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Impact of older donors on hepatic allograft viability
A1234
SSAT ABSTRACTS
GASTROENTEROLOGY, Vol. 108, No. 4
• IMPACT OF O L D E R DONORS O N HEPATIC ALLOGRAFT VIABILITY. D Mital, H Sankary, J Whiting, P Yoster,L M c ~ h ~ J Williams Department of Surgery, Rush Presbyterian St Lukes Medical Center, Chicago, Illinois. As the success of hepatic transplantation improves, there is increasing demand for a limited supply of organs. We recently expanded our static donor pool by utilizing livers from donors of age greater than 60 yrs. The purpose of this study is to determine whether the use of older donors, as a source of hepatic allografts, adversely affects the outcome following transplantation. In a retrospective analysis indices of graft viability were compared in two groups of patients: The first group included 214 adult patients who received grafts from donors of age less than 60 yrs(CONT). The second group included 47 patients who received organs from donors of age greater than 60 yrs of age(OLD). RESULTS: Donor age was significantly greater in the OLD(60-80 yrs) group when compared to CONT(67~5 vs 30±15 yrs,P<.ool). Donor characteristics including hospital stay, preservation time, dopamine dose, total bilirubin, ALT, alkaline phosphatase were similar between groups. Recipients had similar distributions of diseases, ages, Child's classification, rejection episodes and immunosuppression. G~
C~
pRBC
pEAK
pEAK
HOSP
S~V
PRIM
TB~
TRANSF.
~T
T B~
STAY
%
NO~
1
CO~
10±3
20±15
1210±1080
19~14
28±31
80
1.8%
.93±3.5
OLD
10±2
22±16
1406~1588
18~14
38±43
~6
4.2%
.43±.67
• Values ,~ean±SD. All p values a r e g r e a ~ e r than .05, Square where a p p r o p r i a t e
t Ce~ts, ~ i
DISCUSSION: i) Despite a slight increase in the incidence of primary nonfunction and length of hospital stay, parameters of allograft viability and recipient outcomes were not adversely affected when older donors were used. 2) Since the upper age limit for liver donation is still not defined, we should strive to consider all potential referrals, regardless of age.
•
THE EFFECT OF LOVASTATIN AS AN ADJUVANT TO MITOMYCIN-C ON THE GROWTH OF MCA-38 CELLS IN VITRO AND IN VIVO. T.J. Morris. S.L. Palm. H. Bourdaaes. H. Menchacs, V. Michalek, L. Ft~rcht, H. Buchwald. Dept. of Surgery, University of Minnesota, Minneapolis, MN. Cholesterol is essential for cell growth. Due to their rapid growth rate and abnormal synthesis regulation, tumor cells may be more affected by cholesterol deprivation than normal cells. We evaluated the effect of Iovastatin, an HMG-CoA reductase inhibitor alone and as an adjuvant to mitomycin-C on the growth of Murine Colon Adenocarcincma (MCA-38) ceils in vitro and in vivo. MCA38 cells were grown in DMEM+10% FBS at standard conditions. 60mm culture plates (25/group) were inoculated: 2.5xl 0 s MCA-38 cells and treated with Iovastatin and mitomycin. On each of days 1-5, 5 plates per group were harvested and counted in triplicate. Group Lovastatin Mitomvcin-C Day 5 growth (xl 06)
I
0
0
1.93 -+ .15
II 1 I~M 0 1.23 + .19" [11 2 p.M 0 .61 + .04" IV 0 250 nM 1.07 + .09* V 1 p.M 250 nM .54 _+.05"1 VI 2 #M 250 nM .28 -+ .02"1 *p<.01 vs. control; tp<.01 vs. mitomycin-C alone 100 C57B1/6 mice were inoculated with 5x105 MCA-38 cells injected into the spleen, and treated with oral Iovastatin, intraperitoneal mitcmycin-C, or a combination of both drugs. After 13 days the animals were sacrificed, and the fiver and spleen were excised Group Treatment Spleen Tumor Wt _+SEM Liver nodules I Control 1.21 + .07 37 + 8 II Mitcmycin 2 pg/d .94 + .06* 34 + 6 Ill Lovastatin 40 mg/k/d 1.19 + .07 42 _+5 IV Lov 40 mg/k/d+Mit 2 lig/d .97 + .06" 19 + 4* *p<0.05 vs. control Lovastatin acts alone and as an adjuvant to mitomycin-C in slowing the in vitro growth of MCA-38 cells. Currently, in this model, Iovastatin alone had no significant effect on tumor growth in vivo, but the combination of lovastatin plus mitomycin-C significiantly reduced the growth of metastatic liver nodules.
GLU(3OSE INDUCED ISLET HYPEREMIA IS MEDIATED BY NITRIC OXIDE. S_.~.Moldovan,E. Livinqston, P. Guth, K. Zhans, R. Kleinman, F.(3. BrunicardL Depts. of Surgery, VAMC-West Los Angeles, U(SLA School of Medicine, Los Angeles, CA. We hypothesized that hyperglycemia increases islet blood flow and that the increase is mediated by the vasodilator nitric oxide. To test this hypothesis, the in rive transit time of a constant volume bolus of FIT(:; albumin across a selected islet capillary was measured at baseline and following hyperglycemic stimulation. The pancreas of fasted, anesthetized, male BALB/c mice was exteriorized and maintained at 370(3. Systemic'blood pressure was monitored continuously throughout the experiment. An islet capillary was identified and viewed under an Olympus microscope utilizing, monochromatic, transmitted light with the image observed via a closed-circuit television system with a final magnification of 1230:(. Under fluorescent light, a o.o2ml bolus of 2% FITC albumin was injected i.a, and the pulse of FITC albumin (BASELINE) through the islet capillary was videorecorded. Following equilibration, either 300mg/g glucose(G) or normal saline control(NS) was given i.v.; five minutes later, a second bolus of FITC albumin was injected i.a. and the pulse of FITC albumin (RESPONSE) was videerecorded. Plasma glucose was measured before and after the glucose infusion. In another group of mice the experiment with G was repeated in the presence of a constant pancreatic suffusion of 0.3raM nitro-L arginine methyl esther(L-NAME). Slow motion video analysis of the pixil intensity over time of the pulse of FITC albumin through the islet capillary resulted in a highly reproducible estimation of islet capillary flow, defined as transit time(seconds): TRANSIT TIME(sec) INFUSATE BASELINE RESPONSE G 0.6+,039 0.48+.030" n=11 n=11 NS 0.6+.051 0.66+.051# n=11 n=11 G + L-NAME 0.6+.054 0.6+.033# n=10 n=10 * P< .05 vs. basal via paired t-test; # P<.05 vs. G via unpaired t-test These data demonstrate that: a) infusion of G results in a signifidant decrease in islet capillary transit time with no change in systemic blood pressure, b) LNAME negated the effect of G infusion on transit time. We conclude that during in vivo microscopy of the mouse islet 1) hyperglycemia results in increased islet capillary flow and 2) nitric oxide regulates glucose induced islet hyperemia.
O E N T E R I C SMOOTH M U S C L E C O N T R A C T I L E P R O P E R T I E S A F T E R S M A L L B O W E L T R A N S P L A N T A T I O N (SBTx) IN RATS. M.M. Murr, M.G. Sarr, Dept. of Surgery, M a y o Clinic, Rochester, M N How SBTx alters enteric smooth m u s c l e function is not understood. AIM: To study c o n t r a c t i l e p r o p e r t i e s of enteric smooth m u s c l e a f t e r SHTx. METHODS: 5 groups of Lewis rats (n~8 each) w e r e studied: naive controls (NC); o p e r a t e d controls at I w k (OCl) and 8 w k (OC8) a f t e r intestinal t r a n s e c t i o n / r e a n a s t o m o s i s of p r o x i m a l j e j u n u m and distal ileum; and o r t h o t o p i c S B T x at i w k (TXl) and 8 w k (TX8). C o n t r a c t i l e a c t i v i t y was evaluated in c i r c u l a r m u s c l e strips s t r e t c h e d to ~ in organ chambers. Data are mean±SEM. RESULTS: S p o n t a n e o u s a c t i v i t y (Spon Act) i n c r e a s e d in TXl, TX8 and OCl (Table) (p~.01). F r e q u e n c y of c o n t r a c t i o n s doubled in OCl (p~.001) but was u n c h a n g e d in other groups. Under non-adrenergic n e n - c h o l i n e r g i c (NANC) conditions, s p o n t a n e o u s a c t i v i t y increased in TXI and OC1 (p~.005) without change in frequency. T e t r o d o t o x i n (TTX i0"~) increased amplitude and frequency in all groups. B e t h a n e c h o l (Be; 3xl0"S-3xl0"~M) i n c r e a s e d and n o r e p i n e p h r i n e (NE; I0"6-I0"4M) d e c r e a s e d dosed e p e n d e n t l y amplitude and frequency of contractions in all groups; effective c o n c e n t r a t i o n s (ED) did not differ between groups. CONCLUSIONS: SBTx increases c o n t r a c t i l e a m p l i t u d e of circular m u s c l e due, in part, to d o w n r e g u l a t i o n of NA_NC nerves but not via m u s c a r i n i c or a d r e n e r g i c hypersensitivity. The increase in c o n t r a c t i l e a c t i v i t y in OCI is secondary to i n c r e a s e d frequency and not amplitude. Support: NIH DK39337 C o n t r a c t i l e activityt .... GrD Sport Act NANC TTX* B e ( E D s ~ _ ~ N E (ED~51 NC 1.6±0.2 2.6±0.4 3.5±1.0 4.6±0.1 4.7±012 OCI 2.9±0.3* 5.5±0.5* 4.8±0.8 4.5±0.1 5.0±0.1 OC8 1.7±0.2 2.6±0.2 2.5±0.3 4.5±0. I 4.9±0.1 TXI 3.1±0.3" 4.4±0.3* 4.0±0.5 4.8±0.1 5.2±0.2 TX8 2.7±0.3, 4.7±0.3 4.9±0.8 4.6±0.1 5.3±0.2 tg.sec/mg; $neg log (M); *differs from NC (p~.01)
SSAT ABSTRACTS
GASTROENTEROLOGY, Vol. 108, No. 4
• IMPACT OF O L D E R DONORS O N HEPATIC ALLOGRAFT VIABILITY. D Mital, H Sankary, J Whiting, P Yoster,L M c ~ h ~ J Williams Department of Surgery, Rush Presbyterian St Lukes Medical Center, Chicago, Illinois. As the success of hepatic transplantation improves, there is increasing demand for a limited supply of organs. We recently expanded our static donor pool by utilizing livers from donors of age greater than 60 yrs. The purpose of this study is to determine whether the use of older donors, as a source of hepatic allografts, adversely affects the outcome following transplantation. In a retrospective analysis indices of graft viability were compared in two groups of patients: The first group included 214 adult patients who received grafts from donors of age less than 60 yrs(CONT). The second group included 47 patients who received organs from donors of age greater than 60 yrs of age(OLD). RESULTS: Donor age was significantly greater in the OLD(60-80 yrs) group when compared to CONT(67~5 vs 30±15 yrs,P<.ool). Donor characteristics including hospital stay, preservation time, dopamine dose, total bilirubin, ALT, alkaline phosphatase were similar between groups. Recipients had similar distributions of diseases, ages, Child's classification, rejection episodes and immunosuppression. G~
C~
pRBC
pEAK
pEAK
HOSP
S~V
PRIM
TB~
TRANSF.
~T
T B~
STAY
%
NO~
1
CO~
10±3
20±15
1210±1080
19~14
28±31
80
1.8%
.93±3.5
OLD
10±2
22±16
1406~1588
18~14
38±43
~6
4.2%
.43±.67
• Values ,~ean±SD. All p values a r e g r e a ~ e r than .05, Square where a p p r o p r i a t e
t Ce~ts, ~ i
DISCUSSION: i) Despite a slight increase in the incidence of primary nonfunction and length of hospital stay, parameters of allograft viability and recipient outcomes were not adversely affected when older donors were used. 2) Since the upper age limit for liver donation is still not defined, we should strive to consider all potential referrals, regardless of age.
•
THE EFFECT OF LOVASTATIN AS AN ADJUVANT TO MITOMYCIN-C ON THE GROWTH OF MCA-38 CELLS IN VITRO AND IN VIVO. T.J. Morris. S.L. Palm. H. Bourdaaes. H. Menchacs, V. Michalek, L. Ft~rcht, H. Buchwald. Dept. of Surgery, University of Minnesota, Minneapolis, MN. Cholesterol is essential for cell growth. Due to their rapid growth rate and abnormal synthesis regulation, tumor cells may be more affected by cholesterol deprivation than normal cells. We evaluated the effect of Iovastatin, an HMG-CoA reductase inhibitor alone and as an adjuvant to mitomycin-C on the growth of Murine Colon Adenocarcincma (MCA-38) ceils in vitro and in vivo. MCA38 cells were grown in DMEM+10% FBS at standard conditions. 60mm culture plates (25/group) were inoculated: 2.5xl 0 s MCA-38 cells and treated with Iovastatin and mitomycin. On each of days 1-5, 5 plates per group were harvested and counted in triplicate. Group Lovastatin Mitomvcin-C Day 5 growth (xl 06)
I
0
0
1.93 -+ .15
II 1 I~M 0 1.23 + .19" [11 2 p.M 0 .61 + .04" IV 0 250 nM 1.07 + .09* V 1 p.M 250 nM .54 _+.05"1 VI 2 #M 250 nM .28 -+ .02"1 *p<.01 vs. control; tp<.01 vs. mitomycin-C alone 100 C57B1/6 mice were inoculated with 5x105 MCA-38 cells injected into the spleen, and treated with oral Iovastatin, intraperitoneal mitcmycin-C, or a combination of both drugs. After 13 days the animals were sacrificed, and the fiver and spleen were excised Group Treatment Spleen Tumor Wt _+SEM Liver nodules I Control 1.21 + .07 37 + 8 II Mitcmycin 2 pg/d .94 + .06* 34 + 6 Ill Lovastatin 40 mg/k/d 1.19 + .07 42 _+5 IV Lov 40 mg/k/d+Mit 2 lig/d .97 + .06" 19 + 4* *p<0.05 vs. control Lovastatin acts alone and as an adjuvant to mitomycin-C in slowing the in vitro growth of MCA-38 cells. Currently, in this model, Iovastatin alone had no significant effect on tumor growth in vivo, but the combination of lovastatin plus mitomycin-C significiantly reduced the growth of metastatic liver nodules.
GLU(3OSE INDUCED ISLET HYPEREMIA IS MEDIATED BY NITRIC OXIDE. S_.~.Moldovan,E. Livinqston, P. Guth, K. Zhans, R. Kleinman, F.(3. BrunicardL Depts. of Surgery, VAMC-West Los Angeles, U(SLA School of Medicine, Los Angeles, CA. We hypothesized that hyperglycemia increases islet blood flow and that the increase is mediated by the vasodilator nitric oxide. To test this hypothesis, the in rive transit time of a constant volume bolus of FIT(:; albumin across a selected islet capillary was measured at baseline and following hyperglycemic stimulation. The pancreas of fasted, anesthetized, male BALB/c mice was exteriorized and maintained at 370(3. Systemic'blood pressure was monitored continuously throughout the experiment. An islet capillary was identified and viewed under an Olympus microscope utilizing, monochromatic, transmitted light with the image observed via a closed-circuit television system with a final magnification of 1230:(. Under fluorescent light, a o.o2ml bolus of 2% FITC albumin was injected i.a, and the pulse of FITC albumin (BASELINE) through the islet capillary was videorecorded. Following equilibration, either 300mg/g glucose(G) or normal saline control(NS) was given i.v.; five minutes later, a second bolus of FITC albumin was injected i.a. and the pulse of FITC albumin (RESPONSE) was videerecorded. Plasma glucose was measured before and after the glucose infusion. In another group of mice the experiment with G was repeated in the presence of a constant pancreatic suffusion of 0.3raM nitro-L arginine methyl esther(L-NAME). Slow motion video analysis of the pixil intensity over time of the pulse of FITC albumin through the islet capillary resulted in a highly reproducible estimation of islet capillary flow, defined as transit time(seconds): TRANSIT TIME(sec) INFUSATE BASELINE RESPONSE G 0.6+,039 0.48+.030" n=11 n=11 NS 0.6+.051 0.66+.051# n=11 n=11 G + L-NAME 0.6+.054 0.6+.033# n=10 n=10 * P< .05 vs. basal via paired t-test; # P<.05 vs. G via unpaired t-test These data demonstrate that: a) infusion of G results in a signifidant decrease in islet capillary transit time with no change in systemic blood pressure, b) LNAME negated the effect of G infusion on transit time. We conclude that during in vivo microscopy of the mouse islet 1) hyperglycemia results in increased islet capillary flow and 2) nitric oxide regulates glucose induced islet hyperemia.
O E N T E R I C SMOOTH M U S C L E C O N T R A C T I L E P R O P E R T I E S A F T E R S M A L L B O W E L T R A N S P L A N T A T I O N (SBTx) IN RATS. M.M. Murr, M.G. Sarr, Dept. of Surgery, M a y o Clinic, Rochester, M N How SBTx alters enteric smooth m u s c l e function is not understood. AIM: To study c o n t r a c t i l e p r o p e r t i e s of enteric smooth m u s c l e a f t e r SHTx. METHODS: 5 groups of Lewis rats (n~8 each) w e r e studied: naive controls (NC); o p e r a t e d controls at I w k (OCl) and 8 w k (OC8) a f t e r intestinal t r a n s e c t i o n / r e a n a s t o m o s i s of p r o x i m a l j e j u n u m and distal ileum; and o r t h o t o p i c S B T x at i w k (TXl) and 8 w k (TX8). C o n t r a c t i l e a c t i v i t y was evaluated in c i r c u l a r m u s c l e strips s t r e t c h e d to ~ in organ chambers. Data are mean±SEM. RESULTS: S p o n t a n e o u s a c t i v i t y (Spon Act) i n c r e a s e d in TXl, TX8 and OCl (Table) (p~.01). F r e q u e n c y of c o n t r a c t i o n s doubled in OCl (p~.001) but was u n c h a n g e d in other groups. Under non-adrenergic n e n - c h o l i n e r g i c (NANC) conditions, s p o n t a n e o u s a c t i v i t y increased in TXI and OC1 (p~.005) without change in frequency. T e t r o d o t o x i n (TTX i0"~) increased amplitude and frequency in all groups. B e t h a n e c h o l (Be; 3xl0"S-3xl0"~M) i n c r e a s e d and n o r e p i n e p h r i n e (NE; I0"6-I0"4M) d e c r e a s e d dosed e p e n d e n t l y amplitude and frequency of contractions in all groups; effective c o n c e n t r a t i o n s (ED) did not differ between groups. CONCLUSIONS: SBTx increases c o n t r a c t i l e a m p l i t u d e of circular m u s c l e due, in part, to d o w n r e g u l a t i o n of NA_NC nerves but not via m u s c a r i n i c or a d r e n e r g i c hypersensitivity. The increase in c o n t r a c t i l e a c t i v i t y in OCI is secondary to i n c r e a s e d frequency and not amplitude. Support: NIH DK39337 C o n t r a c t i l e activityt .... GrD Sport Act NANC TTX* B e ( E D s ~ _ ~ N E (ED~51 NC 1.6±0.2 2.6±0.4 3.5±1.0 4.6±0.1 4.7±012 OCI 2.9±0.3* 5.5±0.5* 4.8±0.8 4.5±0.1 5.0±0.1 OC8 1.7±0.2 2.6±0.2 2.5±0.3 4.5±0. I 4.9±0.1 TXI 3.1±0.3" 4.4±0.3* 4.0±0.5 4.8±0.1 5.2±0.2 TX8 2.7±0.3, 4.7±0.3 4.9±0.8 4.6±0.1 5.3±0.2 tg.sec/mg; $neg log (M); *differs from NC (p~.01)