- Email: [email protected]
143 Outcome of steatotic graft in living-related liver transplantation
Category 1: Liver Transplantation/Surgery/Acute Liver Failure
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not statistically higher in steatosis group. Overall-QOL was not affected by fibrosis, ductopenia or cirrhosis. Nevertheless, GHP score was lower in Fibrosis patients (p=0.02) and well-being score was lower in ductopenia patients (p=0.05). These finding could be related to HCV infection (p<0.001), HBV recurrence (p=0.01) and ABO mismatch (p<0.05). In conclusion, long-term post-LT graft macrovesicular steatosis and/or fibrosis, could affect some domains of recipient QOL 10 years after LT.
groups receiving NH4+ (P<0.05) and brain water (P<0.05). Tacrolimus did not influence cerebral perfusion pressure. Conclusion: Our results shows that tacrolimus prevents cerebral vasodilatation and ameliorates intracranial hypertension in PCA rats receiving NH4+ infusion. These results indicate that calcineurin is involved in the development of high CBF and brain edema in FHF, and encourage a clinical trial in patients with FHF awaiting liver transplantation.
141 COLD ISCHEMIA-REPERFUSION INJURY OF RAT ISOLATED
143 OUTCOME OF STEATOTIC GRAFT IN LIVING-RELATED LIVER
LIVERS - IMMUNOHISTOCHEMICAL STUDY
M. Kopecky 1 , V. Semecky 1 , P. Nachtigal 1 , I. Tilser 2 . 1 Dept. Biological and Medical Sciences, Faculty of Pharmacy, Charles University, Hradec Kralove,; 2 Dept. Pharmacology and Toxicology, Faculty of Pharmacy, Charles University, Hradec Kralove, Czech Republic The aim of this study was to describe the histological changes of liver structure in isolated cold-stored and reperfused livers and to introduce some immunohistochemical methods into study of this phenomenon. The livers of female Wistar rats (n=5 in each group) were isolated, stored in UW solution for 3h, 24h or 48h in 4 C, then warm up to 37 C for 30min and reperfused for 90min under a recirculation regimen. The liver specimens were fixed in Baker’s or Bouin’s fixatives, paraffin-embedded and the sections were stained with hematoxylin & eosin or methyl green-pyronin. PCNA, vimentin and prohibitin were detected using immunohistochemistry, and the TUNEL assay was performed. We observed that the sinusoidal endothelial cells (SEC) were injured more and earlier than hepatocytes. In the 48h group 51.47±5.32% of SEC and 2.86±0.95% of hepatocytes were damaged. The PCNA staining was highest in control and 3h groups while TUNEL positive cells were observed predominantly in the livers from 24h and 48h groups. Surprisingly, the injury of SEC (and macrophage activation) was well detected in vimentin-stained liver sections where the staining of SEC diminished as these cells detached from the hepatocytes. In the 48h group, a strong prohibitin expression was found in a few hepatocytes while it decreased in a majority of liver cells when compared with controls. In conclusion, the histological and immunohistochemical methods used in this work are useful tools for assessing the cold ischemia-reperfusion injury of the liver. Supported by FRVS 2978/2003 and by the Czech Ministry of Education 11600002.
142 TACROLIMUS AMELIORATES CEREBRAL VASODILATATION AND INTRACRANIAL HYPERTENSION IN THE RAT WITH PORTACAVAL ANASTOMOSIS AND HYPERAMMONEMIA
F.S. Larsen, T. Dethloff, B.A. Hansen. Dept. Hepatology, Rigshospitalet, Copenhagen, Denmark Background and aims: Arterial hyperammonemia and cerebral vasodilatation correlate with cerebral herniation in fulminant hepatic failure (FHF). Tacrolimus is a calcineurin inhibitor that increases cerebrovascular tone. In this study we determined if tacrolimus prevents cerebral vasodilatation and high ICP in rats with a portacaval anastomosis (PCA) challenged to high arterial ammonia (NH4+) concentration. Methods: Four groups of mechanically ventilated rats, with 6-9 rats in each group, were investigated within 48 hours after construction of a PCA. Four groups of the rats received either infusion of NH4+ (55µmol/kg·h) or saline for ∼180 min. Two groups of the rats receiving either NH4+ or saline had i.v. tacrolimus (0.4 mg/kg) or vehicle (veh) 15 min before start of NH4+ or saline infusion. Cerebral blood flow was monitored by a laser Doppler probe in brain cortex. ICP was monitored by placement of a catheter in the cerebrospinal fluid. Brain water was determined by the gravimetric technique. Results: ICP increased in rats with PCA receiving NH4+ infusion from 1.8±0.8 to 7.8±0.8 mmHg (P<0.05), compared to controls (from 1.5±0.3 to 2.4±0.8 mmHg (NS). Tacrolimus prevented an increase in CBF in
TRANSPLANTATION
J.H. Lee, Y.J. Oh, M.S. Choi, K.C. Koh, S.W. Paik, B.C. Yoo. Division of Gastroenterology, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, Korea Background: Steatotic liver graft is known to be a risk factor for cadaveric liver transplantation due to reduced tolerance against ischemia-reperfusion injury. However, there is not enough data regarding the role of steatosis in living-related liver transplantation (LRLT), where cold ischemic time is relatively short. Thus, we aimed to evaluate the impact of hepatic graft steatosis on the outcome of LRLT. Methods: Liver wedge biopsy was performed on 150 consecutive LRLT donors at the time of liver resection. Donor liver grafts were classified by the degree of macrovesicular fat deposition into four groups as follow: minimal (n=89), 0-9%, mild (n=35), 10-19%, moderate I (n=18), 20-29%, moderate II (n=8), 30-60%. Parameters used for evaluation of recipients were initial graft function (peak bilirubin and ALT levels within the 14th postoperative day) and patient survival. Parameters used for evaluation of donors were the peak ALT levels, hospital stay, and postoperative morbidity. Result: There was no significant difference in the peak bilirubin (minimal, 11.1 ± 10.6; mild, 14.2±14.5; moderate I, 9.6±8.0; moderate II, 14.4±14.4 mg/dL) and ALT levels (minimal, 723±1550; mild, 924±1511; moderate I, 524±320; moderate II, 1003±1115 IU/L) of the recipients among groups (p=0.41 and 0.75, respectively). Ninety-day and 1-year survivals of the recipients were comparable among groups. There was no significant difference in peak ALT levels, hospital stay, or postoperative morbidity of donors among groups. Conclusion: A steatotic liver graft up to the moderate level (< 60% steatosis) may be tolerated in LRLT.
144 LONG-TERM RIBAVIRIN MONOTHERAPY SIGNIFICANTLY DELAYS FIBROSIS PROGRESSION IN OLT RECIPIENTS WITH RECURRENT HEPATITIS C
R. Lionetti 1 , G. Palmieri 3 , S. Battista 3 , A. Petrolati 1 , D. Di Paolo 1 , C. Ciceroni 2 , L. De Luca 2 , G. Tisone 2 , M. Angelico 1 . 1 Gastroenterology Unit, Tor Vergata University, Rome, Italy; 2 Liver Transplantation Center, Tor Vergata University, Rome, Italy; 3 Histopathology Department, Tor Vergata University, Rome, Italy Background: Recurrent hepatitis C after OLT is associated with rapid fibrosis progression. HCV eradication with antivirals is difficult in this setting. Aim: To assess whether long-term ribavirin (RBV) monotherapy affects liver inflammation and fibrosis progression. Methods: 15 OLT recipients with HCV recurrent disease and a followup >4 years without any treatment received RBV monotherapy, up to the maximum tolerated dose, for 3 years. In each patient 6 yearly biopsies were assessed for grading and staging (Ishak), 3 before and 3 during treatment. Variations of >2 points for grading and of >1 point for staging in the last pre-treatment biopsy vs that after 3 years of RBV (end-oftreatment) were considered as significant histological changes (improvement/deterioration). Results: Mean ribavirin dose was 353±74 mg/day. Mean grading score was 5.2±1.9 before and 4.3±1.4 after RBV (p<0.07). Fibrosis score im-
49
not statistically higher in steatosis group. Overall-QOL was not affected by fibrosis, ductopenia or cirrhosis. Nevertheless, GHP score was lower in Fibrosis patients (p=0.02) and well-being score was lower in ductopenia patients (p=0.05). These finding could be related to HCV infection (p<0.001), HBV recurrence (p=0.01) and ABO mismatch (p<0.05). In conclusion, long-term post-LT graft macrovesicular steatosis and/or fibrosis, could affect some domains of recipient QOL 10 years after LT.
groups receiving NH4+ (P<0.05) and brain water (P<0.05). Tacrolimus did not influence cerebral perfusion pressure. Conclusion: Our results shows that tacrolimus prevents cerebral vasodilatation and ameliorates intracranial hypertension in PCA rats receiving NH4+ infusion. These results indicate that calcineurin is involved in the development of high CBF and brain edema in FHF, and encourage a clinical trial in patients with FHF awaiting liver transplantation.
141 COLD ISCHEMIA-REPERFUSION INJURY OF RAT ISOLATED
143 OUTCOME OF STEATOTIC GRAFT IN LIVING-RELATED LIVER
LIVERS - IMMUNOHISTOCHEMICAL STUDY
M. Kopecky 1 , V. Semecky 1 , P. Nachtigal 1 , I. Tilser 2 . 1 Dept. Biological and Medical Sciences, Faculty of Pharmacy, Charles University, Hradec Kralove,; 2 Dept. Pharmacology and Toxicology, Faculty of Pharmacy, Charles University, Hradec Kralove, Czech Republic The aim of this study was to describe the histological changes of liver structure in isolated cold-stored and reperfused livers and to introduce some immunohistochemical methods into study of this phenomenon. The livers of female Wistar rats (n=5 in each group) were isolated, stored in UW solution for 3h, 24h or 48h in 4 C, then warm up to 37 C for 30min and reperfused for 90min under a recirculation regimen. The liver specimens were fixed in Baker’s or Bouin’s fixatives, paraffin-embedded and the sections were stained with hematoxylin & eosin or methyl green-pyronin. PCNA, vimentin and prohibitin were detected using immunohistochemistry, and the TUNEL assay was performed. We observed that the sinusoidal endothelial cells (SEC) were injured more and earlier than hepatocytes. In the 48h group 51.47±5.32% of SEC and 2.86±0.95% of hepatocytes were damaged. The PCNA staining was highest in control and 3h groups while TUNEL positive cells were observed predominantly in the livers from 24h and 48h groups. Surprisingly, the injury of SEC (and macrophage activation) was well detected in vimentin-stained liver sections where the staining of SEC diminished as these cells detached from the hepatocytes. In the 48h group, a strong prohibitin expression was found in a few hepatocytes while it decreased in a majority of liver cells when compared with controls. In conclusion, the histological and immunohistochemical methods used in this work are useful tools for assessing the cold ischemia-reperfusion injury of the liver. Supported by FRVS 2978/2003 and by the Czech Ministry of Education 11600002.
142 TACROLIMUS AMELIORATES CEREBRAL VASODILATATION AND INTRACRANIAL HYPERTENSION IN THE RAT WITH PORTACAVAL ANASTOMOSIS AND HYPERAMMONEMIA
F.S. Larsen, T. Dethloff, B.A. Hansen. Dept. Hepatology, Rigshospitalet, Copenhagen, Denmark Background and aims: Arterial hyperammonemia and cerebral vasodilatation correlate with cerebral herniation in fulminant hepatic failure (FHF). Tacrolimus is a calcineurin inhibitor that increases cerebrovascular tone. In this study we determined if tacrolimus prevents cerebral vasodilatation and high ICP in rats with a portacaval anastomosis (PCA) challenged to high arterial ammonia (NH4+) concentration. Methods: Four groups of mechanically ventilated rats, with 6-9 rats in each group, were investigated within 48 hours after construction of a PCA. Four groups of the rats received either infusion of NH4+ (55µmol/kg·h) or saline for ∼180 min. Two groups of the rats receiving either NH4+ or saline had i.v. tacrolimus (0.4 mg/kg) or vehicle (veh) 15 min before start of NH4+ or saline infusion. Cerebral blood flow was monitored by a laser Doppler probe in brain cortex. ICP was monitored by placement of a catheter in the cerebrospinal fluid. Brain water was determined by the gravimetric technique. Results: ICP increased in rats with PCA receiving NH4+ infusion from 1.8±0.8 to 7.8±0.8 mmHg (P<0.05), compared to controls (from 1.5±0.3 to 2.4±0.8 mmHg (NS). Tacrolimus prevented an increase in CBF in
TRANSPLANTATION
J.H. Lee, Y.J. Oh, M.S. Choi, K.C. Koh, S.W. Paik, B.C. Yoo. Division of Gastroenterology, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, Korea Background: Steatotic liver graft is known to be a risk factor for cadaveric liver transplantation due to reduced tolerance against ischemia-reperfusion injury. However, there is not enough data regarding the role of steatosis in living-related liver transplantation (LRLT), where cold ischemic time is relatively short. Thus, we aimed to evaluate the impact of hepatic graft steatosis on the outcome of LRLT. Methods: Liver wedge biopsy was performed on 150 consecutive LRLT donors at the time of liver resection. Donor liver grafts were classified by the degree of macrovesicular fat deposition into four groups as follow: minimal (n=89), 0-9%, mild (n=35), 10-19%, moderate I (n=18), 20-29%, moderate II (n=8), 30-60%. Parameters used for evaluation of recipients were initial graft function (peak bilirubin and ALT levels within the 14th postoperative day) and patient survival. Parameters used for evaluation of donors were the peak ALT levels, hospital stay, and postoperative morbidity. Result: There was no significant difference in the peak bilirubin (minimal, 11.1 ± 10.6; mild, 14.2±14.5; moderate I, 9.6±8.0; moderate II, 14.4±14.4 mg/dL) and ALT levels (minimal, 723±1550; mild, 924±1511; moderate I, 524±320; moderate II, 1003±1115 IU/L) of the recipients among groups (p=0.41 and 0.75, respectively). Ninety-day and 1-year survivals of the recipients were comparable among groups. There was no significant difference in peak ALT levels, hospital stay, or postoperative morbidity of donors among groups. Conclusion: A steatotic liver graft up to the moderate level (< 60% steatosis) may be tolerated in LRLT.
144 LONG-TERM RIBAVIRIN MONOTHERAPY SIGNIFICANTLY DELAYS FIBROSIS PROGRESSION IN OLT RECIPIENTS WITH RECURRENT HEPATITIS C
R. Lionetti 1 , G. Palmieri 3 , S. Battista 3 , A. Petrolati 1 , D. Di Paolo 1 , C. Ciceroni 2 , L. De Luca 2 , G. Tisone 2 , M. Angelico 1 . 1 Gastroenterology Unit, Tor Vergata University, Rome, Italy; 2 Liver Transplantation Center, Tor Vergata University, Rome, Italy; 3 Histopathology Department, Tor Vergata University, Rome, Italy Background: Recurrent hepatitis C after OLT is associated with rapid fibrosis progression. HCV eradication with antivirals is difficult in this setting. Aim: To assess whether long-term ribavirin (RBV) monotherapy affects liver inflammation and fibrosis progression. Methods: 15 OLT recipients with HCV recurrent disease and a followup >4 years without any treatment received RBV monotherapy, up to the maximum tolerated dose, for 3 years. In each patient 6 yearly biopsies were assessed for grading and staging (Ishak), 3 before and 3 during treatment. Variations of >2 points for grading and of >1 point for staging in the last pre-treatment biopsy vs that after 3 years of RBV (end-oftreatment) were considered as significant histological changes (improvement/deterioration). Results: Mean ribavirin dose was 353±74 mg/day. Mean grading score was 5.2±1.9 before and 4.3±1.4 after RBV (p<0.07). Fibrosis score im-