Al150 AASLD ABSTRACTS GASTROENTEROLOGY, Vol. 108, No. 4

• VARIATIONS OF VIRAL LOAD AND GENOTYPE~S AFTER IFN THERAPY IN CHRONIC HEPATITIS C. I Portal, M Boarli~ge*. P.Halfon*, V Gerolami, C Wartelle, H Khiri, PP Gamier, P Castellani, M Antoni, D Botta-Fridlund, AP Gauthier, G Cartouzon. Liver unit and Dept of Biochemistry, H6pital de la ~ n , O H 6 p i t a l Saint Josuph,*Lab Alpha Bio, Marseille, France.

Aim: To study the influence of interferon therapy on the viral load and the get~ype in patients with chronic hepatitis C non responders (NR) and msponder-mlapsers (RR). Methods: Forty patients with a histologically proven chronic hepatitis C confirmed HCV RNA positive by RT-nested PCR were assigned 3M fiw of alfaIFN for at least 24 weeks and followed for at least 24 months aider completion of treatment. Five patients had a complete and sustained response (HCV RNA negative at the end of follow-up), 23140 (57°5) pts responded and relapsed (RR) and 12 pts (3005) did not respond (NR) to IFN. HCV RNA was retrospactivel][ assessed by bDNA signal amplification (Chiton; detection limit of 3.5 103 ¢q/mL) before treatment and 4-6 months after completion of treatment in NR and RR. HCV genotypes were assessed by Line Immunoprobe Assay method (LIPA i n n o ~ c a , Beigium) before and after tmatmenL Direct sequencing of the 5'NC region was performed in case of genotype modification. Results: (1) The mean viremia significantly decreased in RR after treatment (2.78 vs 1.39 105eq; p=0.01) whereas it significantly increased in NR (3.55 vs 10.98 105eq; p=0.01).(2)Variations of mean viremia in relation to 8enotype and response to IFN ~ given in table i.

Genotvue l a l b 2# 3# 4

RR NR RR NR CR RR CR RR NR RR Patients(n) 2 4 5 6 1 4 4 10 2 2 viremia*preT 9 9.2 5.1 38.9 3.5 22 4.3 45.8 78.1 24.7 viremia*postT3.7 99.2 20.8131.5 - 10.8 16 66.1 3.7 * HCV RNA 10,,5. ea/ml

(3) Genotype changed dutin 8 treatment in 2 RR. One patient 8enotype la became la & 3a;one patient geantype lb became lb &la. Conclusions: £1 )Viremia significantly increases during IFN therapy in NR whereas in responders-relapsers the viral load significantly decreases. (2) In n ~ viremia increases only in genotype la and lb patients. In RR, viremia decreases in all genotypes except lb suggesting that a standard ~.girnen o f IFN not only has no effect in patients with genotype lb but may be detrimemal.(3) Modification o f genotype during IFN therapy is uncommon.

• NUCLEATION BEHAVIOUR OF VARIOUS CHOLESTEROL (XOL) CRYSTALS IN BILE FROM GALLSTONE PATIENTS. P Portincasa t'4, KI van Erpecum ], A Jansen ~ W Renooij-", M Gadellaa t, GP vanBerge-Henegouweo t. Depts. Gastroenterology ~ & Surgery -~, Utrecht University. Dept. Surgery, St. Antonius Hosp., Nieuwegein 3, The Netherlands. Clinica Medica 1, Bari University, Italy ~

In addition to classical platelike (PL) monohydrate xol crystals, arcs/needles (AN), spirals (SPI), and tubules (TU) xol crystals were recently observed in model and human biles (JCI, 90:1156-61, 1992 and Gastro, 106.'A1004, 1994). It is unknown which factors determine crystal shape and what is the relevance for gallstone formation. Aims: 1) to relate nucleation of various xol crystals to bile composition, stone type/number, 2) to compare the effect of ultrafihraticn and ultracentrifugation on crystal nucleation and bile composition. Methods: Gallbladder bile was Obtained from: 1) 50 xol stone pts. II) 5 pigment stone pts, and 111) 12 UDCA-treated xol stone pts (10mg/Kg/day for > 3wks beforesurgery bile enrichment with UDCA 42.2+8.2%). Unfiltered (U), crystal- free ultrafiltered (UF, 0.22/~m millipore) and ultracentrifuged isotropic (UC, 100,000g-2hr ~) bile were kept for lipid analysis and nucleation behaviour (polarizing microscopy during 21 days and semiquantitative scale: 0= no crystals -4+ = > 10 crystals / field). R~ults: ultrafihration nor ultracentrifugaticn did infltlence xol, phuspholipid or bile salt concentration (n= 14). PL, aggregates of PL (AG) and AN nucleated significantly faster and more extensively in UC compared to UF bile. However, TU and SP1 were significantly more prevalent in UF bile (n= 16). Xol stone pts (untreated): in fresh U-bile PL, AG and AN were seen in 90%, 36% and 18% reap., whereas SPI and TU were ahvays absent. AN, PL and AG were persistent forms and their score progressively increased during 2l days. Rate/exteot of nucleath~n of these forms in UF-bile was significantly (I)<0.05) higher for multiple than solitary stone pts. SPI and TU were transient, with higher score for multiple stones. SPI/TU occurrence was associated with higher % deoxycholic acid (28.7 + 3.2 % vs 19.2 +- 3.2 %, p = 0.033) and significantly higher speed/extent of nucleation of all crystal forms, regardless stone number. UDCA-treated and pigment stone pts never nucleate d any crystal form in UF bile. Whereas pigmen t stone pts never had crystals in their fresh U- bile. 7 out of 12 UDCA-treated pts had atypical smooth xol crystals (10-200/zm). In 2 cases these crystals dissolved after 15 and 20 days. Conclusions: 1) PL, AG and AN are persistent forms with high rate/extent of nucleation in multiple xol stone pts. 2) TU and SPI are transient forms. Their presence is associated with rapid nucleation of all forms and a high % deoxycholic acid. 3) UDCA-treated patients often have atypical smooth crystals.

• RAPID ABDOMINAL DECOMPRESSION BY TOTAL PARACENTESIS (TP) AMELIORATES HEART FUNCTION AND SUPPRESSES SYMPATHETIC NERVOUS SYSTEM (SNS) OVERACTIVITY IN CIRRHOTICS WITH TENSE ASCITES. M.Pozzi, G.Osculati*, P.Colombo, G.B.Bolla °, A.Lanfranchi, V.Pecci, L.Roffi, S.DeCeglia*, S.Maggiolini*, M.C. Colombo, A.Pipemo, B.M.Cattaneo, G.Grassi and G.Mancia. Cattedra di Medicina Intema, Divisione di Medicina I, *UCC, Ospedale S.Gerardo, Monza, °Centro di Fisiologia Clinica ed Ipertensione, Universit~ degli Studi,Milano, Italy.

BACKGROUND SNS overactivity along with RAAS stimulation are key features in cirrhotics developing ascites. These homeostatic responses counteract the central volume underfilling and arterial bypotension that characterize these patients. Tense ascites further impairs heart function. Plasma norepinephrine (NE) and epinephrine (E) concentrations are an index of SNS activation. TP is a first- line, cost-effective and safe procedure (whe n followed by therapeutic plasma expansion) to treat tense ascites. AIM to study plasma catecholamine concentration and heart volume changes in patients submitted to rapid TP for tense ascites. METHODS 22 CHILD C patients (15M, 7F) with tense ascites, underwent rapid TP (250 mL/min) followed by i.v. Albumin (6gr/liter ascites). Echocardiography was performed at baseline (A), after TP (B), 24h (C) and 15 days (D) later. Plasma samples for NE, E, Aldosterone Concentration (PAC) and Renin Activity (PRA) were collected concurrently, and halfway (50%) during TP (linear interpolation). All measurements were performed with patients supine for 24h. No diuretics were allowed 4 days before and 2 days after TP. RESULTS 10.9±4.7 liters of ascites were drained. Right ventricle end diastolic diameter (RVdd, ram) and left ventricle stroke volume (LVsv, mL) increased (p<O.01, ANOVA, and t-test A vs B, respectively) while PRA, PAC (p<0.03), NE (NS) and E (NS) decreased at C (ANOVA does not include D for all data).

A 50% B C D RVdd 29.3±5 (19) 32.9±2.3 (18) 32.8±2.5 (16) 32.7=~1.7 (7) LVsv 65.6±14(19) 69.8±15.6 (18) 64±13(16) 62.1±8(7) NE 662=~211 (9) 629±203(9) 583±174 (9) 485±149 (8) 343±173 (5) E 149:~71 (9) 117-~69 (9) 104±57 (9) 85±40 (8) 24a:5 (5)

PRA 13:t-2 (20) 10.2±1.3 (20) 9.1±1 (20) 9±1.6 (20) 12.1a-1.2(17) PAC 1208a-338 (20) 1072±240 (20) 916±281 (20) 676~-164 (20) 881±183 f17)

CONCLUSIONS These results indicate that TP ameliorates the filling and emptying patterns of the heart. SNS deactivation (cardiopulmonary receptors activation?) and RAAS suppression (improved renal perfusion?) follow. Albumin infusion prevents circulatory derangement. Most changes can be observed early during TP thus indicating that even minor reductions in intraabdominal pressure by paracentcsis are clinically relevant in patients with tense ascites.

EVIDENCE FOR THE INVOLVEMENT OF STEROL CARRIER PROTEIN (SCP-2) IN A RAPID, MICROTUBULE-, AND GOLGI- INDEPENDENT TRANSPORT OF NEWLY-SYNTHESIZED CHOLESTEROL TO BILE. Puqliel!i L., Rigotti A., Amigo L., Nu~ez L., Santos M., Greco A.V., Nervi F. Departamentos de Gastroenterologia, Centro para la Prevencion y Tratamiento del Cancer Digestivo, y de Biologia Celular y Molecular, Pontificia Universidad Catolica; Santiago, Chile and Istituto di Medicina Interna, UniversitY. Cattolica del S. Cuore, Roma, italia.

Newly synthesized cholesterol represents 10-20% of total biliary cholesterol and is originated from the endoplasmic reticulum of hepatocytes. The cellular mechanisms involved in the transport of newly synthesized cholesterol into bile have not been identified. We have previously shown (Gastroenterology 1994; 106: A966) that the hepatic transport of biliary newly synthesized cholesterol in the rat is mediated by two different mechanisms: one of 10 min, microtubule- and Golgi-independent and one of 20 min, microtubule- and Go!gi- dependent. In vitro studies have suggested that sterol carrier proteim2 (SCP-2) may be involved in the intracellular transport of cholesterol. Aims and Methods : To assess the possibility that SCP-2 is involved in the transport of biliary newly synthesized cholesterol, fresh liver homogenates were obtained from rats with normal and increased hepatic cholesterol synthesis and SCP-2 was quantitated by immunoblotting. To test the possibility that the rapid (10 rain), microtubule- and Golgi-independent transport of newly synthesized cholesterol is specifically mediated by SCP-2 we decided to follow the transfer of fiewly synthesized cholesterol to the plasma membrane in cultured fibroblasts using cholesterol oxidase as a probe. Normal and SCP-2 deficient fibroblasts were pulse-chased with [14C]acetate and several drugs were used to test the involvement of a microtubule- or Golgi-dependent mechanism. FinallY, antisense oligonucleotides corresponding to the 5' region of SCP-2 coding sequence were added to the cultured cells. Results : Immunoblots from fresh liver homogenates showed that SCP-2 was 1.5-2.0 fold over the control values in rats with increased synthesis of hepatic cholesterol. The max!mal values of newly synthesized cholesterol in the plasma membrane of normal fibroblasts were found at 10 min and then it decreased rapidly: Microtubule- or Golgi-disruptive drugs did not affect cholesterol transport. In contrast, in SCP-2 deficient fibroblasts the maximal values of [14C]cholesterol in plasma membrane were reached at 20 min and this transport was drastically reduced by both microtubule- or Golgi-disruptive drugs. Antisense treatment resulted in a significant decrease in cellular levels of SCP-2 and the rapid (10 rain) transport of newly synthesized cholesterol to plasma membrane was significantly reduced. Conclusions : These results in cultured cells (1) confirm the involvement of two different mechanisms in the transport of newly synthesized cholesterol and (2) demonstrate the involveme0t of SCP-2 in the rapid (10 min), microtubule-, and Golgi- independent one. T~he similarities with the results obtained in the transport of newly synthesized cholesterol into bile in the rat and the evidence that animals with increased synthesis of hepatic cholesterol possess increased amount of SCP-2, suggest that SCP-2 is playing a role in the transport of newly synthesized cholesterol into bile. (M~nistero degli Eeted, Italia and Fondscyt 0620).