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1. The effect of gases on the recovery of human red blood cells
( S It. Y t~BI O L O f ~ Y
Vol. 6, No+ 6, 1970
ABSTRACTS FROM SEVENTII ANNUAL MEETING 1, T h e Effect of Ga~c~ on the Recovery of Hum a n Red Blood Ceils. F. C!. WESSLING, ~lll. AND P. L. Bt,.tCKSnZ,u¢, 3t~, (University of Minnvsota, ~Xlimw:q)olis, Minnesota). Th{: freezing of blood in...several type,s of small t~l~t.s we,- studied. The }deed was equilibrated with X'alit~ns gases before freezing. The (tam'tge during free.zing does not aPl,:.ar to depend Ul,On the gas its,If hut upon the change in volume that the gas prodtwes in the. cell. Heat. transfer calculations iu~ticaie that the d'tmage does not appear to be em~svtl hy supercooling or by intraeellular ice forn l a t i o l l h i l t t.,y all a s yet undetermined mechanism. 2. '|'he Effect of Pressure on the Freezing of Red Blood Cells. Fm~uF.lllc I¢. AUL~areN* .hNO Pt~m,Y I.. Bt.ACKSHE.ta, Ja. (Department of Engineering~ U'niversity of Minnesota, Minneapolis, Minnesota). As red blood cells are frozen at atmospheric presm~r+, hemolysis starts to occur ~:!ightly be.low tt~e freezing point and continues to a teml)erature of --60°C. ,"3im+epressure lowers the freezing point of aqlwcms solutions, it was the authors' hyt)othesis that, pressure might be used to reduce the width of this hemolytic temperature region, provided that the colder side of the region was not also de|,re,sell. This experiment was conducted to deterntitm how l~r<:s.~ure aJIected the colder side of that tetnl)eraturo range which causes rapid hemolysis of blood. Whole dog blood was frozen quickly in a snlall Teflon tube which was phmged into isopentane at --93°C. The tube was then quickly transferred to a precooled pressure chamber which was then sealed and pressurized to a ~est pre,.~sure. The pressure elmmber and sample were then warmed to a test temperature and incubated for 15 rain. The chamber was then recooled to --70°C and depressurized. Tlw. sample tube was withdrawn and thawed in wat+rr at 37°C. The level of hemolysis was then determined. It was found that lip tO 1250 aim increasing the pressure deerea.~ed the whlth of the hemolytic temt,eraturo region by both depressing the warmer side and elevating Lho. eohler side. At that pressure the hemolytic region extends from --10°C to -37°C, lcs:s lhnn one-half that of atmospheric pressure. Further increasing the prexsure wits found to increase the width of the hemolytic region. A,~ a follow-up experiment whole blood waz frozen at, a rate of 100°C per rain :it, various pre~ures. A t 1 utm hemolysis was complete but at 1250 arm the level of hemolysis was reduced to 10% ; furlher inere'~sing the pre~ure inorea~M the level of hemolysis.
571
It, has been .~'eport+'d lbat freezing Idood at 1 alto tit a rail: of approximately 7800°C per aniu will tmnluce 1{1% hr.molysis. Thus. the advantages of prv.-_'suro trey.zing an. obvious, 10q hemolysi~ at I00°C per rain vs. 10% henmb'sis 't! 7800°C per min. 3. High liydrostatie l~ressure.in(lueed Variations in Supercooling and Freezing Tempera. lures of Water and Certain Aqueous Solutions, x ~'IAXI.M].), PEIiSlDSKY (Institute of Medical Sciences, l"u'ific Medical C c l l | e r , Sell Francisco, Californ in). Our sttMio.-." of the effects of high hydrostatic pressllrt~ on tissue cryopreserw~tion revealed uliex.. pet:ted irregularitie.,~ in the rate 6f freezing point depre~iou in a certain range of prc~,sure. Since Bridgman's and more rt:eent studies concern only pure water and are re|her sehemalic, we f¢.h that detailed investigation of tlie~e relations in certain •tqueous solutions of CD'ohiological importance would bc of inh~rest. Waist, solutions of nonMeetrotytes such as allmethyl sulfoxidc and polyvinylp3"rrolidone, and some electrolytes were studied. Plx;ssuro was wlrietl from atmospheric to 40,000 p~s.i.; temperature and voltlnK~ changes during phase tr:msition at different pressures were measur&t. Of !mri.ieular interest was the extent of supercooling and freezing tern.. peratures. When plotted against pre~ure these to+mperatun~+s produced compound curves reprosenting thermttt l~rofiles of supercooling and freezing. Thermal profiles for water show that. ,+ut~ercooling disappears at certain points be!ween 24,000 and 30,0t30 p,s,i., and inenm~es above 30,000 p.s.i+ Freezing also shows irregularity within thi~ ~,r~sure range. Above 30,000 p+si. tWO freezings occ~ir at differeni, temperatures. The fir~st is ,as~eiated with volume decrease, the seeond with Volume increase. ComparaMe thermal l/mJfiles wifli el~,~c2roJ 13"~e~ and nonelecirolytes show s o m e similar ;and some dissimilar features, mid will be illustrated. It is pomible that, at i h e ~ pressures, several transient, metastable forms of ice e+xizt w!iid: affdet the thermal profile of mipereooling anti fr~zini+, 4+ Superhyperbarie Cryoprcservat|on o f tleart Valve~Y aMAXI~ D. Pt::RSlIJSK~- ,~0 ~r/LLIAM J, Kt:rt'rIt* (Institute 'of Medical Sciences, Padfie Medical Center, San Francis~x~. California). There are two main advantages which eryopr~:~r.. ration under high hydrostatic pressures offers: I) t Supported by Nadonal Gran~ 5 Ii0t i1E12668,01.
Institutea of H ~ d t h
Vol. 6, No+ 6, 1970
ABSTRACTS FROM SEVENTII ANNUAL MEETING 1, T h e Effect of Ga~c~ on the Recovery of Hum a n Red Blood Ceils. F. C!. WESSLING, ~lll. AND P. L. Bt,.tCKSnZ,u¢, 3t~, (University of Minnvsota, ~Xlimw:q)olis, Minnesota). Th{: freezing of blood in...several type,s of small t~l~t.s we,- studied. The }deed was equilibrated with X'alit~ns gases before freezing. The (tam'tge during free.zing does not aPl,:.ar to depend Ul,On the gas its,If hut upon the change in volume that the gas prodtwes in the. cell. Heat. transfer calculations iu~ticaie that the d'tmage does not appear to be em~svtl hy supercooling or by intraeellular ice forn l a t i o l l h i l t t.,y all a s yet undetermined mechanism. 2. '|'he Effect of Pressure on the Freezing of Red Blood Cells. Fm~uF.lllc I¢. AUL~areN* .hNO Pt~m,Y I.. Bt.ACKSHE.ta, Ja. (Department of Engineering~ U'niversity of Minnesota, Minneapolis, Minnesota). As red blood cells are frozen at atmospheric presm~r+, hemolysis starts to occur ~:!ightly be.low tt~e freezing point and continues to a teml)erature of --60°C. ,"3im+epressure lowers the freezing point of aqlwcms solutions, it was the authors' hyt)othesis that, pressure might be used to reduce the width of this hemolytic temperature region, provided that the colder side of the region was not also de|,re,sell. This experiment was conducted to deterntitm how l~r<:s.~ure aJIected the colder side of that tetnl)eraturo range which causes rapid hemolysis of blood. Whole dog blood was frozen quickly in a snlall Teflon tube which was phmged into isopentane at --93°C. The tube was then quickly transferred to a precooled pressure chamber which was then sealed and pressurized to a ~est pre,.~sure. The pressure elmmber and sample were then warmed to a test temperature and incubated for 15 rain. The chamber was then recooled to --70°C and depressurized. Tlw. sample tube was withdrawn and thawed in wat+rr at 37°C. The level of hemolysis was then determined. It was found that lip tO 1250 aim increasing the pressure deerea.~ed the whlth of the hemolytic temt,eraturo region by both depressing the warmer side and elevating Lho. eohler side. At that pressure the hemolytic region extends from --10°C to -37°C, lcs:s lhnn one-half that of atmospheric pressure. Further increasing the prexsure wits found to increase the width of the hemolytic region. A,~ a follow-up experiment whole blood waz frozen at, a rate of 100°C per rain :it, various pre~ures. A t 1 utm hemolysis was complete but at 1250 arm the level of hemolysis was reduced to 10% ; furlher inere'~sing the pre~ure inorea~M the level of hemolysis.
571
It, has been .~'eport+'d lbat freezing Idood at 1 alto tit a rail: of approximately 7800°C per aniu will tmnluce 1{1% hr.molysis. Thus. the advantages of prv.-_'suro trey.zing an. obvious, 10q hemolysi~ at I00°C per rain vs. 10% henmb'sis 't! 7800°C per min. 3. High liydrostatie l~ressure.in(lueed Variations in Supercooling and Freezing Tempera. lures of Water and Certain Aqueous Solutions, x ~'IAXI.M].), PEIiSlDSKY (Institute of Medical Sciences, l"u'ific Medical C c l l | e r , Sell Francisco, Californ in). Our sttMio.-." of the effects of high hydrostatic pressllrt~ on tissue cryopreserw~tion revealed uliex.. pet:ted irregularitie.,~ in the rate 6f freezing point depre~iou in a certain range of prc~,sure. Since Bridgman's and more rt:eent studies concern only pure water and are re|her sehemalic, we f¢.h that detailed investigation of tlie~e relations in certain •tqueous solutions of CD'ohiological importance would bc of inh~rest. Waist, solutions of nonMeetrotytes such as allmethyl sulfoxidc and polyvinylp3"rrolidone, and some electrolytes were studied. Plx;ssuro was wlrietl from atmospheric to 40,000 p~s.i.; temperature and voltlnK~ changes during phase tr:msition at different pressures were measur&t. Of !mri.ieular interest was the extent of supercooling and freezing tern.. peratures. When plotted against pre~ure these to+mperatun~+s produced compound curves reprosenting thermttt l~rofiles of supercooling and freezing. Thermal profiles for water show that. ,+ut~ercooling disappears at certain points be!ween 24,000 and 30,0t30 p,s,i., and inenm~es above 30,000 p.s.i+ Freezing also shows irregularity within thi~ ~,r~sure range. Above 30,000 p+si. tWO freezings occ~ir at differeni, temperatures. The fir~st is ,as~eiated with volume decrease, the seeond with Volume increase. ComparaMe thermal l/mJfiles wifli el~,~c2roJ 13"~e~ and nonelecirolytes show s o m e similar ;and some dissimilar features, mid will be illustrated. It is pomible that, at i h e ~ pressures, several transient, metastable forms of ice e+xizt w!iid: affdet the thermal profile of mipereooling anti fr~zini+, 4+ Superhyperbarie Cryoprcservat|on o f tleart Valve~Y aMAXI~ D. Pt::RSlIJSK~- ,~0 ~r/LLIAM J, Kt:rt'rIt* (Institute 'of Medical Sciences, Padfie Medical Center, San Francis~x~. California). There are two main advantages which eryopr~:~r.. ration under high hydrostatic pressures offers: I) t Supported by Nadonal Gran~ 5 Ii0t i1E12668,01.
Institutea of H ~ d t h