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Chemical and hematological changes in stored CPD blood.

Blood was drawn from ten healthy volunteer donors into citrate-phosphate-dextrose (CPD) anticoagulant and placed on the quarantine shelf of the blood bank refrigerator. Plasma dextrose, sodium, potassium, chloride, bicarbonate, GOT, LDH, and hemoglobin as well as WBC, hematocrit, MCV, MCHC, whole blood pH, and ammonia were measured on all samples initially and at one, two, seven, 14, 21, and 28 days of storage at 4 C. Whole blood lactate also was analyzed serially on five of the units. An additional 27 units of CPD bank blood (two to 21 days of age), routinely processed, handled, and stored by the blood bank, were submitted to the same analyses on the day of administration to the patient. Five of these processed units, 21 days old, were resampled at 28 days. Results of the analyses are presented and discussed. The most pronounced changes were seen for dextrose, potassium, bicarbonate, lactate, LDH, ammonia, and hemoglobin. Plasma dextrose and bicarbonate declined in concentration while potassium, lactate, LDH, ammonia, and hemoglobin rose with storage. In general, changes in the regularly processed, singly sampled bank units were greater than those observed in the specially processed, quarantined units sampled serially. This study indicates that routine transportation, processing, and handling of bank blood may lead to increased biochemical alteration.

Ammonia

Compatibility of common intravenous solutions with CPD blood.

Blood anticoagulated with CPD was mixed with lactated Ringer's solution, 5 per cent aqueous dextrose, 5 per cent dextrose in 0.225 per cent saline, 5 per cent dextrose in 0.9 per cent saline, and 0.9 per cent saline solution in varying concentrations and incubated at room temperature and 37 C. Clots formed in the blood-lactated Ringer's mixture after five minutes at a citrate:calcium molar ratio of 4:1 or lower. Aqueous dextrose-blood mixtures showed immediate clumping with gross hemolysis after 30 minutes incubation. Blood mixed with 5 per cent dextrose and 0.225 per cent saline hemolyzed within ten minutes incubation at 37 C. No hemolysis occurred in blood mixed with 5 per cent dextrose in 0.9 per cent saline or with 0.9 per cent saline. Traces of solutions labeled with Evans blue dye remained in intravenous administration tubing even 30 minutes after a simulated transfusion was begun. Lactated Ringer's solution and 5 per cent dextrose in 0.225 per cent saline should not be administered concurrently with blood. Lactated Ringer's solution may also be harmful when used to start transfusions as it rapidly produces clots when mixed with CPD blood.

Anticoagulants

Adenine metabolism during and after exchange transfusions in newborn infants with CPD-adenine blood.

CPD-adenine blood (adenine in a final concentration of 0.25 mmol/1) was used in exchange transfusions in four newborn infants. The amount of adenine in one exchange transfusion ranged from 27 to 29 mumol per kg bodyweight. The maximum P-adenine concentration during the exchange transfusions ranged from 4 to 8 mumol/1 but decreased to pretransfusion levels 20 minutes after the exchange transfusions. During a 24-hour period following the exchange transfusions, the total urine excretion of adenine and 2,8-dihydroxyadenine corresponded to 0.5 to 1.3 per cent of the given adenine dose calculated on a molar basis. Accumulated data indicate that CPD-adenine blood can be used even in repeated exchange transfusions in newborn infants.

Adenine

Additional studies concerning the metabolism of packed erythrocytes in CPD adenine.

Studies concerning the relationship between glucose level, hematocrit to which cells were packed (within three hours of initial collection) and adenosine triphosphate (ATP) concentration were undertaken in CPD supplemented with adenine (0.25 mM final concentration). It was found that apparently adequate ATP levels could be maintained in 90 per cent hematocrit packed units at 42 days only if glucose was present in amounts 1.5 times or greater than that provided by CPD. Less tight packing of the units to 70 and 80 per cent hematocrit maintained ATP at greater than 50 per cent of the initial level for a full 42 days of storage when 1.25 times the usual glucose concentration was present in the initial anticoagulant. No difference in 2,3-diphosphoglycerate or pH was found in any of the modified media under study.

Adenine

Changes in agglomeration of human red blood cells in liquid storage in CPD media.

A procedure which may distinguish between old and new CPD blood units in liquid state is described. It is based on the observation of increased tendency to reversible agglomeration in old erythrocytes in liquid preservation. Erythrocytes clump together when they are mixed with low ionic strength solutions in pH range of 5.2 to 6.5. We found that liquid-stored erythrocytes show an augmented tendency to agglomerate in 0.24M sucrose, pH 7.2. The tendency increases with storage so that in the fourth week, more than 70% of the units show agglomeration under these conditions. The addition of minute amounts of sodium chloride may prevent agglomeration. As the cells age, higher salt concentration is required to prevent agglomeration. A short incubation of washed cells with adenosine may reverse the tendency of outdated erythrocytes to agglomerate, concomitantly with reestablishment of initial ATP level. However, depletion of the ATP of fresh cells with fluoride does not induce agglomeration. A 20 hour incubation of units at 37 C with CPD revealed an increased sensitivity of older units, with low ATP and positive agglomeration. This test may help in distinguishing between outdated and younger units in the blood bank.

Adenosine

Hemoglobin function in stored blood. XIX. Inosine maintenance of 2,3-DPG for 35 days in a CPD-adenine preservative.

This study establishes that 10 mM inosine is a sufficient additive to maintain 2,3-DPG levels in blood for five weeks of storage in CPD-adenine. No previous experiments were done with CPD-adenine (0.25 mM) using a design which would give statistical proof of the optimal concentration of inosine needed for maintenance of normal hemoglobin function (2,3-DPG) for five weeks of blood bank storage.

Adenine

Erythrocyte agglomeration and survival studies in citrate-phosphate-dextrose (CPD) units.

We have previously reported increased tendency to agglomeration in CPD-stored erythrocytes. In the present study we investigated the deterioration pattern of fresh units, as detected by agglomeration, free hemoglobin levels, ATP levels, osmotic fragility, and certain enzymatic activities. When the negative test was converted to a positive one, the 24-h survival values of these units were determined by autotransfusion. A positive agglomeration occurred while viability was still above 70%. Fresh units had high ATP levels and negative agglomeration while old units had low ATP levels and positive agglomeration. However, in intermediate-storage units, no definite correlation between ATP and agglomeration was found. Agglomeration seems to reflect the degree of changes in stored erythrocytes. However, further studies are needed before this test may serve for prediction of post-transfusion viability.

Adenosine Triphosphate

Potassium load in CPD-preserved whole blood and two types of packed red blood cells.

The potassium load of transfused blood must be minimized. We have compared the total plasma potassium content of units of CPD-preserved stored whole blood (SWB), stored packed cells (SPC), and packed cells prepared from stored whole blood (WB-PC). Plasma potassium concentrations, unit weights, and hematocrits of 20 units of SWB, 27 units of SPC, and 20 units of WB-PC of various ages were measured. During the 21-day storage period, total plasma potassium content per unit increased in units of SPC at the same rate as in units of SWB, because plasma potassium concentration increased in SPC at three times the rate of SWB. The values for total plasma potassium per unit at 14 and 21 days in mEq/unit were: SWB, 4.4, 5.8; SPC 3.1, 4.4; and WB-PC 1.9, 2.5. Thus, SPC units may contain substantial amounts of plasma potassium when stored for two to three weeks. However, removal of most of the remaining supernatant plasma from SPC units just prior to administration provides a readily available supply of low potassium blood while allowing maximum conservation of scarce blood resources.

Blood Preservation

Reversal of the storage lesion of CPD bank blood: a problem in clinical medicine.

The effect of phosphate buffer on the course of pH, ATP, and 2,3-PDG of CPD red blood cells stored at three temperatures was observed. Basic phosphate at an equilibrated level of 10 mM (as iP) maintained pH above 7.00 and ATP and 2,3-DPG above 70 per cent of initial value in cells stored at 37 C for 24 hours. In contrast however, at 25 and 4 C no buffering was obtained with basic phosphate concentrations up to 50 mM, but values for both ATP and 2,3-DPG were higher in phosphate treated aliquots than in controls throughout storage. When the pH of blood stored at 4 C was adjusted into the range 7.15 to 7.25 with tromethamine and the level of iP raised to 10 mM by addition of Na2HPO4 on day seven, it was found that ATP and 2,3-DPG levels were maintained at 90 and 120 per cent, while control levels fell to 60 and 12 per cent, respectively at 21 days. The process described parallels the normal repair of damaged red blood cells of bank blood that occurs in vivo following transfusion.

Adenosine Diphosphate

Plasma adenine and cellular ATP in red cell concentrates collected and stored in modified CPD at 4 C.

Eight units of blood were drawn into modified CPD containing 25 per cent higher glucose and 17.3 mg adenine (0.25 mM in blood). Red blood cell concentrates (RCC) were prepared to a mean hematocrit (Hct) of 70, the cells stored at 4 C, and plasma adenine and red blood cell adenosine triphosphate (ATP) were measured weekly for 42 days. The removal of plasma in the preparation of RCC reduced by 39 per cent the available adenine. As a result measurable plasma adenine was depleted by 21 days. The loss of ATP in RCC occurs at a significantly faster rate than in whole blood stored under the same conditions. When red blood cells are stored at higher HCT or for periods longer than 35 days, increased anticoagulant adenine levels are recommended.

Adenine

Discocyte--echinocyte reversibility in blood stored in CPD over a period of 56 days.

Blood collected with CPD and stored was examined with optical (LM) and scanning electron microscopy (SEM) before and after reversal of echinocytes into discocytes. Reversal was achieved by incubation of the red blood cells at 37 C in an adenosine containing medium. The transformation of discocytes into echinocytes occurred rapidly during the first three weeks of storage. A shape/density relationship was observed in the various fractions. The denser cells were found to have the more advanced echinocytic changes. After incubation with adenosine, most of the cells reversed into discocytes and early stages of stomatocytes. When spheroechinocytes I and II were present, they reversed into spherostomatocytes. No echinocytogenic property was found in plasma during 5 weeks of storage at 4 C and no immediate reversion of echinocytes was obtained in fresh plasma, therefore the initial discocyte--echinocyte transformation was explained by intracellular changes. The data from the various fractions showed that the more dense cells were more spherocytic. We suggest that removal of this part of the population of stored red blood cells might improve the survival of transfused cells.

Adenosine

Histamine-releasing effect of a corticotrophin derivative. II. Mechanism of action of histamine release by C 44 680-Ba, compared with that of Cpd. 48/80, dextran and triton.

The mechanism of the histamine-liberating action of the synthetic polypeptide C 44 680-Ba, an alkyl-prolyl derivative of beta 1-19 corticotrophin, was investigated and compared with those of Compound 48/80, dextran, Melittin and Triton X-100. It was found that the release of histamine from rat peritoneal cells induced by the polypeptide is dependent on temperature, pH, calcium ions and energy-providing processes. In regard to these criteria, the mode of action of this histamine liberator resembles that of Compound 48/80 but is quite distinct from that of the unspecific substance Triton X-100.

Adrenocorticotropic Hormone

CPD in Canada.

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Blood Preservation