Citrate Phosphate Dextrose Adenine Solution Research Articles

Hematologic and biochemical changes in caprine whole blood stored in CPDA-1 for 28 days

Parasitic anemia is a common affliction of goats in the United States that, in severe cases, requires a blood transfusion. There is currently limited data published on blood storage in small ruminants, goats in particular. With an increase in pet goats as well as a population of show and breeding animal owners, there is an increasing demand for blood transfusions in these patients to treat their anemia. The primary objective of this study was to determine biochemical and hematologic changes in caprine whole blood stored in citrate phosphate dextrose adenine solution (CPDA-1) over 28 days, particularly those that would impact transfusion efficacy, including packed cell vol­ume (PCV) and percent hemolysis in excess of 1%.

Caprine whole blood appears sufficiently stable after 21 days in storage to be suitable for transfusion.

The objective of this study was to determine hematologic changes of stored caprine whole blood in citrate phosphate dextrose adenine solution over a 28-day period. Ten 250-mL bags of whole blood were collected from 10 female Boer goats from Louisiana State University's Division of Laboratory Animal Medicine herd. 10 healthy blood donor goats were selected, and 250 mL of whole blood was drawn from each and stored at 2.78 °C. At the time of collection and every 7 days for a total of 28 days, samples were obtained from the blood bags to determine biochemical and hematologic values of collected blood. Only 5 of the 10 donors had baseline blood bag samples obtained for biochemical evaluation on day 0. At the end of 28 days, the remaining blood was submitted for aerobic and anaerobic culture. Blood values remained within suitable limits for transfusion and below 1% hemolysis for up to 21 days in most samples. Packed cell volume did not change significantly from day 0 to day 28. Lactate significantly increased over the 28 days, though not as dramatically as expected on the basis of other blood storage studies. pH decreased due to anticoagulant acidity but did not drop below 7. Cultures were negative on all blood bags. Changes over time are similar to that in other species, and caprine blood appears biochemically and hematologically stable for up to 21 days in storage. In vivo trials are needed for safety and efficacy.

The Contribution of Storage Medium and Membranes in the Microwave Dielectric Response of Packed Red Blood Cells Suspension

During cold storage, packed red blood cells (PRBCs) undergo slow detrimental changes that are collectively termed storage lesion. The aging of the cells causes alterations in the composition of the storage-medium in the PRBC unit. In this paper, we present the comparison of the dielectric response of water in the primary (fresh) storage medium (citrate phosphate dextrose adenine solution, CPDA-1) versus the storage medium from three expired units of PRBCs. Dielectric response of the water molecules has been characterized by dielectric spectroscopy technique in the microwave frequency band (0.5–40 GHz). The dominant phenomenon is the significant increase of the dielectric strength and decrease the relaxation time τ for the samples of the stored medium in comparison with the fresh medium CPDA-1. Furthermore, we demonstrated that removing the ghosts from PRBC hemolysate did not cause the alteration of the dielectric spectrum of water. Thus, the contribution associated with water located near the cell membrane can be neglected in microwave dielectric measurements.

Effects of L-carnitine on the erythrocytes of stored human blood.

This study aimed to assess the effects of L-carnitine on oxidative stress in human erythrocytes during storage. Using antioxidants as components of blood storage solutions may combat the effects of storage-induced oxidative stress on erythrocytes. Blood from male adults was stored at 4°C for 55 days in citrate phosphate dextrose adenine solution, without L-carnitine (Control) and with L-carnitine as an additive (at concentrations of 10, 30 and 60 mM-Experiments). Every fifth day, erythrocyte markers (morphology, count, haemoglobin, haemolysis and osmotic fragility), antioxidant defences (antioxidant enzymes and total antioxidant capacity) and oxidative stress markers (superoxides, lipid peroxidation and protein oxidation products) were analysed. Oxidative damage was observed in controls (day 25 onwards) and in experiments (day 35 onwards). L-carnitine (10 and 30 mM) protected erythrocytes from damage up to day 35 by maintaining haemoglobin and lipid peroxidation, assisting antioxidant enzymes and increasing antioxidant capacity by elevating sulfhydryls and ascorbic acid. L-carnitine was beneficial in prolonging storage up to 55 days but could not prevent oxidative damage completely in terms of haemolysis and osmotic fragility. L-carnitine ameliorated oxidative stress, but combinations with other antioxidants may provide comprehensive protection to erythrocytes during storage.

Clinical Experience of Autologous Blood Transfusion in Neurosurgery: Prospective Study in Central India

To evaluate the benefits of intraoperative autologous blood transfusion in intracranial procedures and to conserve precious homologous blood due to shortage of donor and associated complications. This was a prospective study performed at Bansal Hospital, Bhopal. Predetermined autologous blood was collected in a well-labeled bag containing citrate phosphate dextrose adenine solution after induction of general anesthesia. Then appropriate amount of crystalloid solution was transfused in to the body. All collected autologous blood was transfused back to the patient at the end of the surgery or during the surgery if excessive blood loss occurred. Demographic data, hemodynamic changes (mean arterial pressure, heart rate) before and after donation, complications, and any additional homologous blood requirement were noted. Pre- and postoperative hemoglobin and hematocrit values were compared. In total, 32 patients were included in this study. In our study, mean age was 48.87 years; male-to-female ratio was 1:1.4. The mean amount of autologous blood collected was 461 mL, and the mean amount of blood loss during surgery was 1048 mL. In our study, there was no statistically significant difference was found in mean arterial pressure and heart rate before and after autologous blood collection (P > 0.05). When we compared pre- and postoperative mean hemoglobin and hematocrit levels, there was a statistically significant difference present (P<0.05); this was due to the fact that many patients had meningiomas (15 of 32). Additional homologous blood was required only in 25% of cases (8/32). Of 8 patients, 5 were again cases of deep-seated meningiomas. No complications were observed during or after autologous blood collection. Autologous blood transfusion is a safe, effective, and affordable method of blood transfusion in patients undergoing intracranial surgery. Complications associated with homologous blood transfusion can be avoided with autologous blood transfusion.

A method to preserve low parasitaemia Plasmodium-infected avian blood for host and vector infectivity assays.

BackgroundAvian malaria vector competence studies are needed to understand more succinctly complex avian parasite-vector-relations. The lack of vector competence trials may be attributed to the difficulty of obtaining gametocytes for the majority of Plasmodium species and lineages. To conduct avian malaria infectivity assays for those Plasmodium spp. and lineages that are refractory to in vitro cultivation, it is necessary to obtain and preserve for short periods sufficient viable merozoites to infect naïve donor birds to be used as gametocyte donors to infect mosquitoes. Currently, there is only one described method for long-term storage of Plasmodium spp.—infected wild avian blood and it is reliable at a parasitaemia of at least 1 %. However, most naturally infected wild-caught birds have a parasitaemia of much less that 1 %. To address this problem, a method for short-term storage of infected wild avian blood with low parasitaemia (even ≤0.0005 %) has been explored and validated.MethodsTo obtain viable infective merozoites, blood was collected from wild birds using a syringe containing the anticoagulant and the red blood cell preservative citrate phosphate dextrose adenine solution (CPDA). Each blood sample was stored at 4 °C for up to 48 h providing sufficient time to determine the species and parasitaemia of Plasmodium spp. in the blood by morphological examination before injecting into donor canaries. Plasmodium spp.—infected blood was inoculated intravenously into canaries and once infection was established, Culex stigmatosoma, Cx. pipiens and Cx. quinquefasciatus mosquitoes were then allowed to feed on the infected canaries to validate the efficacy of this method for mosquito vector competence assays.ResultsStorage of Plasmodium spp.—infected donor blood at 4 °C yielded viable parasites for 48 h. All five experimentally-infected canaries developed clinical signs and were infectious. Pathologic examination of three canaries that later died revealed splenic lesions typical of avian malaria infection. Mosquito infectivity assays demonstrated that Cx. stigmatosoma and Cx. pipiens were competent vectors for Plasmodium cathemerium.ConclusionsA simple method of collecting and preserving avian whole blood with malaria parasites of low parasitaemia (≤0.0005 %) was developed that remained viable for further experimental bird and mosquito infectivity assays. This method allows researchers interested in conducting infectivity assays on target Plasmodium spp. to collect these parasites directly from nature with minimal impact on wild birds.

Intraoperative autologous blood transfusion of peritoneal blood during laparotomy for ectopic pregnancy: prospective study.

The purpose of this study was to evaluate the efficacy and safety of intraoperative autologous blood transfusion during laparotomy for hemoperitoneum in ectopic pregnancy and also safety of homologous blood transfusion along with autologous blood transfusion. Fresh blood, from peritoneal cavity, was collected for autotransfusion in sterile dish, filtered through eight layers of sterile gauze pieces, and collected in a sterile bowl. The collected blood was transferred into blood infusion bag containing citrate phosphate dextrose adenine solution in the proportion of five parts of blood to one part of citrate solution. Mean volume of autologous blood transfused in patients without homologous transfusion was 573±328. Mean preoperative hemoglobin was 4.95±1.5, and postoperative hemoglobin was 6.85±1.3. Hence, rise in hemoglobin was 1.9 g%. Autologous blood volume transfused in 29 patients (who required homologous blood transfusion) was 488±216. Preoperative hemoglobin was 4.35±1.94. The result was compared with other studies. Intraoperative autologous blood transfusion enabled the performance of laparotomy in hemodynamically unstable ectopic pregnancy patients without availability of homologous blood transfusion. Homologous blood transfusion is compatible with autologous blood transfusion.

Hemolytic characteristics of oxygenators during clinical extracorporeal membrane oxygenation.

A connection was previously reported between the hemolytic characteristics associated with oxygenators and the pressure drop measurements in the blood chamber under experimental conditions simulating their use in cardiopulmonary bypass. We examined this association during extracorporeal membrane oxygenation (ECMO) conditions. Three oxygenators for ECMO or pediatric cardiopulmonary bypass (Menox EL4000, Dideco Module 4000, and Mera HPO-15H) were evaluated. Fresh blood from healthy Dexter strain calves anticoagulated with citrate phosphate dextrose adenine solution was used. The blood flow was fixed at 1 L/min, similar to that in ECMO. The Normalized Index of Hemolysis for Oxygenators (NIHO) has been modified according to the American Society of Testing and Materials standards, as was previously reported. The NIHO value was the lowest in the Menox (0.0070+/-0.0009) and increased from Menox to Dideco (0.0113+/-0.0099) to Mera (0.0164+/-0.0043); however, there were no significant differences among the oxygenators. This NIHO value has a close correlation to the pressure drop. In conclusion, this evaluation method is also applicable to comparison of the biocompatibility performance of different types of clinically available oxygenators for ECMO.

HEMOLYTIC CHARACTERISTICS OF CLINICALLY ACCEPTED OXYGENATORS DURING ECMO

Specific aim; A connection was previously reported between the hemolytic characteristics associated with oxygenators and the pressure drop measurements in the blood chamber under experimental conditions simulating their use in cardiopulmonary bypass (CPB). At this time, we examined this association during extracorporeal membrane oxygenation (ECMO) condition. Method; Three oxygenators for ECMO or pediatric CPB (Menox EL4000®, Dideco Module 4000®, and Mera HPO-15H®) were evaluated. Fresh blood from healthy Dexter strain calves anti-coagulated with citrate phosphate dextrose adenine solution was used. The blood flow was fixed at 1 L/min similar to ECMO. The Normalized Index of Hemolysis for Oxygenators (NIHO) has been modified according to the American Society of Testing and Materials (ASTM) standards as we previously reported. Result; The NIHO value was the lowest in the Menox (0.0070 ± 0.0009) and increased from Menox <Dideco (0.0113 ± 0.0099) <Mera (0.0164 ± 0.0043); however, there were no significant differences among the oxygenators. This NIHO value has a close correlation to the pressure drop. Conclusion; This evaluation method is also applicable to compare the biocompatibility performance of different types of clinically available oxygenators for ECMO usage.

Decreased Hemolysis and Lipid Peroxidation in Blood during Storage in the Presence of Nicotinic Acid

Background and Objectives: There is increase in lipid peroxidation with consequent increase in hemolysis when blood is stored in di–(2–ethyl hexyl)phthalate (DEHP) plasticized bags. Studies carried out by us and others have indicated the ability of red cells to synthesize NAD⁺ from added nicotinic acid. Apart from the role of NAD⁺ in glycolysis, NADPH is required for reduction of oxidized glutathione to its reduced form by glutathione reductase. Reduced glutathione is an important antioxidant, which protects cell membrane from oxidative damage. Reduced glutathione is also involved in the regeneration of vitamin E, another important membrane antioxidant. In view of these, a study was undertaken to find out the effect of addition of nicotinic acid to the citrate–phosphate–dextrose–adenine (CPDA) solution on lipid peroxidation and integrity of red cells when whole blood is stored in DEHP plasticized bags. Materials and Methods: Blood was collected in Penpol blood storage bags (which is a DEHP plasticized bag) in CPDA solution in the presence and absence of nicotinic acid. Various parameters of lipid peroxidation and membrane stability – level of malondialdehyde (MDA), conjugated dienes, vitamin E, reduced glutathione, plasma Hb and K⁺, levels of adenosine triphosphate (ATP) and 2,3–diphosphoglycerate (2,3–DPG) were studied in the blood samples after various periods. Results: Plasma Hb and K⁺ concentrations were significantly lower in the presence of added nicotinic acid both after 28 and 42 days. Concentration of MDA and conjugated dienes was lower and the levels of reduced glutathione and vitamin E higher in the presence of nicotinic acid. ATP levels were not significantly different, but 2,3–DPG levels were higher. pH of the blood was nearer to 7.0 in the presence of nicotinic acid, while leaching out of DEHP into the blood was significantly lower. Conclusion: Inclusion of nicotinic acid in the CPDA solution has a beneficial effect in that (1) it reduces plasma Hb and K⁺; (2) reduces lipid peroxidation and increases antioxidant protection; (3) maintains pH nearer to 7.0, and (4) decreases the leaching out of DEHP into the blood.

Decreased Hemolysis and Lipid Peroxidation in Blood during Storage in the Presence of Nicotinic Acid

Abstract Background and Objectives: There is increase in lipid peroxidation with consequent increase in hemolysis when blood is stored in di‐(2‐ethyl hexyl)phthalate (DEHP) plasticized bags. Studies carried out by us and others have indicated the ability of red cells to synthesize NAD+ from added nicotinic acid. Apart from the role of NAD+ in glycolysis, NADPH is required for reduction of oxidized glutathione to its reduced form by glutathione reductase. Reduced glutathione is an important antioxidant, which protects cell membrane from oxidative damage. Reduced glutathione is also involved in the regeneration of vitamin E, another important membrane antioxidant. In view of these, a study was undertaken to find out the effect of addition of nicotinic acid to the citrate‐phosphate‐dextrose‐adenine (CPDA) solution on lipid peroxidation and integrity of red cells when whole blood is stored in DEHP plasticized bags. Materials and Methods: Blood was collected in Penpol blood storage bags (which is a DEHP plasticized bag) in CPDA solution in the presence and absence of nicotinic acid. Various parameters of lipid peroxidation and membrane stability – level of malondialdehyde (MDA), conjugated dienes, vitamin E, reduced glutathione, plasma Hb and K+, levels of adenosine triphosphate (ATP) and 2,3 diphosphoglycerate (2,3‐DPG) were studied in the blood samples after various periods. Results: Plasma Hb and K+ concentrations were significantly lower in the presence of added nicotinic acid both after 28 and 42 days. Concentration of MDA and conjugated dienes was lower and the levels of reduced glutathione and vitamin E higher in the presence of nicotinic acid. ATP levels were not significantly different, but 2,3‐DPG levels were higher. pH of the blood was nearer to 7.0 in the presence of nicotinic acid, while leaching out of DEHP into the blood was significantly lower. Conclusion: Inclusion of nicotinic acid in the CPDA solution has a beneficial effect in that (1) it reduces plasma Hb and K+; (2) reduces lipid peroxidation and increases antioxidant protection; (3) maintains pH nearer to 7.0, and (4) decreases the leaching out of DEHP into the blood.