504A

1589

A A S L D A B S T R A C T S HEPATOLOGY O c t o b e r 1995

CUNICAL SIGNIFICANCE OF SERUM HYALURONATE CONCEN- TRATIONS IN PATIENTS WITH ALCOHOLIC UVER DISEASE. S Urashima, M Tsutsumi, S Takase, Y Ueshima, H Kawahara: Div. of Gastroent., Dept. of Int. Med., Kanazawa Med. Univ., Uchinada, Ishikawa, Japan.

It has been reported that serum hyaluronate concentrations are increased in various liver diseases, especially in alcoholic liver disease (ALD). The aim of this study was to clarify whether serum hyaluronate concentrations reflect the severity of the histopathologic changes of ALD. Methods: Serum hyaluronate concentrations of patients at different stages of ALD (n=39) and non-ALD (n=63) were measured by a sandwich enzyme=binding assay. The amount of type IV collagen and laminin in the liver biopsy specimens was also measured using one-step sandwich enzyme immunoassay kits for human serum type IV collagen or laminin levels with monoclonal antibodies with modification. Results: Serum hyaluronate concentrations in patients with liver disease were higher than the cut-off value (33.4 /~ g/ml), and. increased significantly (p<0.001) in parallel with the progression of hepatic fibrosis. Serum hyaluronate concentrations in patients actively drinking with ALD were significantly higher (p<0.001) than those in patients with non-ALD. After 4 weeks o f abstinence, the elevated serum hyaluronate concentrations fell to the levels of non-ALD. A significant correlation between serum hyaluronate and hepatic type IV collagen and laminin content was present patients actively drinking with ALD, but not non-ALD. Conclusions: There are two reasons for the elevation of serum hyaluronate concentrations in ALD; one reversible when ALD patients stop drinking and the other unreversible associated with hepatic fibrosis. The reversible phase of hyaluronale elevation is probably due to reversible endothelial damage caused by alcohol.

1590 CELL TO CELL HCV TRANSMISSION BY CHRONICALLY INFECTED TOFE CELL LINE TO HUMAN HEPATOMA CELLS 2.2.15 M.B. Valli+., L. Ber.tolioi $., A. Ponzetto°., S. Iacovacci+., F. Fal2etti&., and G. Carl~ni +. Institute Of Expt, Med. +:and C~ll.Biol. ~, C.N.R. Roma; Institute of Hematology% Univ. Perugia; Div. Gastroent. Hosp. Holinette, ° Torino, Italy

We previously demonstrated the susceptibility of TOFE human B cell line, bone marrow -derived, to HCV infection. In these cells (TOFE-HCV) viral replication and production, assayed by PCR techniques, occurred for more than 6 months. To verify whether these ceils were able to work as infectious centers and transmit the virus to hepatic cells by cooultivation, we cocultivated TOFE-HCV with 2.2.15 hepatoma cells. In this assay we took advantage: a) of the ability of TOFE to loosely attach to the substrate represented by the monolayer of 2.2.15 recipient cells and b) of the neomycin resistance and unsusceptibility of 2.2.15 cells to free-virus infection. In the third day of cocultivation growth medium was removed and replaced with a selective medium containing neomycin at a lethal dose for TOFE cells. 45 to 60 days later, cells became PCR-positive for HCV RNA, while they were PCR- negative when tested for EBV DNA, TOFE cells being EBNA positive and containing EBV DNA. The lack of detection by PCR of EBV DNA in the drug resistant 2.2.15 cells, indicate that no residual TOFE-HCV cells were present at these times in the coltures. Therefore the presence of HCV RNA in 2.2.15 cell extracts indicate a cell to cell HCV transmission.

1591 TERLIPRESSIN ENHANCES FREE WATER CLEARANCE IN EXPERIMENTAL CIRRHOSIS IN THE RAT. M.Van de Casteele. G.Van Roev. F.Nevens. J.Fevery. Lab. of Hepatology, Catholic University of Leuven, B-3000 Leuven, Belgium.

We investigated the renal effects ofterlipressin (terlipr), a synthetic vasupressin analogue, in CCl4/phenobarbital cirrhosis. Methods: 3 groups of male Wistar rats, 6 normal (N), 7 cirrhotic without (C) end 4with ascites (A), were given tedipr subcut.20 ttg/kg t.i.d, or saline in a cross-over way during a salt load (9 g NaCI/! in drinking water). Rats were followed up in metabolic cages. Portal und Arterial Pressure were measured later en in the freely moving rat, before and 30 rain after a single dose ofterlipr SC. Re,nits: expressed as mean + SEM without terlipr end mean % change+SEM with terlipr. *p<0:05 **p<0.0l ***p<O.001 (paired t-test)

N plaf N tedinr C nlae C terliar A Dine A terlior Urnsm 1744+180 -8%+11 2107+419 -220/0+8 * 2327+661-40e/0+8 *

(mosm/ks) Na balance 2.1+0.4 -220/0+26 1.5+0.7 -70/0+7 3.0+0.4 -540/0+13"

(meq Na/24 h) Fluid balanea 19+8 -20/0+42 13+11 +50/0+38 23+2 -1100/0+6"

(ml/24 h) Freewtrclear-88+6 -8%+7* -111+6 +320/0+11" -96+5 +270/0+7*

(ml/24 h) N basal N tedier C basal C terlior A basal A terlinr

PP (ram Hg) 5+1 -60°/0+1 * 12+4 °00 -680/0+2* 15+5 °00 -670/0+6** SP(mmHg) 139+9 +20%+4* 122+16 +220/0+1"* 106+4 °0 +230/0+4*** °°p<0.01 °°°p<0.001 unpaired t-test between basal values (Anova significant at 5% level) Cenclnsiens: l.We cenfirm that portal vein and arterial pressure in cirrhosis are vary sensitive to vasopressin activity (VI receptor). 2. In normal animals, free water clearance decreases slightly due to an ADH effect (V2 receptor). In cirrhosis, however, terliprassin enhances free water clearance and induces diuresis if ascites is present; 24 h salt excretion is not altered.

1592 URSODEOXYCHOLIC ACID (UDCA) REDUCES PROTEIN LEVELS AND THEIR NUCLEATION PROMOTING ACTIVITY IN GALLBLADDER BILE OF CHOLF~'I'EROL STONE PATIENTS. KJ v~m Eroecum. P Portincasa. E Eekhardt, GP vanBcr2e~Henenouwen. AK Groen=. Departments of Gastroenterology Universitai Hospital Utrecht and Amsterdam*, The Netherlands.

Apart from decreased cholesterol saturation index (CSI), decreased levels of nucleation promoting proteins in bile might contribute to efficacy of UDCA as gallstone dissolving agent. We therefore determined concentrations of various proteins and their nucleation promoting activity in gallbladder bile of 13 cholesterol gallstone patients (pts) treated during three weeks before cholecystectomy with UDCA (I0 mg/kg/day) and 13 comparable untreated pts. Solitary/multiple stone ratio (3/10 and 3/10) was the same in both groups. All pts had well concentrated bile (TLCo > 5g/dl) and at most minimal gallbladder wall inflammation. Total protein concentration in gallbladder bile (mean4.SE: 2.84-0.6 vs 6.74-1.3 mg/ml: P=0.008) and concanavalin A binding fraction (0.164-0.03 vs 0.424-0.07: P=0.003) were strongly decreased in the UDCA treated group. Total protein concentration in gallbladder bile and associated Concanavalin A binding fraction correlated strongly (R=0.94, P<0.0001). Significant decreases were also found for gallbladder bile cd acid glycoprotein, haptoglobin, IgA, IgG but not IgM concentration. Mucin concentration also tended to b e lower in the UDCA group (P=0.1). Gallbladder bile total protein CR=0.54, P=0.0047) and mucin correlated significantly with CSI but not with bile salt hydrophobicity. Fresh unprocessed bile contained typical cholesterol monohydrate crystals in 85% of untreated pts but atypical smooth cholesterol crystals in 77% of UDCA-treated pts. Nucleation time in filtered bile was 6.1_+1.6 and >22 days resp. Crystal- lization promoting activity of the Concanavalin A binding fraction (nephe- lometry) was also decreased in the UDCA treated group (P=0.01). Conco- mitant microscopical examination revealed significant decrease of nucleation of plates and their aggregates, arcs, needles, tubules but not spirals. Conclusion: UDCA strongly decreases levels of pronucleatlng bile proteins and their nucleation promoting activity.