Effect of Plasma Removal on Blood Stored in ACD with Adenine

An evaluation of the effects of plasma in blood preservation was carried out on units of blood from which the plasma had been removed at different times during storage and on other units in which the plasma volume had been replaced by saline. Blood was either collected in ACD and stored for 14 or 21 days, or collected in ACD supplemented with adenine and stored for 21 or 42 days. These units were given in homologous transfusions to human volunteers. Autologous transfusions were used for separate groups studied at the same time.
Stored units reflected progressive chemical deterioration with increasing storage. The elevation of plasma hemoglobin concentration was observed to be greater in units stored as packed cells.
Average posttransfusion survival values for blood stored for 21 or 42 days were higher for adenine-supplemented blood than for blood in ACD alone, Autologous transfusions with ACD-adenine blood stored for 21 or 42 days survived for the same length of time as did homologous transfusions. Packed cells with plasma removed shortly after collection had the poorest survival.
The similarity of values obtained for cells without plasma suggested that plasma was not particularly involved with the preservation of blood for up to 21 days of storage as tested by red blood cell survival. Improved preservation obtained with adenine was observed even when plasma was removed. Hence, the usefulness of plasma for component therapy may well outweigh its role in blood storage.     

Uptake and Retention of Adenine Moiety by Stored Human Red Blood Cells

Tritium-labeled adenine was added to freshly collected human blood and at selected storage intervals the distribution of the adenine moiety between plasma and erythrocytes was determined. The retention of the radioactive label by erythrocytes was studied by repeated extractions with isotonic saline. The results show that the adenine moiety, which is utilized for net synthesis of various nucleotides, does not appear to be in a steady-state equilibrium between plasma and red cells. With increasing time of storage, the equilibrium is shifted toward the erythrocytes which reach a maximum radioactive content at 21 days of storage. Specific association of the adenine moiety with red cell populations of different density, investigated by density distribution of erythrocytes with polyester mixtures of known specific gravity, shows that cell populations of low density (young cells) have a high specific activity.     

Effects of Adenine on Clotting Factors in Fresh Blood, Stored Blood, and Stored Fresh Frozen Plasma

The addition of small quantities of adenine to whole blood may prolong the useful shelf life of bank blood. The present study was undertaken to evaluate the effects of adenine on the clotting factors in blood containing ACD and CPD. Units of whole blood were collected in ACD, ACD-adenine, CPD and CPD-adenine, and each was stored 42 days under standard blood bank conditions. Samples of fresh frozen plasma containing these anticoagulants were stored three to four months at -30 C. Assays of Factors V, VIII (AHF), IX (PTC), X (Stuart Factor), fibrinogen, and prothrombin were performed on fresh blood, stored blood and stored fresh frozen plasma. The presence of small quantities of adenine did not appear to produce any appreciable alteration in the activity of the clotting factors in fresh blood. Further, adenine did not appear either to improve or worsen the survival of the procoagulants in whole blood stored 42 days or in fresh frozen plasma stored three to four months. There was significant deterioration of Factors V and VIII in whole blood stored 42 days in ACD, ACD-adenine, CPD, and CPD-adenine, but the degree of storage loss was independent of the anticoagulant employed. Factor X, fibrinogen, and prothrombin remained stable in blood stored 42 days regardless of the anticoagulant used, but Factor IX activity increased during storage possibly as the result of contact activation. Fresh frozen plasma stored three to four months showed a uniform slight loss of Factor VIII in all four anticoagulants, but Factors V, IX, X, fibrinogen, and prothrombin remained stable in stored fresh frozen plasma regardless of the anticoagulant employed.     

Metabolism of Autologous and Homologous IgG in Rheumatoid Arthritis

The metabolism of radioiodinated IgG was studied in 20 patients with rheumatoid arthritis and 11 normal controls using autologous IgG and homologous IgG pooled from normal donors. Fractional catabolic rates in the controls were 4.44% of the autologous- and 4.29% of the homologous-labeled protein per day. The corresponding rates in the rheumatoid patients were 9.67% of the autologous- and 8.64% of the homologous-labeled protein per day. Extravascular catabolism occurred only in the rheumatoid group and accounted essentially for the entire increased catabolism of IgG observed in these patients. 10 patients were especially hypercatabolic, with fractional catabolic rates for autologous IgG greater than 10%. Moreover, they catabolized their autologous IgG significantly faster than the homologous IgG (12.6 vs. 9.9%). The increment of catabolism of autologous over homologous IgG also occurred in the extravascular compartment. These highly hypercatabolic patients had a significantly increased number of manifestations of extra-articular disease.     

Some in Vitro Effects of Adenine Added to Stored Blood

When adenine was added to freshly collected ACD blood and the blood incubated at 37 C. for one or two days, the concentration of ATP and adenine nucleotides (AMP + ADP + ATP) was higher than in the same blood without adenine. The addition of inosine with adenine was even more effective. Adenine was used by the red cells and the effective amounts of adenine to be added varied with the experimental conditions, for example, length of incubation. When adenine or adenine and inosine were incubated with outdated ACD blood, the ATP and adenine nucleotide concentrations rose, indicating that synthetic abilities were still present in red cells which had deteriorated energetically as a result of storage in citrate in the cold. When outdated red cells were returned to a more normal p H, the ATP levels increased. When adenine addition was included along with neutralization, the effects were remarkable, the ATP and adenine nucleotide levels going even above normal fresh blood levels. These experiments suggest that added adenine enhances ATP synthesis and allows formulation of a scheme for the participation of adenine in the metabolic reactions of stored cells.     

The In Vivo Survival of Coombs Positive Autologous Erythrocytes Produced by Agglomeration

Coombs positive erythrocytes were observed following the preservation of autologous, human erythrocytes using glycerol, slow freezing, and agglomeration. In 22 of 48 autotransfusions, the preserved erythrocytes were Coombs positive when tested with an anti-gamma + non-gamma globulin serum. In ten of these 22 units, the erythrocytes were tested with both an anti-7S gamma globulin serum and an anti-β₁C globulin serum. All of the erythrocytes in these ten units reacted positively with the anti-β₁C globulin serum, and only one unit reacted equivocally with the anti-7S gamma globulin serum.
The reduction in ionic strength during glycerolization allowed for the uptake of the complement fraction β₁C by the erythrocytes. There was a significant correlation between the Coombs test and the per cent of initial ionic strength of the erythrocyte resuspension medium after glycerolization.
There was no significant correlation between the Coombs test and the 24-hour and seven-day chromium survivals.     

The Viability of Human Blood Stored in Phosphate Adenine Media

The addition of .03 M phosphate ion and adenine to stored blood produces a profound effect on erythrocyte ATP. At the end of 50–65 days storage, the ATP content of such blood is approximately 40% higher than that of blood supplemented with adenine alone. Despite the large increment of ATP, there was little, if any, difference in the viability of blood stored in these solutions. Thus, after 50 days storage, the ATP content is no longer the limiting factor in maintaining erythrocyte viability.     

Comparison Studies of Whole Blood Stored in ACD and CPD and with Adenine

Using 70% 24-hour posttransfusion survival as one of the criteria of preservation, ACD and CPD anticoagulant solutions with and without adenine were tested after storage for 28, 35, or 42 days. At 28 days, all solutions had average survivals of 70% or better. The average survival values for the ACD and CPD solution groups were less than 70% at 35 and 42 days. However, both solutions with adenine had a group average value over 70% even after 35 or 42 days of storage. The average survival values between the two anticoagulants alone, were not significantly different at each time period. The two anticoagulants when adenine was included had increased survival and there was no statistical difference between the levels. In comparing two units obtained from the same subject, survival percentages were significantly higher in almost every recipient when the adenine-supplemented stored blood was used. Other chemical determinations did not show significant alterations and no toxic effects were observed in the recipients.
Since all units containing adenine had survival values greater than 70% in the 28- and 35-day periods, these units would appear to have been effectively preserved and blood stored under these conditions could be used in routine transfusions, reserving units stored 42 days for emergency use.     

Specific Agglutinability of Erythrocytes from Whole Blood Stored at 4 C

When whole blood is supplemented with adenine, the erythrocytes are suitable for transfusion after at least 35 days of storage at 4C, but information is not available as to whether such cells remain satisfactory for blood typing and pre transfusion tests for evidence of incompatibility. Accordingly, we designed experiments to evaluate by AutoAnalyzer the specific agglutinability of erythrocytes obtained from whole blood stored at 4C. Seven normal volunteer donors of both sexes, aged between 22 and 31 years, were chosen and bled periodically so that, over a three-day testing period, all blood specimens from each donor could be evaluated. Each donor provided at each bleeding sufficient blood to permit aliquots of 20 ml to be stored as clotted blood and 8 ml to be stored as citrated whole blood. Four kinds of citrate solution were used: ACD Formula A, CPD, ACD supplemented with adenine, and CPD supplemented with adenine. All clotted specimens were tested after 0, 1, 2, 3, 4, and 5 weeks of storage, while all citrated specimens were tested after 0, 1, 2, 3, 4, 5, 6, 8, and 10 weeks of storage. Six blood group systems were used for evaluation with the following reagents: anti-A or anti-B, anti-Rhl (Rh_o or D), anti-K2 (k or Cellano), anti-Jk^a, anti-Fy^a or anti-Fy^b, and anti-M. Each reagent was used at a dilution to support from 20 to 80 per cent agglutination with freshly drawn blood, and each reagent was tested under two conditions: low ionic and normal ionic. The results disclosed loss of specific agglutinability in association with the time of storage of whole blood at 4 C. The onset and degree of loss depended both on the kind of test used (normal ionic tests showed loss 2.8 weeks earlier than did low ionic tests) and the condition in which the blood was stored (loss occurred sooner and then progressed more with clotted blood than with citrated blood). Average losses of 25, 50, and 75 per cent were observed at 1.1, 2.1, and 4.0 weeks of storage for normal ionic tests of clotted blood, and at 4.1, 6.0, and 7.6 weeks for similar tests of citrated blood. Loss of specific agglutinability during storage was significantly less for the red blood cells of some donors than for others, but systematic differences between specific blood-typing tests were not observed. ACD and CPD solutions behaved similarly, while adenine supplementation exerted a slight preservative effect after 6, 8, and 10 weeks of storage. Loss of specific hemagglutinability of stored cells was not due to loss of specific antigenic sites because stored cells were as effective as fresh cells for the absorption of anti-B, anti-Rhl, and anti-M. Loss might, however, be related to an increase in the accumulation of IgG, fibrinogen, and albumin at the surfaces of erythrocytes.     

Effect of Prednisolone on Incompatible Blood Transfusions

In control experiments small volumes of incompatible, chromium⁵¹ tagged red cells were given to recipients. Several days later the studies were repeated 30 minutes after the intravenous administration of prednisolone. Prednisolone was found completely ineffective in prolonging the survival of small amounts of red cells in the presence of incompatible antibodies of the ABO and Rh systems.     

Observations on the Preservation of Autologous Human Erythrocytes Using Glycerol, Slow-Freeze Technic and Agglomeration

An evaluation was made of 43 autotransfusions of 10 cc aliquots of chromium labeled human erythrocytes preserved as full units with glycerol, the slow-freeze technic, and agglomeration. These units were stored in the frozen state at −80 C for up to seven and a half months. An evaluation was also made of five autotransfusions, using this same technic, but with storage of two units at a combination of −80 C and −20 C and three units at −20 C alone for approximately five months. The agglomeration process produced Coombs positive erythrocytes. The data showed that the storage period of approximately five months at a combination of −80 C and −20 C and at −20 C alone produced unacceptable in vivo survival and excessive in vitro loss of hemoglobin. Optimum in vitro and in vivo results were produced when 250 cc of isotonic saline was used instead of 75–100 cc to disaggregate the agglomerated erythrocytes which were frozen and stored at −80 C. When 250 cc of isotonic saline was used to disaggregate the agglomerated erythrocytes, which were frozen and stored at −80 C up to seven and a half months, the cells could be stored in saline or, after centrifugation, in autologous plasma for up to 11 days at 4 C with 24-hour posttransfusion survival of 65 per cent or greater. The in vitro loss related to preservation, the total supernatant hemoglobin on the day of transfusion, and the mechanism of the immediate removal of the nonviable erythrocytes are reported.     

输血对肿瘤术后预后的影响

输血对肿瘤术后预后的影响于俊秀，刘永慧（山东肥城矿务局中心医院外科肥城２７１６０８）ＥｆｆｅｃｔｏｆＢｌｏｏｄＴｒａｎｓｆｕｓｉｏｎｓｏｎＰｒｏｇｎｏｓｉｓａｆｔｅｒＴｕｍｏｒＥｘｃｉｓｉｏｎＹｕＪｕｎｘｉｕ，ＬｉｕＹｏｎｇｈｕｉ（ＴｈｅＣｅｎｔｒａ...     

Hemoglobin Function of Blood Stored at 4 C in ACD and CPD with Adenine and Inosine

Normal hemoglobin function depends on adequate erythrocyte levels of 2,3‐diphosphoglycerate (2,3‐DPG), a compound that is poorly maintained during blood bank storage in acid‐citrate‐dextrose (ACD). Since 2,3‐DPG is better maintained at the higher pH afforded by citrate‐phosphate‐dextrose (CPD), hemoglobin function was compared during storage in CPD and ACD. Further, hemoglobin function was studied in CPD blood containing adenine and inosine, compounds that provide metabolic energy and thus prolong the shelf‐life of blood, because they also effect the levels of 2,3‐DPG during storage. Hemoglobin function, expressed as the P₅₀ (the P₀₂ at 50 per cent oxygenation, an inverse but direct measure of oxygen affinity) is considerably better maintained during storage in CPD than in ACD. The hemoglobin function or P₅₀ of blood stored in CPD‐adenine is not maintained as well as blood stored in CPD without adenine, but the oxyhemoglobin dissociation curves show only a small difference when compared to the difference between ACD and CPD. Blood stored in CPD‐adenine with inosine, present initially or added at day 25, allows higher P₅₀ values late in storage, thus providing better hemoglobin function for more of the storage period.     

Complement-mediated Defect in Clearance and Sequestration of Sensitized, Autologous Erythrocytes in Rodent Malaria

We investigated the ability of malaria-infected and normal mice to clear particulate immune complexes consisting of autologous erythrocytes sensitized with either IgG or complement.     

Transfusion of Long Stored Whole Blood or Washed Red Blood Cells Incubated with Adenine and Inosine

Full unit autotransfusions of long stored ACD blood, incubated with adenine and inosine at 37 C., were given to healthy young male volunteers. Red cells of blood stored for 35 days showed, after regeneration, a significant increase in ATP and a 24-hour posttransfusion survival of 78.8 per cent (70.9–85.9%); red cells of blood stored for 42 days, thus regenerated, showed a similar increase in ATP and a 24-hour posttransfusion survival of 75.6 per cent (71.5–80.6%). These results were not significantly different from those obtained with 10-ml token autotransfusions of blood similarly treated, the posttransfusion survival of red cells in token transfusions being 78.8 per cent for blood stored 35 days prior to regeneration with adenineinosine and 74 per cent for blood stored for 42 days prior to regeneration. Available data on toxicity of adenine and inosine have been critically reviewed: Chance of direct toxic effects with the small amounts involved may be dismissed when few transfusions are involved; however, uric acid overload must be considered when multiple transfusions are required within a short period of time. A single washing with saline-glucose solution reduces by 90 per cent the concentration of un-metabolized adenine and inosine, and of the product of their metabolism, hypoxanthine. The washing procedure involves a loss of only 0.55 per cent of the total red blood cell population; washing additionally reduces the amount of free hemoglobin. Washing has no effect on the ATP or red blood cell viability, and is recommended when multiple transfusions of cells treated with adenine and inosine are required in a short period of time.     

Lytic Effect of Mycobacillin and Its Derivatives on Erythrocytes

Mycobacillin as well as its O-acetyl (tyrosyl and seryl) and ester derivatives is known to have antifungal activity. The antibiotic also possesses hemolytic activity. This latter property is not appreciably altered by partial or complete esterification of its free carboxyl groups. On the other hand, acetylation of the molecule at the two tyrosyl hydroxyl groups nullifies selectively the hemolytic activity.     

The Effect of the Addition of Adenine and Nucleosides at the Beginning of Storage on the Concentrations of Phosphates of Human Erythrocytes during Storage in Acid-Citrate-Dextrose and Citrate-Phosphate-Dextrose

Bloods collected in ACD and CPD preservatives, supplemented with varying amounts of adenine, inosine, adenosine, and guanosine, were stored anaerobically and aerobically at 4 C for six weeks. At two-week intervals, phosphorylated carbohydrate intermediates of washed red cells were assayed by ion-exchange chromatography. The 2,3-diphospho-glycerate values were higher in the CPD or CPD-supplemented than in the ACD bloods which were at lower pH levels. Generally, the ATP and the sum of adenylate (ΣAd) concentrations were maintained at higher levels for longer periods when bloods were collected in more acidic solutions. The highest ATP and ΣAd levels were observed in ACD-adenine (.075, 0.10 and 0.25 mmole/100 ml blood) and in CPD-adenine (0.5 mmole) anaerobically-stored bloods. Relatively high ATP and ΣAd levels were maintained in blood collected in ACD-inosine -+ adenine + guanosine and in inosine-adenine preservatives. Lower values were obtained with ACD-inosine, ACD-adenosine and ACD-adeno-sine + adenine, in the order listed. The ADP values remained relatively constant during storage. An appreciable increase in AMP concentration was observed only in the adenine-supplemented bloods. The IMP concentration increased in all stored bloods and was particularly marked in the presence of adenosine and least in the adenine-supplemented bloods.     

微波对红细胞免疫功能的影响

体外实验证明，微波输出功率８０Ｗ，辐射时间２分钟，对红细胞免疫功能有明显的促进作用．２０例正常人静脉血微波辐射后ＲＢＣ－ＣＲ＿１花环率显著提高（Ｐ＜０．０１），ＲＢＣ－ＩＣ花环率则无变化（Ｐ＞０．０５）．肿瘤、风湿性心脏病、冠心病及肺疾病患者静脉血共８０例，微波辐射后ＲＢＣ－ＣＲ＿１及ＲＢＣ－ＩＣ花环率均明显提高．实验提示，微波的这种生物学效应为非热效应，临床上应根据不同个体选择不同的微波输出功率和辐射时间，才能充分发挥微波的生物学效应，提高患者的红细胞免疫功能．     

Comparison of Blood Stored in ACD and in a Solution Containing Heparin, Phosphate, Glucose and Adenine

Duplicate blood samples were obtained from six healthy young males, one aliquot was collected in ACD and one aliquot in a solution of heparin-glucose-phosphate-adenine (HGPA). All were stored under blood bank conditions for a total of four weeks. The HGPA samples maintained their ATP and nucleotide adenine levels much better than the ACD samples and had generally higher p H's and faster rates of glycolysis. Their osmotic fragility and plasma K⁺ levels were similar to those of the ACD samples. Because there was considerably more hemolysis in the HGPA samples than in the ACD samples and in the absence of red cell survival data, the HGPA system is not recommended at this time as a substitute for the ACD system but as a comparative system to reveal metabolic shortcomings of the ACD storage system.     

In Vivo Survival and Supernatant Hemoglobin of Autologous, Deglycerolized, Resuspended Erythrocytes Processed Using Centrifugation

Autologous human erythrocytes were frozen at −80 C and stored for periods of three to eight months, using centrifugation to add and remove the intracellular additive, glycerol. The length of storage had no adverse effect on the 24-hour and seven-day posttransfusion survivals. Acceptable 24-hour posttransfusion survival was observed in the nine subjects studied. The mechanism of removal of the nonviable erythrocytes immediately following infusion was not associated with hemoglobinemia. The total supernatant hemoglobin in the unit on the day of transfusion and the total in vitro loss related to preservation are reported.