G Cassidy's research while affiliated with University of Massachusetts Boston and other places

BACKGROUND: Preoperative bleeding time (BT) does not correlate with postoperative bleeding in patients subjected to surgical procedures. A significant positive correlation has been reported between the BT 2 hours after cardiopulmonary bypass surgery and the nonsurgical blood loss during the first 4 hours after bypass surgery. This study was done to investigate the effect of Hct and platelet count on the BT measurement in normal, healthy men and women.STUDY DESIGN AND METHODS: To assess the relative effect of RBCs and platelets on the BT, 22 healthy male and 7 healthy female volunteers were subjected to the removal of 2 units of RBCs (360 mL), followed by the return of the platelet-rich plasma (PRP) from both units and the infusion of 1000 mL of 0.9-percent NaCl. Four of the men and all seven women received their RBCs 1 hour after their removal. Shed blood levels of thromboxane B2 (TXB2), 6-keto prostaglandin F1, and peripheral venous Hct were measured. BTs were measured in 15 men and 13 women before and after a plateletpheresis procedure to collect 3.6 × 1011 platelets per unit.RESULTS: The 2-unit RBC apheresis procedure produced a 60-percent increase in the BT associated with a 15-percent reduction in the peripheral venous Hct and a 9-percent reduction in the platelet count. The plateletpheresis procedure produced a 32-percent decrease in the platelet count, no change in peripheral venous Hct, and no change in the BT. After the removal of 2 units of RBCs, the shed blood TXB2 level decreased significantly. Reinfusion of 2 units of RBCs restored the BT and restored the TXB2 level to the baseline levels.CONCLUSION: The acute reduction in Hct produced a reversible platelet dysfunction manifested by an increase in BT and a decrease in the shed blood TXB2 level at the template BT site. Return of the RBCs restored both the BT and the shed blood TXB2 level to normal. The platelet dysfunction observed with the reduction in Hct was due in part to a reduction in shed blood TXB2 and other, unknown mechanisms.

The RBC injury that occurs during collection of the first few milliliters of blood into the pH 5.0 ACD (NIH, Formula A) is referred to as the lesion of collection. The RBC injury was evaluated by labeling the ACD RBCs with (51)Cr and measuring the 24-hour posttransfusion survival. The effect of the acidification of ACD blood on the in vivo elution of (51)Cr from the RBC has not been reported. Baboon blood was collected in heparin and in ACD and CP2D at different ratios of blood to anticoagulant. ACD blood with a pH of 5.7 to 6.9 was labeled with (51)Cr. Heparinized blood with a pH of 7.4 was labeled with biotin-X-N-hydroxysuccinimide (NHS). ACD blood with a pH of 5.9 was labeled with both (51)Cr and biotin-X-NHS. The RBC volumes, 24- and 48-hour posttransfusion survivals, and lifespans (T50) were measured. The RBC volume of ACD blood with a pH ranging from 5.7 to 6.9 was not affected by (51)Cr labeling. (51)Cr-labeled ACD blood with a pH of 6.9 had an RBC volume that was significantly greater than that seen in heparinized blood with a pH of 7.4 labeled with biotin-X-NHS. In vivo elution of (51)Cr from the RBCs prepared from the ACD blood with a pH of 5.7 to 6.9 and in vivo elution of biotin-X-NHS from RBCs prepared from ACD blood with a pH of 5.9 were associated with reductions in 24- and 48-hour posttransfusion survivals and T50. The anticoagulant and the pH of the medium in which the RBCs were labeled with (51)Cr or biotin-X-NHS affected in vivo elution of the label from the RBCs and may reduce posttransfusion survival values.

Preoperative bleeding time (BT) does not correlate with postoperative bleeding in patients subjected to surgical procedures. A significant positive correlation has been reported between the BT 2 hours after cardiopulmonary bypass surgery and the nonsurgical blood loss during the first 4 hours after bypass surgery. This study was done to investigate the effect of Hct and platelet count on the BT measurement in normal, healthy men and women. To assess the relative effect of RBCs and platelets on the BT, 22 healthy male and 7 healthy female volunteers were subjected to the removal of 2 units of RBCs (360 mL), followed by the return of the platelet-rich plasma (PRP) from both units and the infusion of 1000 mL of 0.9-percent NaCl. Four of the men and all seven women received their RBCs 1 hour after their removal. Shed blood levels of thromboxane B(2) (TXB(2)), 6-keto prostaglandin F(1 alpha), and peripheral venous Hct were measured. BTs were measured in 15 men and 13 women before and after a plateletpheresis procedure to collect 3.6 x 10(11) platelets per unit. The 2-unit RBC apheresis procedure produced a 60-percent increase in the BT associated with a 15-percent reduction in the peripheral venous Hct and a 9-percent reduction in the platelet count. The plateletpheresis procedure produced a 32-percent decrease in the platelet count, no change in peripheral venous Hct, and no change in the BT. After the removal of 2 units of RBCs, the shed blood TXB(2) level decreased significantly. Reinfusion of 2 units of RBCs restored the BT and restored the TXB(2) level to the baseline levels. The acute reduction in Hct produced a reversible platelet dysfunction manifested by an increase in BT and a decrease in the shed blood TXB(2) level at the template BT site. Return of the RBCs restored both the BT and the shed blood TXB(2) level to normal. The platelet dysfunction observed with the reduction in Hct was due in part to a reduction in shed blood TXB(2) and other, unknown mechanisms.

Transfusion-associated GVHD results from the presence of viable lymphocytes in transfused allogeneic blood components. Viable immunocompetent lymphocytes have been detected in RBCs that were frozen with glycerol and washed before transfusion. The study reported here assessed the effect of irradiation on human RBCs frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. In vitro and in vivo testing was done on human RBCs that were frozen with 40-percent (wt/vol) glycerol at -80 degrees C, with some units exposed to 2500 cGy of gamma radiation and others not irradiated, and that, after thawing and washing, were stored in a sodium chloride-glucose solution at 4 degrees C for 3 days before autologous transfusion. The glycerol-frozen RBCs treated with 2500 cGy before deglycerolization had a mean freeze-thaw-wash recovery of 87 percent and a mean 24-hour posttransfusion survival of 86 percent after storage for 3 days at 4 degrees C in a 0.9-percent NaCl and 0.2-percent glucose solution. For the nonirradiated units, the mean freeze-thaw-wash recovery was 85 percent and the mean 24-hour posttransfusion survival was 83 percent. These data show similar, acceptable results for RBCs frozen with 40-percent (wt/vol) glycerol at -80 degrees C and treated in the frozen state with 2500 cGy of gamma radiation and for RBCs that were not irradiated, all of which were washed and then stored in a sodium chloride-glucose solution for 3 days before autologous transfusion.

BACKGROUND : Transfusion‐associated GVHD results from the presence of viable lymphocytes in transfused allogeneic blood components. Viable immunocompetent lymphocytes have been detected in RBCs that were frozen with glycerol and washed before transfusion. STUDY DESIGN AND METHODS : The study reported here assessed the effect of irradiation on human RBCs frozen with 40‐percent (wt/vol) glycerol and stored at –80°C. In vitro and in vivo testing was done on human RBCs that were frozen with 40‐percent (wt/vol) glycerol at –80°C, with some units exposed to 2500 cGy of gamma radiation and others not irradiated, and that, after thawing and washing, were stored in a sodium chloride‐glucose solution at 4°C for 3 days before autologous transfusion. RESULTS : The glycerol‐frozen RBCs treated with 2500 cGy before deglycerolization had a mean freeze‐thaw‐wash recovery of 87 percent and a mean 24‐hour posttransfusion survival of 86 percent after storage for 3 days at 4°C in a 0.9‐percent NaCl and 0.2‐percent glucose solution. For the nonirradiated units, the mean freeze‐thaw‐wash recovery was 85 percent and the mean 24‐hour posttransfusion survival was 83 percent. CONCLUSION : These data show similar, acceptable results for RBCs frozen with 40‐percent (wt/vol) glycerol at –80°C and treated in the frozen state with 2500 cGy of gamma radiation and for RBCs that were not irradiated, all of which were washed and then stored in a sodium chloride‐glucose solution for 3 days before autologous transfusion.

This study was designed to assess the effects of changes in storage temperature of frozen RBCs such as might occur during a malfunction of the -80 degrees C mechanical freezer or during shipment. Fifteen participants donated blood for autologous transfusion of RBCs; all RBCs were frozen with 40-percent (wt/vol) glycerol. Five subjects received RBCs that were stored at -80 degrees C alone before transfusion. Five subjects received RBCs that were stored initially at -80 degrees C, then at -40 degrees C for 4 weeks, and finally at -80 degrees C before transfusion. Five subjects received RBCs that were stored at -80 degrees C, then at -20 degrees C for 2 weeks, and finally at -80 degrees C before transfusion. After deglycerolization, the RBCs were stored at 4 degrees C in a sodium chloride-glucose solution for 3 days before transfusion. No significant differences were observed in freeze-thaw recovery, freeze-thaw-wash recovery, 24-hour posttransfusion survival, index of therapeutic effectiveness, or RBC ATP levels. Greater hemolysis and reduced RBC K+ levels were observed in the units stored at -80 degrees C/-40 degrees C/-80 degrees C and in those stored at -80 degrees C/ -20 degrees C/-80 degrees C compared with the units stored at -80 degrees C alone, but these differences did not affect the 24-hour posttransfusion survival. The results of this study indicated that RBCs frozen with 40-percent (wt/vol) glycerol can be stored at -40 degrees C for 4 weeks or at -20 degrees C for 2 weeks between periods of frozen storage at -80 degrees C with satisfactory results.

Red blood cells (RBC) were collected either by a manual method using a 16-gauge needle or by an apheresis procedure using an 18-gauge needle, and were stored at 4 degrees C in a solution of CP2D (anticoagulant)/AS-3 (Nutricel) for 56 days. The purpose was to compare the outcome of the autotransfused red cells collected by both techniques. Five healthy male volunteers were studied on two occasions. The autotransfusions of the manual and apheresed RBC resulted in a mean 24-h post-transfusion survival of 71%, a normal mean 51Cr RBC life span, a 2,3 DPG level that was less than 10% of normal, and 0.6% haemolysis. Whether collected manually or by apheresis, the outcomes were similar for RBC stored at 4 degrees C for 56 days in CP2D/AS-3.

Red blood cells (RBC) were collected either by a manual method using a 16-gauge needle or by an apheresis procedure using an 18-gauge needle, and were stored at 4 °C in a solution of CP2D (anticoagulant)/AS-3 (Nutricel) for 56 days. The purpose was to compare the outcome of the autotransfused red cells collected by both techniques. Five healthy male volunteers were studied on two occasions. The autotransfusions of the manual and apheresed RBC resulted in a mean 24-h post-transfusion survival of 71%, a normal mean 51Cr RBC life span, a 2,3 DPG level that was less than 10% of normal, and 0·6% haemolysis. Whether collected manually or by apheresis, the outcomes were similar for RBC stored at 4 °C for 56 days in CP2D/AS-3.

Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4 degrees C to only 24 hours. Healthy male volunteers who met AABB requirements for blood donors (n = 38) were studied. A volume of 450 mL of blood was collected into CPDA-1. The RBC concentrates were stored at 4 degrees C for 3 to 6 days before being frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. The RBCs were deglycerolized, resuspended in 0.9-percent sodium chloride and 0.2-percent glucose (SG) solution or SG solution supplemented with AS-1, AS-3, or AS-5, and stored in the resuspension medium at 4 degrees C for 14 days. The mean +/- SD freeze-thaw-wash process recovery was 90.0 +/- 4.0 percent for all 38 units. The mean 24-hour posttransfusion survival value was 79 percent for deglycerolized RBC stored at 4 degrees C for 7 days in SG alone, SG plus AS-3, or SG plus AS-5. Deglycerolized RBC that were stored at 4 C for 14 days in SG supplemented with AS-1, AS-3, or AS-5 had a mean 24-hour posttransfusion survival of 74 percent. After 7 days of storage of deglycerolized RBCs in SG alone, the mean hemolysis was 3. 7 percent. After 14 days of storage of deglycerolized RBCs in SG supplemented with AS-1, AS-3, or AS-5, the mean hemolysis was 2.5 percent. The levels of hemolysis did not correlate with the 24-hour posttransfusion survival values.

BACKGROUND: A study was done to assess the quality of RBCs stored at 4°C in AS-1, AS-3, or AS-5 for 42 days before biochemical modification and freezing. STUDY DESIGN AND METHODS: RBCs were stored at 4°C for 42 days in AS-1, AS-3, or AS-5 and then biochemically modified with pyruvate, inosine, phosphate, and adenine solution (Rejuvesol), frozen with 40-percent (wt/vol) glycerol, and stored at –80°C for at least 2 months. The RBCs were deglycerolized by the use of a cell washer (Haemonetics 115), and stored for 24 hours at 4°C in a 0.9-percent sodium chloride and 0.2-percent glucose solution before the autologous transfusion. RESULTS: The mean freeze-thaw-wash recovery process produced RBC recovery values of 85 percent, with the mean 24-hour posttransfusion survival at 75 percent, and the mean index of therapeutic effectiveness at 64 percent for the RBCs stored at 4°C in AS-1, AS-3, or AS-5 for 42 days before biochemical modification and freezing. All the units exhibited normal or slightly higher than normal 2,3 DPG levels after deglycerolization and postwash storage at 4°C for 24 hours. CONCLUSION: RBCs stored in AS-1, AS-3, or AS-5 at 4°C for 42 days and then biochemically modified with pyruvate, inosine, phosphate, and adenine and glycerolized, frozen, washed, and stored at 4°C for 24 hours before autologous transfusion had acceptable in vitro and in vivo measurements.