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Biological and biophysical considerations of INA bacteria
ABSTRACTS,
26th ANNUAL
rocytes by the mentioned method allows the maintenance of the native structure of the spectrin net after thawing. Hence, when the temperature is lowered, the strength of association between membrane skeleton components and phospholipid molecules increases, which is manifested in an increase the mechanical stability of osmotically shrunken cells. However, the process of cytoskeleton protein stabilization is manifested only under conditions of cell dehydration when their aggregate state and spatial distribution change, as well as hemoglobin transition into ion gel. Therefore, the mechanism of higher cell resistance may be associated with the cytoplasm dehydration which changes the structure and function of membrane skeleton proteins. 163. Mechanisms of Neuromediator Secretion and Reception upon Cooling. G. A. BABIYCHLJK (Institute for Problems of Cryobiology and Cryomedicine of the UkrSSR Academy of Sciences, Kharkov, USSR).
MEETING
phase reaction in changing the EEG synchronization being maintained upon B-AR blockage. WORKSHOP
VII-BIOLOGY
OF
ICE
NUCLEATION
164. Introduction to Biology and Molecular Biology of Ice Nucleation. M. BURKE (Oregon State University, Corvallis, Oregon). 165. Ecology of INA Bacteria. C. UPPER. 166. Molecular Biology of INA Bacteria. G. WARREN (DNA Plant Technology Corporation, Oakland, California). 167. Biological and Biophysical Considerations of INA Bacteria. S. LINDOW (University of California, Berkeley, California). SESSION
The receptor-ligand interrelations are important for interpretation of the effect of craniocerebral hypothermia (CCH) on organisms. Narcotized rats were subjected to CCH using a device for controlled cooling. Experiments in viva were performed by superfusion of various brain regions. Preparations of synaptic membranes were obtained from cerebral cortex and hypothalamus for experiments in vitro. After 15 min CCH, the secretion of [3H]noradrenaline ([3H]NA) in animals with rectal temperatures near 34°C sharply increased in the hypothalamus preoptical region. Lowering the body temperature to 32°C decreased the spontaneous and K+-induced secretions of [3H]NA. The level of [3H]serotonin decreased both in cortex and in hypothalamus at these temperatures. Of interest seems the appearance of the u,-adrenoreceptor (AR) population with high affinity for ligand, with kinetic parameters of a,-AR remaining unchanged. As a result of great NA ejection with onset of cooling, the desensitization of B-AR occurred along with a decrease in the activity of adenylate cyclase. The intensification of EEG synchronization upon CCH requires increasing the NA secretion in anterior hypothalamus. In this case the synchronous effects of hypothalamus are transmitted to cortex through AR. It has been found that superfusion of hypothalamus of uncooled animals with B-AR blockers intensifies the EEG spatial synchronization which is depressed, however, by a-AR blockers, thus indicating a possibility of transmitting the synchronous effects of hypothalamus through the action of NA on a-AR and the spatially desynchronous ones on B-AR. Under CCH with a-AR blockage, the depression is followed by the intensification of synchronization of biorhythms of neocortex that is believed to be associated with the appearance of a second active population of a-AR upon CCH and a one-
593
XIIdRGAN
PRESERVATION
II
168. Comparison of Renal Tubule Survival in Vitro during Cold Storage with Various Solutions. K. A. ANDEREGG, K. G. M. BROCKBANK, AND D. A. SENS (Medical University of South Carolina, Charleston, South Carolina). The objective of this study was to compare the ability of currently marketed kidney preservation solutions (University of Wisconsin solution [UW], Collins 2, and Euro-Collins) and salt solutions (normal saline, phosphate-buffered saline, and phosphate-buffered saline plus 140 mM sucrose) to preserve confluent human kidney proximal tubule cells in vitro at WC. The cultures were refrigerated for l-5 days. Cell numbers in experimental groups were determined by morphometric analysis of fixed toluidine blue-stained cell monolayers. Viability was assessed by quantitation of Neutral Red and 2-[3H]deoxyglucose uptake. The results demonstrate that Collins 2 was superior to both UW and Euro-Collins, while less than 33% of the cells survived 24 hr in the salt solutions. Survival in the organ preservation solutions deteriorated rapidly after 2 days of refrigeration. Cell survival was better in UW solution than in Euro-Collins, supporting previous studies on whole kidney preservation. These results suggest that proximal tubule cultures may be an appropriate model for testing cold storage solutions, prior to experiments utilizing intact kidneys and subsequent transplantation. 169. Comparison of Euro-Collins Solution and RPS-2 as Potential Carriers for Vitrification Solutions Using the Rabbit Renal Autograft Model. B. S. KHIRABADI, G. M. FAHY, S. E. ALI, AND P. MEHL (Transplantation Laboratory, American Red Cross, Rockville, Maryland).
26th ANNUAL
rocytes by the mentioned method allows the maintenance of the native structure of the spectrin net after thawing. Hence, when the temperature is lowered, the strength of association between membrane skeleton components and phospholipid molecules increases, which is manifested in an increase the mechanical stability of osmotically shrunken cells. However, the process of cytoskeleton protein stabilization is manifested only under conditions of cell dehydration when their aggregate state and spatial distribution change, as well as hemoglobin transition into ion gel. Therefore, the mechanism of higher cell resistance may be associated with the cytoplasm dehydration which changes the structure and function of membrane skeleton proteins. 163. Mechanisms of Neuromediator Secretion and Reception upon Cooling. G. A. BABIYCHLJK (Institute for Problems of Cryobiology and Cryomedicine of the UkrSSR Academy of Sciences, Kharkov, USSR).
MEETING
phase reaction in changing the EEG synchronization being maintained upon B-AR blockage. WORKSHOP
VII-BIOLOGY
OF
ICE
NUCLEATION
164. Introduction to Biology and Molecular Biology of Ice Nucleation. M. BURKE (Oregon State University, Corvallis, Oregon). 165. Ecology of INA Bacteria. C. UPPER. 166. Molecular Biology of INA Bacteria. G. WARREN (DNA Plant Technology Corporation, Oakland, California). 167. Biological and Biophysical Considerations of INA Bacteria. S. LINDOW (University of California, Berkeley, California). SESSION
The receptor-ligand interrelations are important for interpretation of the effect of craniocerebral hypothermia (CCH) on organisms. Narcotized rats were subjected to CCH using a device for controlled cooling. Experiments in viva were performed by superfusion of various brain regions. Preparations of synaptic membranes were obtained from cerebral cortex and hypothalamus for experiments in vitro. After 15 min CCH, the secretion of [3H]noradrenaline ([3H]NA) in animals with rectal temperatures near 34°C sharply increased in the hypothalamus preoptical region. Lowering the body temperature to 32°C decreased the spontaneous and K+-induced secretions of [3H]NA. The level of [3H]serotonin decreased both in cortex and in hypothalamus at these temperatures. Of interest seems the appearance of the u,-adrenoreceptor (AR) population with high affinity for ligand, with kinetic parameters of a,-AR remaining unchanged. As a result of great NA ejection with onset of cooling, the desensitization of B-AR occurred along with a decrease in the activity of adenylate cyclase. The intensification of EEG synchronization upon CCH requires increasing the NA secretion in anterior hypothalamus. In this case the synchronous effects of hypothalamus are transmitted to cortex through AR. It has been found that superfusion of hypothalamus of uncooled animals with B-AR blockers intensifies the EEG spatial synchronization which is depressed, however, by a-AR blockers, thus indicating a possibility of transmitting the synchronous effects of hypothalamus through the action of NA on a-AR and the spatially desynchronous ones on B-AR. Under CCH with a-AR blockage, the depression is followed by the intensification of synchronization of biorhythms of neocortex that is believed to be associated with the appearance of a second active population of a-AR upon CCH and a one-
593
XIIdRGAN
PRESERVATION
II
168. Comparison of Renal Tubule Survival in Vitro during Cold Storage with Various Solutions. K. A. ANDEREGG, K. G. M. BROCKBANK, AND D. A. SENS (Medical University of South Carolina, Charleston, South Carolina). The objective of this study was to compare the ability of currently marketed kidney preservation solutions (University of Wisconsin solution [UW], Collins 2, and Euro-Collins) and salt solutions (normal saline, phosphate-buffered saline, and phosphate-buffered saline plus 140 mM sucrose) to preserve confluent human kidney proximal tubule cells in vitro at WC. The cultures were refrigerated for l-5 days. Cell numbers in experimental groups were determined by morphometric analysis of fixed toluidine blue-stained cell monolayers. Viability was assessed by quantitation of Neutral Red and 2-[3H]deoxyglucose uptake. The results demonstrate that Collins 2 was superior to both UW and Euro-Collins, while less than 33% of the cells survived 24 hr in the salt solutions. Survival in the organ preservation solutions deteriorated rapidly after 2 days of refrigeration. Cell survival was better in UW solution than in Euro-Collins, supporting previous studies on whole kidney preservation. These results suggest that proximal tubule cultures may be an appropriate model for testing cold storage solutions, prior to experiments utilizing intact kidneys and subsequent transplantation. 169. Comparison of Euro-Collins Solution and RPS-2 as Potential Carriers for Vitrification Solutions Using the Rabbit Renal Autograft Model. B. S. KHIRABADI, G. M. FAHY, S. E. ALI, AND P. MEHL (Transplantation Laboratory, American Red Cross, Rockville, Maryland).