- Email: [email protected]
Functional liver plasma flow after orthotopic liver transplantation (OLT)
Category 1: Liver transplantation, surgery, acute liver failure 7 HEPATOCYTE TRANSPLANTATION AS A TREATMENT FOR GLYCOGEN STORAGE DISEASE TYPE IA Maurizio Muraca I , Giorgio Gerunda I , Daniele Neri I , Mafia Teresa Vilei 1, Anna Granato 1, Paolo Feltracco z, Gianpiero Giron 2, Alberto B. Burlina 3. 1Department of Medical and Surgical Sciences,
Padova; 2Department of Anesthesiology and Pharmacology, Padova; 3Department of Pediatrics, Padova, Italy Hepatocyte transplantation has been used in some patients with liver-based inherited metabolic disorders. We treated with this procedure a 47 year-old woman with glycogen storage disease IA (GSDIA). Despite of oral cornstarch every three hours, she suffered of frequent hypoglycemia, hypertriglyceridemia (20-22 mmol/L, hyperuricemia (0.75 mmol/L) and hyperlacticedemia (4-6.5 mmol/L). Her liver was enlarged and irregular, with multiple adenomas. Compliance with dietary treatment was increasingly difficult because of the poor quality of life. Both the patient and her family gave informed written consent for the procedure, after permission from the local Ethics Committee. Two billion viable hepatocytes were infused in 230 min via one catheter, while heparin (20 UI/kglhr) was infused via the other catheter. Basal portal pressure was 15 mm Hg and it increased to 31 mm Hg during infusion, while portal flow decreased from 14 to 6 cm/s. A triple immunosuppression regime was initiated with mycophenolate mofetil, tacrolimus and steroids. Metabolic stability was achieved in 3 days and the patient was placed on a normal diet; she was able to tolerate at least 7 hours fasting with normal blood glucose levels (ranging from 70 to 110 mg/dl), normal lactate and free fatty acids. Blood triglycerides stabilized around 10-15 mmol/L. Blood urate and lactate were not affected. The metabolic improvement is persisting after one year under tacrolimus immunosuppression. The present observation indicates that hepatocyte transplantation can improve the metabolic defect and quality of life in GSDIA.
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HYPERURICAEMIA, GOUT AND CARDIOVASCULAR RISK AFTER LIVER TRANSPLANTATION
David Neal 1, Brian Tom 2, Alexander Gimson I , Paul Gibbs 3, Graeme Alexander i. 1Department of Medicine, Addenbrooke's NHS
Trust, Cambridge; 2Centrefor Applied Medical Statistics, Department of Public Health and Primary Care, Cambridge; ~University Department of Surgery, Addenbrooke's NHS Trust, Cambridge, UK Background: Hyperuricaemia and gout are recognised complications of renal and cardiac transplantation. In contrast the development of hyperuricaemia following liver transplantation has received less attention. Elevated serum uric acid has been cited as a risk factor for cardiovascular disease in the general population. To evaluate the prevalence of hyperuricaemia and its association with other cardiovascular risk factors we reviewed the case records of 134 consecutive liver transplant recipients with a median follow up of 49 months (range 6-92 months). Results: 47% had hyperuricaemia after liver transplant. Peak uric acid correlated significantly with corresponding serum creatinine (r = 0.694). 6% of patients developed an acute episode of gout. Hypertension, hypercholesterolaemia and a body mass index greater than 25 kg/metre squared were present in 53, 46 and 48% of hyperuricaemic patients respectively and in 47, 54 and 52% of patients with normal serum urate. None of these differences were significant. Cardiovascular events comprised 1 myocardial infarct and 1 incident angina, each patient having hyperuricaemia. Two patients sustained ischaemic strokes, one of whom had hyperuricaemia. Conclusions: There is an important association between liver transplantation and hyperuricaemia. Gout is a significant cause of morbidity but occurs less frequently than after renal or cardiac transplant. There was no association between hyperuricaemia and other cardiovascular risk factors. Too few cardiac events occurred to draw any conclusions about an association with elevated serum uric acid.
41
11261 CONSEQUENCES OF SEVERE ALCOHOLIC RELAPSE AFTER LIVER TRANSPLANTATION (L'r) FOR ALCOHOLIC CIRRHOSIS (AC) Geor~es-Philippe Pageaux 1, Michael Bismuth 1, Pascal Perney 2, Valerie Coste 3, Samir Jaber t , Jean-Michel Fabre l, Pierre Blanc l , Jacques Domergue I , Dominique Larrey I . ~Federation
medico-chirurgicale hepatogastroenterologie, Hopital Saint Eloi, Montpellier; 2Medecine Intern E, Hopital Saint Eloi, Montpellier; 3Anatomo-pathologie, Hopital Gui de Chauliac, Montpellier, France Aim: to distinguish the types of alcohol consumption after LT for AC and to assess the consequences of heavy drinking. Methods: pts transplanted for AC, surviving more than 3 months, were studied. Alcoholic relapse diagnosis was based upon patient's and family members's reports performed with interview, liver enzyme tests, graft biopsy, and use of urine alcohol test. Abstinence was defined by absence of any alcohol drink, social drinking by less than 14 units/week, heavy drinking by more than 14 units/week or periods of time with more than 4 units/day. Results: 128 pts were studied. Mean follow-up was 53.8 months. Overall survival was 83%. 88 pts were considered abstinent, 12 pts social drinkers, and 28 pts heavy drinkers. Among these, overall survival was 77%. Three deaths were related to alcoholic relapse: stop immunosuppression 2, brain hematoma 1. The complications related to alcoholic relapse were: bad compliance to immunosuppressive drugs 2, myocardiopathy 1, ascites 1. 28 pts relapsed during the first year following LT. 23 had at least 1 biopsy of the graft after alcoholic relapse, with a total of 44 biopsies: normal 8, isolated fatty infiltration 20, fatty infiltration and fibrosis 10, alcoholic steatohepatitis 3, cirrhosis 1, chronic hepatitis 2. Fifteen heavy drinkers agreed for inclusion into alcohol specialized treatment programs and 7 of them became abstinent. Conclusion: after LT for AC, 22% were heavy drinkers. Among these, mortality and complications related to alcohol relapse were 11% and 15%, respectively.
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FUNCTIONAL LIVER PLASMA FLOW AFTER ORTHOTOPIC LIVER TRANSPLANTATION (OLT)
C. Pollet 1, F. Bar t , S. Battista I , D. Peola I , A. Gariboldi 2, M.C. Bucchi 1, M. Salizzoni 3, E. Cerutti 3, G. Molino 1. 1Division of General Medicine 6;
2Laboratory of Clinical Chemistry; 3Liver Transplant Unit, San Giovanni Battista Hospital, Turin, Italy Several physiopathological events may significantly modify the hemodynamic features in the early phases post-OLT, determining anatomic or functional alterations of liver plasma flow. The aim of our study was to measure functional liver plasma flow (FLPF) in the short-term after OLT, correlating it with ischemia times of the transplanted liver and the age of donors, and to evaluate changes related to the onset of rejection. FLPF, evaluated by the hepatic clearance of D-sorbitol, was determined in 26 patients undergoing OLT (7 females, 19 males; mean age 46 years, range 19-61 years) in the 1st, 3rd, 7th and 14th day after surgery. The test was conducted by intravenous infusion of a 3% pyrogen-free water solution of D-sorbitol, at a rate of 30 mg/min for 3 hours, followed by steady-state blood and urine sampling in order to calculate the hepatic clearance. Ten of the patients developed clinical and anatomopathological signs of rejection in the study period, resolved after potentiation of immunosuppressive therapy. Mean values of FLPF in the four post-OLT evaluations were higher than those previously described in healthy subjects (1453 + 749, 1211 4- 688, 1222 4666 and 1220 + 380 mL/min in the 1st, 3rd, 7th and 14th day, respectively, vs. 911 4- 137 mL/min). No correlations were found between 1st day FLPF and ischemia times (total and warm) of the transplanted organ or donors' age. Mean percent variations of FLPF found in patients who developed rejection and in those without signs of rejection are shown in the table. Hemodynamic alterations without surgical and/or vascular complications may justify significantly higher mean values of FLPF found in the short-
Poster Sessions
42
Rejection No rejection p (Student t-test)
N 10 16
FLPF 3rd vs 1st -30.0 4- 29.1% +5.6 4- 38.2% 0.019
FLPF 7th vs 1st -24.4 4- 28.5% +5.3 4- 34.8% 0.033
FLPF 14th vs 1st -21.5 4- 27.0% +19.3 4- 26.3% <0.001
term after OLT with respect to healthy subjects. FLPF in the 1st day after surgery, considered as reference value, seems to be influenced neither by ischemia times of the transplanted organ nor by donors' age. Since FLPF significantly decreases in the short-term in the group of patients with signs of rejection with respect to the other group, D-sorbitol clearance test may be a useful tool to show early FLPF falls related to the development of liver rejection.
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HYPOXlA INDUCIBLE FACTOR 1 (HIF 1) IS OVER EXPRESSED IN ACUTE REJECTION FOLLOWING LIVER TRANSPLANTATION
Toby Richards, Debbie Stroka, Daniel Candinas. Liver Unit, Queen
Elizabeth Hospital Edgbaston, Birmingham, UK Introduction: HIF 1 is an oxygen responsive heterodimeric transcription factor regulated by stabilisatiun of the alpha subunit, it is the master regulator of oxygen homeostasis. HIF 1 induces the expression of multiple genes to mediate the molecular adaptation to hypoxic stress including genes involved in oxygen delivery (VEGF, EPO) and cellular metabolism (GLUT 1, LDH). HIF 1 is also upregulated in the pathological processes of inflammarion and during tumor development. Aim: To assess HIF 1 expression in acute liver allograft rejection Methods: Graft biopsies from 18 adult patients were taken one week following liver transplantation, all were assessed by one pathologist for rejection. Immunohistochemistry with a monoclonal antibody against HIF 1 alpha was performed on frozen sections from these biopsies. Two blinded observers scored positive staining. Results: HIF 1 alpha expression was increased in specimens showing acute rejection compared with those showing no rejection (p < 0.02). The severity of rejection did not influence the pattern of HIF 1 alpha expression. HIF 1 alpha expression mostly localised in hepatocytes with occasional expression in sinusoidal cells, there was no zonal pattern of HIF 1 alpha expression. Macrophages (CD 68+) did not express HIF 1 alpha. Conclusion: The increased expression of HIF 1 alpha in hepatocytes may represent an adaptive response to acute rejection involving a novel mechanism of oxygen dependent gene expression.
~ ' 9 " ] ADEFOVIR THERAPY FOR HBV INFECTION AFTER LIVER TRANSPLANTATION (LT) Bruno Roche, Anne-Made Roque-Afonso, Cyrille Feray, Jean-Charles Duclos-Vallee, Henri Bismuth, Didier Samuel.
Hepato-Biliary Center, Paul Brousse Hospital Villejuif, France HBV infection of liver graft is characterized by severe outcome leading to graft failure and is associated with high level of HBV replication. Acquired HBV infection can also occur after LT. Breakthrough or resistance to Lamivudine is common after prolonged use. Nine patients with acquired (n = 8) or recurrent (n = 1) hepatitis B virus infection developed resistance (n = 3) or breakthrough (n = 6) under Lamivudine treatment. Four patients received prior antiviral therapies: ARA-AMP (n = 3), FAMCICLOVIR (n = 3) and GANCICLOVIR (n = 1). Patients (pts) (8 males, 1 females; mean age: 48.8 years 30--67) were treated with Adefovir dipivoxil in a dose of 10 mg daily for a median of 11.1 months (6--17). Lamivudine was continuated in 8 of them. Therapy was initiated at a mean of 60.1 months (18-130) after HBsAg positivation. Pretreatment liver graft histology showed acute hepatitis (n = 1) or chronic active hepatitis (n = 8). Serum HBV DNA was positive in all pts (mean serum level: 7.9 -4- 0.81 log copies/mi), HBeAg in 7. During therapy, serum HBV DNA (PCR), HBsAg and HBeAg were
tested for every 3 months. After 6 months of therapy, - 3 . 9 4- 0.9 Log reductions in HBV DNA levels was observed and negativation of PCR in two cases. No seroconversion to anti-HBe was noted. Seroconversion Ag HBs/Anti-HBs was observed in one patient. Adefovir dipivoxil was well tolerated except in 2 patients in whom the dosage was reduced to 5 mg daily because of renal toxicity. Conclusion: Adefovir dipivoxil is highly effective to reduce HBV replication after LT in patients with resistance or breakthrough under Lamivudine. This therapy is well tolerated.
~0]
VEGF EFFECT ON HEPATOCYTE ENGRAFTMENT AFTER INTRASPLENIC CELL TRANSPLANTATION IN RATS
Hagit Shani 1, Vladislav Tsiperson 1, Gideon Shoshany 2, Gera Neufeld 3, Yaacov Baruch 1.1Live r Unit, Rambam Medical Center and Technion
Medical School Haifa; 2Department of Pediatric Surgery Rambam Medical Center and Technion Medical School Haifa; 3Faculty of Biology, Technion - Israel Institute of Technology Haifa, Israel Introduction: VEGF, a potent angiogenic factor, that increases permeability of blood vessels and promotes hepatocyte growth, may accelerate engraftment and function of transplanted hepatoytes. Aim: VEGF effect on hepatocyte engraftment after intrasplenic cell transplantation in partially hepatectomized (PHP) rats. Methods: Of 18 Lewis syngeneic female rats after 70% PHP, 11 received 240 ng VEGF164, and 7 received saline. Thereafter, intrasplenic transplantation of male hepatocytes (1 x 107) was done. Semi quantitative analysis of PCR product of the SRY region on Y-chromosome was performed. Paraffin embedded sections stained for H+E. Results: Rats were sacrificed as follows: at 24 hrs, 3 in the VEGF group and 2 controls and at 48 hrs, 6 and 4 respectively. By densitometry, the number (Mean 4- SD) of cells in the spleen of VEGF treated group was 1.6 × 103 4- 1.3 compares to saline treated (4.0 x 103 4- 4.9, NS). Transplanted cells were identified only in the liver of the VEGF treated group (1.5 x 103 -4- 0.9) and cells were seen within portal spaces in 4/9 rats until 48 hrs compared to one in saline treated rats at 24 hrs. Cells were attached to each other without signs of damage to blood vessels or adjacent hepatocytes. Conclusions: VEGF164 may enhance hepatocyte engraftment after transplantation. Significant number of cells repopulate the liver within the portal spaces when treated with VEGE
~I-]
SHORT-TERM MECHANICAL HYPERVENTILATION DOES NOT REDUCE CEREBRAL OXIDATIVE METABOLISM IN PATIENTS WITH FULMINANT HEPATIC FAILURE (FHF)
Gitte I. Strauss t , Kirsten Moeller 2, Gitte M. Knudsen 3, Fin S. Larsen 1.
1Department of Hepatology A-2121, Rigshospitalet, Copenhagen; 2Department of Infectious Diseases, Rigshospitalet, Copenhagen; 3Neurobiology Research Unit, Rigshospitalet, Copenhagen, Denmark Spontaneous hyperventilation in patients with FHF could be a compensatory mechanism to reduce intracranial pressure and intracellular acidosis. It remains, however, unknown to which P a C t 2 level FHF patients should be hyperventilated, as cerebral blood flow (CBF) may be compromised. The aim of this study was to determine if cerebral oxidative metabolism in patients with FHF is reduced during acute mechanical hyperventilation. Patients: Fifteen patients with FHF (age 43 + 10 yr.) were enrolled in the study. Investigations were performed within 24 h after appearance of stage 4 hepatic encephalopathy. All patients were kept on mechanical ventilation. Methods: Measurements were performed during normoventilation and during acute mechanical hyperventilation. CBF was measured by the 133Xenon washout technique. Blood samples were withdrawn simultaneously from the radial artery and the internal jugular bulb. Results: Mechanical hyperventilation decreased arterial carbon dioxide tension from 4.9 4- 0.5 to 3.8 4- 0.3 kPa, and CBF from 39 4- 8 to 32 45 mL/100 g/min (p < 0.05). The cerebral metabolism of glucose remained
Padova; 2Department of Anesthesiology and Pharmacology, Padova; 3Department of Pediatrics, Padova, Italy Hepatocyte transplantation has been used in some patients with liver-based inherited metabolic disorders. We treated with this procedure a 47 year-old woman with glycogen storage disease IA (GSDIA). Despite of oral cornstarch every three hours, she suffered of frequent hypoglycemia, hypertriglyceridemia (20-22 mmol/L, hyperuricemia (0.75 mmol/L) and hyperlacticedemia (4-6.5 mmol/L). Her liver was enlarged and irregular, with multiple adenomas. Compliance with dietary treatment was increasingly difficult because of the poor quality of life. Both the patient and her family gave informed written consent for the procedure, after permission from the local Ethics Committee. Two billion viable hepatocytes were infused in 230 min via one catheter, while heparin (20 UI/kglhr) was infused via the other catheter. Basal portal pressure was 15 mm Hg and it increased to 31 mm Hg during infusion, while portal flow decreased from 14 to 6 cm/s. A triple immunosuppression regime was initiated with mycophenolate mofetil, tacrolimus and steroids. Metabolic stability was achieved in 3 days and the patient was placed on a normal diet; she was able to tolerate at least 7 hours fasting with normal blood glucose levels (ranging from 70 to 110 mg/dl), normal lactate and free fatty acids. Blood triglycerides stabilized around 10-15 mmol/L. Blood urate and lactate were not affected. The metabolic improvement is persisting after one year under tacrolimus immunosuppression. The present observation indicates that hepatocyte transplantation can improve the metabolic defect and quality of life in GSDIA.
~-~
HYPERURICAEMIA, GOUT AND CARDIOVASCULAR RISK AFTER LIVER TRANSPLANTATION
David Neal 1, Brian Tom 2, Alexander Gimson I , Paul Gibbs 3, Graeme Alexander i. 1Department of Medicine, Addenbrooke's NHS
Trust, Cambridge; 2Centrefor Applied Medical Statistics, Department of Public Health and Primary Care, Cambridge; ~University Department of Surgery, Addenbrooke's NHS Trust, Cambridge, UK Background: Hyperuricaemia and gout are recognised complications of renal and cardiac transplantation. In contrast the development of hyperuricaemia following liver transplantation has received less attention. Elevated serum uric acid has been cited as a risk factor for cardiovascular disease in the general population. To evaluate the prevalence of hyperuricaemia and its association with other cardiovascular risk factors we reviewed the case records of 134 consecutive liver transplant recipients with a median follow up of 49 months (range 6-92 months). Results: 47% had hyperuricaemia after liver transplant. Peak uric acid correlated significantly with corresponding serum creatinine (r = 0.694). 6% of patients developed an acute episode of gout. Hypertension, hypercholesterolaemia and a body mass index greater than 25 kg/metre squared were present in 53, 46 and 48% of hyperuricaemic patients respectively and in 47, 54 and 52% of patients with normal serum urate. None of these differences were significant. Cardiovascular events comprised 1 myocardial infarct and 1 incident angina, each patient having hyperuricaemia. Two patients sustained ischaemic strokes, one of whom had hyperuricaemia. Conclusions: There is an important association between liver transplantation and hyperuricaemia. Gout is a significant cause of morbidity but occurs less frequently than after renal or cardiac transplant. There was no association between hyperuricaemia and other cardiovascular risk factors. Too few cardiac events occurred to draw any conclusions about an association with elevated serum uric acid.
41
11261 CONSEQUENCES OF SEVERE ALCOHOLIC RELAPSE AFTER LIVER TRANSPLANTATION (L'r) FOR ALCOHOLIC CIRRHOSIS (AC) Geor~es-Philippe Pageaux 1, Michael Bismuth 1, Pascal Perney 2, Valerie Coste 3, Samir Jaber t , Jean-Michel Fabre l, Pierre Blanc l , Jacques Domergue I , Dominique Larrey I . ~Federation
medico-chirurgicale hepatogastroenterologie, Hopital Saint Eloi, Montpellier; 2Medecine Intern E, Hopital Saint Eloi, Montpellier; 3Anatomo-pathologie, Hopital Gui de Chauliac, Montpellier, France Aim: to distinguish the types of alcohol consumption after LT for AC and to assess the consequences of heavy drinking. Methods: pts transplanted for AC, surviving more than 3 months, were studied. Alcoholic relapse diagnosis was based upon patient's and family members's reports performed with interview, liver enzyme tests, graft biopsy, and use of urine alcohol test. Abstinence was defined by absence of any alcohol drink, social drinking by less than 14 units/week, heavy drinking by more than 14 units/week or periods of time with more than 4 units/day. Results: 128 pts were studied. Mean follow-up was 53.8 months. Overall survival was 83%. 88 pts were considered abstinent, 12 pts social drinkers, and 28 pts heavy drinkers. Among these, overall survival was 77%. Three deaths were related to alcoholic relapse: stop immunosuppression 2, brain hematoma 1. The complications related to alcoholic relapse were: bad compliance to immunosuppressive drugs 2, myocardiopathy 1, ascites 1. 28 pts relapsed during the first year following LT. 23 had at least 1 biopsy of the graft after alcoholic relapse, with a total of 44 biopsies: normal 8, isolated fatty infiltration 20, fatty infiltration and fibrosis 10, alcoholic steatohepatitis 3, cirrhosis 1, chronic hepatitis 2. Fifteen heavy drinkers agreed for inclusion into alcohol specialized treatment programs and 7 of them became abstinent. Conclusion: after LT for AC, 22% were heavy drinkers. Among these, mortality and complications related to alcohol relapse were 11% and 15%, respectively.
~-7
FUNCTIONAL LIVER PLASMA FLOW AFTER ORTHOTOPIC LIVER TRANSPLANTATION (OLT)
C. Pollet 1, F. Bar t , S. Battista I , D. Peola I , A. Gariboldi 2, M.C. Bucchi 1, M. Salizzoni 3, E. Cerutti 3, G. Molino 1. 1Division of General Medicine 6;
2Laboratory of Clinical Chemistry; 3Liver Transplant Unit, San Giovanni Battista Hospital, Turin, Italy Several physiopathological events may significantly modify the hemodynamic features in the early phases post-OLT, determining anatomic or functional alterations of liver plasma flow. The aim of our study was to measure functional liver plasma flow (FLPF) in the short-term after OLT, correlating it with ischemia times of the transplanted liver and the age of donors, and to evaluate changes related to the onset of rejection. FLPF, evaluated by the hepatic clearance of D-sorbitol, was determined in 26 patients undergoing OLT (7 females, 19 males; mean age 46 years, range 19-61 years) in the 1st, 3rd, 7th and 14th day after surgery. The test was conducted by intravenous infusion of a 3% pyrogen-free water solution of D-sorbitol, at a rate of 30 mg/min for 3 hours, followed by steady-state blood and urine sampling in order to calculate the hepatic clearance. Ten of the patients developed clinical and anatomopathological signs of rejection in the study period, resolved after potentiation of immunosuppressive therapy. Mean values of FLPF in the four post-OLT evaluations were higher than those previously described in healthy subjects (1453 + 749, 1211 4- 688, 1222 4666 and 1220 + 380 mL/min in the 1st, 3rd, 7th and 14th day, respectively, vs. 911 4- 137 mL/min). No correlations were found between 1st day FLPF and ischemia times (total and warm) of the transplanted organ or donors' age. Mean percent variations of FLPF found in patients who developed rejection and in those without signs of rejection are shown in the table. Hemodynamic alterations without surgical and/or vascular complications may justify significantly higher mean values of FLPF found in the short-
Poster Sessions
42
Rejection No rejection p (Student t-test)
N 10 16
FLPF 3rd vs 1st -30.0 4- 29.1% +5.6 4- 38.2% 0.019
FLPF 7th vs 1st -24.4 4- 28.5% +5.3 4- 34.8% 0.033
FLPF 14th vs 1st -21.5 4- 27.0% +19.3 4- 26.3% <0.001
term after OLT with respect to healthy subjects. FLPF in the 1st day after surgery, considered as reference value, seems to be influenced neither by ischemia times of the transplanted organ nor by donors' age. Since FLPF significantly decreases in the short-term in the group of patients with signs of rejection with respect to the other group, D-sorbitol clearance test may be a useful tool to show early FLPF falls related to the development of liver rejection.
~-]
HYPOXlA INDUCIBLE FACTOR 1 (HIF 1) IS OVER EXPRESSED IN ACUTE REJECTION FOLLOWING LIVER TRANSPLANTATION
Toby Richards, Debbie Stroka, Daniel Candinas. Liver Unit, Queen
Elizabeth Hospital Edgbaston, Birmingham, UK Introduction: HIF 1 is an oxygen responsive heterodimeric transcription factor regulated by stabilisatiun of the alpha subunit, it is the master regulator of oxygen homeostasis. HIF 1 induces the expression of multiple genes to mediate the molecular adaptation to hypoxic stress including genes involved in oxygen delivery (VEGF, EPO) and cellular metabolism (GLUT 1, LDH). HIF 1 is also upregulated in the pathological processes of inflammarion and during tumor development. Aim: To assess HIF 1 expression in acute liver allograft rejection Methods: Graft biopsies from 18 adult patients were taken one week following liver transplantation, all were assessed by one pathologist for rejection. Immunohistochemistry with a monoclonal antibody against HIF 1 alpha was performed on frozen sections from these biopsies. Two blinded observers scored positive staining. Results: HIF 1 alpha expression was increased in specimens showing acute rejection compared with those showing no rejection (p < 0.02). The severity of rejection did not influence the pattern of HIF 1 alpha expression. HIF 1 alpha expression mostly localised in hepatocytes with occasional expression in sinusoidal cells, there was no zonal pattern of HIF 1 alpha expression. Macrophages (CD 68+) did not express HIF 1 alpha. Conclusion: The increased expression of HIF 1 alpha in hepatocytes may represent an adaptive response to acute rejection involving a novel mechanism of oxygen dependent gene expression.
~ ' 9 " ] ADEFOVIR THERAPY FOR HBV INFECTION AFTER LIVER TRANSPLANTATION (LT) Bruno Roche, Anne-Made Roque-Afonso, Cyrille Feray, Jean-Charles Duclos-Vallee, Henri Bismuth, Didier Samuel.
Hepato-Biliary Center, Paul Brousse Hospital Villejuif, France HBV infection of liver graft is characterized by severe outcome leading to graft failure and is associated with high level of HBV replication. Acquired HBV infection can also occur after LT. Breakthrough or resistance to Lamivudine is common after prolonged use. Nine patients with acquired (n = 8) or recurrent (n = 1) hepatitis B virus infection developed resistance (n = 3) or breakthrough (n = 6) under Lamivudine treatment. Four patients received prior antiviral therapies: ARA-AMP (n = 3), FAMCICLOVIR (n = 3) and GANCICLOVIR (n = 1). Patients (pts) (8 males, 1 females; mean age: 48.8 years 30--67) were treated with Adefovir dipivoxil in a dose of 10 mg daily for a median of 11.1 months (6--17). Lamivudine was continuated in 8 of them. Therapy was initiated at a mean of 60.1 months (18-130) after HBsAg positivation. Pretreatment liver graft histology showed acute hepatitis (n = 1) or chronic active hepatitis (n = 8). Serum HBV DNA was positive in all pts (mean serum level: 7.9 -4- 0.81 log copies/mi), HBeAg in 7. During therapy, serum HBV DNA (PCR), HBsAg and HBeAg were
tested for every 3 months. After 6 months of therapy, - 3 . 9 4- 0.9 Log reductions in HBV DNA levels was observed and negativation of PCR in two cases. No seroconversion to anti-HBe was noted. Seroconversion Ag HBs/Anti-HBs was observed in one patient. Adefovir dipivoxil was well tolerated except in 2 patients in whom the dosage was reduced to 5 mg daily because of renal toxicity. Conclusion: Adefovir dipivoxil is highly effective to reduce HBV replication after LT in patients with resistance or breakthrough under Lamivudine. This therapy is well tolerated.
~0]
VEGF EFFECT ON HEPATOCYTE ENGRAFTMENT AFTER INTRASPLENIC CELL TRANSPLANTATION IN RATS
Hagit Shani 1, Vladislav Tsiperson 1, Gideon Shoshany 2, Gera Neufeld 3, Yaacov Baruch 1.1Live r Unit, Rambam Medical Center and Technion
Medical School Haifa; 2Department of Pediatric Surgery Rambam Medical Center and Technion Medical School Haifa; 3Faculty of Biology, Technion - Israel Institute of Technology Haifa, Israel Introduction: VEGF, a potent angiogenic factor, that increases permeability of blood vessels and promotes hepatocyte growth, may accelerate engraftment and function of transplanted hepatoytes. Aim: VEGF effect on hepatocyte engraftment after intrasplenic cell transplantation in partially hepatectomized (PHP) rats. Methods: Of 18 Lewis syngeneic female rats after 70% PHP, 11 received 240 ng VEGF164, and 7 received saline. Thereafter, intrasplenic transplantation of male hepatocytes (1 x 107) was done. Semi quantitative analysis of PCR product of the SRY region on Y-chromosome was performed. Paraffin embedded sections stained for H+E. Results: Rats were sacrificed as follows: at 24 hrs, 3 in the VEGF group and 2 controls and at 48 hrs, 6 and 4 respectively. By densitometry, the number (Mean 4- SD) of cells in the spleen of VEGF treated group was 1.6 × 103 4- 1.3 compares to saline treated (4.0 x 103 4- 4.9, NS). Transplanted cells were identified only in the liver of the VEGF treated group (1.5 x 103 -4- 0.9) and cells were seen within portal spaces in 4/9 rats until 48 hrs compared to one in saline treated rats at 24 hrs. Cells were attached to each other without signs of damage to blood vessels or adjacent hepatocytes. Conclusions: VEGF164 may enhance hepatocyte engraftment after transplantation. Significant number of cells repopulate the liver within the portal spaces when treated with VEGE
~I-]
SHORT-TERM MECHANICAL HYPERVENTILATION DOES NOT REDUCE CEREBRAL OXIDATIVE METABOLISM IN PATIENTS WITH FULMINANT HEPATIC FAILURE (FHF)
Gitte I. Strauss t , Kirsten Moeller 2, Gitte M. Knudsen 3, Fin S. Larsen 1.
1Department of Hepatology A-2121, Rigshospitalet, Copenhagen; 2Department of Infectious Diseases, Rigshospitalet, Copenhagen; 3Neurobiology Research Unit, Rigshospitalet, Copenhagen, Denmark Spontaneous hyperventilation in patients with FHF could be a compensatory mechanism to reduce intracranial pressure and intracellular acidosis. It remains, however, unknown to which P a C t 2 level FHF patients should be hyperventilated, as cerebral blood flow (CBF) may be compromised. The aim of this study was to determine if cerebral oxidative metabolism in patients with FHF is reduced during acute mechanical hyperventilation. Patients: Fifteen patients with FHF (age 43 + 10 yr.) were enrolled in the study. Investigations were performed within 24 h after appearance of stage 4 hepatic encephalopathy. All patients were kept on mechanical ventilation. Methods: Measurements were performed during normoventilation and during acute mechanical hyperventilation. CBF was measured by the 133Xenon washout technique. Blood samples were withdrawn simultaneously from the radial artery and the internal jugular bulb. Results: Mechanical hyperventilation decreased arterial carbon dioxide tension from 4.9 4- 0.5 to 3.8 4- 0.3 kPa, and CBF from 39 4- 8 to 32 45 mL/100 g/min (p < 0.05). The cerebral metabolism of glucose remained