection of the graft with histological signs of acute hepatitis in five cases and transition to chronic hepatitis in one patient. Treatment with rIFNa did not prevent or reduce HBV replication. Reinfection of the graft with HDV was demonstrated by PCR in four patients co-infected with HDV. During treatment with rIFNa liver biopsy specimens from three reinfected patients were transiently negative for HDV antigen but not for HDV RNA, and the sera from two patients were transiently negative for HDV RNA. The data indicate that rIFNa can reduce HDV replication in reinfected liver allografts.

Introduction: Studies investigating impaired glucose tolerance or diabetes mellitus (DM) in transplantation mostly use fasting glucose for determination of DM. We prospectively analyzed the impact of fasting glucose in comparison to advanced methods in recipients and donors of livingdonor liver transplants (LD-LTX) for determination of impaired glucose tolerance. Methods: Recipients and donors without overt diabetes mellitus were investigated longitudinally before and on day 10, month 6 and month 12 after right lobectomy or liver transplantation of the right lobe. Insulin sensitivity (SI) was assessed by a computer-assisted analysis of a frequently sampled intravenous glucose tolerance test with 300 mg/kg BW glucose 50% (Konrad et al., 1999). (%cell responsiveness (first and second phase of pancreatic (i-cell secretion, Phi 1 and Phi 2) was determined by a c-peptide modeling analysis (SAAM 11 software). Conventional parameters (fasting glucose, insulin, c-peptide, HOMA-IR score) were correlated with modeling results. Results: Donors developed insulin resistance by day 10 (S1 2.65&0.41 10-4 min-1 pU ml-1 vs. 4.90&0.50 10-4 min-1 pU ml-1 in control, p < 0.01; Phi 2 increased which was normalized again by month 6 and 12. In contrast to healthy donors, recipients were insulin resistant prior to transplantation (SI 1.91+0.29 10-4 min-l pU ml-I, p i 0 . 0 I ) . There was no significant correlation between SI and fasting glucose, but significant correlations to fasting insulin, c-peptide, and HOMA-IR (p i 0.001). Discussion: Impaired glucose tolerance in cirrhosis was normalized by liver transplantation of the right lobe. Liver resection of the right lobe in donors leads to acute insulin resistance. Impaired glucose tolerance could not be predicted by fasting glucose. The reason for this is relative undernourishment of recipients before transplantation. Additional parameters such as fasting insulin, c-peptide, or HOMA-IR are necessary. Transplantution Surgery, Churit4, Berlin; ’Institute ,ft)r Mrtuholic

Patients with chronic liver disease are at higher risk of hepatitis B (HB) virus infection before and after liver transplantation, and theycommonly have a suboptimal immune response to HB vaccines. In this randomized trial, we compared the immunogenicity ofprimary vaccination with 2 doses of an experimental adjuvanted HB vaccine (adjuvant system 04 containing aluminium andmonophosphoryl lipid A [HB-AS04] ) to that of 3 double doses of a licensed HB vaccine in 93 liver transplant candidates. Dependingon the waiting list for liver transplantation, a booster dose of HB-AS04 or double booster dose of the licensed HB vaccine was givenbefore or after surgery, at 6 to 12 months after initiation of the vaccination course. The percentage of subjects with seroprotectiveanti-HB surface antibody concentrations 1 month after booster was twice as high in the HB-AS04 group (60.0%), vs. patients in thecomparator group (32.0%) (

Background: Members of the IGF-family regulate growth and development. IGF-II is a growth factor for HCC. Its expression, as detected by Northern analysis, is enhanced in cirrhotic nodules close to HCC. Methods: We studied the expression pattern of IGF-II RNA on 8 cirrhotic livers \HBV: n=4; HCV: n=3; alcoholic: n=2), using a double-labeled [3 S]-UTP/GTP riboprobe for IGF-II RNA. This was combined with immunostaining for the IGF-IIR (mab 2C2). Results: IGF-II RNA was expressed homogeneously over all hepatocytes of regenerative nodules, with weak signals over mesenchymal cells and no signal over bile duct epithelia. IGFIIR was found over all hepatocytes, but prominent over periportal hepatocytes in lobular zone I. Conclusions: Since comparable levels of IGF-II RNA are expressed over all hepatocytes and the IGF-IIR protein predominates on periportal hepatocytes, autoand paracrine stimulation of neoplastic growth in preneoplastic lesions of HCC appears to be mediated by an upregulated IGF-IIR. ENDbSCOPlC AND ULTRASOUND CRITERIA PREDICTING LESS DEVELOPMENT OF PORTALHYPERTENSIVE GASTROPATHY IN HEPATIC CIRRHOSIS C. Gheorahe. L. Gheorahe, S. Griaoruta, A. ODroiu, Center of Gastroenterology Fundeni Hospital, Bucharest, Romania