April 1995 AASLD A l 1 4 7

NATURAL ALFA-INTERFERON (na-IFN) THERAPY IN PATIENTS WITH CHRONIC HEPATITIS C NON-RESPONDERS TO RECOMBINANT ALFA-INTERFERON (ra-IFN). A. Picciotto, E Bardellini, P. Borro, M. Pireddu, N. Sinelli, G. Celle. DI.M.I. Gastroenterology Unit, University of Genoa, Italy.

AIM: Biochemical and virological evaluation of nct-IFN therapy in patients with chronic hepatitis C (CAH-C) who failed to respond tO rcx-IFN. MATERIALS AND METHODS: Thirty-six patients of both sexes (ages 20-65 yrs) with biopsy-proven CAH-C, non-responders to a first cycle of rcx-lFN (3 MU three times a week for 6 months) underwent a second cycle of nct-IFN at the same dose for 6 months. All patients were followed-up for 6 months after the end of the second cycle of n~x-IFN. ALT was tested every month until the end of follow-up. HCV-RNA was detected by "nested" PCR (5' non-coding region) in the patients' sera at baseline and after the end of both ra-IFN and na-IFN therapy. RESULTS: Six patients (16.6%) normalized ALT levels at the end of na-IFN therapy. However, all of them relapsed during follow-up period. ALT and HCV- RNA data are reported below.

r~x-IFN ncx-IFN

PTS BEFORE AFTER BEFORE AFTER ALT HCV-RNA ALT HCV-RNA ALT HCV-RNA ALT HCV-RNA

C.V. 145 POS 109 POS 93 POS 36 POS C.A. 195 POS 57 NEG 371 POS 40 NEG L.M. 324 POS 400 POS 136 NEG 10 NEG D.M. 168 POS 97 POS 93 NEG 39 NEG P.E. 307 POS 67 POS 248 POS 40 NEG C.S. 126 76 175 38

CONCLUSIONS: na-IFN can induce a biochemical and virological response in a part of patients non-responders to ra-IFN. Higher doses and/or a longer course of na-IFN may reduce the relapse rate occurring in these patients.

SYNTHETIC RAB3A EFFECTOR DOMAIN PEPT1DES STIMULATE INOSITOL 1,4,5-TRISPHOSPHATE PRODUCTION IN HEPATOCYTES. A. Piiper, D. Stryjek-Kaminska, J. Stein, S. Zeuzem. l)¢pamnmt of Medicine, University of Frankfurt a. M., Germany.

Small molecular weight GTP-binding proteins of the Rab family regulate vesicular transport within the cell. Peptides corresponding to the effector domain of Rab3A protein have been shown to stimulate exocytosis in various cells. However, the mechanism of Rab3 peptide- induced stimulation of exocytosis remained unknown. We have previously shown that a modified peptide corresponding to the effector domain of the small molecular weight GTP-binding protein Rab3A, Rab3A AL, stimulates inositol 1,4,5-trisphosphate (1,4,5-lP3)- production and amylase release in digitonin-permeabilized pancreatic aeini. In the present study we investigated the effect of Rab3 effector domain peptides on 1,4,5-IP3-production in isolated permeabilized hepatoeytes. Hepatocytes were isolated from rat by EDTA-perfusion method and 1,4,5-IP3-produetion in digitnnin-permeabilized cells was measured using a radioimmnno assay. In digitonin-permeabilized rat hepatocytes an effector domain peptide of Rab3A (7.5 nM) caused rapid accumulation of 1,4,5-IP 3. Rab3A effector domain peptide (7.5 taM) increased 1,4,5-IP 3 level from 2.34-1.5 to 6.8:1:1.8 pmol/mg protein. By contrast, a scrambled peptide of Rab3A had no effect demonstrating that the Rab3A effector domain response was specific. In intact cells Rab3A effector domain peptides had no effect. Similar responses were observed in digitonin-permeabilized Swiss 3T3 fibroblasts and SH-SY5Y neuroblastoma cells. These data suggest that Rab3 effector domain peptide-induced accumulation of 1,4,5-1P 3 is a widespread phenomenon, suggesting regulation of phosphoinositide- specific phospholipase C by Rab3-1ike proteins.

Supported by the Deutsche Forschungsgemeinsehafl (Ze 237/4-1)

TRANSJUGULAR APPROACH TO DIAGNOSIS AND TREATMENT OF LIVER DISEASES. A Pifiar, F Marcos, H Pallarts, F Ferngndez, M Hassan, JM Herrerfas, M Jimtnez. Dept of Gastroenterology, University of Seville, Spain.

INTRODUCTION: The development of the therapy for portal hypertension and liver failure is increasing the interest in the efficiency of the tecniques of transjugular access to the liver.

MATERIALS AND METHODS: We used suprahepatic cathe- terism using transjugular access in 44 patients (M/F 59%/41%, mean age 40.6, range 21-65).

RESULTS: VSH catheterism was possible in 42 out of 44 patients (95.1%) and it was not possible in 2. In 40 out of 42 cases (95.2%) a valorable hepatic biopsy was obtained, 58 % were a cylindrical shape and 37% were fragmented biopsies. Insufficient material was obtained from 2 patients, in one of them the flebographic study pointed to a diagnosis of Budd-Chiari's syndrome. The average length of the material obtained was from 0 .9+0 .6 cm. Several connected but different procedures were carried out (VSH flebography in 40/42 patients, manometry of VSH in 41/42 patients and retrograde portography in 39/42 examinations). We found 10 complications: 1 broken subeapsular hematoma, 1 perforation of Glisson's capsule, 1 hematoma in the place of venopuncture,3 crisis of supraventriular taquicardia,4 episodes of abdominal pain.

Concerning those patients who had biopsies taken, 65 % of them had histological material wich was of a diagnostic value in itself, and the other 35 % had a diagnostic value in the overall clinical context. With the connected techniques portal hypertension was apparent in 58% of the patients and alterations in portal circulation in 9%.Biopsy and the simultaneous examinations had a direct effect on the treatment in 29.

CONCLUSIONS: The tecniques of reaching the liver using transjugular access show themselves to be an efficient study method, which allows us to obtain a biopsy in a subgroup of patients in whom other tecuiques are not safe and it allows other connected vascular and manometric studies to be carried out, which have a definitive impact on the patients' hardling.

• CHOLANGIOCARCINOMA IN PATIENTS WITH PRIMARY SCLEROSING CHOLANGITIS - A POPULATION-BASED STUDY. D. Pinczowski 1.2 A. Ekbom ~, ~Dept. of Cancer Epidemiology Un vers ty Hospital, Uppsa a, Sweden; 2Dept. of Surgery, Stdersjukhuset, Stoc "kholha, Sweden.

Background: Cholungiocarcinoma following a diagnos s of primary sclerosing cholangths has been reported in several studies and cholangiocarcinoma has been found in up to 30% of patients subjected to liver transplant due to primary sclerosing cholangitis. Due to the difficulties to distinguish primary sclerosing cho angitis from other chronic liver disorders such as primary b ary cirrhosis and chronic acute hepatitis there have been difficulties in assessing the risk for cho angiocarcinoma in patients with primary sclerosing cholungitis. The introduction of endoscopic retrograde cholangiopancreatography (ERCP) in the late 1970'ies was a break- through that lead to a much higher specificity in the diagnosis.

Method: In order to analyse and quantif~ the risk oT cholangiocarcinoma among patients withprimary sclerosing cholangitis we conducted an extensive search in the archives in 7different hospitals in central Sweden between the years 1979-91, identifying all patients subjected to an ERCP (approximately 5,000). Through the ERCP charts we could identify all cases of primary scleros ng cholangitis thus creating a population-based cohort of all incndent and prevalent cases w th n a defined-population. We identified 123 patients with an endoscopc diagnosis of primary selerosing eholangitis. All the patient charts were identified-and we retrieved infonnatmn about medical history including a prior diagnosis of ulcerative colitis and if and when the patient had been subjected to choleetomy. Through linkage with the Swedish Cancer and Death Registry all cases of cholangiocarcinoma were identified as well as the date of the diagnosis of the cancer and the date of death.

Results: There were 12 patients with cholangiocarcinoma d agnosed up to 10 years after the diagnosis of primary sclerosing cholangit s In 8 of those 12oat ents the diagnosis ofcholungiocarcinoma was made within 2 years after the diagnos s of primary sc erosing cqaolangitis 78 patients w th a mean a e of 39 5 veers had a pnor-d[iagnosis o~" ulcerative colitis of which 20 patients ~ d been su~bject~d to a cholectomy before the diagnosis of primary sclerosing cholangitis. In this group there were 3 cases of cholangiocarcinoma diagnosed in this subgroup of which Shad been subjected to a cholectomy prior to the diagnosis o f primary sclerosing cholangitns. 45 patients, with a mean age of 53 9 years did not have a diagnosis oT ulceratwe colitis at the diagnosis of primary sclerosihg cholangitis although 10 patients had a subsequent dmgnosis ofuleerative co it s among which no cases of cholungiocarcinoma occurred. Among the patients w thout prior diagnosis of ulceratwe colitis who later were diagnosed wRh cho ang ocarc noma the mean age at the time of diagnosis ofprimary sclerosing cholangitis was 54.0 years.

Conclusion: We conclude that patients wRh primary sclerosing seem to constitute two populations. One group, associated with ulceratwe colitis are on average 15 years younger at diagnosis than patients with primary scleros ng ch~ ang t s without a history oY ulcerattve colitis. Moreover, coleetomy among patients with a prior ulcerative colitis do not seem to protect against the risk of cholangiocarcinoma.