Intraoperative cell salvaging: ex vivo evaluation of two swab-washing methods.

OBJECTIVE: To compare erythrocyte recovery by a cell salvage device between swab-washing by manual agitation or filtration. SAMPLE: 12 recently expired units of canine packed RBCs. PROCEDURE: The packed RBC units underwent quality analysis before donation from a pet blood bank. Each unit was volume-expanded with anticoagulant and subsequently divided into 2 equal aliquots used to soak surgical swabs before washing. Two different swab-washing techniques were evaluated-standard swab-washing-manual agitation (SW-MA) and swab-washing-filtration (SW-F)-with a novel prototype device. The resulting bloody fluid was processed using the Cell Saver Elite Autotransfusion System (Haemonetics). The volume, manual PCV, CBC, and RBC mass, calculated as the product of the volume and PCV, were measured before and after salvaging. Last, the RBC mass recovery was recorded as a percentage. RESULTS: The RBC mass recovered from SW-MA and SW-F averaged 85.73% and 83.99%, respectively. There was no significant difference in RBC recovery between the 2 methods (P = .52). CLINICAL RELEVANCE: SW-MA and SW-F recovered a similar quantity of RBCs from blood-soaked swabs in an ex vivo setting.

Effect of ascorbic acid on storage of Greyhound erythrocytes.

OBJECTIVE: To assess changes in biochemical and biophysical properties of canine RBCs during cold (1° to 6°C) storage in a licensed RBC additive solution (the RBC preservation solution designated AS-1) supplemented with ascorbic acid. SAMPLE: Blood samples from 7 neutered male Greyhounds; all dogs had negative results when tested for dog erythrocyte antigen 1.1. PROCEDURES: Blood was collected into citrate-phosphate-dextrose and stored in AS-1. Stored RBCs were supplemented with 7.1mM ascorbic acid or with saline (0.9% NaCl) solution (control samples). Several biochemical and biophysical properties of RBCs were measured, including percentage hemolysis, oxygen-hemoglobin equilibrium, and the kinetic rate constants for O2 dissociation, carbon monoxide association, and nitric oxide dioxygenation. RESULTS: Greyhound RBCs stored in AS-1 supplemented with ascorbic acid did not have significantly decreased hemolysis, compared with results for the control samples, during the storage period. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, ascorbic acid did not reduce hemolysis during storage. Several changes in stored canine RBCs were identified as part of the hypothermic storage lesion.

The 24-hour posttransfusion survival of baboon red blood cells preserved in citrate phosphate dextrose/ ADSOL (CPD/AS-1) for 49 days.

The purpose of this study was to evaluate the baboon as an animal model for evaluating red blood cell (RBC) preservation by comparing the 24-h posttransfusion survival of baboon RBCs preserved in citrate phosphate dextrose/ADSOL (CPD/AS-1) solution at 4 degrees C for 49 days to that of human RBCs preserved under similar conditions. CPD/AS-1 originally was approved by the Food and Drug Administration for 49-day storage of RBCs, but this period subsequently was reduced to 42 days. Adult male baboons (Papio anubis and P. cynocephalus) were autotransfused with RBCs that had been harvested using CPD and that had been resuspended and stored in AS-1 solution at 4 degrees C for as long as 49 days. The 24-h posttransfusion survival was measured using the 51Cr/125I-albumin method. The 24-h posttransfusion survival (mean +/- standard deviation) was 74% +/- 7% for seven units of CPD/AS-1-treated RBCs stored for 35 days, 65% +/- 15% for 12 units stored for 42 days, and 43% +/- 16% for seven units stored for 49 days. The mean 24-h posttransfusion survival rate for autologous baboon RBCs stored in CPD/AS-1 at 4 degrees C for 35 days (74%) was similar to that for autologous human RBCs stored in a similar manner. Further storage for 42 and 49 days resulted in lower values for baboon RBCs compared with human RBCs.

Comparison of 4 blood storage methods in a protocol for equine pre-operative autologous donation.

OBJECTIVE: To compare viability of equine whole blood stored by 4 different methods, and to establish optimal storage protocols for an equine autologous blood donation program. STUDY DESIGN: In vitro study of stored equine whole blood. Animals- Six healthy adult horses. METHODS: Blood from each horse was collected into 4 different containers: glass bottles containing acid-citrate-dextrose solution (ACD), plastic bags containing ACD, citrate-phosphate-dextrose (CPD), and CPD with supplemental adenine (CPDA-1). Blood was stored for 5 weeks and sampled at 2-day intervals. Standard hematologic and biochemical variables were evaluated, and adenosine-5-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) concentrations were measured and normalized to total hemoglobin content. RESULTS: Plasma hemoglobin, % hemolysis, lactate, potassium, ammonia, and lactate dehydrogenase (LDH) increased, whereas glucose concentration and pH decreased in all stored blood over 5 weeks. There was a temporal increase in hemolysis with all storage methods, but the increase was greatest in glass bottles. Lactate and ammonia were highest in CPD and CPDA-1 samples, indicating more active red blood cell (RBC) metabolism. 2,3-DPG concentrations decreased during storage, but were optimally preserved with CPDA-1. ATP concentrations were significantly higher for blood stored in CPDA-1, and were lowest in glass bottles. CONCLUSIONS: Hematologic and biochemical values measured for blood stored in CPDA-1 are suggestive of improved RBC viability compared with other storage methods. With the exception of ATP, results from stored equine blood were similar to those reported for other species. CLINICAL RELEVANCE: Commercial CPDA-1 bags appear to be the optimal storage method for equine whole blood.

Evaluation of an additive solution for preservation of canine red blood cells.

The effect of an additive preservative solution on canine red blood cell posttransfusion viability (PTV) and on selected canine red blood cell biochemical parameters was studied. One unit (450 mL) of blood was collected from 6 clinically normal dogs into the anticoagulant citrate phosphate dextrose, centrifuged, and the plasma removed. The red blood cells were then suspended in 100 mL of a saline, adenine, dextrose, and mannitol solution and stored at 4 degrees C. Aliquots were removed for study at 1, 10, 20, 30, 37, and 44 days. The 24-hour PTV of autologous red blood cells was determined using a sodium chromate (51Cr) label. Red blood cell concentrations of 2,3-diphosphoglycerate (2,3-DPG), adenosine-5'-triphosphate (ATP), and pH were also determined. Canine red blood cell PTV, pH, ATP, and 2,3-DPG concentrations decreased during storage (P < .05). The PTV decreased from 94% using day 1 red blood cells to 80% and 75% using day 37 and day 44 red blood cells, respectively (P < .05). Although the mean PTV of the day 44 stored units equaled the Food and Drug Administration (FDA) minimum standard for human red blood cells, the PTV was substandard in 75% of the day 44 units. The FDA standard was exceeded in 83% of the day 37 units. It was concluded that 37-day-old canine red blood cells preserved with a saline, adenine, dextrose, and mannitol solution are of acceptable quality for transfusion.

Relationship of serum and plasma copper and ceruloplasmin concentrations of cattle and the effects of whole blood sample storage.

The present study was conducted to determine whether plasma and serum copper and ceruloplasmin concentrations in cattle are different and whether transport of samples with storage on ice before centrifugation affects the measurements. Mean copper and ceruloplasmin values were higher in plasma than in serum. Linear regressions were plasma copper (microgram/ml) = 1.200 serum copper -0.032 (r2 = 0.99), serum ceruloplasmin (mg/dl) = 14.0 serum copper + 2.34 (r2 = 0.48), and plasma ceruloplasmin (mg/dl) = 18.2 plasma copper + 2.1 (r2 = 0.43). The percentage of copper associated with ceruloplasmin was less in serum (55%) than in plasma (66%). Storage of blood samples on ice for 3 days decreased serum copper value by 3.5%. Linear regressions to correct for storage effects were corrected serum copper = 1.11 stored serum copper -0.04 (r2 = 0.94) and corrected plasma copper = 1.22 stored plasma copper -0.17 (r2 = 0.86). A cuproprotein may be involved in the blood clotting process, and some ceruloplasmin and its copper are apparently trapped in the fibrin clot, causing less copper in serum, compared with that in plasma. The difference between plasma and serum copper concentrations of calves was slightly increased by dietary copper supplementation.

Comparison of selenium levels and glutathione peroxidase activity in bovine whole blood.

Blood glutathione peroxidase activity and selenium levels were found to correlate well, indicating that glutathione peroxidase activity can be used to assess blood selenium levels in beef cattle. The glutathione peroxidase activity of blood is less stable than is the selenium concentration but when blood was stored at 4 degrees C, the glutathione peroxidase activity remained constant for seven days.

Freeze-preserved baboon red blood cells: effects of biochemical modification and perfusion in vitro.

Nonrejuvenated and rejuvenated baboon RBC were freeze-preserved with 40% (w/v) glycerol at -80 C. To prepare rejuvenated RBC, a 50-ml solution containing pyruvate, inosine, glucose, phosphate, and adenine was used and RBC were incubated with this solution before glycerolization and freezing. Appropriate steps were taken to minimize osmotic damage to the RBC during glycerolization and deglycerolization. Nonrejuvenated and rejuvenated cryopreserved RBC had freeze-thaw recovery values of 98%, freeze-thaw-wash recovery values of 92%, and 24-hour post-transfusion survival values of 85%. Some units of cryopreserved RBC were autotransfused after thawing, washing, and storage at 4 C for 24 hours. Other units were perfused in vitro before autotransfusion. After 24 hours of postwash storage, the RBC were concentrated by centrifugation and suspended in a plasma protein fraction and/or an electrolyte solution, and then were exposed to extracorporeal perfusion. Serious adverse effects were not observed on posttransfusion survival, function, or hemolysis in nonrejuvenated or rejuvenated baboon RBC as a result of perfusion in vitro.

A new storage medium for canine blood.

A solution consisting of ascorbate phosphate, citric acid, sodium citrate, sodium phosphate, and dextrose was developed to extend the shelf life of canine blood stored for transfusion. The 24-hour poststorage viability remained above 70% for 6 weeks of storage at 4 C. The concentration of 2,3 diphosphoglycerate remained constant for 3 weeks, then declined slowly. After 6 weeks of storage, the 2,3 diphosphoglycerate content was still sufficiently high to allow adequate dissociation of oxygen from oxyhemoglobin in vivo. It was concluded that blood stored up to 6 weeks in this solution would be safe to use for transfusion.