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

Gabrio, Beverly Wescott; Donohue, Dennis M.; Huennekens, F.M.; Finch, Clement A.

doi:10.1172/jci103322

Cited by 61 publications

(27 citation statements)

References 12 publications

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“…(28). Since improved preservation of stored red cells has been reported after addition of certain purine nucleosides (29)(30)(31), the effects of adenosine and inosine (0.03 to 10 mM) were studied in relation to the development of agglutinability.…”

Section: Stndies Of the Serum Factormentioning

confidence: 99%

Agglutination of Stored Erythrocytes by a Human Serum. Characterization of the Serum Factor and Erythrocyte Changes*

Ozer¹,

Chaplin²

1963

J. Clin. Invest.

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There have been four previous reports of human sera causing agglutination of stored but not of fresh red cells (1-4). The sera are of special interest for two reasons. Since the agglutinins are active against the patients' own stored cells as well as against cells from other donors, one must consider whether such patients represent examples of autoimmune disease. Of more immediate practical importance, the sera provide a new means for studying the red-cell storage defect, since the agglutinins detect in the erythrocyte membrane a hitherto unrecognized alteration that progresses during in vitro storage concurrently with deterioration in cellular metabolism.Characterization of the previously reported stored-cell agglutinins has been limited. The present communication describes detailed studies of a serum causing strong agglutination of stored human red cells. The serum factor responsible for the agglutination was shown to be a gamma macroglobulin. Studies of red cells during storage in vitro indicate that development of agglutinability is closely related to the alterations in erythrocyte glucose metabolism occurring during storage. Also included are the results of sixteen studies of erythrocyte survival in vivo that are designed to examine the relationship of agglutinability to nonviability of the stored cells. METHODSThe patient was a 71-year-old woman whose chief complaints were abdominal swelling and progressive weakness. The principal positive physical findings were pallor and hepatosplenomegaly. Laboratory findings of special interest were: hemoglobin, 9.7 g per 100 ml; reticulocytes, 4.2%o; serum bilirubin concentration, nor-* Work supported by U. S. Public Health Service research grant G-2918 (C5) from the National Cancer Institute, Bethesda, Md. mal; total serum protein concentration, 7 g per 100 ml (albumin, 2.7 g, globulin, 4.3 g). Survival in vivo of Cr51-labeled, f resh, compatible, red cells of normal donors was moderately severely reduced, judged by redcell life-span (Cr51 tt, about 8 days) based on daily blood sampling for 1 week. Exploratory laparotomy revealed moderate hepatic enlargement, diffuse mesenteric and retroperitoneal lymph-node enlargement, and massive splenomegaly. Splenectomy was performed, and the patient's postoperative course was uneventful. Two units of blood were transfused before and two additional units during surgery. Although the patient was improved at the time of discharge and required no further transfusions, she died at home 7 months later of undetermined cause; necroscopy was not performed. The spleen removed at operation weighed 2,050 g and showed hyperplasia of the white pulp, with impaired follicle formation; each Malpighian corpuscle was surrounded by a margin of hyperplastic lymphoid cells. Lymph nodes revealed intact architecture, but were filled out with lymphoid elements without follicle formation. A liver sample from biopsy was not remarkable, and aspirated bone marrow revealed only generalized hyperplasia. A specific pathological diagnosis was not made. Stndies...

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Section: Stndies Of the Serum Factormentioning

confidence: 99%

Agglutination of Stored Erythrocytes by a Human Serum. Characterization of the Serum Factor and Erythrocyte Changes*

Ozer¹,

Chaplin²

1963

J. Clin. Invest.

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“…Hemolysis during storage was determined by the measurement of plasma hemoglobin as the pyridine he- ' adjusted to pH 10 with NH4OH, according to the methods described previously (1). After identification of the purine-containing compounds with the use of an ultraviolet light (Mineralite), the materials were eluted from the paper with water and measured spectrophotometrically.…”

Section: Methodsmentioning

confidence: 99%

“…Since both hypoxanthine and inosine were present in the plasma of blood stored in ACDI or ACDA (1,4,8), the relative amounts of each substance could not be estimated directly from light absorption measurements alone. However, after the amount of hypoxanthine had been determined by means of xanthine oxidase, it was possible from the light absorption data on the plasma filtrates, to correct the extinction values at 249 m'& (peak light absorption of hypoxanthine, millimolar extinction coefficient 10.5) for hypoxanthine concentration.…”

Section: Methodsmentioning

confidence: 99%

“…Washington and King County Blood Bank) (Submitted for publication May 23, 1956; accepted November 29,1956) It has been shown previously that the addition of inosine to ACD preservative 2 prolongs the effective period of in vitro storage of erythrocytes (1). Inosine is utilized by the red cell after a phosphorolytic cleavage to ribose-l-phosphate and hypoxanthine, mediated by a nucleoside phosphorylase (2,3).…”

mentioning

confidence: 99%

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Erythrocyte Preservation. VIII. Metabolic Degradation of Nucleosides In Vitro and In Vivo1

Hennessey¹,

Finch²,

Gabrio³

1957

J. Clin. Invest.

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It has been shown previously that the addition of inosine to ACD preservative 2 prolongs the effective period of in vitro storage of erythrocytes (1). Inosine is utilized by the red cell after a phosphorolytic cleavage to ribose-l-phosphate and hypoxanthine, mediated by a nucleoside phosphorylase (2, 3). Ribose-l-phosphate enters the "aerobic shunt pathway" of glucose metabolism after conversion to ribose-5-phosphate, and a subsequent effect is a generation of ATP 8 and the resultant maintenance of the energy reserve of the red cell (4). Inasmuch as the red cell lacks the enzyme, xanthine oxidase (5), it is evident that the other cleavage product, hypoxanthine, is not metabolized further but remains in the red cell and the plasma during storage.The present investigation is concerned with the rate of conversion of inosine to hypoxanthine by the red cells and the ratio of these substances in the plasma of blood stored in ACDI, as well as the in vivo metabolism of inosine after infusion. Related studies on the nucleosides, adenosine and guanosine, will be presented also. METHODSHuman blood was collected in ACD, and the nucleoside, dissolved in 0.9 per cent NaCi, was added with sterile precautions.Inosine and adenosine were obtained from Schwarz Laboratories; hypoxanthine and guanosine from Nutritional Biochemicals Corporation. adjusted to pH 10 with NH4OH, according to the methods described previously (1). After identification of the purine-containing compounds with the use of an ultraviolet light (Mineralite), the materials were eluted from the paper with water and measured spectrophotometrically. These substances were estimated also by quantitative densitometry4 of the paper chromatograms. The hypoxanthine content of the plasma and red cells was determined enzymatically, using xanthine oxidase (7), except that the assays were performed on neutralized. PCA filtrates of the various fractions.Acid filtrates of plasma were prepared in the following way: 2 ml. of plasma were added to 2 ml. of cold 0.6 N PCA, mixed well, and centrifuged at 40 C for 5 minutes at 15,200 X g.5 The precipitate was washed once, with 2 ml. of cold 0.3 N PCA, and the centrifugation was repeated. The supernatant fluid and the washing were, combined and neutralized with cold 20 per cent KOH, followed by centrifugation at 4°C for 10 minutes at 15,200 X g. The KCIO4 precipitate was washed once with 2 ml. of cold distilled water and centrifugation repeated. This second supernatant fluid and the washing were combined for analysis. Acid filtrates of the red cells were prepared in essentially the same way. Approximately 4 ml. of cells were washed twice with an equal volume of cold 0.9 per cent NaCl each time with centrifugation for 20 minutes at 1,700 X g.6 The washings were discarded, since they contained only negligible amounts of ultraviolet absorbing materials.A 50 per cent cell suspension was prepared with 0.9 per cent NaCl. Hematocrit determinations were made on this cell suspension. One ml. of cell suspension was added to 3 ml. of cold 0.6 N P...

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“…In 1955 Gabrio, Donohue, Huennekins and Finch (1) reported that inosine prolonged the period that human blood could be stored at 40 C. in acid citrate dextrose (ACD); their experimental evidence indicated greater than 70 per cent viability of the erythrocytes after six weeks of storage. Because of these encouraging findings, a cooperative study was undertaken in which the posttransfusion survival of erythrocytes from blood stored in ACD and in ACD plus inosine (ACDI) was compared (2).…”

mentioning

confidence: 99%

Effect of Inosine on the Post-Transfusion Survival of Stored Rabbit Erythrocytes*

Shafer¹,

Bartlett²

1960

J. Clin. Invest.

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

Cited by 61 publications

References 12 publications

Agglutination of Stored Erythrocytes by a Human Serum. Characterization of the Serum Factor and Erythrocyte Changes*

Agglutination of Stored Erythrocytes by a Human Serum. Characterization of the Serum Factor and Erythrocyte Changes*

Erythrocyte Preservation. VIII. Metabolic Degradation of Nucleosides In Vitro and In Vivo1

Effect of Inosine on the Post-Transfusion Survival of Stored Rabbit Erythrocytes*

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