George P. Cassidy

Anemia-induced increase in the bleeding time : implications for treatment of nonsurgical blood loss

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.

An Experiment with Glycerol-Frozen Red Blood Cells Stored at –80°C for up to 37 Years

Background and Objectives: Red cells frozen using 40% W/V glycerol are currently FDA approved for frozen storage at –80°C for up to 10 years. Materials and Methods: Red cells frozen with 40% W/V glycerol and stored at –80°C for up to 37 years were thawed, deglycerolized, and stored at 4°C for 24 h. Results: Red cells frozen for up to 37 years had mean freeze-thaw-wash recovery values of 75%, less than 1% hemolysis, and normal ATP, 2,3-DPG and P50 levels, and 60% of normal RBC K+ levels. Conclusions: Red cells frozen with 40% W/V glycerol can be stored at –80°C for up to 37 years with acceptable in vitro results.

The safety and therapeutic effectiveness of human red cells stored at — 80°C for as long as 21 years

Human red cells frozen by various methods have been stored in the frozen state at —80°C for as long as 21 years. This report discusses: red cells frozen with 42 percent weight per volume (wt/vol) glycerol in an ionic medium in a polyvinylchloride (PVC) plastic bag using the Cohn method; red cells frozen with 45 percent wt/vol glycerol in a low ionic medium in a PVC plastic bag using the Huggins method; red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a polyolefin plastic bag using the Meryman‐Hornblower method; and red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a standard 600‐ml or an elongated 800‐ml PVC plastic primary collection bag with an adapter port using the Naval Blood Research Laboratory (NBRL) method. After frozen storage for as long as 21 years by the four methods described above, the thawed red cells were deglycerolized with 50 to 150 ml of 12 percent sodium chloride and 1.5 to 2.0 l of sodium chloride‐glucose or sodium chloride‐glucose‐phosphate solution. After washing and storage at 4°C for 24 hours, the red cells had a mean freeze‐thaw‐wash recovery value of 90 percent, a mean 24‐hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis. When red cells frozen by the NBRL method in the standard 600‐ml or the elongated 800‐ml primary collection bag for as long as 5.7 years were stored after washing at 4°C for up to 3 days, these units had a mean freeze‐thaw‐wash recovery value of 90 percent, a mean 24‐hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis. Cultures done after storage at 4°C for 1 week showed that the red cells remained sterile. The incidence of container breakage for red cells frozen in the standard 600‐ml or elongated 800‐ml primary collection bag was about 3 percent for units subjected to shipment and less than 1 percent for units that were not transported.

Reduced coronary vasoconstrictor activity of hemoglobin solutions purified by ATP-agarose affinity chromatography.

Stroma-free hemoglobin (Hb) solutions are being developed as blood substitutes. We previously described coronary vasoconstrictor activity of Hb solutions prepared by conventional methods. In the present study we assessed the constrictor activity of unmodified and covalently modified Hb solutions purified by ATP-agarose affinity chromatography. The starting material was a red cell lysate, partially purified by ultrafiltration. Coronary constrictor activity was measured as increased perfusion pressure in isolated rabbit hearts perfused at constant coronary flow rate with buffer containing various concentrations of added Hb. The starting material increased perfusion pressure by 35 +/- 7 mmHg at 50 mg/dl. Purified Hb, retained by the affinity column, increased perfusion pressure by only 18 +/- 2 mmHg at 50 mg/dl. Hemoglobin covalently linked to ATP or pyridoxal phosphate, then purified by affinity chromatography, also had less constrictor activity than the starting material. Thus, a substance, removed by affinity chromatography but not by conventional purification, contributes to the vasoconstrictor activity of Hb solutions.

Volume of RBCs, 24‐ and 48‐hour posttransfusion survivals, and the lifespan of 51Cr and biotin‐X‐N‐hydroxysuccinimide (NHS)‐labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on 51Cr and biotin‐X‐NHS elution in vivo

BACKGROUND: 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 51Cr and measuring the 24‐hour posttransfusion survival. The effect of the acidification of ACD blood on the in vivo elution of 51Cr from the RBC has not been reported.

In vitro and in vivo measurements of human RBCs frozen with glycerol and subjected to various storage temperatures before deglycerolization and storage at 4°C for 3 days

BACKGROUND: 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°C mechanical freezer or during shipment.

Posttransfusion survival (24‐hour) and hemolysis of previously frozen, deglycerolized RBCs after storage at 4°C for up to 14 days in sodium chloride alone or sodium chloride supplemented with additive solutions

BACKGROUND: Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4°C to only 24 hours.

In vitro and in vivo measurementsof gamma-radiated, frozen, glycerolized RBCs

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.

24‐hour 51Cr post‐transfusion survival, 51Cr life span and haemolysis of red blood cells stored at 4 °C for 56 days in 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.