Recovery, lifespan, and function of CPD-Adenine (CPDA-1) platelet concentrates stored for up to 72 hours at 4 C

We studied the in vivo recovery, lifespan, and hemostatic effectiveness of CPDA-1 platelet concentrates stored for up to 72 hours at 4 C. A total of 120 CPDA-1 concentrates containing an average (+/− 1 S.D.) of 6.6 +/− 2.0 × 10(10) platelets were prepared. The pH of the units following storage at 4 C was 6.8 +/− 0.2; no unit had a pH below 6.3. Autologous transfusion of six normal volunteers showed that platelets stored at 4 C for 72 hours had an in vivo recovery of 40 +/− 18 per cent and a lifespan of 5.1 +/− 1.5 days. The hemostatic effectiveness of CPDA-1 platelets was determined by platelet counts and template bleeding time measurements in 10 thrombocytopenic patients. Patients receiving 48-hour-stored platelets had a four- to six-hour posttranfusion corrected platelet increment averaging 15,300 +/− 3,200/microliter which was 67 +/− 34 per cent of expected recovery. Four of the five patients transfused with this preparation showed an improved bleeding time. In contrast, three patients receiving 72-hour- stored platelets had a four- to six-hour posttransfusion increment of 5,800 +/− 2,400/microliter that was only 26 +/− 13 per cent of the expected recovery; furthermore, only one of these patients showed any correction of the bleeding time. These data indicate that CPDA-1 platelets are hemostatically effective when stored at 4 C for up to 48 hours.     

The Effects of Continuous—Flow Washing on Stored Red Blood Cells

Whole blood and packed cells stored up to 21 days prior to being washed by a continuous-flow (CF) technic or by batch washing (BW) were found to have an average posttransfusion survival of greater than 70 per cent. CF-washed red blood cells stored in 5 per cent dextrose and 0.9 per cent saline (5% D&S) wash solution for 24 hours had an average survival of 86 per cent. An average plasma protein dilution of 1:25,000 was obtained with the CF technic as compared to 1:600 with the conventional BW technic. A series of bloods were inoculated either with plasma containing hepatitis-associated antigen (HAA), cytomegalovirus (CMV) or poliovirus (PVS) and then washed. CMV could not be detected by plaque assay in the washed red blood cells nor could HAA by immunodiffusion, counterimmunoelectrophoresis or radio-immune assay. The poliovirus-inoculated washed red blood cells had a marked reduction of the virus when washed by the CF technic as compared to the BW cells. Red blood cells washed by the CF technic did not induce a rise in an AHF inhibitor when transfused into a hemophilic patient with a circulating inhibitor.     

Concentrated Antihemophilic Factor (AHF) from Outdated Blood

The antihemophilic factor (AHF) content of bank blood decreases during storage under standard blood bank conditions. In the present study, an average of 58 per cent of the original activity was present after the usual shelf life of 21 days expired. This material was concentrated by a modification of the Pool cryoprecipitation method. Almost 60 per cent of the residual AHF in the outdated plasma was found in the precipitate. The resulting precipitate was resuspended in about 25 ml of the residual plasma. On a volume basis, this had an average of about four times the activity of fresh human plasma. The cold insoluble material resuspended in residual plasma can be used for therapy without further processing. Following intravenous injection into hemophilic patients, high levels of circulating AHF, in excess of 60 per cent of normal, have been achieved without causing circulatory overload.     

Antihemophilic Factor (AHF) Stability in Fresh Frozen Blood Bank Plasma

One hundred randomly chosen units of fresh frozen blood bank plasma were assayed for antihemophilic activity (AHF). A wide range in AHF levels was observed. The variability appears to be due in part to different AHF levels of donors. Most of the units had as much as 30 per cent less AHF than was contained in unfrozen, freshly prepared plasma. There was no trend toward lower values in plasma frozen up to 15 weeks. Many factors contribute to the unpredictable loss of AHF. The effects of slow and rapid freezing on the AHF content of plasma were studied. Freezing of fresh bank plasma in a bath of liquid and in air are compared. Plasma freezes much more rapidly in liquid, with negligible loss of AHF. Freezing is considerably slower in air, and losses of 20 per cent or greater frequently occur.     

Studies of the recovery and the cost of low-glycerol cryopreserved human red blood cells

Red blood cells were equilibrated with 28 per cent (v/v) glycerol and 3 per cent mannitol in 0.65 g/100 ml sodium chloride. The units were frozen by immersion into liquid nitrogen and stored at -160 C. After thawing, they were reconstituted and washed using the IBM 2991 Blood Cell Processor. Freeze-thaw rate curves, the effect of thawing techniques, the effect of varying postthaw washing and processing techniques, estimates of red blood cell losses because of hemolysis, and in vitro recovery were determined. In vivo recovery was determined by 51Cr techniques 24 hours after infusion and Ashby survivals and subsequent life span were measured. Metabolic, scanning electronmicroscopy, cost estimates and quality control studies were done on the reconsituted red blood cells. Recipients were evaluated before and after transfusion for metabolic erythrocyte characteristics and for evidence of hemolysis. The modified method requires less wash solution and less technician time than does the standard low-glycerol method. Two units for the same recipient could be passed through the IBM software with no alteration of cell survival or loss. Revision of the IBM 2991 processing procedure provided excellent recovery of viable previously frozen red blood cells at probably a lower cost.     

Retinal and gastrointestinal disease due to cytomegalovirus in patients with the acquired immune deficiency syndrome: prevalence, natural history, and response to ganciclovir therapy

Of 760 AIDS patients seen at San Francisco General Hospital in 1986, 5.7 per cent had retinitis and 2.2 per cent had gastrointestinal disease caused by cytomegalovirus. We reviewed the records of 44 patients treated with ganciclovir for culture-confirmed cytomegalovirus retinal (31 patients) or gastrointestinal disease (17 patients) or both (four patients) in 1986. Retinitis stabilized or improved during initial treatment with ganciclovir in 22 of 27 (81.5 per cent) patients. Following a median 10-day induction course, 16 patients with retinitis continued to have serial ophthalmologic assessments: eight patients were maintained on treatment and eight had maintenance treatment deferred. Before treatment, the two groups were comparable in age, Karnofsky scores, hematologic assessment, visual acuity, and history with respect to Pneumocystis carinii pneumonia. Retinitis did not progress for a median 53.8 days in the immediate maintenance group compared to 18.8 days for the deferred maintenance group (p = 0.01). In 17 patients with CMV gastrointestinal disease, nine of 14 (64 per cent) had resolution of pain and eight of 11 (73 per cent) had resolution of diarrhea when treated initially with ganciclovir. In both retinitis and gastrointestinal disease patients, ganciclovir decreased recovery of CMV from urine and blood markedly. Ganciclovir also caused a decrease in mean absolute neutrophil counts to about half of baseline values; decreases in mean platelet count and hemoglobin were also noted but were less than 25 per cent. Neutropenia severe enough to require dose adjustment (less than 800 cells/microliters) occurred in 31 per cent of patients receiving maintenance ganciclovir.     

Long Term Frozen Storage of Human Red Blood Cells: Studies in Vivo and in Vitro of Autologous Red Blood Cells Preserved up to Six Years with High Concentrations of Glycerol

Human red blood cells preserved with approximately 45 per cent w/v glycerol, frozen by the slow freeze-thaw technic, and stored at -80 C for up to six years were evaluated by measurements of the posttransfusion survival of autologous ⁵¹chromium-labeled red blood cells, the per cent recovery of the preserved red blood cells, supernatant hemoglobin in the unit, and intracellular potassium levels. The glycerolized red blood cells were washed either with electrolyte solutions using continuous centrifugation, or by dilution with nonelectrolyte solutions and recovery of the red blood cells by agglomeration (Huggins technic). Glycerolized red blood cells stored frozen for more than two years and washed by the Huggins technic had significantly decreased postthaw stability when the de-glycerolized red blood cells were kept after thawing for longer than four hours at 4 C. Glycerolized red blood cells stored frozen for up to six years and washed by continuous centrifugation with electrolyte solutions showed clinically acceptable post-transfusion survival after postthaw storage at 4 C for up to 24 hours. The results of washing glycerolized red blood cells by continuous centrifugation showed that prior dilution of the thawed cells with a ten per cent glycerol solution was vitally important. The method used in washing glycerolized red blood cells may significantly limit the length of time that red blood cells may be stored at 4 C after having been frozen.     

Washing of Red Blood Cells Previously Frozen in Liquid Nitrogen. Experience with the Elutramatic Ultra-Flo System

An automated continuous-flow washing system, the Elutramatic Ultra-Flo system, has been tested for the deglycerolization of red blood cells previously frozen in liquid nitrogen. Packed red blood cells frozen with 100 ml of a 55 per cent glycerol solution were, after thawing, diluted with 175 ml 3.7 per cent sodium chloride solution. The standard wash program was used for the processing. Approximately 2.2 liter of saline solution were used for the washing of one unit of blood. The processing time for two units was 27 minutes in the automated cycles. The mean red blood cell recovery was 81 per cent and the mean residual hemolysis in the washed blood was 0.36 per cent. No important changes of ATP, DPG, and intraerythro-cytic potassium occurred during the processing. The 24-hour posttransfusion red blood cell survival was 91.6 ± 1.8 per cent. For the trained blood bank technician the Elutramatic Ultra-Flo system offers a rapid and convenient method for the deglycerolization of red blood cells previously frozen in liquid nitrogen.     

Cryopreserved red blood cells for pediatric transfusion. Frozen storage of small aliquots in polyvinyl chloride (PVC) plastic bags

Human nonrejuvenated and rejuvenated red bood cells were prepared for cryopreservation and subsequent pediatric transfusion. Glycerol was added to the red blood cells in the primary polyvinyl chloride plastic collection bag to achieve a concentration of 40 per cent W/V. The red blood cells were concentrated by centrifugation, and the supernatant glycerol was discarded. Each glycerolized unit was divided into four equal aliquots in the individual 600-ml bags of a dry quadruple polyvinyl chloride plastic system, and each aliquot was frozen and stored at −80 C. After thawing, sodium chloride solutions were used to wash the aliquots in the IBM Blood Processor 2991-1 or 2991-2 or the Haemonetics Blood Processor 115, and the washed aliquots were stored in a sodium chloride-glucose-phosphate solution at 4 C for 24 hours. Freeze-thaw recovery of the red blood cells was about 97 per cent, and freeze-thaw-wash recovery was about 84 per cent. Twenty-four-hour posttransfusion survival values were about 92 per cent for both nonrejuvenated and indated-rejuvenated red blood cells. Nonrejuvenated red blood cells, those frozen within three to five days of collection without biochemical modification, had normal oxygen transport function at the time of transfusion; rejuvenated red blood cells, those biochemically treated with PIGPA Solution A after three to five days of storage at 4 C, had improved oxygen transport function at the time of transfusion.     

Preferential decrease in thymus dependent lymphocytes during storage at 4 C in anticoagulant

Human lymphocytes stored at 4 C either as leukocyte concentrates (LCs) in citrate-phosphate-dextrose (CPD) or as whole blood anticoagulated with CPD show a rapid and marked decrease in the relative and absolute numbers of thymus derived (T) lymphocytes. Determinations were made on cells recoverable on a Ficoll-Hypaque (F-H) gradient. In evacuated LCs, the relative percentage of T cells dropped to less than 10 per cent within 72 hours with a concomitant increase in the relative percentage of bone marrow derived (B) cells to 80 per cent or more. LCs opened to the air and subsequently stored at 4 C displayed an even more precipitous decline in the relative percentage of T cells, reaching a 10 per cent level within 72 hours. The relative percentage of T cells in CPD-anticoagulated whole blood samples stored at 4 C displayed similar decreases, reaching 20 per cent levels within 24 hours. The change in the relative percentage of T cells at the Ficoll-Hypaque interface was shown to reflect a decrease in the total numbers of T cells placed on the F-H gradient with time, since determinations of T and B cell numbers in NH4Cl-treated whole blood showed a 65 to 80 per cent decrease in the numbers of T cells within 24 hours in anticoagulated whole blood held at 4 C. Thus, it may be inferred that the T cell decrease is mediated via some interaction of anticoagulant, storage time, and some component(s) present in both LCs and whole blood.     

Therapeutic effectiveness and safety of outdated human red blood cells rejuvenated to restore oxygen transport function to normal, frozen for 3 to 4 years at −80 C, washed, and stored at 4 C for 24 hours prior to rapid infusion

Human red blood cell concentrates with hematocrit values of 75 V% were prepared from citrate-phosphate-dextrose (CPD) blood, stored at 4 C for 20 to 28 days, and biochemically modified with a solution containing pyruvate, inosine, glucose, phosphate, and adenine (PIGPA Solution A). The rejuvenated red blood cells were frozen with 40% W/V glycerol in a polyolefin plastic bag and were stored at ?80 C. After three to four years of frozen storage, the units were thawed, washed, and stored at 4 C in a sodium chloride-glucose-phosphate solution for 24 hours prior to transfusion. Red blood cell recovery was 97 per cent after thawing and 90 per cent after washing. An automated differential agglutination procedure (ADA) showed 24-hour survival values of about 80 per cent, and long-term survival values of about 85 days depending on the disease state of the recipient. The red blood cells had normal affinity for oxygen on the day of transfusion. Plasma hemoglobin levels measured immediately after transfusion indicated extravascular removal of nonviable donor red blood cells. There was no increase in the uric acid level during the 24-hour posttransfusion period. A pool of three to ten units of rejuvenated washed previously frozen red blood cells was transfused rapidly to each of 19 anemic elderly patients. The red blood cells which had normal oxygen delivery capacity immediately upon transfusion increased the recipient's red blood cell mass and produced no untoward effects.     

Accelerated immune red cell destruction in the absence of serologically detectable alloantibodies

Two patients are described in whom clinically significant red blood cell alloantibodies could be demonstrated only by in vivo 51chromium (51Cr) survival studies. The first patient had experienced a severe delayed hemolytic transfusion reaction to four units of crossmatch compatible blood. Serial phenotype studies suggested the presence of a serologically undetectable anti-c (hr') antibody. 51Cr survival of c- positive red blood cells was one per cent at 24 hours, while survival of c-negative red blood cells was 80 per cent at 24 hours. The second patient had multiple red blood cell alloantibodies. An anti-c antibody was suspected but could not be convincingly demonstrated by in vitro techniques. 51Cr survival of c-positive red blood cells, however, was 57 per cent at 24 hours and 17 per cent at 48 hours. 51Chromium red blood cell survival studies should be considered whenever an unexplained hemolytic transfusion reaction occurs, or when an expected red blood cell alloantibody cannot be demonstrated by in vitro laboratory studies.     

Therapeutic Effectiveness of Homologous Erythrocyte Transfusions Following Frozen Storage at ‐80 C for up to Seven Years

Cohn‐processed red blood cells that had been stored for as long as seven years at ‐80 C., washed by the ADL procedure and then stored at 4 C for up to 48 hours, showed approximately 90 per cent 24‐hour recovery in vivo by an automated differential agglutination (ADA) technic, recovery in vitro of approximately 90 per cent, and an index of therapeutic effectiveness of approximately 80 per cent. Washing Huggins‐preserved red blood cells with EDTA by the Huggins process produced a significant deterioration (decreased 24‐hour posttransfusion survival and decreased recovery in vitro ) following storage at ‐80 C for as long as three years. In two of seven patients studied the Huggins‐processed red blood cells that had been stored at ‐80 C for 1.8 years and longer and washed by the Huggins procedure showed intravascular destruction of the compatible nonviable red blood cells. Huggins‐preserved red blood cells with EDTA that had been stored at ‐80 C up to 1.6 years showed, following washing with an electrolyte solution in the ADL bowl, a somewhat better 24‐hour ADA survival, better recovery of the preserved red blood cells, lower supernatant hemoglobin concentrations, and higher intracellular potassium levels on the day of washing and resuspension. These findings suggest that Hugginspreserved red blood cells following storage at ‐80 C for one and one half years or more should not be washed by the Huggins dilution/agglomeration procedure.     

Viability and function of outdated human red blood cells after biochemical modification to improve oxygen transport function, freezing, thawing, washing, postthaw storage at 4 C, perfusion in vitro through a bubble oxygenator, and autotransfusion

The quality of transfused blood is especially important during cardiac surgery, and red blood cell viability and function may be adversely affected during perfusion through the artificial blood oxygenator used during extracorporeal bypass. In this study, we administered 10 ml aliquot autotransfusions of rejuvenated red blood cells to 13 healthy volunteers after perfusion through an infant bubble oxygenator for one to three hours. Twenty-three other volunteers received rejuvenated red blood cells that had not been perfused. The red blood cells were biochemically modified after they had reached their outdating period, a process used to increase 2,3 DPG and ATP levels and improve oxygen transport function. The rejuvenated red blood cells were frozen with 40% W/V glycerol, stored frozen at -80 C for about 3 months, thawed, washed, and stored in a sodium chloride-glucose-phosphate solution at 4 C for as long as three days. Freeze-thaw recovery was about 97 per cent, and freeze-thaw-wash recovery about 90 per cent. Twenty-three units were transfused after 1 to 3 days of post-wash storage, and 13 units were perfused through an infant bubble oxygenator for as long as three hours before transfusion. The 24-hour posttransfusion survival values were about 80 per cent and oxygen transport function was either normal or improved whether or not the units were perfused before transfusion.     

Hemoglobin labeled by radioactive lysine; erythrocyte life cycle

A dog, doubly depleted of blood cells and plasma proteins, was fed dl-lysine labeled with C(14) in the epsilon carbon position. In the first 8 hours 28 per cent of the administered C(14) was excreted in the urine; in the first 72 hours, 35 per cent. Twenty-four hours after feeding, 4.2 per cent of the fed C(14) was circulating in the plasma, decreasing to 1 per cent at the end of 17 days. The C(14) content of the blood cells increased from 1 per cent at 24 hours to 5.5 per cent in 5 days and 6.8 per cent in 22 days. Evidence based on the rate of decrease of the C(14) content of circulating blood cells is presented indicating an average life of 115 days for the erythrocyte protein as an entity not interchanging with extracellular constituents. This corresponds closely to the life span of the dog erythrocyte, 112 to 133 days according to the best evidence otherwise available and indicates that this experiment has actually measured the life span of the dog erythrocyte. Following breakdown of blood erythrocytes the protein comprising them is not used preferentially for the formation of new erythrocytes.     

Factors Influencing the 24-Hour Posttransfusion Survival and the Oxygen Transport Function of Previously Frozen Red Cells Preserved with 40 Per Cent W/V Glycerol and Frozen at —80 C

A 6.2 M glycerol solution was added directly to concentrated red blood cells before storage at — 80 C for at least two and one-half years. The glycerol was removed from the thawed red blood cells by one of four different washing procedures, and the washed cells were stored at 4 C for an additional 24 hours before transfusion. Recovery in vitro was about 90 per cent, and the posttransfusion survival was about 85 per cent. The CPD anticoagulant maintained the oxygen transport much better than ACD during storage of the red blood cells at 4 C for one week prior to freezing. Results were similar whether glycerolized red blood cells were washed in reusable stainless steel bowls, disposable polycarbonate bowls, or collapsible disposable polyvinylchloride plastic bags. The composition of the wash solution had no significant effect on the posttransfusion survival or oxygen transport function. When the washed red blood cells were stored in a sodium chloride-glucose-phosphate solution at 4 C for 24 hours before transfusion, the 24-hour posttransfusion survival and oxygen transport function was satisfactory. Freeze-preservation of red blood cells with hematocrits of about 40 V per cent and postthaw storage at 4 C for 24 hours resulted in an accumulation of supernatant hemoglobin and extracellular potassium. At the time of transfusion, the red blood cells were concentrated by centrifugation, the supernatant medium was removed, and the hematocrit adjusted to 70 V per cent.     

老年慢性肾功能衰竭并发急性左心功能衰竭的早期诊断

目的为老年慢性肾功能衰竭(CRF)患者并发急性左心功能衰竭(AHF)的早期诊断和处理提供帮助。方法50例60岁以上CRF患者分为两组,I组为CRF并发AHF31例,Ⅱ组为同期其他疾病并发AHF19例,采用临床调查法对其常见的临床早期症状进行对比分析。结果老年CRF并发AHF时早期症状不典型,更容易延误病情。结论在临床上准确及时诊治可阻止病情的发展,挽救患者的生命。     

Retinal and Gastrointestinal Disease due to Cytomegalovirus in Patients with the Acquired Immune Deficiency Syndrome: Prevalence, Natural History, and Response to Ganciclovir Therapy

Of 760 AIDS patients seen at San Francisco General Hospitalin 1986, 5.7 per cent had retinitis and 2.2 per cent had gastrointestinaldisease caused by cytomegalovirus. We reviewed the records of44 patients treated with ganciclovir for culture-confirmed cytomegalovirusretinal (31 patients) or gastrointestinal disease (17 patients)or both (four patients) in 1986. Retinitis stabilized or improvedduring initial treatment with ganciclovir in 22 of 27 (81.5per cent) patients. Following a median 10-day induction course,16 patients with retinitis continued to have serial ophthalmologicassessments: eight patients were maintained on treatment andeight had maintenance treatment deferred. Before treatment,the two groups were comparable in age, Karnofsky scores, hematologicassessment, visual acuity, and history with respect to Pneumocysticcarinii pneumonia. Retinitis did not progress for a median 53.8days in the immediate maintenance group compared to 18.8 daysfor the deferred maintenance group (p=0.01). In 17 patientswith CMV gastrointestinal disease, nine of 14 (64 per cent)had resolution of pain and eight of 11(73 per cent) had resolutionof diarrhea when treated initially with ganciclovir. In bothretinitis and gastrointestinal disease patients, ganciclovirdecreased recovery of CMV from urine and blood markedly. Gancicloviralso caused a decrease in mean absolute neutrophil counts toabout half of baseline values; decreases in mean platelet countand hemoglobin were also noted but were less than 25 per cent.Neutropenia severe enough to require dose adjustment (<800cells/µl) occured in 31 per cent of patients receivingmaintenance ganciclovir.     

A survey about blood bank policies and procedures

A questionnaire about blood bank policies and procedures was prepared and sent out to 170 hospitals across the nation, 50 per cent responded. There is wide variation in both technical and nontechnical policies and procedures. Serum is separated from the original tube within 8 hours by 68 per cent of the hospitals and in 32 per cent serum is never separated from the original tube. Auto-control is included by 66 per cent of hospitals in at least one phase of the antibody testing, 34 per cent do not include auto-control in any phase. Du testing of the recipient is done in 85 per cent of the hospitals and in 15 per cent Rho type is determined by the immediate spin reactions alone. Slide type is the only method used to determine ABO group of cells of the recipients in 10 per cent of the hospitals, 18 per cent use serum-cell suspension, 10 per cent use the applicator stick method and only 62 per cent use washed saline-cell suspension routinely. Compatiblity testing between a recipient and a donor vary from a one-tube major crossmatch to a three-tube major crossmatch and minor crossmatch.     

Comparison of methods to wash liquid-stored red blood cells and red blood cells frozen with high or low concentrations of glycerol

The efficiency of washing liquid-stored red blood cells and red blood cells frozen with high or low glycerol concentrations was evaluated by measuring the recovery of red blood cells in vitro, supernatant hemoglobin, extracellular potassium and red blood cell potassium levels, supernatant osmolality, residual 125I albumin, glycerol, hypoxanthine, and di-2-ethylhexyl phthalate (DEHP) levels. Four commercial washing systems were studied, three which used sodium chloride solutions with serial or continuous-flow centrifugation and one which used sugar solutions and dilution/agglomeration. Washing was most efficient using sodium chloride solutions in the IBM Blood Processor, an automated serial centrifugation procedure and in the Fenwal Elutramatic, a continuous-flow centrifugation procedure. Less efficient washing was achieved in the Haemonetics Processor 15, a continuous-flow centrifugation procedure and the least efficient washing occurred using the original and modified dilution/agglomeration procedures. To achieve the most efficient washing, three principles must be utilized: concentration of the red blood cells to hematocrit values of 90 per cent, prior to washing or freezing. Liquid-stored red blood cells concentrated to hematocrit values of 90V per cent should be diluted with hypertonic sodium chloride solutions prior to recovery and washing. Red blood cells containing 20 per cent or 40 per cent W/V glycerol should be diluted with hypertonic sodium chloride solutions before recovery and washing. Finally, on-line dilution should be achieved in the washing systems that use continuous-flow centrifugation.