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Biomedical subjects

Gina Ragno

Publications and source records attributed to Gina Ragno.

27 records · Page 2Linked to original sources

The in vitro quality of red blood cells frozen with 40 percent (wt/vol) glycerol at -80 degrees C for 14 years, deglycerolized with the Haemonetics ACP 215, and stored at 4 degrees C in additive solution-1 or additive solution-3 for up to 3 weeks.

BACKGROUND: Red blood cells (RBCs) frozen with 40 percent (wt/vol) glycerol, stored at -80 degrees C (mean temperature; range, -65 to -90 degrees C) for 14 years, deglycerolized in the Haemonetics automated cell processor (ACP) 215 with the 325-mL disposable bowl, and stored at 4 degrees C in additive solution (AS)-1 or AS-3 for 21 days were evaluated. STUDY DESIGN AND METHODS: A total of 106 units of citrate phosphate dextrose adenine-1 RBCs were frozen with 40 percent (wt/vol) glycerol in the original 800-mL polyvinylchloride plastic bag and stored in corrugated cardboard boxes at -80 degrees C for 14 years. The thawed units were deglycerolized with the ACP 215 with a 325-mL disposable bowl and stored in AS-1 or AS-3 at 4 degrees C for 21 days. RESULTS: The freeze-thaw recovery value was 94 +/- 4 percent (mean +/- SD), the freeze-thaw-wash recovery value was 80 +/- 7 percent, and there was no breakage. Thirty-eight units were processed as 19 pairs. Two units of ABO-matched units were thawed, pooled, divided equally into two units, and deglycerolized. One unit was stored in AS-1 and the other in AS-3 at 4 degrees C for 21 days. Units stored in AS-1 exhibited significantly greater hemolysis than those stored in AS-3. CONCLUSIONS: Acceptable results were achieved when RBCs frozen at -80 degrees C for 14 years were deglycerolized in the ACP 215. Deglycerolized RBCs in AS-1 exhibited significantly higher hemolysis than those in AS-3 after storage at 4 degrees C for 7 to 21 days.

Blood Preservation↗

Incidence of breakage of human RBCs frozen with 40-percent wt/vol glycerol using two different methods for storage at -80 degrees C.

BACKGROUND: We reported previously that the incidence of breakage was 34.2 percent when human RBCs were frozen with 40-percent wt/vol glycerol in polyolefin plastic bags stored in aluminum containers at -80 degrees C and subjected to transportation. When human RBCs were frozen with 40-percent wt/vol glycerol at -80 degrees C in PVC plastic bags placed in polyester plastic bags and stored in rigid corrugated cardboard boxes, transportation resulted in a 2.4-percent incidence of breakage. The present study was done to confirm this incidence of breakage. STUDY DESIGN AND METHODS: The Meryman- Hornblower freezing method was compared to the Naval Blood Research Laboratory (NBRL) method of freezing for incidence of bag breakage. Human RBCs frozen by the Meryman-Hornblower method with 40-percent wt/vol glycerol with supernatant glycerol and stored in polyolefin plastic bags in aluminum containers at -80 degrees C were stored at the NBRL from 1974 to 2002. With the NBRL method, human RBCs frozen at -80 degrees C without supernatant glycerol in the 800-mL PVC plastic primary bag inside a polyester plastic bag in a rigid corrugated cardboard box were stored at the NBRL from 1984 to 2002. RESULTS: The incidence of breakage for 532 units of RBCs that had been frozen by the Meryman- Hornblower method and stored in aluminum containers was 47.3 percent for nontransported units. RBCs that had been frozen by the NBRL method and stored in rigid corrugated cardboard boxes exhibited breakage of 2.4 percent for 2424 nontransported units and 6.7 percent for 633 transported units. DISCUSSION: The incidence of breakage was significantly lower for RBCs frozen by the NBRL method than for the RBCs frozen by the Meryman-Hornblower method.

Blood Preservation↗

Effect of WBC reduction and storage temperature on PLTs frozen with 6 percent DMSO for as long as 3 years.

BACKGROUND: PLTs frozen with 6 percent DMSO can be stored at -80 degrees C for 2 years, while those frozen with 5 percent DMSO at -150 degrees C can be stored for at least 3 years. The more rapid deterioration seen in PLTs frozen at -80 degrees C may be due to the presence of granulocytes. The effects of storage temperature and WBC reduction on PLTs frozen with DMSO and the breakage of the PVC plastic bags stored at -80 and -135 degrees C were assessed. STUDY DESIGN AND METHODS: Apheresed PLT-rich plasma (PRP) was either divided into two equal volumes where one volume was WBC-reduced and the other volume was not or filtered or not and then divided into two equal volumes. PLTs frozen with 6 percent DMSO were stored in PVC plastic bags at either -80 or -135 degrees C for as long as 3 years. RESULTS: After 2 years of storage at -80 degrees C, the PLTs exhibited satisfactory freeze-thaw-wash values regardless of whether or not they were WBC-reduced, but after 2 to 3 years of storage at -80 degrees C, the PLTs had significantly reduced freeze-thaw-wash values. Freeze-thaw-wash values were not reduced in PLTs stored at -135 degrees C for up to 3 years. CONCLUSIONS: WBC reduction did not improve freeze-thaw-wash recovery values in PLTs stored at -80 or -135 degrees C for up to 3 years, but reducing the storage temperature from -80 to -135 degrees C did. Breakage of PVC plastic bags stored at -135 degrees C was excessive.

Blood Platelets↗

Comparison of radioisotope methods and a nonradioisotope method to measure the RBC volume and RBC survival in the baboon.

BACKGROUND: RBC volume, 24-hour posttransfusion survival, and life span can be measured with radio-isotopes and nonradioactive procedures. STUDY DESIGN AND METHODS: RBC volume was measured directly with autologous baboon RBCs labeled with biotin-X-N-hydroxysuccinimide (NHS), 51Cr, 99mTc, and 111In-oxine and indirectly from the 125I plasma volume and the total body Hct. Twenty-four-hour posttransfusion survival and life span were measured in autologous fresh baboon RBCs labeled with 51Cr, 111In-oxine, 99mTc, and biotin-X-NHS. RESULTS: Significantly larger RBC volumes were observed when the fresh autologous RBCs were labeled with 51Cr, 111In-oxine, or 99mTc than when they were labeled with the nonradioactive biotin-X-NHS. Twenty-four-hour posttransfusion survival values were significantly lower in the RBCs labeled with 111In-oxine or 99mTc than in the RBCs labeled with 51Cr. CONCLUSIONS: The greater in vivo elution of 51Cr, 111In-oxine, and 99mTc than that of biotin-X-NHS influenced the measurements of RBC volume, 24-hour posttransfusion survival, and life span of the fresh baboon RBCs.

Animals↗

Process for the preparation of pathogen-inactivated RBC concentrates by using PEN110 chemistry: preclinical studies.

BACKGROUND: A pathogen-inactivation process for RBC concentrates is being developed by using PEN110 chemistry (INACTINE, V.I. Technologies). The objective of this study was to characterize the quality of RBCs prepared by using the PEN110 process and to measure the virucidal effect achieved against two viruses. STUDY DESIGN AND METHODS: Virology and RBC studies were conducted with standard RBC units treated with 0.1-percent (vol/vol) PEN110 at 22 degrees C for 6 hours. The quality of PEN110-treated human RBCs was assessed with biochemical and phenotypic variables. The in vivo viability of PEN110-treated RBCs in baboons was studied with the double-label (51)Cr/(125)I method. RESULTS: Decreases in infectious titer by inactivation of greater than a 5 log 50-percent tissue culture infectious doses per mL of bovine viral diarrhea virus (an enveloped RNA virus) and porcine parvovirus (a nonenveloped DNA virus) was observed. RBC hemolysis was less than 1 percent after 42 days of storage, and no changes in RBC antigens were observed. The in vivo viability of PEN110-treated baboon RBCs was unchanged from control. CONCLUSION: The preparation of RBCs by using the PEN110 process achieved a significant viral reduction of two diverse viruses without causing adverse effects to the RBCs. The process appears to be a promising approach, thus justifying further study.

2,3-Diphosphoglycerate↗

Volume of RBCs, 24- and 48-hour posttransfusion survivals, and the lifespan of (51)Cr and biotin-X-N-hydroxysuccinimide (NHS)-labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on (51)Cr 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 (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. STUDY DESIGN AND METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Animals↗

Circulation and hemostatic function of autologous fresh, liquid-preserved, and cryopreserved baboon platelets transfused to correct an aspirin-induced thrombocytopathy.

BACKGROUND: The survival of fresh and preserved platelets has been used primarily to determine their therapeutic effectiveness. The function of the fresh and preserved platelets has been difficult to assess. In stable thrombocytopenic patients, platelet function of fresh and preserved allogeneic platelets is evaluated by the reduction in bleeding time. In this study of healthy male baboons, both the survival and function of autologous fresh, liquid-preserved, and cryopreserved platelets in the correction of an aspirin-induced thrombocytopathy was evaluated. STUDY DESIGN AND METHODS: Five healthy male baboons were studied on eight occasions over a 4-year period. To produce a prolonged bleeding time, the baboon was administered 325 mg of aspirin 18 hours before receiving autologous transfusion. The fresh, liquid-preserved, and previously frozen washed platelets were labeled with (111)In-oxine before autologous transfusion. The autologous, nonaspirinated platelets' ability to reduce the aspirin-induced prolonged bleeding time and increase the shed blood thromboxane B2 level at the template bleeding time site was studied. RESULTS: Platelets stored at 22 degrees C for 48 hours had in vivo recovery values similar to those platelets stored for 18 hours, and they significantly reduced the bleeding time and increased the shed blood thromboxane level after transfusion. Platelets stored at 22 degrees C for 72 hours had in vivo recovery values similar to those platelets stored for 18 hours, but the bleeding time was not corrected after transfusion, although there was a significant increase in the shed blood thromboxane B2 level. The cryopreserved platelets significantly reduced the bleeding time and significantly increased the shed blood thromboxane level after transfusion. Cryopreserved platelets had better in vivo survival and function than the 5-day liquid-stored platelets. CONCLUSIONS: The survival of autologous fresh, liquid-preserved, or cryopreserved platelets did not correlate with their function to reduce an increased bleeding time in baboons treated with aspirin.

Animals↗

Hemostatic defect in baboons autotransfused treated plasma to simulate shed blood.

BACKGROUND: Nonwashed shed blood may contain products of clotting and fibrinolytic, and antifibrinolytic substances. This study was done to determine how autotransfusion of nontreated plasma and plasma treated with urokinase with and without aprotinin affected hemostasis in healthy baboons. METHODS: A 500-mL volume of blood was collected from the baboon, a 250-mL volume of plasma was isolated, and the RBCs were reinfused. Three baboons were autotransfused untreated plasma. Four baboons received plasma that had been treated with 3000 IU/mL urokinase at +37 degrees C for 30 minutes. Eight baboons received plasma that had been treated first with urokinase 3000 IU/mL at +37 degrees C for 30 minutes and then with aprotinin (1000 KIU/mL). Bleeding time, fibrinogen degradation products (FDP), D-dimer, and alpha-2 antiplasmin levels were measured. RESULTS: During the 4-hour period following autotransfusion of the urokinase-aprotinin-treated plasma, the levels of D-dimer and FDP were significantly higher and fibrinogen and alpha-2 antiplasmin levels were significantly lower than those levels seen after the autotransfusion of nontreated plasma. FDP and D-dimer levels showed significant positive correlations with prothrombin time (PT) and activated partial thromboplastin time (aPTT). A significant negative correlation was observed between thrombin time (TT) and fibrinogen level. A significant positive correlation was observed between bleeding time and D-dimer level and a significant negative correlation between the bleeding time and the fibrinogen level. CONCLUSIONS: The infusion of a volume of urokinase or urokinase-aprotinin treated autologous plasma equivalent to 15% of the blood volume was not associated with a bleeding diathesis in healthy baboons.

Animals↗