Regeneration of 2,3-diphosphoglycerate (DPG) was determined following transfusion of DPG-depleted group O red cells into group A recipients. Blood from five donors was stored in the adenine-containing solutions CPDA-1, AS-1 or AS-3 for 35 d at 4 degrees C. Post-transfusion red cell DPG and ATP were measured in separated group O red cells over a 7 d period. The studies confirmed rapid in vivo DPG regeneration with greater than or equal to 50% of the maximum level being achieved within 7 h. An average of 95% of the recipients' pre-transfusion DPG level was achieved by 72 h and by 7 d mean (+/- SEM) DPG levels relative to recipient's pre-transfusion DPG averaged 84% (+/- 13%), 92% (+/- 17%) and 84% (+/- 21%) for CPDA-1, AS-1 and AS-3 red cells, respectively. Results were comparable to those previously reported for blood stored in ACD for 15-20 d (Valeri & Hirsch, 1969; Beutler & Wood, 1969). The immediate regeneration rate, V, closely approximated first order regeneration kinetics with AS-3 red cells exhibiting double the rate of CPDA-1 red cells (P less than 0.001). AS-1 red cells exhibited an intermediate rate of regeneration which was not significantly different compared to either CPDA-1 or AS-3 (P greater than 0.05). V exhibited a significant (P less than 0.05) positive correlation with ATP levels 5-7 h post-infusion. ATP regeneration of the infused cells was rapid with a mean increase of 1.2 mumol/g Hb above post-storage levels being achieved 1 h following transfusion.
Two techniques for the preparation of platelet concentrate (PC), the standard platelet-rich plasma (PRP) and buffy coat (BC) methods, were compared in nine paired studies with regard to platelet harvest, white cell (WBC) contamination, and PC quality after 5 days of 22 degrees C storage. Platelet harvest using the BC method averaged approximately 56 percent of the whole blood level (6.2 x 10(10)/concentrate), which was less than the 76 percent achieved with the PRP-PC method (8.7 x 10(10)/concentrate). An additional 5 units collected into an experimental siphon bag for BC-PC processing showed improved platelet harvest (6.7 x 10(10)/concentrate, or approx. 70% of whole blood). WBCs remaining in the BC-PC averaged 0.19 x 10(8) per unit compared to 3.6 x 10(8) per unit for PRP-PC. Buffy coat processing produced red cell (RBC) units with 50 percent of the WBC contamination of conventionally prepared units (9.8 +/- 6.2 x 10(8)/unit vs. 18.9 +/- 7.1 x 10(8)/unit). The siphon bag further reduced WBC levels in the AS-3 RBC units (6.4 +/- 3.7 x 10(8)/unit). In vitro studies performed on Days 1 and 5 after collection showed no significant differences in platelet metabolic and biologic function or cell integrity. Beta-thromboglobulin and surface glycoprotein levels, indicators of platelet activation and membrane alteration, respectively, did not differ significantly in the PRP-PC and BC-PC; nor was lactate production higher in PRP-PC, despite the substantially higher WBC counts. Autologous in vivo platelet viability determinations were performed by using concurrent transfusion of 111In-labeled freshly drawn platelets and 51Cr-labeled stored platelets.(ABSTRACT TRUNCATED AT 250 WORDS)
Red cells washed and stored in a citrate-phosphate-glucose-adenine solution at pH 7.4-7.6 demonstrate excellent maintenance of adenosine triphosphate, elevation of 2,3-diphosphoglycerate well above normal levels for more than 6 weeks, reduced hemolysis and 24-hour in vivo survival comparable to that of cells stored in ADSOL. These results can be attributed in part to a chloride shift in which the washout of intracellular chloride is associated with an influx of OH-, which increases intracellular pH and thereby increases the rate of glycolysis. The phosphate functions primarily as a buffer to maintain both extra- and intracellular pH. Reducing the effective osmolality of the storage solution reduces hemolysis and improves cell morphology.
An in vivo and in vitro study of 42-day saline, adenine, glucose, mannitol (SAGM) red cells (Terumo Corporation, Elkton, Md.) was conducted using 20 volunteers to document in vivo efficacy and analyze the validity of the 99mTc/51Cr technique. In vivo autologous post-transfusion recovery was measured by a double-label procedure involving the use of 99mTc-labeled freshly drawn red cells to quantify recipient blood volume and 51Cr to document both post-transfusion 24-hour recovery (PTR) and red cell survival. As an internal control, 5 of the 20 donors studied had red cell volumes estimated by the 125I-albumin (RISA) plasma volume technique on a separate occasion. For comparison, PTR was also calculated according to the single-label 51Cr protocol in which recipient blood volume was estimated by extrapolation from circulating 51Cr-labeled red cell activity 5-15 min after infusion. After 42 days of storage, PTR averaged 78 +/- 6% with the double-label 99mTc/51Cr technique and 81 +/- 5% with the single-label (51Cr alone), confirming a small difference between the two techniques. Correlation between the two techniques was high, r = 0.81, though the average 3% difference between them was significant (p less than 0.05). Red cell volumes of the 5 donors measured by both the 99mTc-red cell method and the 125I-albumin method exhibited excellent correlation, r = 0.96, and averaged 1,961 +/- 420 ml and 2,048 +/- 381 ml, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
The effect of cotton wool filtration of apheresis platelet concentrates (PCs) on platelet viability and complement activation was evaluated by two laboratories. PCs were prepared by automated (Lab A, n = 5) or manual (Lab B, n = 5) apheresis. After storage for 1 day, the PC was filtered through cotton wool before transfusion on one occasion and, on the other occasion, filtered through a standard screen filter before transfusion to the same donor. Five paired studies were performed by each laboratory. Except for a small, but significant reduction in mean platelet size, from 7.3 +/- 1.1 to 6.6 +/- 0.9 microns 3, after cotton wool filtration, no effect of filtration on various tests of in vitro platelet function and morphologic integrity was found. As demonstrated by autologous radiolabeled studies, no effect of cotton wool filtration on platelet viability was found by Laboratory B, while Laboratory A found a slight increase in the percentage of recovery from 59 +/- 4 to 68 +/- 13 percent, and a small reduction in survival, from 8.2 +/- 0.9 to 7.7 +/- 0.5 days after cotton wool filtration (p less than 0.05). Cotton wool filtration was associated with a slight increase in C3a levels found in manual apheresis PCs. Neither laboratory found any effect of cotton wool filtration per se on the recipients' white cell (WBC) counts or C3a and C5a levels after transfusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Studies were carried out to examine whether a single additive solution could support both platelet and red cell storage. An ionically balanced electrolyte solution fortified with citrate, glucose and bicarbonate was used. This solution has previously been shown to provide good platelet viability with storage for up to 7 d. Optimization studies demonstrated that with adenine added to this solution (CSM), it also allowed for satisfactory preservation of in vitro red cell parameters for up to 49 d of storage. Confirmatory paired in vivo post-transfusion studies were carried out in which platelets and red cells obtained from the same donor were processed and stored in CSM on one occasion, and in CPD-plasma (platelets) and AS-1 (red cells) on another occasion. Five paired studies were conducted with platelets stored for 5 d and red cells for 42 d; another five paired studies with platelets stored for 7 d and red cells for 49 d. Except for a slight decrease in platelet survival (P less than 0.05) with storage in CSM, there were no statistically significant differences in post-transfusion recoveries and survivals between test and control media, demonstrating that both platelets and red cells may be satisfactorily stored in a combined single additive solution.
Previous studies with CPDA-1 and Nutricel preserving solutions indicated that red cell properties were affected by small differences in storage temperature. The influence of a 3 degrees C differential on the preservation of the in vitro properties of ADSOL-preserved (AS-1) red blood cells was investigated in a paired study. AS-1 red blood cells were stored at 2.5 and 5.5 degrees C for 42 days. Extent of hemolysis, glucose consumption and pH levels were comparable at the two storage temperatures. Lactate levels were slightly, but significantly higher at 5.5 degrees C. ATP levels were slightly but significantly higher at 2.5 degrees C, only during the later part of the storage period. 2.3-DPG levels were slightly better retained at 2.5 degrees C after 7 days of storage. Holding units of whole blood for either 1 or 8 h at ambient temperature after phlebotomy prior to processing did not influence the types of temperature-dependent changes. The differences as a function of storage temperature were small and appear to be of no practical importance in connection with the storage of AS-1 red blood cells.
A recently developed nonradioisotopic 52Cr technique was used to measure either red cell volume or posttransfusion recovery of stored red cells. The experimental method uses Zeeman electrothermal atomic absorption spectrophotometry to measure red cell chromium. Results from the 52Cr method were compared with those from 51Cr single-label and 125I-albumin/51Cr double-label procedures using 49-day AS-1 red cell concentrates drawn and prepared according to standard procedures. In the first group of five donors, red cell volume was estimated concurrently with both 52Cr-labeled fresh red cells and 125I-albumin. The latter measured plasma volume from which red cell volume was estimated on the basis of the hematocrit (125I red cell volume). 51Cr-labeled stored red cells were transfused to measure posttransfusion recoveries. The correlation between 52Cr and 125I red cell volumes was significant (r = 0.68, p less than 0.01), and, in this group, the differences were not significant (p less than 0.05). Twenty-four-hour posttransfusion recoveries of 51Cr-labeled stored red cells averaged 66 +/- 5 percent when measured with the 125I/51Cr technique and 69 +/- 8 percent when measured with the 52Cr/51Cr method. In the second group of five donors, red cell volume was estimated by the 125I-albumin technique, and the posttransfusion recovery of stored red cells was quantitated by 51Cr- and 52Cr-labeled stored cells simultaneously. In this group, posttransfusion recoveries with 125I/51Cr averaged 73 +/- 7 percent; with 125I/52Cr, they averaged 75 +/- 10 percent. Using the single-label method of calculation, recoveries averaged 76 +/- 7 and 75 +/- 10 percent for the 51Cr and 52Cr methods, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
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