HEPATOLOGY Vol. 22, No. 4, Pt . 2, 1995 A A S L D A B S T R A C T S 4 7 7 A

1481 IN VITRO PERFORMANCE OF LIVERS OBTAINED PROM

HYPEROSMOLAR RATS n Podkameni. I Wolf *. I Banu£vev. S

¢Brtlss. P Boros. CM Miller. Departments of Surgery and Biochemistry ", Mount Sinai School of Medicine NYC, NY 10029

Rapid changes in a donor's fluid and electrolyte balance can influence liver graft function. The aim of this study was to assess the in vitro performance of livers obtained from rats in a state of hyperosmolarity using bile production, levels of ATP and lysosomal enzyme release. STUDY DESIGN:

Hyperosmolarity was induced by deprivation of food and water for 72h (307±2.5 osm compared to 295±3.3 osm, p<0.008). Livers obtained from this group (G3, n=4) were preserved in UW for 8h at 4°C then perfused with Krebs-Henseleit buffer in a recirculating system for 90 min. Livers for normosmolar rats were harvested and preserved as descirbed above (G2, n=4) or irmnediately perfused as control (G1, n=lO). Biopsies were taken for ATP measurement using HPLC. Caval effluent was obtained for lysosomal enzyme measurement at the start of perfusion and at every 30 min. Bile output was also monitored. Data were analysed by ANOVA. RESULTS: Changes in bile output and enzyme levels are presented in the Table. Recovery of ATP production is shown in the Figure.

BII, p~duvlkm Aryl I u~alllle I-Clueullelt~ll!

/Atglmln Utl U/I

M~IttSD t ~ ~o rain o ,d , ~0 m~

IG 1.2~.1 @ U.J~0.1" 13~t

G 0~S&O.l 0./~1.2 i*o.~ 1444~3 lf.lkt271 2

G o.ft:~U.l b 9.41&03 Z~41.2" 147:k46 411t~14~ 3

b:Gl vs S3, p<0.001, a: o vs 90, p<0.04

CONCLUSIONS: Livers obtained from

h y p e r o s m o l a r r a t s h a v e i m p a i r e d

performance when compared to normosmolar,

::: ,o, . . . . . . . . . . . . 4

show signs of hepatocellular injury and are not able to significantly increase ATP production after preservation.

1482 ONCOGENES EXPRESSION AFTER BILE D U C T LIGATION IN RATS T RE AT E D WITH F U R A N L Polimeno. M Barone. A Azzarone. A Iacobellis. O H Zen~. V Subbotin. B Carr. TE Starzl. and A Francavilla. Dept. of Surgery, University of Pittsburgh, Dept. of Gastroenterology, University of Bari.

Bile duct libation (BDL) in rats is an experimental model for inducing a typical bile ductular hy:pe~lasia. The expression of ~rowth control genes in this model is different from that observed m the remnant part of rat liver after 70% partial hepatectomy. In particular the expression of transforming growth factor alpha (TGFalpha) m R N A is reduced in contrast with elevated levels of the oncogene e-erb-B2, and proto-oncogene c-raf (Hepatology, 21(4):1070-78, 1995).

In the present work we evaluate the hepatic expression of growth- control genes in rats operated with BDL and treated with furan, a potent hepatocellular carcinomas inducer. This drug in combination with DDL, induced replacement of most part of the liver with well-differentiated hyperplasia bile ductules. Fiftyfive male Fischer F344 rats were used; fifty of which underwent BDL and one week later divided in ten groups of five animals each. Five groups were treated with furan (45mg/Kg b.w./day by gavage) and five groups treated with vehicle, were used as control. Five normal animals were sacrificed to evaluate m R N A basal levels of the genes studied. Rats (5, BDL-control and 5, BDL-furan) were sacrificed at 1,2,3,4, and 5 weeks after the beginning of furan treatment. Northern blot analysis of m R N A was used to examine hepatic expression of the genes of c-myc, c-Ha ras, c-erb-B2, c-rat and transforming growth factor beta 1 (TGFbl) . The results show a clear difference of hepatic gene expression between the two groups of rats. Despite the typical feature reported by us in BDL there are profound changes in the oncogene expression of T G F b l and c-erb- B2 and c-rat which start to appear at the first week of treatment and reach the maximum expression at 3-4 weeks. The changes for c-erb-B2 confirm the importance of this oncogene in normal and pathological bile duct protiferatiorL

1483 ASSOCIATION BETWEEN MYALURONIC ACID UPTAKE AND

ENDOTHELIN-I LEVEL IN PRESERVED RAT LIVERS

E L P o n a . I P o ~ u . T S u e h i ~ . l B ~ v e v . T G u i n e . P B o ~ s . C M M m e r .

D ~ i s m n o f L i v e r T r a n s p l a n t , M o u ~ S i n a i S e h ~ l o f M ~ i c m e , N Y C , N Y , l ~

Cold storage is the standard technique for liver allograft preservation.Hepatic sinusoidal endothelial cells (SEC) are the primary target for preservation-perfusion injury. Based on the rapid clearance by SEC through high-affinity receptors hyaluronic acid (HA) uptake has been proposed as a reliable functional assay. Involvement of endothelins (ET-s) in SEC damage has also been suggested. In this study we sought to determine the value of exogenous HA uptake and perfusate ET- 1 level in the assessment of cold preservation injury in isolated perfused rat livers. Study design: 90 Sprague-Dawley rat livers were randomly divided into eight

i cF

m , ~81 111-8010-16 2 1 - 1 15-o FIA uDtnke

groups (non-preserved [NP], cold- flushed [CF], and I/2, 4, 6, 8, 24 and 48 h stored at 4°(: in UW), then perfused for 120 rain in a reeirculating system with Krebs- Hanseleit solution containing HA at 500 ng/ml. Perfusate samples were collected at 0, 5, 15, 30, 60, 90 and 120 min, later assayed for I-IA

and ET-1 content by ELISA (Chugai Seiyaku Inc. and Wako Pure Chemical Industries, LTD, Tokyo, Japan). Data were analyzed by ANOVA. Results: The mean HA uptake of the NP group after 120 min was considered as 100% and used to assign the other livers to 5 categories (>85, 85-50, 50-25, 25-5, 5-0 %) regardless of preservation time (Figure). The El" values measured in the 0 min samples showed a steady increase as the HA uptake was decreasing but no level of significance was attained. Condusions: HA uptake is a reliable assay for the assessment of SEC function. It remains relatively uncompromised up to 4 h of preservation. A moderate reduetlon is observed between 4 and 8 h, followed by a quick deterioration. ET-1 aceumulalion during cold storage is highly variable and seems to correlate with preservation time and profound impairment in HA uptake.

1484 APOLIPOPROTEIN (Apo) E4 GENOTYPE IS ASSOCIATED WITH INCREASED RISK OF CHOLESTEROL (XOL) GALLSTONE RECURRENCE AFTER SHOCK-WAVE LITHOTRIPSY (ESWL). P Portineasa 1"3. KI van Emecum I . P van de Meeber~ 1. GM Dallin~a-Thie 2. TWA de Bruin 2. GP vanBerae-Hene~ouwen t. Dept. Gastroenterology I & Internal Medicine 2, University Hospital Utrecht, The Netherlands. Istituto di Clinica Medica I, University Medical School Bari, Italy 3.

ESWL is effective in selected gallstone patients but stone recurrence is a major drawback. Apo E4 genotype is associated with increased intestinal XOL absorption and hepatic secretion, which might increase risk of recurrence. We investigated factors potentially influencing gallstone clearance and recurrence in 84 patients with stone dissolution after ESWL+bile salt therapy for initial solitary (n=55) or multiple (n=29) radiolucent stones. APOE genotyping & postprandial gallbladder motility (sonography, mixed meal 30g fat) were studied in a representative subgroup of patients (n= 50). Median follow-up after ESWL was 36 (range 4.5-67) too. Median gallstone clearance time was 8.7 (range 0.2- 30) too. Factors enhancing gallstone clearance were: apoE4 genotype, small initial gallstone size & number, effective fragmentation, & good gallbladder emptying (Cox-regression model, P<0.002). 30 patients had stone recurrence after 18.6 (range 1.0-50) too. Cumulative recurrence rate (by Kaplan-Meiar life- table analysis) was 15% at lyr increasing to 60% after 5.5 yrs and was greater in homozygous and heterozygous patients for apoE4 allele compared to patients not expressing the apoE4 allele (at 58mo. 82.8+ 13.6% vs 35.2+ 10.0%, log- rank test, P=0.0371). Recurrence rate at short-term follow-up was smaller in patients with initial solitary stones than in those with multiple stones, but differences disappeared at prolonged follow-up. Patients with recurrence had impaired gallbladder emptying (increased fasting & residual volume), compared to patients with no recurrence or controls. Patients with a small postprandial residual gallbladder volume (< 6ml) had a lower probability of stone recurrence than those with large (>6ml) residual gallbladder volume (45.2-t-23.0% vs 78.0+ 17.0%, P=0.020). Biliary pain was more frequent with recurrence (55% vs 13%, P=0.001). Conclusions: apoE4 genotype is associated with increased speed of gallstone clearance (probably explained by high stone XOL come•t) but high risk of recurrence (probably explained by strongly increased hepatic XOL secretion) after ESWL. Effective gallbladder emptying is important for fast clearance and prevention of recurrence. Patients with initial solitary stones have a decreased early -but not long-term- gallstone recurrence rate.