- Email: [email protected]
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Vol. 7, No. 2 2003
that ATP regulates mitochondrial Ca2+ uptake via a receptor-like mechanism. Methods. Rat livers were perfused with UW solution via the portal vein, harvested and homogenized. Mitochondria were separated from liver homogenate by differential centrifugation and mitochondrial Ca2+ uptake was determined by using 45Ca2+. After incubation at 37C in the presence 0.2 M 45Ca2+, 1mM pyruvate and 1 mM malate, the mitochondrial suspension was filtered and Ca2+ uptake was calculated from the radioactivity of the filters. A non-hydrolysable analog of ATP, AMPPNP (100 M), Ruthenium Red (inhibitor of mitochondrial Ca2+ uniporter, RR, 10 M) and P2Y receptor antagonist reactive blue-2 (RB-2, 300 M) were added to the incubation medium to determine their influence on Ca2+ uptake. The experiment was repeated on 3 animals and each measurement was performed in triplicates. Statistical analysis was performed using Student’s t-test, with p0.05 taken as significant. Results: 1. ATP analog AMP-PNP significantly activates mitochondrial Ca2+ uptake. 2. RR completely inhibits Ca2+ uptake by itself and in combination with AMP-PNP. 3. P2Y inhibitor RB-2 completely inhibits mitochondrial Ca2+ uptake in combination with AMP-PNP. Conclusions. 1. The effect of AMP-PNP suggests that intracellular ATP at concentration lower then normal activates mitochondrial Ca2+ uptake. 2. Inhibition of this effect by RR suggests that intracellular ATP activates Ca2+ uptake via the calcium uniporter. 3. The ability of RB-2 to cancel the effect of AMP-PNP and depress Ca2+ uptake below baseline suggests that ATP influences Ca2+ uptake via a P2Y receptor mechanism.
43 Biliary Complications in 96 Right Lobe Living Donor Liver Transplants Giovanni Varotti, Gabriel E Gondolesi, Luis Munoz, Sander Florman, Thomas M Fishbein, Sukru Emre, Myron E Schwartz, Charles Miller, Mount Sinai School of Medicine, New York, NY
Abstracts
271
Hemangioendotheliomas are exceedingly rare liver tumors that are often multi-focal and incompatible with resection. Liver transplantation has been anecdotally utilized in patients but significant series have not been reported. Methods: We retrospectively reviewed our transplant tumor database for all hemangioendothelioma patients who underwent liver transplantation. Results: Eighteen patients received liver transplants for the diagnosis of hemangioendotheliomas. The mean age of the group was 33.3 13.2 years (range 3-57 yrs) and was comprised of 12 females and 6 males. There were 16 Caucasians, 1 African American, and 1 Asian. The diagnosis was made by preoperative biopsy and the presence of lymph node invasion confirmed at the time of explantation. Antibody induction was utilized in 3 patients while IL-2 receptor antibody was utilized in 2 other patients. Maintenance immunosuppression consisted of prednisone (n18), cyclosporine (n10), tacrolimus (n8), azathioprine (n7), and Mycophenolate mofeltil (n4). Post-operative adjuvant chemotherapy was given to three patients (2 patients with node + disease). Mean follow-up was 27.2 35.8 months (range 6 to 110 months). Node positive disease was encountered in 3 patients. Recurrent disease was observed in three patients (16.7%), two of these had node + disease. The overall patient survival was 77% at 3-years. When compared the survival in the node + group was 33% with the 2 deaths occurring in the first six months post-op. While in contrast, survival in the node negative group was 75%, p0.001. The mean survival was 9.2 5.4 months for those with recurrent disease. Conclusions: Transplantation for hemangioendotheliomas can be performed safely with excellent 3-year survival. Unfortunately, node positive disease portends a high-risk for recurrence, which significantly diminishes patient survival.
45 Does Estrogen Protect Against Reperfusion Injury in the Liver? Carson Cunningham, M K Ghanta, Xinje Mu, Kevin N Boykin, Gazi B Zibari, LSUHSC Shreveport, Shreveport, LA
Introduction: Biliary reconstruction during right lobe living donor liver transplantation (RL LDLT) is the most technically challenging aspect of the procedure. This study documents the incidence, nature, and outcome of biliary complications following RL LDLT. Method: Between June 1999 and January 2002, 96 RL LDLT were performed in our center (91 adults; 5 children). We retrospectively reviewed the records of these recipients, noting the number of bile duct anastomosis; type of reconstruction; incidence, timing, treatment, and outcome of leaks and strictures; and patient survival. Results: Multiple ducts ( 2) were found in 58 grafts (60.4%). Roux-en-Y reconstruction was performed in 53 cases (55.2%), duct-to-duct in 39 (40.6%), and both in 4 cases (4.2%). Thirty-nine recipients (40.6 %) had biliary complications: 21 patients had leaks and 22 had strictures (4 patients had both). Six patients had multiple biliary leaks requiring multiple operations. Patients with 2 biliary anastomosis had a higher incidence of bile leaks (19% vs. 5.5%, pNS). The incidence of leaks was higher with Roux-en-Y compared to duct to duct (18.2% vs 7.3%; pNS); the opposite was for strictures (16.3% vs 31.7%; pNS). Leaks occur at a median of 12 days after transplant, while strictures presented at a median of 178.5 days. Freedom from biliary complications was 55% at 2 years. Two-year survival for patients with and without biliary leaks was 65% and 85%, respectively (p0.07). Overall 2-year patient and graft survival was 81% and 77%, respectively. Conclusion: The bile duct is still the Achilles’ Heel of the liver transplant, with a complication rate of 40% in RL LDLT. Leaks seem to be more common with multiple anastomosis and Roux-en-Y reconstructions; duct-to-duct reconstruction is more prone to stricture. Bile leaks require aggressive treatment to improve outcome.
Organ ischemia followed by reperfusion injury impacts many areas of surgical care. Ongoing research is directed at limiting or preventing reperfusion injury. Prior studies have shown female rats to have decreased lung injury after hemorrhagic shock followed by resuscitation. The purpose of our study was to investigate whether estrogen would protect against reperfusion injury in the murine liver. Twenty male C57BL/6 mice were divided into 2 groups: control and experimental. Each group underwent midline laparotomy and ligation of the vascular pedicle of the left lateral lobe of the liver for one hour. After one hour of ischemia, the left lateral lobe was allowed to reperfuse for 5 hours. After 5 hours of reperfusion, the carotid artery was cannulated and 0.5 cc of blood withdrawn. The abdomen was then re-opened and a biopsy of the left lateral lobe was taken to be stained with TUNEL reagent. The animal was then sacrificed. Mice in the experimental group were given a subcutaneous injection of estrogen twenty-four hours prior to laparotomy. Mean AST in the control group was 2368. Mean AST in the experimental group was 514. There was not a statistically significant difference between these two groups (p.1365). Mean ALT in the control group was 3767. Mean ALT in the experimental group was 861. Again, there was not a statistically significant difference between these groups (p.2025). Tissue specimens from neither the control group nor the experimental group had TUNEL positive cells indicating the start of apoptosis. Although mean transaminase levels between the control and experimental groups are not equal, the difference was not statistically significant.
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Liver Transplantation for Hemangioendothelioma Manish Gupta, Todd Merchen, Thomas M Beebe, Thomas G Gross, Rita R Alloway, Michael J Hanaway, Jennifer Trofe, Steve Rudich, M R First, E S Woodle, Joseph F Buell, The University of Cincinnati, Cincinnati, OH
Impact of Donor and Recipient Risk Factors on Survival and Quality of Life After Liver Transplantation Derek E Moore, Irene Feurer, C W Pinson, Vanderbilt University Medical Center, Nashville, TN
that ATP regulates mitochondrial Ca2+ uptake via a receptor-like mechanism. Methods. Rat livers were perfused with UW solution via the portal vein, harvested and homogenized. Mitochondria were separated from liver homogenate by differential centrifugation and mitochondrial Ca2+ uptake was determined by using 45Ca2+. After incubation at 37C in the presence 0.2 M 45Ca2+, 1mM pyruvate and 1 mM malate, the mitochondrial suspension was filtered and Ca2+ uptake was calculated from the radioactivity of the filters. A non-hydrolysable analog of ATP, AMPPNP (100 M), Ruthenium Red (inhibitor of mitochondrial Ca2+ uniporter, RR, 10 M) and P2Y receptor antagonist reactive blue-2 (RB-2, 300 M) were added to the incubation medium to determine their influence on Ca2+ uptake. The experiment was repeated on 3 animals and each measurement was performed in triplicates. Statistical analysis was performed using Student’s t-test, with p0.05 taken as significant. Results: 1. ATP analog AMP-PNP significantly activates mitochondrial Ca2+ uptake. 2. RR completely inhibits Ca2+ uptake by itself and in combination with AMP-PNP. 3. P2Y inhibitor RB-2 completely inhibits mitochondrial Ca2+ uptake in combination with AMP-PNP. Conclusions. 1. The effect of AMP-PNP suggests that intracellular ATP at concentration lower then normal activates mitochondrial Ca2+ uptake. 2. Inhibition of this effect by RR suggests that intracellular ATP activates Ca2+ uptake via the calcium uniporter. 3. The ability of RB-2 to cancel the effect of AMP-PNP and depress Ca2+ uptake below baseline suggests that ATP influences Ca2+ uptake via a P2Y receptor mechanism.
43 Biliary Complications in 96 Right Lobe Living Donor Liver Transplants Giovanni Varotti, Gabriel E Gondolesi, Luis Munoz, Sander Florman, Thomas M Fishbein, Sukru Emre, Myron E Schwartz, Charles Miller, Mount Sinai School of Medicine, New York, NY
Abstracts
271
Hemangioendotheliomas are exceedingly rare liver tumors that are often multi-focal and incompatible with resection. Liver transplantation has been anecdotally utilized in patients but significant series have not been reported. Methods: We retrospectively reviewed our transplant tumor database for all hemangioendothelioma patients who underwent liver transplantation. Results: Eighteen patients received liver transplants for the diagnosis of hemangioendotheliomas. The mean age of the group was 33.3 13.2 years (range 3-57 yrs) and was comprised of 12 females and 6 males. There were 16 Caucasians, 1 African American, and 1 Asian. The diagnosis was made by preoperative biopsy and the presence of lymph node invasion confirmed at the time of explantation. Antibody induction was utilized in 3 patients while IL-2 receptor antibody was utilized in 2 other patients. Maintenance immunosuppression consisted of prednisone (n18), cyclosporine (n10), tacrolimus (n8), azathioprine (n7), and Mycophenolate mofeltil (n4). Post-operative adjuvant chemotherapy was given to three patients (2 patients with node + disease). Mean follow-up was 27.2 35.8 months (range 6 to 110 months). Node positive disease was encountered in 3 patients. Recurrent disease was observed in three patients (16.7%), two of these had node + disease. The overall patient survival was 77% at 3-years. When compared the survival in the node + group was 33% with the 2 deaths occurring in the first six months post-op. While in contrast, survival in the node negative group was 75%, p0.001. The mean survival was 9.2 5.4 months for those with recurrent disease. Conclusions: Transplantation for hemangioendotheliomas can be performed safely with excellent 3-year survival. Unfortunately, node positive disease portends a high-risk for recurrence, which significantly diminishes patient survival.
45 Does Estrogen Protect Against Reperfusion Injury in the Liver? Carson Cunningham, M K Ghanta, Xinje Mu, Kevin N Boykin, Gazi B Zibari, LSUHSC Shreveport, Shreveport, LA
Introduction: Biliary reconstruction during right lobe living donor liver transplantation (RL LDLT) is the most technically challenging aspect of the procedure. This study documents the incidence, nature, and outcome of biliary complications following RL LDLT. Method: Between June 1999 and January 2002, 96 RL LDLT were performed in our center (91 adults; 5 children). We retrospectively reviewed the records of these recipients, noting the number of bile duct anastomosis; type of reconstruction; incidence, timing, treatment, and outcome of leaks and strictures; and patient survival. Results: Multiple ducts ( 2) were found in 58 grafts (60.4%). Roux-en-Y reconstruction was performed in 53 cases (55.2%), duct-to-duct in 39 (40.6%), and both in 4 cases (4.2%). Thirty-nine recipients (40.6 %) had biliary complications: 21 patients had leaks and 22 had strictures (4 patients had both). Six patients had multiple biliary leaks requiring multiple operations. Patients with 2 biliary anastomosis had a higher incidence of bile leaks (19% vs. 5.5%, pNS). The incidence of leaks was higher with Roux-en-Y compared to duct to duct (18.2% vs 7.3%; pNS); the opposite was for strictures (16.3% vs 31.7%; pNS). Leaks occur at a median of 12 days after transplant, while strictures presented at a median of 178.5 days. Freedom from biliary complications was 55% at 2 years. Two-year survival for patients with and without biliary leaks was 65% and 85%, respectively (p0.07). Overall 2-year patient and graft survival was 81% and 77%, respectively. Conclusion: The bile duct is still the Achilles’ Heel of the liver transplant, with a complication rate of 40% in RL LDLT. Leaks seem to be more common with multiple anastomosis and Roux-en-Y reconstructions; duct-to-duct reconstruction is more prone to stricture. Bile leaks require aggressive treatment to improve outcome.
Organ ischemia followed by reperfusion injury impacts many areas of surgical care. Ongoing research is directed at limiting or preventing reperfusion injury. Prior studies have shown female rats to have decreased lung injury after hemorrhagic shock followed by resuscitation. The purpose of our study was to investigate whether estrogen would protect against reperfusion injury in the murine liver. Twenty male C57BL/6 mice were divided into 2 groups: control and experimental. Each group underwent midline laparotomy and ligation of the vascular pedicle of the left lateral lobe of the liver for one hour. After one hour of ischemia, the left lateral lobe was allowed to reperfuse for 5 hours. After 5 hours of reperfusion, the carotid artery was cannulated and 0.5 cc of blood withdrawn. The abdomen was then re-opened and a biopsy of the left lateral lobe was taken to be stained with TUNEL reagent. The animal was then sacrificed. Mice in the experimental group were given a subcutaneous injection of estrogen twenty-four hours prior to laparotomy. Mean AST in the control group was 2368. Mean AST in the experimental group was 514. There was not a statistically significant difference between these two groups (p.1365). Mean ALT in the control group was 3767. Mean ALT in the experimental group was 861. Again, there was not a statistically significant difference between these groups (p.2025). Tissue specimens from neither the control group nor the experimental group had TUNEL positive cells indicating the start of apoptosis. Although mean transaminase levels between the control and experimental groups are not equal, the difference was not statistically significant.
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Liver Transplantation for Hemangioendothelioma Manish Gupta, Todd Merchen, Thomas M Beebe, Thomas G Gross, Rita R Alloway, Michael J Hanaway, Jennifer Trofe, Steve Rudich, M R First, E S Woodle, Joseph F Buell, The University of Cincinnati, Cincinnati, OH
Impact of Donor and Recipient Risk Factors on Survival and Quality of Life After Liver Transplantation Derek E Moore, Irene Feurer, C W Pinson, Vanderbilt University Medical Center, Nashville, TN