Acid Citrate Dextrose Solution Research Articles

Quetiapine, Olanzapine and Haloperidol Affect Human Plasma Lipid Peroxidation in vitro

Objective: Oxidative injury in schizophrenia may be caused not only by pathophysiological processes but partly also by treatment with antipsychotics. The purpose of the present study was to examine and to compare the effects of quetiapine (QUE), olanzapine (OLA) and haloperidol (HAL), at final concentrations corresponding to doses used for treatment of acute episodes of schizophrenia, on plasma lipid peroxidation in vitro, measured by the level of thiobarbituric acid-reactive substances (TBARS). Methods: Blood from 30 healthy volunteers was collected into ACD (citric acid/citrate/dextrose) solution. The drugs in form of active substances were dissolved in 0.01% dimethyl sulfoxide, added to plasma at the final concentrations [QUE (175 and 275 ng/ml), OLA (20 and 40 ng/ml), HAL (4 and 20 ng/ml)] and incubated for 1 and 24 h at 37°C. The level of TBARS was measured spectrophotometrically (according to the Rice-Evans method, 1991). Results: The comparative study in vitro showed that QUE causes a decrease in the TBARS level in plasma, whereas HAL increases the plasma TBARS level. After 24 h of incubation of plasma with QUE or HAL (at lower and higher concentrations),thedifferences in TBARS levels between the drugs were significant (p = 5.9 × 10^–4, p = 2.2 × 10^–5, respectively). Conclusion: QUE and OLA, contrary to the prooxidative action of HAL, did not induce oxidative stress; moreover, QUE has antioxidant properties.

Parameters affecting the yield of DNA from human blood

An examination of variables affecting the yield of DNA from blood was undertaken in order to improve sample processing and to evaluate alternative methods of mailing blood samples for DNA analysis. A rapid, high-yield method was developed for the isolation of high-molecular-weight DNA from fresh and frozen blood. In addition, the following observations were made: (1) Of the anticoagulants examined, acid citrate dextrose (ACD) solution B was found to be superior to EDTA and heparin for preserving yields of DNA during incubation at room temperature. If DNA is isolated from frozen blood, high yields of undegraded DNA are achieved after incubation at 23°C for 5 days with ACD solution B. (2) High yields of undegraded DNA are obtained from blood stored with ACD solution B for at least 1 day at 42°C, 5 days at 0°C, or 1 month at −20°C. (3) Three cycles of freezing and thawing may have little if any affect on the yield of DNA. The results indicate that blood for DNA extraction may be mailed in an ambient temperature container and, in many cases, sent by first-class mail rather than by overnight delivery services.

Statistical analyses of multielement electronprobe data correlated with morphological changes in stored human erythrocytes

Methods are described for preparing human erythrocytes for energy dispersive electronprobe X-ray microanalysis in the transmission electron microscope and for the subsequent statistical analysis of data by univariate and multivariate analysis of variance combined with canonical variate analysis. Since no element is independent of others in the chemical make-up of a cell, multielement analysis of all the available data confers upon an individual cell a unique identity. The statistical approach described in this paper allocates an identity to each cell which is then tested for conformity with others of the same sample. Human blood was stored for up to 29 days in acid citrate dextrose solution at +4°C. Monolayers of washed erythrocytes were prepared on carbon-coated, celloidin films on TEM specimen grids, quenched in liquid nitrogen and subsequently freeze-dried. Fifty erythrocytes were analysed at 0, 7, 14, 23 and 29 days and the X-ray emissions (per 100 s) of sodium, phosphorus, sulphur, potassium, iron and a specified background value measured. Chemical data processed by the statistical routine indicated the presence of ‘outliers’—cells which possessed statistically significant differences in their chemical composition from others of the sample. These data were correlated with alterations in erythrocyte shape examined by scanning electron microscopy. Discocytes, prevalent in stored blood on day 0 were characterized by low Na and high P, S, K and Fe. With storage, discocytes were replaced by spheroechinocytes (stages I, II and III) and, later, spherocytes—these cells being present in greatest numbers on days 7, 7–14, 23 and 29, respectively. Individual erythrocytes gained Na and lost K, P, S and Fe during this period. Loss of erythrocyte mass was demonstrated by a reduction in the background radiation.

Amphotericin-B promotes leukocyte aggregation of nylon-wool-fiber- treated polymorphonuclear leukocytes

Severe pulmonary reactions have been reported in patients receiving leukocyte transfusion and amphotericin-B. To study the interaction of amphotericin-B with polymorphonuclear leukocytes (PMN), purified human PMN were incubated with 200 mg of nylon wool fiber for 60 min either in the absence or presence of 2 mM EDTA. PMN were recovered in acid citrate dextrose solution and were suspended in balanced salt solution for determination of their aggregation properties. The cells exposed to nylon wool fibers without EDTA aggregated in response to concentration as low as 1.25 micrograms/ml of amphotericin-B. Cells initially treated with EDTA, however, failed to aggregate. Serum from a patient treated with amphotericin-B aggregated PMN exposed to nylon wool fiber but not control cells, whereas serum taken before amphotericin was given without effect on the PMN treated with nylon wool fiber. Amphotericin-B at 5 micrograms/ml failed to potentiate the release of beta- glucocuronidase or lactic dehydrogenase by PMN treated by nylon wool beyond that seen with exposure to the fibers alone. Rabbit peripheral blood was similarly incubated with nylon wool fibers and the recovered PMN were infused into recipient rabbits that had received 1 mg/kg of amphotericin-B intravenously 1 hr prior to the infusion of the leukocytes. Rabbits were sacrificed 30 min after transfusion of PMN, and their lungs were excised for histologic sectioning. Those rabbits receiving a combination of amphotericin-B and 4 x 10(7) nylon-wool- fiber-treated PMN had evidence of pulmonary hemorrhage and accumulation of leukocytes in the pulmonary vasculature whereas those animals who received such cells alone had normal appearing lung tissue. In summary, amphotericin-B at concentrations achievable in vivo enhanced the aggregation of PMN damaged by incubation with nylon fiber with subsequent accumulation of the phagocytes in pulmonary tissue.

Amphotericin-B Promotes Leukocyte Aggregation of Nylon-Wool-Fiber-Treated Polymorphonuclear Leukocytes

AbstractSevere pulmonary reactions have been reported in patients receiving leukocyte transfusion and amphotericin-B. To study the interaction of amphotericin-B with polymorphonuclear leukocytes (PMN), purified human PMN were incubated with 200 mg of nylon wool fiber for 60 min either in the absence or presence of 2 mM EDTA. PMN were recovered in acid citrate dextrose solution and were suspended in balanced salt solution for determination of their aggregation properties. The cells exposed to nylon wool fibers without EDTA aggregated in response to concentrations as low as 1.25 µg/ml of amphotericin-B. Cells initially treated with EDTA, however, failed to aggregate. Serum from a patient treated with amphotericin-B aggregated PMN exposed to nylon wool fiber but not control cells, whereas serum taken before amphotericin was given was without effect on the PMN treated with nylon wool fiber. Amphotericin-B at 5 µg/ml failed to potentiate the release of beta-glucocuronidase or lactic dehydrogenase by PMN treated by nylon wool beyond that seen with exposure to the fibers alone. Rabbit peripheral blood was similarly incubated with nylon wool fibers and the recovered PMN were infused into recipient rabbits that had received 1 mg/kg of amphotericin-B intravenously 1 hr prior to the infusion of the leukocytes. Rabbits were sacrificed 30 min after transfusion of PMN, and their lungs were excised for histologic sectioning. Those rabbits receiving a combination of amphotericin-B and 4 × 107 nylon-wool-fiber-treated PMN had evidence of pulmonary hemorrhage and accumulation of leukocytes in the pulmonary vasculature whereas those animals who received such cells alone had normal appearing lung tissue. In summary, amphotericin-B at concentrations achievable in vivo enhanced the aggregation of PMN damaged by incubation with nylon fiber with subsequent accumulation of the phagocytes in pulmonary tissue.

Left Ventricular Pump Failure associated with Diffuse Coagulation in Experimental Intraoperative Autotransfusion

Autotransfusion of blood from the peritoneal cavity of pigs under a regimen of acid citrate dextrose solution 1:10 v/v for reservoir anticoagulation or systemic heparinization with 300 Iu/kg did not affect cardiac performance. Rapid deterioration of left ventricular pump performance was observed when low dose systemic heparin 100 Iu/kg was used. Evidence suggests that this is related to diffuse coagulation.

Oxygen dissociation after transfusion of blood stored in ACD or CPD solution

In 20 patients undergoing open-heart surgery, 2,3-diphosphoglycerate (2,3-DPG) concentrations, oxygen affinity of hemoglobin (Po2 at half saturation of hemoglobin with oxygen [P50]), hemoglobin concentration, and pH were measured repeatedly. Measurements were made before and at various times after open-heart surgery and replacement of blood loss with blood stored in acid-citrate-dextrose (ACD) or citrate-phosphate-dextrose (CPD) solutions for less than 72 hours (10 cases per group). Infusion of ACD blood caused P50 and 2,3-DPG concentration to decrease significantly after the operation. The infusion of blood stored in CPD did not significantly increase the oxygen affinity. No significant changes in hemoglobin concentration or pH were observed immediately after the operation in either group. To compensate for the increased oxygen affinity, there must be a rise in cardiac output or more likely a decrease in venous Po2. The transfusion of CPD blood, therefore, is more favorable in terms of oxygen supply, particularly in patients who have had cardiac surgery.

The rhesus monkey as a model for evaluation of the preservation of stored whole blood.

Blood was collected into Acid Citrate Dextrose solution (ACD) from a group of 20 large, adult rhesus monkeys. In ten of the experiments, the blood was labeled immediately with radioisotopes and reinfused into the donor monkeys. The other ten blood units were stored at 4 C for 21 days, then tabled and infused into the donor monkeys. In vitro determinations were made immediately after collection and at the end of the storage period and compared with the results obtained in stored human blood.The ten fresh blood infusions had a mean survival of 101.3 per cent with an average half time of 10.4 days. The 21 (lay‐old bloods had a mean survival of 73.7 per cent (S.D. = 5.1%) with an average half time of 11.3 days. The average red blood cell survival of 123 human blood units stored 21 days and reinfused into their donors had an average red cell survival of 81.0 per cent (S.D. = 8.6%). The determinations in rhesus blood and human blood were comparable.These data strongly suggest that in vivo studies done in the rhesus monkey may be substituted for those performed in man to evaluate red cell preservation.

輸血用血液中の白血球系細胞の生存期間の検討

I studied on the survival time of leucocytes in whole blood preserved in acid citrate dextrose (ACD) solution at 4°C and found that mononuclear leucocytes were rapidly killed. These conclusions were based on the loss of mitosis in tissue cultures at 37°C during the 10 days of storage, and a decrease in survival rate in neutral red vital staining during the 14 days of storage.

Hemolytic Anemia Caused By Penicillin

To the Editor:— Although washed normal human erythrocytes suspended in saline are rapidly hemolyzed when exposed to 25 mg/ml of cephalothin, no hemolysis was detected after exposure of whole blood mixed with anticoagulant acid citrate dextrose solution to 25 mg/ml of cephalothin for four hours. It seems likely that binding of cephalothin by plasma proteins inhibits its hemolytic action. The half-life of autologous 51 Crtagged cephalothin-treated erythrocytes was five days in a man with autoimmune hemolytic anemia. His hematocrit reading (16%) approximated that (19%) of the patient with antipenicillin antibodies at the time the survival studies were done. The proportions of erythrocytes, plasma proteins, and cephalothin were similar in the cephalothintreated erythrocyte survival studies of the two patients. It seems unlikely that the much shorter cephalothin-treated erythrocyte half-life of 135 minutes in the patient with antipenicillin antibodies was due to direct damage of the erythrocytes by cephalothin. However, the possibility

Studies on Aggregation and Adhesion of Platelets

SummaryA modification of the Wright method for the evaluation of the aggregation and adhesive properties of platelets is described; the test is relatively simple and can be performed in a clinical laboratory.Using this procedure, the characteristics of platelets acidified to a pH of 6.5 from their normal pH by addition of small amounts of acid citrate dextrose (ACD) solution to platelet-rich plasma were evaluated. Aggregation and adhesive properties were decreased but returned to normal when the acidified platelets were resuspended in nonacidified platelet-poor plasma. These results confirm the transfusion and isotopie labeling survival studies using acidified platelets and indicate no significant injury of the platelets as exhibited by their aggregation and adhesive characteristics which are considered to be indicative of hemostatic function.

Prevention of citrate intoxication during exchange transfusions.

CITRATE INTOXICATION, which can result from the use of large amounts of stored blood containing the anticoagulant, acid-citrate-dextrose (ACD) solution, has been shown to be a danger in experimental animals 1-4 and in clinical cases. 5 Infusions of the ACD solution alone into laboratory animals can produce the same untoward effects. The citrate itself, low pH, lack of ionized calcium, excess potassium, and alterations in the potassium: calcium ratio have each been considered the dangerous factor. Foote, Trede, and Maloney 6 successfully used ACD blood to prime heart-lung machines after giving sufficient calcium to the patient to reverse the alterations in the electrocardiogram. Bunker and associates 7 were able to reverse the effects of sodium citrate by the intravenous administration of calcium chloride, but they found that the increase in ionized calcium in the blood could not be predicted on the basis of the amount of calcium administered. When the

Variations in Levels of Transaminases and Lactic Dehydrogenase in Bank Blood

Abstract Results of the assay of transaminases and lactic dehydrogenase in blood, proposed as a screening procedure for subclinical hepatitis in blood donors, show that the method of sampling influences the analytic results. Determinations of GOT and GPT may prove useful if made on specimens separated from cells soon after collection. Plasma in contact with cells from blood preserved in acid citrate dextrose solution appears to yield good results up to 21 days. Lactic-dehydrogenase levels are of limited value for this screening.

Prolonged survival of viable, mitotically competent mononuclear leukocytes in stored whole blood.

Earlier investigations on the survival of leukocytes in whole blood preserved in acid citrate dextrose (ACD) solution at 4°C. and in tissue cultures at 37°C. indicated that granulocytes were rapidly killed, presumably as a result of the toxicity of citrate.1 2 3 4 These conclusions were based on the loss of motility, a decrease in phagocytic ability and a marked decrease in the number of granulocytes. Mononuclear leukocytes had been noted to persist in bank blood for several weeks, but owing to the lack of technics for the assessment of the viability of these cells, it was generally assumed that they rapidly became . . .

Effect of citrate on ultrafiltrable and ionic calcium concentrations in serum.

The effect of citrate on the concentration of ultrafiltrable and ionic calcium in blood serum and bovine serum albumin (BSA) solutions was studied. Theoretical equations were derived which predict the fractions of total calcium (in simultaneous equilibrium with protein and citrate) which are ultrafiltrable and ionized. The predictions were made directly from concentrations of total protein, calcium and citrate, and pH. Ultrafiltration experiments on purified BSA, rabbit serum containing added citrate, and human blood-bank serum with acid citrate dextrose solution, yielded data which were in good agreement with predicted values. Additional experiments were performed in order to obtain an empirical expression relating pH to the dissociation of calcium proteinate in normal human serum. Submitted on July 3, 1961

Glucose-6-phosphate dehydrogenase activity in Papuans.

DURING a recent investigation into hæmolytic and other anæmias in Papuans the glucose-6-phosphate dehydrogenase (G-6-P D) activity of erythrocytes was estimated in 45 cases. Blood was collected into heparin, and on several occasions into acid–citrate dextrose solution, and dispatched by air to Melbourne, arriving the same day. The G-6-P D activity was estimated in the Biochemistry Department of the Peter MacCallum Clinic, Melbourne, by the method described by Hsia1. By this method normal readings are 150–210 units/100 ml. The enzyme appears to be relatively stable for at least a week, provided the cells remain intact. The highest values were obtained in cases of thalassæmia major, acute hæmolytic anæmia of undetermined cause, and congenital hæmolytic ansemia of unknown cause. There was no significant difference in the results obtained, whether A.C.D. solution or heparin was used as an anti-coagulant. The twelve subjects used as controls all gave readings within the accepted range of normality.

A Note on the Determination of Some Alkali Salts of Organic Acids by Ion Exchange*

The present official method of analysis for preparations containing alkali salts of organic acids is time consuming. A cation exchange resin was used to convert the salts to the corresponding free acid which was then titrated directly with sodium hydroxide. The method was applied to four parenteral preparations, i. e., anticoagulant sodium citrate solution, anticoagulant acid citrate dextrose solution, sodium lactate injection, and sodium salicylate ampuls. The citric acid in A. C. D. solution was determined by direct titration before passing the solution through the resin column. Dextrose does not interfere in the ion exchange assay procedure. The ion exchange assay procedure is rapid, with an accuracy equal to or better than that achieved by the official method.

AN ACID CITRATE-DEXTROSE SOLUTION WITH LOW WATER VOLUME AND LOW DEXTROSE CONCENTRATION 1

AN ACID CITRATE-DEXTROSE SOLUTION WITH LOW WATER VOLUME AND LOW DEXTROSE CONCENTRATION 1