Red blood cell concentrates treated with the amustaline (S-303) pathogen reduction system and stored for 35 days retain post-transfusion viability: results of a two-centre study.

BACKGROUND AND OBJECTIVES: Pathogen reduction technology using amustaline (S-303) was developed to reduce the risk of transfusion-transmitted infection and adverse effects of residual leucocytes. In this study, the viability of red blood cells (RBCs) prepared with a second-generation process and stored for 35 days was evaluated in two different blood centres. MATERIALS AND METHODS: In a single-blind, randomized, controlled, two-period crossover study (n = 42 healthy subjects), amustaline-treated (Test) or Control RBCs were prepared in random sequence and stored for 35 days. On day 35, an aliquot of 51 Cr/99m Tc radiolabeled RBCs was transfused. In a subgroup of 26 evaluable subjects, 24-h RBC post-transfusion recovery, mean life span, median life span (T50 ) and life span area under the curve (AUC) were analysed. RESULTS: The mean 24-h post-transfusion recovery of Test and Control RBCs was comparable (83·2 ± 5·2 and 84·9 ± 5·9%, respectively; P = 0·06) and consistent with the US Food and Drug Administration (FDA) criteria for acceptable RBC viability. There were differences in the T50 between Test and Control RBCs (33·5 and 39·7 days, respectively; P < 0·001), however, these were within published reference ranges of 28-35 days. The AUC (per cent surviving × days) for Test and Control RBCs was similar (22·6 and 23·1 per cent surviving cells × days, respectively; P > 0·05). Following infusion of Test RBCs, there were no clinically relevant abnormal laboratory values or adverse events. CONCLUSION: RBCs prepared using amustaline pathogen reduction meet the FDA criteria for post-transfusion recovery and are metabolically and physiologically appropriate for transfusion following 35 days of storage.

Multi-institutional randomized control study of haemolysis in stored red cell units prepared manually or by an automated system.

BACKGROUND: The haemolysis level at the end of storage is a performance parameter for RBC preparations. In the evaluation of new devices or new processes for processing blood, it is relevant to evaluate whether the haemolysis is linked to (1) specific characteristics of the blood donor, or (2) the nature of the blood-processing methodologies. MATERIALS AND METHODS: As part of the validation of a new automated whole blood processing system compared to the current manual methods, randomized, paired crossover studies were conducted evaluating measures of blood component quality, including RBC haemolysis over 42 days of storage. RESULTS: The association between haemolysis and the individual subject was evaluated by modelling haemolysis with independent predictors of treatment (control and test processing) and leucocyte reduction as fixed factors with donor and laboratory as random effects in a mixed-effects ANOVA model. It was found that the day 42 haemolysis values were strongly dependent on the donor subject, with an intraclass correlation coefficient of 0.81. CONCLUSIONS: The data reported in this study suggest a link between the specific whole blood donor and the haemolysis levels observed in red-blood-cell units stored refrigerated for 42 days. Additional research to identify possible donor characteristics associated with haemolysis during storage is warranted.

Buffering and dilution in red blood cell storage.

BACKGROUND: Red blood cell (RBC) storage solutions work in a narrow pH range between 7.2 and 6.4. While keeping RBC within that pH range, ATP production can be increased by buffering or dilution. STUDY DESIGN AND METHODS: In the first study, 12 units of packed CP2D RBCs were pooled in groups of four, re-aliquoted, and added to one of four additive solutions (ASs): AS-3, 110 mL; EAS-61, 170 mL; EAS-78, 170 mL; or EAS-81, 110 mL. EAS-78 and -81 contain bicarbonate. Units were sampled approximately weekly for 10 weeks for biochemical measures. In the second study, 12 volunteers donated RBCs for measures of (51)Cr in vivo recovery after 6 or 8 weeks of storage in EAS-81. RESULTS: RBCs stored in the higher-volume or buffered ASs had higher RBC ATP concentrations. The combination had an additive effect. Hemolysis was reduced in dilute ASs and less so with buffering. RBCs stored for 8 weeks (n=6) in EAS-81 exhibited 87+/- 2 percent 24-hour (51)Cr in vivo recovery and 0.4+/- 0.2 percent hemolysis. CONCLUSIONS: It is possible to store RBCs for 8 weeks in buffered conventional volume ASs. Combining buffering and increased AS volume improves stored RBC characteristics further.

Studies in red blood cell preservation 10. 51Cr recovery of red cells after liquid storage in a glycerol-containing additive solution.

The purpose of the present study was to compare the 24-hour recovery of red blood cells stored for 9 weeks in a hypoosmolar additive solution containing 150 mM glycerol to cells stored in Adsol. Seven units of packed red cells were split into 2 aliquots. To one sample, 100 ml of the experimental additive solution (EAS 25) was added, and to the other, 50 ml of Adsol. At the end of the storage period the cells were labeled with 51Cr. A double chromium technique was used to make it possible to perform comparative autologous studies in the same donor. The 24-hour 51Cr recovery value for EAS 25 was 73.0 +/- (SD) 4.2% and for Adsol 60.9 +/- 7.1. At 9 weeks the adenosine triphosphate levels were not significantly better compared to Adsol but the other in vitro measurements were better. New approaches to the study of red cell preservation are suggested.

Studies in red blood cell preservation: 9. The role of glutamine in red cell preservation.

Previous studies have demonstrated that an additive solution containing ammonium chloride (NH4+) and phosphate (Pi) in addition to adenine, glucose and mannitol would support red blood cell (RBC) in vitro characteristics and in vivo 24-hour viability after storage for 9 weeks. The purpose of the present study was to determine if NH4+ generated by the action of glutaminase on glutamine could be substituted for added NH4+ salts. Packed RBCs were stored with equal volumes of adenine, glucose, mannitol, and citrate containing additive solutions with 10 mM glutamine (EAS 31) or with 10 mM glutamine and either 10 (EAS 36) or 20 mM (EAS 37) Pi. One aliquot was stored with Adsol. The mean ATP levels of the RBCs stored in the glutamine plus phosphate EASs were 132 (10 mM Pi) and 144% (20 mM Pi) of the initial levels at 28 days, and at 84 days remained at 48 and 56%, respectively. The ATP levels of the RBC stored in Adsol were 105 and 25% at 28 and 84 days of storage, respectively. Percentage hemolysis and vesiculation was significantly lower (p < 0.01) for RBCs stored in glutamine and glutamine plus phosphate as compared to RBCs stored in Adsol. The levels of NH4+ were 22 to 34% higher in the EASs than in Adsol at the end of 84 days of storage, suggesting that glutamine is broken down by glutaminase to generate NH4+. The mean corpuscular volumes (MCVs) of RBCs in EASs 36 and 37 were substantially higher than in Adsol throughout the course of storage (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of hypotonicity, glutamine, and glycine on red cell preservation.

BACKGROUND: Red cells (RBCs) stored in hypo-osmolar additive solutions with the same concentrations of adenine, dextrose, mannitol, and sodium chloride and varied amounts of ammonium, phosphate, glycerol, and glutamine were better preserved than RBCs in the standard additive solution (Adsol). Cell swelling occurred in all the experimental additives. This observation prompted the evaluation of glutamine and glycine alone, as well as a combination of glutamine and glycine, all of which have been described as producing swelling of rat liver cells. STUDY DESIGN AND METHODS: Aliquots of RBCs were stored at 4 degrees C in Adsol or experimental additive solutions (EASs) all containing adenine, 2 mM; dextrose, 110 mM; mannitol, 55 mM; and sodium chloride, 50 mM. EAS 42 had, in addition, glutamine, 10 mM; glycine 5 mM, and phosphate, 20 mM. EAS 43 had glutamine, 10 mM; glycine, 10 mM; and phosphate 20 mM. EAS 44 had glutamine, 10 mM; EAS 45 had glutamine, 10 mM, and phosphate, 20 mM, and EAS 46 had only glycine, 10 mM. At intervals, measurements were made of mean corpuscular volume, mean corpuscular hemoglobin concentration, morphology, ATP, hemolysis, supernatant potassium, ammonia, pH, and microvesicles shed. RESULTS: The initial mean corpuscular volumes were larger in all EASs than in Adsol, but the greatest difference was between EASs 44 and 46 (108 fL) and Adsol (86 fL) (p < 0.001). The morphology scores were significantly better in all the EASs (p < 0.04). The ATPs were significantly greater in all the EASs (p < 0.001), and highest in those with phosphate. potassium leakage and hemolysis were less in the EASs (p < 0.001). The ammonia levels higher in all the EASs than in Adsol, with the exception of EAS 46. During storage, the extracorpuscular and intracorpuscular pH levels were essentially identical. The shedding of microvesicles was greatly reduced in all the EASs. CONCLUSION: Cell swelling induced in RBCs after collection appears to improve preservation. Ammonia and phosphate enhance RBC ATP maintenance. Glycine decrease the formation of ammonia by RBCs stored in a hypotonic medium.

A feasibility evaluation of an automated blood component collection system platelets and red cells.

BACKGROUND: The purpose of these studies was to evaluate the functional properties of blood components collected with an automated collection system. STUDY DESIGN AND METHODS: Single-donor platelets (n = 44) and packed red cell (RBC) units (n = 10) were collected. In vitro and in vivo assays were used to assess the function of single-donor platelet components stored for 5 days and of packed RBC units after storage for 42 days at 4 degrees C. RESULTS: Adverse events observed in the 44 study subjects were minor. The mean 24-hour recovery value for the packed RBC units stored for 42 days was 83.6 +/- 5.4 percent, with a mean percentage of hemolysis on Day 42 at 0.46 +/- 0.19 percent. The 25 patients receiving platelet components achieved a mean corrected count increment of 15.1 +/- 10.4 x 10(3). All platelet concentrates had less than 1 x 10(6) total white cells. CONCLUSION: Both in vitro and in vivo testing for the packed RBCs collected and stored for 42 days met the standards for both hemolysis and percentage of 51Cr 24-hour RBC recovery. The in vitro results and transfusion data on white cell-reduced platelet components transfused to thrombocytopenic patients were comparable to those on available platelet components.

A fully automated solid-phase radioimmunoassay for triiodothyronine.

We report the first solid-phase radioimmunoassay for triiodothyronine with use of antibody-coated tubes, which is designed specifically for the fully automated radioimmunoassay instrument, Micromedic Systems' "Concept 4 Automatic Radioassay." Antisera to triiodothyropropionic acid/bovine serum albumin were raised in rabbits, purified by ammonium sulfate precipitation, and coated onto polypropylene tubes. Analytical recovery of exogenous triiodothyronine added to sera from normal men and women and pregnant women was quantitative. Intra-assay and inter-assay coefficients of variation are 4-5 and 6-9%, respectively. Correlation coefficients (r) for comparison of sample values with those obtained by three commercial laboratories were 0.95, 0.84, and 0.91. The sensitivity of the assay is 0.5 microng/liter. The assay can be performed either manually or be fully automated on the "Concept 4."

Successful storage of RBCs for 9 weeks in a new additive solution.

BACKGROUND: This study explored the effect of storing packed RBCs suspended in 200 mL of an alkaline, hypotonic, experimental additive solution (EAS 61). STUDY DESIGN AND METHODS: Packed RBC units prepared from RBCs collected from healthy donors in CPD were stored for 8 (n = 10) and 9 (n = 10) weeks under blood bank conditions after the addition of 200 mL of EAS 61 (adenine, 2 mM:; dextrose, 110 mM:; mannitol, 55 mM:; NaCl, 26 mM:; Na(2)HPO(4), 12 mM:). Standard methods were used for in vitro assays. The 24-hour in vivo autologous recoveries were measured with (51)Cr. RESULTS: Mean +/- SD recoveries at 8 and 9 weeks were 81 +/- 7 and 77 +/- 7 percent. After 9 weeks, the ATP of the RBCs was 81 percent of the initial value, hemolysis was 0.35 percent, supernatant potassium was 46 mEq per L, and the morphologic index was 94.1. CONCLUSION: Packed RBCs suspended in 200 mL of EAS 61 can be stored satisfactorily for 9 weeks. Longer RBC storage should reduce outdating, increase availability of transfusions in remote locations, and improve the efficiency of autologous donor programs.

Successful storage of RBCs for 10 weeks in a new additive solution.

BACKGROUND: The effect of storing packed RBCs suspended in 300 mL of an alkaline, experimental additive solution (EAS 64) was explored. STUDY DESIGN AND METHODS: RBC units prepared from blood collected from healthy donors into CPD were WBC reduced and stored for 10 weeks under blood bank conditions after the addition of 300 mL of EAS 64 (adenine, 2 mM:; dextrose, 50 mM:; mannitol, 20 mM:; NaCl, 75 mM:; Na(2)HPO(4), 9 mM:). For comparison, non-WBC-reduced units from the same donors were stored in a different additive solution (AS-1, Baxter Healthcare) for 6 weeks. Standard methods were used for the in vitro assays. The 24-hour in vivo recoveries were measured by using (51)Cr- and (99m)Tc-labeled RBCs. RESULTS: Mean recovery in the EAS 64 units after 10 weeks was 84 +/- 8 percent, the same as in the AS-1 units stored for 6 weeks. For EAS 64 and AS-1 units, respectively, the ATP of the RBCs was 85 percent and 64 percent of the initial value, hemolysis was 0.43 percent and 0.63 percent, supernatant potassium was 24 mEq per L and 44 mEq per L, and the morphologic index was 98 and 71. CONCLUSION: RBCs suspended in 300 mL of EAS 64 can be stored satisfactorily for 10 weeks. Longer RBC storage should reduce outdating, increase availability of transfusions in remote locations, and improve the efficiency of autologous donor programs.

The role of electrolytes and pH in RBC ASs.

BACKGROUND: Experimental additive solutions (EASs) containing saline, adenine, glucose, mannitol and disodium phosphate can support RBCs for 9 or 10 weeks if used in 200- or 300-mL volumes. The effects of variations in the electrolyte composition and volume of EASs were explored. STUDY DESIGN AND METHODS: In three four-arm studies, 24 RBC units were pooled in groups of 4 and realiquoted as test units to ensure that all donors were equally represented in each study arm. In Study 1, units were stored for 11 weeks in EAS containing 0, 10, 20, or 30 mmol per L of sodium bicarbonate. In Study 2, units were stored for 9 weeks in EAS containing 26, 50, 100, or 150 mmol per L of sodium chloride. In Study 3, units were stored in 100 or 200 mL of AS-3 or EAS-61. RBC ATP concentrations and hemolysis were measured weekly. RESULTS: Increasing the sodium bicarbonate content of EASs increased the pH throughout storage and increased RBC ATP concentrations in the later phases of storage, but it had no effect on hemolysis. Increased sodium chloride content of EASs led to lower RBC ATP concentrations and increased hemolysis. In EAS-61, RBC ATP concentrations were increased throughout storage, and hemolysis was lower than that of RBCs stored in AS-3. CONCLUSION: RBC ATP synthesis is highly dependent on the pH of the AS. Hemolysis is affected by the salt content and volume of the AS.

The effects of phosphate, pH, and AS volume on RBCs stored in saline-adenine-glucose-mannitol solutions.

BACKGROUND: RBC ATP concentrations are the most important correlate of RBC viability. Tests were performed to determine whether increased AS volume, pH, and phosphate content increased stored RBC ATP concentrations. STUDY DESIGN AND METHODS: In three studies, packed RBCs were pooled in groups of 3 or 4 units and realiquoted as combined units to reduce intradonor differences. Pooled units were stored in the licensed ASs, AS-1 or AS-5, which contain saline, adenine, glucose, and mannitol (SAGM), or in experimental ASs (EASs) containing SAGM and disodium phosphate. Ten pools were stored in AS-1 at RBC concentrations equivalent to 100, 200, or 300 mL of AS. Six pools were stored in 100, 200, 300, or 400 mL volumes of EAS-61. Ten pools were stored in 100 mL of AS-5, 200 mL of EAS-61, or 300 mL of EAS-64. RBC ATP concentration and other measures of RBC metabolism and function were measured weekly. RESULTS: RBC ATP concentrations decreased sooner with storage in increasing volumes of AS-1. In EAS-61 and EAS-64, RBC ATP concentrations initially increased and stayed elevated longer with increasing AS volume. CONCLUSIONS: The addition of disodium phosphate to SAGM AS increases the RBC ATP concentrations. Reducing storage Hct appears to have a separate beneficial effect in reducing hemolysis.

RBC storage for 11 weeks.

BACKGROUND: Increasing the length of RBC storage can increase both RBC availability and quality. This work addresses 11-week RBC storage in experimental ASs (EASs). STUDY DESIGN AND METHODS: Three studies were performed. In the first, 24-hour in vivo recovery of (51)Cr-labeled autologous RBCs was measured in nine volunteers after storage of their RBCs for 11 weeks in EAS 67. In the second study, 4 units of blood were divided and stored in aliquots with an EAS containing 0, 15, 30, or 45 mmol per L of mannitol; then hemolysis, RBC morphology, and microvesicle protein were measured. In the third study, 6 full units were stored for 12 weeks in the EAS containing 30 mmol per L of mannitol, with weekly sampling for morphologic and biochemical measures of RBC quality. RESULTS: RBCs stored for 11 weeks in EAS-67 had a mean 24-hour in vivo recovery of 79 +/- 5 percent, but the hemolysis was 1.35 +/- 0.68 percent. Increasing mannitol content of the EAS reduced hemolysis but increased microvesiculation. EAS-76, with 30 mmol per L of mannitol allowed 11-week storage with 0.48 +/- 0.10 percent hemolysis at 11 weeks and 0.62 +/- 0.14 percent hemolysis at 12 weeks. CONCLUSION: It is possible to store RBCs for 11 weeks in EAS with greater than 75 percent recovery and less than 1 percent hemolysis.