- Email: [email protected]
16. Myocardial performance after perfusion with DMSO for cardiac preservation
298
ANNUAL M E E T I N G ABSTRACTS
adjacent vitreous. The values obtained in the rabbit experiments and the gelaline solution correlate(l well with the theoretical va.hws found using the thermo/lynanli(: formula. The adhesive str6ngtl~ of the bond exceeded 1000 g. Sliding required less force th'm tumbling in cataract e.xtraction in a rabbit. A stmly of the temperature gradients demonstrate the safet.y of eryopexy and cryoextraelion. The sup'~rior adhesive strengtt: of a cryoprobe an(t the advantages of sliding during cataract extraction are well demonstrated.
14. The Effect of Hypothermia on Lung Elastic Properties of the Dog. EmL BLAIR (Departmen( of Surgery, University of MarylmM School of Medicine, Baltimore, Maryl,'md). Progressively deeper cooling results in prol)ortionate degrees of respiratory dysfunction, apparently due to peril)heral chemo/'eptor and central depression. Since struclures with viseoelaslic properties are subject to temperature alterations, an evaluation of lung mech'mics was undertaken. Studies were done in open chest preparations with ventilatory support to eliminate the thoracic component. Static eomplianee, hysteresis, stress relaxalion, and elastic work of breathing were ev'duated fi'om data obtained from measurement of the tran~pulmonary pressure, hmg volume, and air flow. Pressure-volume loops were constructed from data obtained by the syringe increment technique with the respirator disconnected. Three groups were studied: 3) normothermie controls; 2) hypothermia alone, 30 to 30°C; 3 ~, Normothermie artificial p~rfusion controls; 4) hypothermia wilh artificial perfusion, 30 to 10°C. Results were as follows. Controls: 3) elastic properties were unchanged over 3 to 4 hrs; 2) hypothermia: normal limits through 20°C with decrease at 30°C; 3) bypass eontrols:"normaI limits te 2 hrs bypass with ~ome impvirment postbypass; and 4) hypothermia with bypass: no change at 30 or 25°C, except prolonged bypass (1+ hrs); impairment at 20 and 10~C. Upon rewarming discrepancies appeared in animals with unchanged properties during hypothermie periods and persisted in those with impairment during hypothermia. Alterations in lung mechanical (elqstie) properties do not contribute to dysfunction during hypothermia except at profound levels (30°C). Total bypass, if prolonged, eomplic, ates matters effeeting a distm-banee at more moderate levels.
15. Regional Deep Hypothermia and AboveKnee Amputations for Infection Complieat-
ing Isehemic Gangrene. J F. G0mxmN* aNl) d. C. Rosvxt~mm (D,,par(ment of Sm'~ery, Manmeo Valley /tospi(al, Tole(t(), Ohio) A study of 72 consecutive al)ove-knee ampulslions for ischemic gangrene was un(h,rtaken to (letermine the 1)enefits of routine l)reol)erative r~,,ciomd deep hyl)othormia. Solid carbon dioxide (Dry Ice), with an inherent temlwrature of -45°C, was applied to 39 exlremities preoperativ'qy. The control groll 1) consistvcl of 33 amputalions p,',rform,,d without refrig(,ration. Mortality in th:, ('oniroI group was 27~. The mortality of pa(ients un(lergoing preol)erative refrigeration was 30¢;~ (P of (lifferenee, 0.05). Thirt:~en patients had sc,psis complicating gangrene in the control group. Seventeen patients had Spl)sis in {he ~sroul) with regional (leep 1W1)othc,rmia. The mortality among the patients with infection was 46% in the mltreated gl'ouD. Only 6% of lmtients whose infected isehomie legs were fi'ozen (lied (P of difference, <0.0i). Puhnonary embolus, myocardial faihwe, and pneumonia wer/~ flip tiles( frequent /'aliSON of dealh. T]w incidence of wound infection was 30% in the nonrefrigerated l~atients and 30% in the treat:'d group. Stlllnp necrosis and (lehtyed healing oeem'r.'d in 13% of the extremities which were refrigerated but in only 3% of the extremi*ies nol refrigerated. Three serious complications of refrigeration oeeurred; all were caused by faulty technique in freezing the limb. In ~his series of amputations, sepsis was causally rehtted to mort,tlity and morbidity. There was no relationship between the presenet~ of diabetes and morlality or morbidity. Sincd refrigeration produced a reduelmn in mortality and wound infections in pati:,nts with isehemie gangrene complicated by infection, it is eoncluded tlmt regional deep hypothermia should be utilized routinely in these |)ati;-u~s. Dr)" Ice was foun(t to be an effective anti .~,:.onomicai means of producing refrigeration of the involved extremity.
16. Myocardial Performance after Pcrfusion with DMSO for Cardiac Preservation. I-I]~DIIICK B. BARNER, MAx .JELLINCK,* AND KI':NNETI-t E. Moor~:* (Department of Surgery, St. I,ouis University ScllooI of Medicine, St. Louis, Missouri). Attempts at preservation of the heart for transplantation by freezing with the aid of agents suc,h as dimethyI sulfoxide have not clarified the. specific (:fleets of these, agents on the heart wi~houl freezing. After perfusion of the canine heart with DMSO, we have assessed the status of the myoeardium by study of enzymes and electrical ac-
A N N U A L MI~ETING A B S T R A C T S t ivity an,l t)y light and electron microscopy. These stu(lies hart, t)eon tmrformed up to 8 weeks after l)erfusion. Dogs weighing 14 to 18 kg are supporled extracorlmreally and cooled to 25°C. The heart is iso-. l;tlo~t in .+ilu and lhe niyocar(lium perfuscd via a left v,'ntricular or aortic root eannula with iced laclttle(l Ringer's solulion until arrested. Perfusion is con|inuod with 200 to 260 ml of 15% DMSO in siandard dex(ran at 5 to 10°O over a 10- to 12rain interval. Myocardial blood flow is restored
and tim animal warmed to 35"C. The heart is defibrilhded and allowed to resume its circulatory role. The initial 6 animals have survived at least 1 week before saerifice. Additional studies under vm'ied experimental conditions are in progress. Under the conditions specified, we have found no sl~eeific abnornmliiy of enzymatic activity, the oleclrocar(liogram is not significantly changed and there is a normal pattern by light microscopy and ele('l ron microscopy.
Cryophylaetie 17, The Comparative Cryoproteetive Effect. of MG ~* as Assayed by Pharmacological Method, AtlM.~xD M. KAROW, Jm, ME'rL~' O. UxaL,* OLivrm C,'dlltlER, JR.,* C. i-{, DaNIFLS,* AND ~?'ATTS It. W1~m~ (l)epartmont of P h a r m a c o l o g y , The University of MissisMppi School of Mr_duroc, Jackson, Mississippi, aml Delmrtment of Surgery, The University of Texas Southwestern Medi,:al School, Dalla,.,' . ~ Texas). Whole isolated hearts of rats, guinea pigs, and r'dfifits were frozen at - 2 0 ° C for 20 rain. Three different vryoprotective agents were tested" 15% (1.9 st) dimethyl sulfoxide in Ringer's solution, 6% high molecular weight dextran (average mol. wt. 70,000) in saline and 1.3% (6.6 X 10-~ .~t) MgCI,_..6H.oO in Ringer's solution. Also 3 whole isolated dog hearts perfused with 5% (4.2 × 10-~ .,,I) MgSO~ were frozen in cold baths at - 2 W C for 30 rain and 45 rain and --30°C for 30 rain. Hearts after thawing in e'tch of the experimental groups Juc.'~m . . d,,' atrial and ventricular cont~'ac'hon, ss. I~,'tt hearts after thawing were then treated sucoos,dvely with 10 -'~ ~f isoproteranol, 10"~ a~ MJ1999 (a beta-adrenergic blocking agent), and 10-~ 5~ ........ . . . ~,atm of these drugs elicited typical pharnmeologic responses, suggesting minimal functional damage to membranes, and receptor sites. The i$rol)osed mechanism of action for Mg '+ is st'tbilizalion of proteins and eerlain other macromolecules during freezing and prevention of bound water exiracomn - . t" leading to (!cnaturation. 18, Comparison o f Various Agents as Protective Agents Against Freezing In"jury o f Platelets," IsAac DJgRASSI,* ALBERT I~OY, SIDNET FARBER,* JOtlN CAVINS, AND EDMUND KbEIN* (Children's Cancer Research Foundation and The Department of Pathology, Harvard Medical School, at The Children's HospitM, Boston, Massachusetts). =This investigation was supported by Grant AT (30-1) 1753 fi'om ghe Atomic Energy Commission.
299
Agents
In previous comnmnications, we have reported on the prescrv,ttion of both animal and human plateleis. A combination of dimethyl sulfoxide and dextrose protected a significant number of these platelets from freezing injury as mehsured by a variety of in vitro criteria iucluding 1) morphological appearance, 2) ability to promote clot retraction of platelet poor "native" plasma., 3) circulation in r i v e 4) normalization of prolonged bleeding time, and 5) promotion of hemostasis. However, when used clinically only 30 to 50% of the increase seen with fresh platclets was obtained when comparable numbers of the frozen preparation were used. In order to improve the viability of platelets preserved by freezing, a number of other additives were studie.d as cryophylactic agents. R,tt platelet; concentrates were prepared and frozen in liquid nitrogen at I°C per min with added dimethyl sulfoxide, dimethyl acetamide, glycerol or propylene glycol, alone or combined with detxrose. After 5 to 14 clays of storage at - ! 9 6 ° C these preparations were infused into rats rendered thromlmc _ ~ .... ,~ ,,~u, .~-iiTactlatlon. Of these agents, alone or in. v~rious combinations and concentrations, 5% dimethyl acetamide plus 5% dextrose proved to be superior to 5% D M S O plus 5% dextrose in its ability to protect rat platelets from the effects of strorage at these temperatures. Propylene glycol and glycerol were not con~dered to be very promising cryophylactic agents for p]atelets on the basis of the present studies. 19. Studies Intertidal
on the Freezing Resistance o f Mollusks2 ~.OBERT J. WILLIAMS
aFrom the Bureau of Medicine and Surgery~ Navy Department. Research Task MR-005.020001.07. Supported in part by NASA Contract tt-63. The opinions or assertions contained herein are the private ones of tile author and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large.
ANNUAL M E E T I N G ABSTRACTS
adjacent vitreous. The values obtained in the rabbit experiments and the gelaline solution correlate(l well with the theoretical va.hws found using the thermo/lynanli(: formula. The adhesive str6ngtl~ of the bond exceeded 1000 g. Sliding required less force th'm tumbling in cataract e.xtraction in a rabbit. A stmly of the temperature gradients demonstrate the safet.y of eryopexy and cryoextraelion. The sup'~rior adhesive strengtt: of a cryoprobe an(t the advantages of sliding during cataract extraction are well demonstrated.
14. The Effect of Hypothermia on Lung Elastic Properties of the Dog. EmL BLAIR (Departmen( of Surgery, University of MarylmM School of Medicine, Baltimore, Maryl,'md). Progressively deeper cooling results in prol)ortionate degrees of respiratory dysfunction, apparently due to peril)heral chemo/'eptor and central depression. Since struclures with viseoelaslic properties are subject to temperature alterations, an evaluation of lung mech'mics was undertaken. Studies were done in open chest preparations with ventilatory support to eliminate the thoracic component. Static eomplianee, hysteresis, stress relaxalion, and elastic work of breathing were ev'duated fi'om data obtained from measurement of the tran~pulmonary pressure, hmg volume, and air flow. Pressure-volume loops were constructed from data obtained by the syringe increment technique with the respirator disconnected. Three groups were studied: 3) normothermie controls; 2) hypothermia alone, 30 to 30°C; 3 ~, Normothermie artificial p~rfusion controls; 4) hypothermia wilh artificial perfusion, 30 to 10°C. Results were as follows. Controls: 3) elastic properties were unchanged over 3 to 4 hrs; 2) hypothermia: normal limits through 20°C with decrease at 30°C; 3) bypass eontrols:"normaI limits te 2 hrs bypass with ~ome impvirment postbypass; and 4) hypothermia with bypass: no change at 30 or 25°C, except prolonged bypass (1+ hrs); impairment at 20 and 10~C. Upon rewarming discrepancies appeared in animals with unchanged properties during hypothermie periods and persisted in those with impairment during hypothermia. Alterations in lung mechanical (elqstie) properties do not contribute to dysfunction during hypothermia except at profound levels (30°C). Total bypass, if prolonged, eomplic, ates matters effeeting a distm-banee at more moderate levels.
15. Regional Deep Hypothermia and AboveKnee Amputations for Infection Complieat-
ing Isehemic Gangrene. J F. G0mxmN* aNl) d. C. Rosvxt~mm (D,,par(ment of Sm'~ery, Manmeo Valley /tospi(al, Tole(t(), Ohio) A study of 72 consecutive al)ove-knee ampulslions for ischemic gangrene was un(h,rtaken to (letermine the 1)enefits of routine l)reol)erative r~,,ciomd deep hyl)othormia. Solid carbon dioxide (Dry Ice), with an inherent temlwrature of -45°C, was applied to 39 exlremities preoperativ'qy. The control groll 1) consistvcl of 33 amputalions p,',rform,,d without refrig(,ration. Mortality in th:, ('oniroI group was 27~. The mortality of pa(ients un(lergoing preol)erative refrigeration was 30¢;~ (P of (lifferenee, 0.05). Thirt:~en patients had sc,psis complicating gangrene in the control group. Seventeen patients had Spl)sis in {he ~sroul) with regional (leep 1W1)othc,rmia. The mortality among the patients with infection was 46% in the mltreated gl'ouD. Only 6% of lmtients whose infected isehomie legs were fi'ozen (lied (P of difference, <0.0i). Puhnonary embolus, myocardial faihwe, and pneumonia wer/~ flip tiles( frequent /'aliSON of dealh. T]w incidence of wound infection was 30% in the nonrefrigerated l~atients and 30% in the treat:'d group. Stlllnp necrosis and (lehtyed healing oeem'r.'d in 13% of the extremities which were refrigerated but in only 3% of the extremi*ies nol refrigerated. Three serious complications of refrigeration oeeurred; all were caused by faulty technique in freezing the limb. In ~his series of amputations, sepsis was causally rehtted to mort,tlity and morbidity. There was no relationship between the presenet~ of diabetes and morlality or morbidity. Sincd refrigeration produced a reduelmn in mortality and wound infections in pati:,nts with isehemie gangrene complicated by infection, it is eoncluded tlmt regional deep hypothermia should be utilized routinely in these |)ati;-u~s. Dr)" Ice was foun(t to be an effective anti .~,:.onomicai means of producing refrigeration of the involved extremity.
16. Myocardial Performance after Pcrfusion with DMSO for Cardiac Preservation. I-I]~DIIICK B. BARNER, MAx .JELLINCK,* AND KI':NNETI-t E. Moor~:* (Department of Surgery, St. I,ouis University ScllooI of Medicine, St. Louis, Missouri). Attempts at preservation of the heart for transplantation by freezing with the aid of agents suc,h as dimethyI sulfoxide have not clarified the. specific (:fleets of these, agents on the heart wi~houl freezing. After perfusion of the canine heart with DMSO, we have assessed the status of the myoeardium by study of enzymes and electrical ac-
A N N U A L MI~ETING A B S T R A C T S t ivity an,l t)y light and electron microscopy. These stu(lies hart, t)eon tmrformed up to 8 weeks after l)erfusion. Dogs weighing 14 to 18 kg are supporled extracorlmreally and cooled to 25°C. The heart is iso-. l;tlo~t in .+ilu and lhe niyocar(lium perfuscd via a left v,'ntricular or aortic root eannula with iced laclttle(l Ringer's solulion until arrested. Perfusion is con|inuod with 200 to 260 ml of 15% DMSO in siandard dex(ran at 5 to 10°O over a 10- to 12rain interval. Myocardial blood flow is restored
and tim animal warmed to 35"C. The heart is defibrilhded and allowed to resume its circulatory role. The initial 6 animals have survived at least 1 week before saerifice. Additional studies under vm'ied experimental conditions are in progress. Under the conditions specified, we have found no sl~eeific abnornmliiy of enzymatic activity, the oleclrocar(liogram is not significantly changed and there is a normal pattern by light microscopy and ele('l ron microscopy.
Cryophylaetie 17, The Comparative Cryoproteetive Effect. of MG ~* as Assayed by Pharmacological Method, AtlM.~xD M. KAROW, Jm, ME'rL~' O. UxaL,* OLivrm C,'dlltlER, JR.,* C. i-{, DaNIFLS,* AND ~?'ATTS It. W1~m~ (l)epartmont of P h a r m a c o l o g y , The University of MissisMppi School of Mr_duroc, Jackson, Mississippi, aml Delmrtment of Surgery, The University of Texas Southwestern Medi,:al School, Dalla,.,' . ~ Texas). Whole isolated hearts of rats, guinea pigs, and r'dfifits were frozen at - 2 0 ° C for 20 rain. Three different vryoprotective agents were tested" 15% (1.9 st) dimethyl sulfoxide in Ringer's solution, 6% high molecular weight dextran (average mol. wt. 70,000) in saline and 1.3% (6.6 X 10-~ .~t) MgCI,_..6H.oO in Ringer's solution. Also 3 whole isolated dog hearts perfused with 5% (4.2 × 10-~ .,,I) MgSO~ were frozen in cold baths at - 2 W C for 30 rain and 45 rain and --30°C for 30 rain. Hearts after thawing in e'tch of the experimental groups Juc.'~m . . d,,' atrial and ventricular cont~'ac'hon, ss. I~,'tt hearts after thawing were then treated sucoos,dvely with 10 -'~ ~f isoproteranol, 10"~ a~ MJ1999 (a beta-adrenergic blocking agent), and 10-~ 5~ ........ . . . ~,atm of these drugs elicited typical pharnmeologic responses, suggesting minimal functional damage to membranes, and receptor sites. The i$rol)osed mechanism of action for Mg '+ is st'tbilizalion of proteins and eerlain other macromolecules during freezing and prevention of bound water exiracomn - . t" leading to (!cnaturation. 18, Comparison o f Various Agents as Protective Agents Against Freezing In"jury o f Platelets," IsAac DJgRASSI,* ALBERT I~OY, SIDNET FARBER,* JOtlN CAVINS, AND EDMUND KbEIN* (Children's Cancer Research Foundation and The Department of Pathology, Harvard Medical School, at The Children's HospitM, Boston, Massachusetts). =This investigation was supported by Grant AT (30-1) 1753 fi'om ghe Atomic Energy Commission.
299
Agents
In previous comnmnications, we have reported on the prescrv,ttion of both animal and human plateleis. A combination of dimethyl sulfoxide and dextrose protected a significant number of these platelets from freezing injury as mehsured by a variety of in vitro criteria iucluding 1) morphological appearance, 2) ability to promote clot retraction of platelet poor "native" plasma., 3) circulation in r i v e 4) normalization of prolonged bleeding time, and 5) promotion of hemostasis. However, when used clinically only 30 to 50% of the increase seen with fresh platclets was obtained when comparable numbers of the frozen preparation were used. In order to improve the viability of platelets preserved by freezing, a number of other additives were studie.d as cryophylactic agents. R,tt platelet; concentrates were prepared and frozen in liquid nitrogen at I°C per min with added dimethyl sulfoxide, dimethyl acetamide, glycerol or propylene glycol, alone or combined with detxrose. After 5 to 14 clays of storage at - ! 9 6 ° C these preparations were infused into rats rendered thromlmc _ ~ .... ,~ ,,~u, .~-iiTactlatlon. Of these agents, alone or in. v~rious combinations and concentrations, 5% dimethyl acetamide plus 5% dextrose proved to be superior to 5% D M S O plus 5% dextrose in its ability to protect rat platelets from the effects of strorage at these temperatures. Propylene glycol and glycerol were not con~dered to be very promising cryophylactic agents for p]atelets on the basis of the present studies. 19. Studies Intertidal
on the Freezing Resistance o f Mollusks2 ~.OBERT J. WILLIAMS
aFrom the Bureau of Medicine and Surgery~ Navy Department. Research Task MR-005.020001.07. Supported in part by NASA Contract tt-63. The opinions or assertions contained herein are the private ones of tile author and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large.