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Cholate handling by a bioartificial liver (BAL)
HEPATOLOGY Vol. 22, N o . 4, P t . 2, 1 9 9 5
1465
AASLD
ABSTRACTS
ELEVATED LEVELS OF NATRIURETIC HORMONES IN PATIENTS WITH LIVER CIRRHOSIS, F. PATANE' (1), C. FERRI (2), A. PERRONE (2), C. BELLINI (2), A. MAGRINI (3) and C. PUOTI (3). (1) Dept. of Exp. Med. and Pathol. and (2) Inst. of I Clin. Med., University of Rome "La Sapienza", and (3) Liver Unit, Health District RM-H, Marino (Rome),
1466
Deficient production of endogenous Natduretic Hormones (NH) [Digoxin-like Substance (DLS) and Atrial Natduretic Factor (ANF)] has been suggested as a possible mechanism of water end sodium retention in cirrhotic patients with ascites. AIM OF THE STUDY: to evaluate plasma levels of DLS, ANF and aldosterone (PA) in 15 normotensive cirrhotic patients without ascites, 14 cirrhotic patients with ascites (10 with slight to moderate ascites, and 4 with massive ascites) and 28 normotensive healthy subjects. RESULTS: [t] Plasma levels of DLS were significantly lower in controls (18.3i-6.2 pg/mL) than in cirrhotic patients without ascites (68.2:1:28.5 pg/mL, p<0.05) and with ascites (80.7-+21.9 pg/mL, p<0.05). [2] Among patients with ascites DLS significantly correlated with plasma renin activity (PRA: r=0.85, p<0.005) and aldosterone (PA: r=-0.76, p<0.005). [3] ANF levels were significantly lower in healthy subjects (33.9+14.5 pg/mL) than in patients without ascites (122.0+36.1 pg/mL, p<0.05) and in subjects with ascites (128.2+43.8 pg/mL, p
CHOLATE HANDLING BY A BIOARTIFICIAL LIVER (BAL) P Pazzi, AD Moscioni, EMorsiani, J Rozga, AA Demetriou. Dept. o f Surgery and Liver Support Unit, Cedars-Sinai Medical Center, Los Angeles, CA, USA. A BAL, consisting of a hollow fiber module Containing isolated, microcarrierattached porcine hepatocytes, has been shown to reduce serum bile acid levels (BA) in patients with fulminant hepatic failure. We investigated BAL handling of cholie acid. The BAL, loaded with 2.2 x 108 porcine hepatoeytes, was perfused with human plasma containing either 1 laCi of [24-14C]eholate (CA) or 1 laCi of [24-14C]tauroeholate (TCA) and supplemented to 100 pmol with unlabelled CA or TCA, respectively. Samples were collected from plasma and from extrafiber space during a 5-hr perfusion. At the end of the perfusion, hepatocytes were collected for radioactivity and protein determinations, and for examination by transmission electron microscopy (TEM). BAs were separated into 4 fractions (U-~mconjugated, G=glyco- and T=Tanro-conjugated, S=Sulphated) on prepacked silica-based strong anion exchange cartridges; radioactivity was determined by liquid scintillation counting. Data observed during perfusion with CAenriched plasma (expressed as % of total radioactivity detected in each fraction) are reported in the table (mean4-SEM, n=9). ~<0.001 ANOVA Fraction ITime 0 1 2 3 "4 5 U [97.5~0.2 89.74-0.9 81.4=t=111 78.7±2.1 73.5±0.9 71.9±2.7' G 1.4±0.4 9.0±0.7 17.2±1.1 20.5±1.9 25.3±1.3 26.9~2:8' T 0.5±0.1 0.7±0.3 1.1±0A 1.7±0.8 0.7±0.3 0.6+0.1 S 0.4-4-0.1 0.6-4-0.2 0.8±0.1 0.6-4-0.1 0.5-4-0.1 0.54-0.1 After 3-hr, G and S fractions in the extrafiber space were significantly higher than in plasma. During perfusion with TCA-enriched plasma, a relative decrease in T and an increase in G fractions were observed. CA was accumulated by hepatocytes to a three-fold lower level than TCA (3.44-1.8 vs. 10.3±4.5 nmol/mg cell protein), but a significant proportion of radioactivity (26.2±7.3%) was detected in G-fraction. TEM examination of mierocarrier-attaehed porcine hepatoeytes showed ultrastruetural features indicative of typical epithelial cell polarity, the presence of junctional complexes and intact bile eanalieuli as well. On TEM, Golgi complexes and small vesicles were present in the pericanalicular cytoplasm, suggesting the maintenance of a synthetic activity. Conclusions: These results confirm the ability of the BAL to clear BAs from the circulation: porcine hepatucytes accumulate CA and TCA, and are able to conjugate substantial amount of CA.
D O E S N 'T THERAPY
OF
INFLUENCE CHRONIC
RESPONSE HEPATITIS
B.
T0 JL
PAYEN*. J IZOPET**. K IMANI*. L ALRIC***.K BARANGE*. M DUFFAUT***. JP VZNRL*. JP PASCAL*. J PUEL**. *Service d'H6pato-gastroent~rologie et **Service de Virologie;CHU Toulouse-Purpan 31059 Toulouse Cedex, ***Service de M~decine Interne A, CHU Toulouse-Ranguell 31054 Toulouse Cedex-France. The aim of our study was to compare biochemical and virological response to interferon therapy (IFN) in patients with chronic active hepatitis B (CHHBV) with or without cirrhosis. 95 patients with CHHBV were treated by IFN (5MU tiw for 24 weeks) between November 90 and June 94. All patients were positive for HBs Ag, HBVDNA and had histologically proved CHHBV. Among these patients 23 (24%) had histologically proved compensed cirrhosis: groupl, the noncirrhotic patients constituted the group If. Following data were compared between the 3 groups: age, sex, serum ALT activity and level of viraemia (Genestix®) before treatment, Knodell score, biochemical response (complete normalization of ALT) , virological response :typeI:HBVDNA(-),without seroconversion;typeII: HBVDNA (-) ,with aeroconveraion e, typeIII :HBVDNA (-) ,with aeroconveraion e,a. Biochemical and virological relapse post treatment, side effect occurrence. The mean follow up was 26.6+12 months. R e s u l t s : we didn't find any difference between the 2 groups for : sex ratio, serum alt activity, viraemia level, side effect occurrence. Cirrhosis doesn 't influence response to IFN therapy respectively (groupl v3 group If) biochemical response: (48%vs39%, NS) , virological response type I: (17%vsli%, NS) , type Ii : (17%vs26%, NS) , type III: (4%vs5%, NS) ; or relapse : biochemical: (63%vs46%, NS) , virological : (44% vs50%, NS). The mean follow up post treatement was:16-+ll vS 17--+15 months, NS. C o n c l u s i o n s : IFN (SMU, TIW, 24 weeks) is well tolerated in chronic hepatitis B patients with cirrhosis. Cirrhosis doesn't influence biochemical, virological response or relapse in this patients.
Italy.
1467
CIRRHOS i S INTERFERON
473A
1468
FEMUR FRACTURE TRAUMA TO DONORS INCREASES GRAFT SURVIVAL AFTER O R T H O T O P I C RAT LIVER TRANSPLANTATION. XX Pen~._ RT Currin. T Huynh.* CC Baker.* RG Thurmant and JJ Lemasters. Departments of Cell Biology & Anatomy, *Surgery and tPharmacology, University of North Carolina, Chapel Hill. BACKGROUND: Kupffer cell activation after liver storage and transplantation is implicated in primary non-function and initial poor function of liver grafts following orthotopic liver transplantation. Donor variables also influence graft success, and we recently showed that Kupffer cell activation by donor treatment with small amounts of bacterial lipopolysaeeharide increases graft injury and failure substantially (Pang et al., Hepatology 20, 136A). We also observed that blunt femur fracture trauma causes alterations in Kupffer cells, evoking a more antimierobial and less proinflammatory phenotype (Huynh et aL, FASEB J. 9, A571). Since blunt trauma and soft tissue injury typically occur in livers harvested from accident victims, our AIM was to determine what effect donor trauma has on the outcome oforthotopic liver transplantation. METHODS: Under ether anesthesia, male donor Lewis rats underwent closed femur fracture while sham animals received anesthesia only. After 24 hours, livers were harvested, stored in cold UW solution for 24 or 30 hours and transplanted into syngeneic rats using a modified Kamada cuff technique with arterialization. A randomized prospective blinded study design was used. RESULTS: After 24 hours of storage, 4 of 6 grafts from rats subjected to trauma survived 30 days, whereas 2 of 5 grafts from sham-operated rats had long-term survival. Mean survival time was 489 + 147 (S.E.) hours for the femur fracture group vs. 380 + 155 hours for the sham group (p=0.3). In these small groups, the difference of survival was not significant. To test the hypothesis that donor trauma improves graft survival, we stored livers 30 hours when most grafts from normal animals will fail. After longer storage, 5 of 10 livers from traumatized rats survived for 30 days, whereas only 2 of 10 livers from sham-operated rats survived. Mean survival was 340 -+ 103 hours for the femur fracture group vs. 115 _+72 hours for the sham group (p<0.05, l-tailed). CONCLUSION: Soft tissue trauma to liver donors may improve graft survival after orthotupic rat liver transplantation by inducing a more antimicrobial and less proinflammatory Kupffer cell phenotype.
1465
AASLD
ABSTRACTS
ELEVATED LEVELS OF NATRIURETIC HORMONES IN PATIENTS WITH LIVER CIRRHOSIS, F. PATANE' (1), C. FERRI (2), A. PERRONE (2), C. BELLINI (2), A. MAGRINI (3) and C. PUOTI (3). (1) Dept. of Exp. Med. and Pathol. and (2) Inst. of I Clin. Med., University of Rome "La Sapienza", and (3) Liver Unit, Health District RM-H, Marino (Rome),
1466
Deficient production of endogenous Natduretic Hormones (NH) [Digoxin-like Substance (DLS) and Atrial Natduretic Factor (ANF)] has been suggested as a possible mechanism of water end sodium retention in cirrhotic patients with ascites. AIM OF THE STUDY: to evaluate plasma levels of DLS, ANF and aldosterone (PA) in 15 normotensive cirrhotic patients without ascites, 14 cirrhotic patients with ascites (10 with slight to moderate ascites, and 4 with massive ascites) and 28 normotensive healthy subjects. RESULTS: [t] Plasma levels of DLS were significantly lower in controls (18.3i-6.2 pg/mL) than in cirrhotic patients without ascites (68.2:1:28.5 pg/mL, p<0.05) and with ascites (80.7-+21.9 pg/mL, p<0.05). [2] Among patients with ascites DLS significantly correlated with plasma renin activity (PRA: r=0.85, p<0.005) and aldosterone (PA: r=-0.76, p<0.005). [3] ANF levels were significantly lower in healthy subjects (33.9+14.5 pg/mL) than in patients without ascites (122.0+36.1 pg/mL, p<0.05) and in subjects with ascites (128.2+43.8 pg/mL, p
CHOLATE HANDLING BY A BIOARTIFICIAL LIVER (BAL) P Pazzi, AD Moscioni, EMorsiani, J Rozga, AA Demetriou. Dept. o f Surgery and Liver Support Unit, Cedars-Sinai Medical Center, Los Angeles, CA, USA. A BAL, consisting of a hollow fiber module Containing isolated, microcarrierattached porcine hepatocytes, has been shown to reduce serum bile acid levels (BA) in patients with fulminant hepatic failure. We investigated BAL handling of cholie acid. The BAL, loaded with 2.2 x 108 porcine hepatoeytes, was perfused with human plasma containing either 1 laCi of [24-14C]eholate (CA) or 1 laCi of [24-14C]tauroeholate (TCA) and supplemented to 100 pmol with unlabelled CA or TCA, respectively. Samples were collected from plasma and from extrafiber space during a 5-hr perfusion. At the end of the perfusion, hepatocytes were collected for radioactivity and protein determinations, and for examination by transmission electron microscopy (TEM). BAs were separated into 4 fractions (U-~mconjugated, G=glyco- and T=Tanro-conjugated, S=Sulphated) on prepacked silica-based strong anion exchange cartridges; radioactivity was determined by liquid scintillation counting. Data observed during perfusion with CAenriched plasma (expressed as % of total radioactivity detected in each fraction) are reported in the table (mean4-SEM, n=9). ~<0.001 ANOVA Fraction ITime 0 1 2 3 "4 5 U [97.5~0.2 89.74-0.9 81.4=t=111 78.7±2.1 73.5±0.9 71.9±2.7' G 1.4±0.4 9.0±0.7 17.2±1.1 20.5±1.9 25.3±1.3 26.9~2:8' T 0.5±0.1 0.7±0.3 1.1±0A 1.7±0.8 0.7±0.3 0.6+0.1 S 0.4-4-0.1 0.6-4-0.2 0.8±0.1 0.6-4-0.1 0.5-4-0.1 0.54-0.1 After 3-hr, G and S fractions in the extrafiber space were significantly higher than in plasma. During perfusion with TCA-enriched plasma, a relative decrease in T and an increase in G fractions were observed. CA was accumulated by hepatocytes to a three-fold lower level than TCA (3.44-1.8 vs. 10.3±4.5 nmol/mg cell protein), but a significant proportion of radioactivity (26.2±7.3%) was detected in G-fraction. TEM examination of mierocarrier-attaehed porcine hepatoeytes showed ultrastruetural features indicative of typical epithelial cell polarity, the presence of junctional complexes and intact bile eanalieuli as well. On TEM, Golgi complexes and small vesicles were present in the pericanalicular cytoplasm, suggesting the maintenance of a synthetic activity. Conclusions: These results confirm the ability of the BAL to clear BAs from the circulation: porcine hepatucytes accumulate CA and TCA, and are able to conjugate substantial amount of CA.
D O E S N 'T THERAPY
OF
INFLUENCE CHRONIC
RESPONSE HEPATITIS
B.
T0 JL
PAYEN*. J IZOPET**. K IMANI*. L ALRIC***.K BARANGE*. M DUFFAUT***. JP VZNRL*. JP PASCAL*. J PUEL**. *Service d'H6pato-gastroent~rologie et **Service de Virologie;CHU Toulouse-Purpan 31059 Toulouse Cedex, ***Service de M~decine Interne A, CHU Toulouse-Ranguell 31054 Toulouse Cedex-France. The aim of our study was to compare biochemical and virological response to interferon therapy (IFN) in patients with chronic active hepatitis B (CHHBV) with or without cirrhosis. 95 patients with CHHBV were treated by IFN (5MU tiw for 24 weeks) between November 90 and June 94. All patients were positive for HBs Ag, HBVDNA and had histologically proved CHHBV. Among these patients 23 (24%) had histologically proved compensed cirrhosis: groupl, the noncirrhotic patients constituted the group If. Following data were compared between the 3 groups: age, sex, serum ALT activity and level of viraemia (Genestix®) before treatment, Knodell score, biochemical response (complete normalization of ALT) , virological response :typeI:HBVDNA(-),without seroconversion;typeII: HBVDNA (-) ,with aeroconveraion e, typeIII :HBVDNA (-) ,with aeroconveraion e,a. Biochemical and virological relapse post treatment, side effect occurrence. The mean follow up was 26.6+12 months. R e s u l t s : we didn't find any difference between the 2 groups for : sex ratio, serum alt activity, viraemia level, side effect occurrence. Cirrhosis doesn 't influence response to IFN therapy respectively (groupl v3 group If) biochemical response: (48%vs39%, NS) , virological response type I: (17%vsli%, NS) , type Ii : (17%vs26%, NS) , type III: (4%vs5%, NS) ; or relapse : biochemical: (63%vs46%, NS) , virological : (44% vs50%, NS). The mean follow up post treatement was:16-+ll vS 17--+15 months, NS. C o n c l u s i o n s : IFN (SMU, TIW, 24 weeks) is well tolerated in chronic hepatitis B patients with cirrhosis. Cirrhosis doesn't influence biochemical, virological response or relapse in this patients.
Italy.
1467
CIRRHOS i S INTERFERON
473A
1468
FEMUR FRACTURE TRAUMA TO DONORS INCREASES GRAFT SURVIVAL AFTER O R T H O T O P I C RAT LIVER TRANSPLANTATION. XX Pen~._ RT Currin. T Huynh.* CC Baker.* RG Thurmant and JJ Lemasters. Departments of Cell Biology & Anatomy, *Surgery and tPharmacology, University of North Carolina, Chapel Hill. BACKGROUND: Kupffer cell activation after liver storage and transplantation is implicated in primary non-function and initial poor function of liver grafts following orthotopic liver transplantation. Donor variables also influence graft success, and we recently showed that Kupffer cell activation by donor treatment with small amounts of bacterial lipopolysaeeharide increases graft injury and failure substantially (Pang et al., Hepatology 20, 136A). We also observed that blunt femur fracture trauma causes alterations in Kupffer cells, evoking a more antimierobial and less proinflammatory phenotype (Huynh et aL, FASEB J. 9, A571). Since blunt trauma and soft tissue injury typically occur in livers harvested from accident victims, our AIM was to determine what effect donor trauma has on the outcome oforthotopic liver transplantation. METHODS: Under ether anesthesia, male donor Lewis rats underwent closed femur fracture while sham animals received anesthesia only. After 24 hours, livers were harvested, stored in cold UW solution for 24 or 30 hours and transplanted into syngeneic rats using a modified Kamada cuff technique with arterialization. A randomized prospective blinded study design was used. RESULTS: After 24 hours of storage, 4 of 6 grafts from rats subjected to trauma survived 30 days, whereas 2 of 5 grafts from sham-operated rats had long-term survival. Mean survival time was 489 + 147 (S.E.) hours for the femur fracture group vs. 380 + 155 hours for the sham group (p=0.3). In these small groups, the difference of survival was not significant. To test the hypothesis that donor trauma improves graft survival, we stored livers 30 hours when most grafts from normal animals will fail. After longer storage, 5 of 10 livers from traumatized rats survived for 30 days, whereas only 2 of 10 livers from sham-operated rats survived. Mean survival was 340 -+ 103 hours for the femur fracture group vs. 115 _+72 hours for the sham group (p<0.05, l-tailed). CONCLUSION: Soft tissue trauma to liver donors may improve graft survival after orthotupic rat liver transplantation by inducing a more antimicrobial and less proinflammatory Kupffer cell phenotype.