Erythrocyte Preservation. I. The Relation of the Storage Lesion to in Vivo Erythrocyte Senescence 1

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Previous investigations on erythrocyte preservation (1-4) have demonstrated a marked effect of purine nucleosides on the metabolism of the red cell throughout storage. Attention has been focused upon the effect of adenosine in the maintenance of glucose metabolism and of high energy phosphate compounds, such as adenosine triphosphate (ATP). As a result, not only are the physical and chemical characteristics of the erythrocyte preserved, but the storage period, during which satisfactory post-transfusion viability is maintained, is extended to approximately twice that observed in acid-citrate-dextrose (ACD) preservative alone. It was postulated that the effect of adenosine was the result of an initial enzymatic conversion to inosine by a deaminase and the subsequent splitting of inosine to yield hypoxanthine and ribose-l-phosphate, mediated by a nucleoside phosphorylase (4, 5). Thus, it appeared that inosine might be the actual substrate for the initial enzymic reaction, and hence the preferred substance for viability studies. In order to evaluate a new preservative, it is desirable to correlate 1) the chemical characteristics of the stored red cells, 2) the ability of the cells to survive after transfusion, and 3) possible toxic effects of the preservative. Studies employing acid-citrate-dextroseinosine (ACDI) as a blood preservative are presented in this paper, as well as further investigations on the mechanism of action of purine nucleosides. METHODSProcedures for the determination of phosphate partition, glucose, and osmotic fragility of erythrocytes have been described elsewhere (3,6).Inosine, adenosine, deoxyadenosine, and adenine were obtained from Schwarz Laboratories; guanine, xanthine,

Section: Discussionmentioning

confidence: 97%

“…Procedures for the determination of phosphate partition, glucose, and osmotic fragility of erythrocytes have been described elsewhere (3,6).…”

Section: Methodsmentioning

confidence: 99%

Erythrocyte Preservation. Vii. Acid-Citrate-Dextrose-Inosine (Acdi) as a Preservative for Blood During Storage at 4° C1

Gabrio¹,

Donohue²,

Huennekens³

et al. 1956

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“…The techniques for the measurements of erythrocyte phosphate partition, blood glucose, sodium and potassium, osmotic fragility, and aerobic utilization of glucose have been described (1,4). Oxygen content and oxygen capacity of erythrocytes were determined by the method of Van Slyke and Neill (5).…”

Section: Methodsmentioning

confidence: 99%

“…Inasmuch as blood stored in ACD + adenosine is bright red after several weeks of storage, in contrast to the darker color of blood preserved in ACD alone, the oxygen content and capacity 4 of such blood were measured. The results, recorded in Table IV, suggest a decreased oxygen utilization of red cells during storage in blood supplemented with adenosine.…”

Section: Methodsmentioning

confidence: 99%

Erythrocyte Preservation. Vi. The Storage of Blood With Purine Nucleosides 1

Donohue¹,

Finch²,

Gabrio³

1956

Self Cite

The addition of adenosine to stored blood results in a chemical restoration of deteriorated erythrocytes and sustains their viability during continued storage (1,2). This phenomenon has been referred to as the in vitro reversibility of a biochemical lesion of storage. Apart from the implications of this reaction in red cell metabolism (3), it seemed of importance to evaluate the effect of adenosine in the preservation of blood. The present study indicates that adenosine and some related purine nucleosides retard the onset of the storage lesion and extend significantly the period of viability of stored blood. The effective concentration of these supplements, as well as their potential toxicity, is discussed. METHODSThe techniques for the measurements of erythrocyte phosphate partition, blood glucose, sodium and potassium, osmotic fragility, and aerobic utilization of glucose have been described (1,4

“…Alterations in the relationship of the phosphate fractions develop in erythrocytes during storage (10)(11)(12)(13)(14)(15)(16). Similar changes do not occur with the senescence of erythrocytes in vivo (12).…”

mentioning

confidence: 90%

The Phosphate Partition of the Erythrocytes of Normal Newborn Infants and of Infants With Hemolytic Disease

Greenwalt¹,

Ayers²

1956

The mammalian erythrocyte depends on a complex glycolytic mechanism for most of its energy. It is believed that this mechanism controls the transfer of cations across the cell membrane, and serves to maintain the biconcave shape of the red blood cell. Adenosine triphosphate and 2,3-diphosphoglycerate participate in the energy exchanges involved in these processes (1)(2)(3)(4)(5)(6)(7)(8)(9).Alterations in the relationship of the phosphate fractions develop in erythrocytes during storage (10-16). Similar changes do not occur with the senescence of erythrocytes in vivo (12). The deviations from the normal pattern may be reversed following the transfusion of stored blood (14), and by the addition of some of the purine nucleosides in vitro (17)(18)(19)(20)(21)(22)(23)(24) (31,32).The purpose of this study was to find out if the action of specific antibodies on fetal erythrocytes produces any alteration in the phosphate partitioning. The intra-erythrocytic phosphate pattern of normal fetal red cells was studied and compared with that of the red cells of adults. MATERIAL AND METHODSAll the infants studied had been delivered normally but were otherwise unselected. There were ten female and six male babies in the normal group and nine females and one male with erythroblastosis fetalis. The average weight of the normal infants was 7.6 pounds (6.1 to 9.0), and the weight of the infants with hemolytic disease averaged 6.8 pounds (5.1 to 8.4). The blood group of one normal infant was A and that of the mother B.The blood group of two infants with hemolytic disease was AB and their mothers were A2 and B. The other infants were all products of homospecific pregnancies. The medication and anesthesia administered for delivery were similar in the two groups.The cord blood 1 of the normal babies was drawn aseptically from the umbilical vein at the time of delivery. The blood of the infants with hemolytic disease was obtained at the time of the first replacement transfusion within the first eight hours of life. All of the blood specimens were collected into acid-citrate-dextrose solution (ACD) formula A2 in the proportion of 100 ml. of blood to 15 ml. of preservative. The final concentration of glucose added was approximately 320 mg. per cent in whole blood. The specimens reached the laboratory within 30 minutes. Aliquots were distributed into sterile vacuum tubes. The chemical analysis of the fresh specimens was started immediately. The remaining aliquots were stored at 40 to 60 C. Mixing of the specimens during storage was avoided in order to duplicate the usual conditions under which "bank blood" is kept. Each specimen was cultured at the time of use, and the data were discarded if contamination was present.For studying the effect of antibodies added in vitro the plasma was removed from a portion of the blood specimen and replaced with sterile anti-D serum 3 and ACD solution. This mixture and the control specimen were incubated at 37 + 1°C in a water bath, and aliquots of each were distributed into sterile tubes for storage at 4...