HEPATOLOGY Vol. 22, No. 4, P t . 2, 1995 A A S L D A B S T R A C T S 4 1 3 A

1225 MINIMAL PRESERVATION-REPERFUSION INJURY (PRI) IN LIVING RELATED LIVER TRANSPLANTATION (LRLT) - ACCURATE EVALUATION BY GLUTATNION S- TRANSFERASES (GST) H Euawa. H Okaiima. K Tanaka. ¥ Inomata. S Uemoto. K Satomura. M Kawashima° Y Yamaoka. Second Dept. of Surgery, Kyoto University Hospital, Kyoto, Japan.

Ob~ect lve~ TO assess PRI in LRLT, we prospectively collected blood samples and studied the clinical courses of 41 pediatric patients, measuring GST recently reported as a sensitive and specific marker of hepatic parenchymal injury. Meth~e: To assess PRI, we excluded 15 patients undergoing laparotomy within 7 days after LRLT from 56 patients (5 month-old to 17 years) undergoing primary LRLT between September 1993 and October 1994. Serum levels of AST, ALT, and GST were measured daily for 7 days after LRLT. The serum level of GST was determined using Hepkit (Biotrin Int. Ireland). lelultsl All donors (age: 21 to 46) had normal liver function. Cold and warm ischemic time (WIT) was 134±85 (mean±SD) minutes and 48±11 minutes. Intraoperative blood transfusion was 36 to 819 ml/kg (medianl 103 ml/kg). The peak values of ALT, AST, and GST were 55 to 669 IU/L, 66 to 765 IU/L, and 30 to 247 ng/ml (normalz< 5 ng/ml), respectively. The WIT significantly co-related with the peak GST. The GST returned to the normal range significantly (p<0.05) earlier (mean:3.2 days) than ALT (6.4 days) and AST (10.3 days). Con=lemion: I) LRLT presumably contributes to the minimal graft injury which was significantly effected by WIT. 2) The GST demonstrated that the grafts recovered from PRI for 3 days after LRLT. 3) GST could be an useful parameter for postoperative management after liver transplantation.

1226 INDUCTION OF TRANSFORMING GROWTH FACTOR 13 IN CONGENITAL HEPATIC FIBROSIS: POSSIBLE ROLE OF THE FAT STORING CELL

EI-Youssef, M, Yu Y, Edelstein B, and Crawford S. Department of Pediatrics and Pathology. Northwestern University Medical School, Chicago IL.

HYPOTHESIS: Congenital hepatic fibrosis (CHF) is a rare disorder characterized by hepatic fibrosis and varying degree of biliary duct ectaaia. It is a pure form of fibrosis without inflammation and offers a unique opportunity to study this phenomenon in its simplest manifestation. We postulated that the fibrosis of CHF is due to spontaneous activation of the Fat Storing Cell (FSC), the main source of collagen in the liver; and that the biliary epithelium interacts abnormally with the FSC. The FSC is known to produce mainly collagens type III and to be stimulated by TGF [3~. MATERIAL AND METHODS: We retrospectively reviewed the clinical charts and histologic slides from 7 patients diagnosed with CHF. Two normal livers were obtained from the live procurement and distribution system for controls. The slides were deparaffinized and restained using conventional immunohistochemistry methods using antibodies against collagen type III and TGF [~. In separate experiments the cellular localization of TGF I]~ mRNA transcripts were detected by in situ polymerase chain reaction hybridization with biotinylated cDNA probe in paraffin sections of normal and CHF livers. RESULTS: Normal livers as expected had very little staining of collagen type [11 or TGF ~1 • The fibrous tissue in livers of patients with CHF stained prominently for collagen type II1. The strongest staining for TGF 13~ occurred in the ectatic biliary epithelium with mild staining of the island of hepatocytes that are separated by fibrotic tissue. In situ PCR for TGF 131 mRNA confirmed localization to both biliary epithelium and hepatocytes in CHF livers in contrast to normal livers. CONCLUSION: CHF offers a unique opportunity to study pure liver fibrosis. The studies performed suggest activation of FSC, possibly by TGF 131 since collagen type III is the main component of fibrous tissue deposited in this condition. TGF ~1 was prominent and showed the strongest staining Within the ectatic biliary epithelium suggesting an abnormal interaction between the, biliary epithelium and the FSC. Further experiments are planned involving the FSC and biliary epithelium co-culture.

1227 CONTINOUS VENO-VENOUS HEMODIAFILTRATION (CVVHD) USING HEPARIN / ALBUMIN PRECIRCULATION IN ACUTE LIVER FAILURE. Ellis AJ. Lever R. Wendon I. Langley P, and Ro~er Williams. Institute of Liver Studies, Kings' College Hospital and School of Medicine and Dentistry, London SE5 9PJ, UK.

Acute liver failure (ALF) is associated with renal failure in up to 55% of patients and CVVHD is the treatment of choice. Anticoagulation is achieved by the infusion of heparin or epoprustenol (PGI2). Perfusion of extracorporeal circuits is associated with piatelet consumption and both heparin and PGI2 may exacerbate bleeding, commonly seen in FHF with in addition PGI2 causing vasodilatation which may worsen pre-existing hypotenalon. Aim. To minimise side-effects associated with CVVHD, we have developed a method of priming the circuitry with human albumin solution and heparin to avoid the need for continous anticoagulation. Method. 36 dialysis episodes were performed in 22 patients with FHF secondary to paraeetamol toxicity in 19, anti TB Rx, NANB hepatitis, and ischaemia in 1 case respectively with a median age of 38 yrs. Patients were randomised to receive one of three modes of anticoagulation. 22 circuits were heparin/albumin bonded, 8 utilised a heparin infusion (activated clotting time maintained between 150-200 secs) and 6 PGI2 infusion (Sng/kg/min). Heparin/albumin circuits were prepared by circulating for 4 hours 500mi of 4.5% albumin to which 5000IU Na heparin had been added. Haemodynamic measurements were performed at 10 minute intervals for 1 hour, and platelet counts were performed at 0, 30, 60 minutes, and daily thereal~er. Results. CVVHD circuits lasted for a mean of 46 hrs in both the heparin/albumin and heparin infusion patient groups as compared to 41 hrs in those anticoagulated with PGI2. Falls in mean arterial blood pressure during the first 30 minutes of treatment were significantly less in the heparin bonded group (2mmHg) in comparison to heparin infusion [10mmHg (p <0.0001)] and PGI2 [10mmHg(p<0.0002)]. Platelet consumption over the first hour of treatment was less in the heparin bonded group (12 xl0~/l) compared to heparin 31 x109/l (p=0.0014) and PGI2 24 x109/l (p=0.04). Conclusions. Precirculation with heparin/albumin leads to improved biocompatibility of CVVHD treatment in patients with ALF. The more costly usage of prostacycline can be avoided by this simple technique.

1228 SERIAL QUANTITATIVE ANALYSIS OF HEPATITIS C VIRUS RNA IN SERUM DURING INTERFERON THERAPY FOR PREDICTING RESPONSE BY USING AMPLICOR TM ASSAY. H Endo, G Yamada, H Tsu,cleno v M Takatani, F Kishi, M Takahashi I K Manabe, M Mizun% TTsuji. The First Department of Internal Medicine, Okayama University Medical School, Okayama, Japan.

Amplicor TM HCV monitor kit (Amplicor) (Nippon Roche, Japan) was developed to quanti tate the amount of hepatit is C virus RNA (HCV-RNA) using rTth polymerase for reverse transcription polymerase chain reaction (RT-PCR). We investigated its clinical usefulness for predicting response to interferon (IFN) therapy in the patients with chronic hepatitis C by measuring HCV-RNA level before and during therapy. Materials and Methods: Serum from 49 patients with chronic hepatit is C were obtained at the beginning, one week, two weeks after the beginning and the end of IFN therapy. Patients were treated with IFN-~z three times a week for 24 or 36 weeks (total 360 MU). The amount of HCV-RNA was quanti tated by Amplicor. Initial HCV-RNA level before the therapy was also measured by branched DNA probe assay (bDNA) (QuantiplexrMHCV-RNA Assay, Chiron). Results: All 18 responders (37%) had HCV-RNA less than 3 x 10s copies/ml by Amplicor and 2 x 106 Eq/ml by bDNA prior to administration of IFN and had less than 4 x 1 03 copies/ml by Amplicor one week after the beginning o f the therapy. The ratio of responders under those HCV-RNA levels were 45%, 62% and 82%, respectively. Conclusion: The combination of the quantitat ion of HCV-RNA by bDNA at the beginning of the therapy and by Amplicor one week after may be the effective and useful way to predict response to IFN therapy.