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PubMed1979

Complement components detected on normal red blood cells taken into EDTA and CPD.

Normal red blood cells (RBC) from fresh EDTA and CPD blood and from stored CPD blood were examined for the presence of bound subcomponents of C3 and C4. By serologic agglutination tests, only C3d was detectable on the cells. Incubation in compatible fresh normal serum (FNS) at 37 degrees C appeared to increase the amount of 3Cd on the RBC. C3b was serologically detectable only on stored CPD cells and only after incubation in compatible FNS. No. C4 components were detected on the cell surfaces in agglutination tests. Using an indirect labeling technique, small, but significant, amounts of C3d and C4d were found on all three types of untreated cells. C3b was present on stored CPD cells only. The indirect labeling technique showed a significant increase in C3d and C4d on all cells following incubation i- compatible FNS, whereas bound C3b was significantly increased only with stored CPD cells. There was no increase in bound C4b following serum incubation. The average number of C3d molecules per cell on normal EDTA cells was 557 and average Ko was 3.6 x 10(7) l/mol.

Animals↗
PubMed1993

Half-strength citrate CPD combined with a new additive solution for improved storage of red blood cells suitable for clinical use.

Currently used systems for red blood cell (RBC) collection and storage for transfusion have the disadvantage that the RBC 2,3-bisphosphoglycerate (BPG) concentration is depleted within two weeks of storage, resulting in a left-shift of the oxygen dissociation curve and a temporarily impaired capacity to deliver oxygen. We have studied the effects on red cell metabolism, morphology and in vivo recovery of 49-day storage of RBC, with collection in half-strength citrate CPD (0.5CPD) and storage in an additive solution containing citrate, adenine, mannitol, phosphate and glucose (RAS2). Traditional CPD-SAGM was used for comparison. Component preparation was performed after an initial holding period of the whole blood at ambient temperature for 8 h. The BPG concentration in 0.5CPD-RAS2 RBC was 0.633 +/- 0.120 mol (mol Hb)-1 as compared to 0.454 +/- 0.138 mol (mol Hb)-1 in CPD-SAGM RBC which implied a decrease to 67 and 48% of normal concentration, respectively. The mean RBC BPG concentration was maintained at the initial level for 28 days in the new system but decreased to very low levels within 14 days in the controls. The total adenine nucleotides were well maintained in both systems, adenosine triphosphate slightly better in the new system. Hemolysis after 49 days was 0.35 +/- 0.21% in the new system and 0.72 +/- 0.25% in the controls (p < 0.001). The morphology was better maintained in the new system (p < 0.001). The 24-hour posttransfusion survival of 49-day stored RBC was 78.9 +/- 7.1%. The membrane leakage of sodium and potassium was not significantly different in the two systems.(ABSTRACT TRUNCATED AT 250 WORDS)

2,3-Diphosphoglycerate↗
PubMed1976

Oxygenation properties and intraerythrocytic constituents of human blood when stored in different media of ACD and CPD.

In the blood stored in acid-citrate-dextrose solution (ACD blood), the oxygen affinity and red cell 2, 3-diphosphoglycerate (2, 3-DPG) content showed parallel exponential decays with half-lives of 3 to 4 days. In the blood stored in citrate-phosphate-dextrose solution (CPD blood), the two parameters increased during the first 4 days before showing the same decay as that seen in the ACD blood. There was no significant change in the transmembrane pH gradient of the red cells, and thus the intracellular pH at the plasma pH of 7.40 was always in the range of 7.17 +/- 0.02 throughout the period of storage in ACD medium. In both ACD and CPD blood, the Hill exponent n was always normal (approximately 2.8) while the Bohr coefficient (delta log P50/delta pH) rose along with the lapse of time for preservation. The oxygen affinity of the CPD blood was less influenced by the red cell 2, 3-DPG than was that of the ACD blood. This phenomenon was thought to derive from higher concentration of salts within the CPD-stored red cells. The efficiency of blood oxygen transport in ACD and CPD blood was compared.

Blood Preservation↗
PubMed1983

Storage of red cell concentrates in CPD-A2 for 42 and 49 days.

CPD-A2 is a modified CPD blood preservative with adenine, containing 1 1/2 times as much glucose as CPD. Units (450 ml) of blood from 21 normal donors were collected in CPD-A2 in plastic bags and held at room temperature for 8 hr. An 80% red cell concentrate was prepared and this was stored for 42 or 49 days at 4 degrees C, with the containers in either a standing or lying position. Measurements of glucose consumption, red cell ATP, and 2,3-DPG and of plasma hemoglobin, pH, Na+, and K+ were performed on all samples. The size of the "fragile tail" of osmotically fragile red cells was estimated in 12 samples. The poststorage 24 hr viability of their own stored 51Cr-tagged red cells was documented in 19 of the volunteers. At least 4 months after the original donation, a second unit of blood was collected from eight of the donors to make possible intradonor comparison of the biochemical effects of storage position. After 42 days but not after 49 days of storage, red cells in concentrates stored in the lying position had consumed more glucose and had a higher poststorage pH than did cells stored in the standing position. The poststorage 24 hr viability of red cells stored for 42 days averaged 83.6%, with all units exceeding 70% viability. At 49 days the average viability was 69.1%. Although the average viability of cells stored in the lying position for 42 days was higher than that of concentrates stored standing, the difference was not statistically significant at the 5% level. The plasma hemoglobin level showed a weak correlation with viability of stored cells. Red cell ATP levels were correlated with viability only at 42 days' and not at 49 days' storage. Concentrates of red cell collected in CPD-A2 manifested fully satisfactory viability for 42 days. At 49 days storage the results of viability studies were borderline. High plasma hemoglobin values are observed at both 42 and 49 days' storage and may limit the usefulness of red cell concentrates stored for prolonged periods of time.

2,3-Diphosphoglycerate↗
PubMed1993

Novel serine penicillocarboxypeptidase CPD-S3 from Penicillium janthinellum IBT 3991: purification, characterization, and uses in peptide synthesis and modification.

A novel carboxypeptidase (CPD-S3) from Penicillium janthinellum IBT 3991 has been isolated in a two-step purification procedure by cation exchange and affinity chromatography. The enzyme is a serine carboxypeptidase with a denatured molecular mass determined by SDS of 62 kDa of which 32% is carbohydrate. The isoelectric point is 5.1. CPD-S3 exhibits a high stability towards organic solvents and elevated temperatures. Besides the carboxypeptidase activity, CPD-S3 exhibits esterase, amidase, and carboxamidohydrolase activities. CPD-S3 favors substrates of L-configuration with basic amino acid residues in either P1 or P1', and particularly dibasic substrates and medium-sized straight-chain alkyl esters for hydrolysis. In aminolysis of esters, amino acid amides and hydrazines coupled in good yield, but methyl esters poorly, and unlike other carboxypeptidases, free amino acids could not be coupled or transpeptidation effected to form amides. In ester semisynthesis, peptides with neutral, but not basic, residues in P1 could be esterified. The scope of applicability for enzymatic peptide synthesis is limited.

Amides↗
PubMedDate not supplied

Blood preservation XVI packed red cell storage in CPD-adenine.

Interest has been renewed in CPD-adenine as a long-term liquid blood preservative. The question of whether the metabolic product of adenine, 2,8-dioxyadenine was toxic to humans has apparently been resolved by extensive animal and human studies in favor of there being no potential toxicity in the amounts used in blood preservation. Sweden is adopting CPD-adenine (0.25 mM) as its national blood preservative after ten years of clinical experience in trials. They have shown that each additional week of storage time beyond the current three weeks with CPD results in a 50 per cent reduction of wasteage caused by outdating. They are adopting the 35-day time for regular use with 42 days for an emergency reserve supply. However, many units of blood in the U.S. are stored as packed red blood cells and the question has been raised as to whether there is sufficient glucose in the preservative to maintain red blood cell metabolism in the packed cell unit. The present investigation indicates that there is sufficient glucose for 35 days of packed cell storage in CPD-adenine (0.25 mM) but in some units this might be marginal at 42 days of storage.

Adenine↗
PubMedDate not supplied

Blood storage XXIII: 2,3-DPG maintenance for six weeks in a CPD-adenine-inosine preservative with and without methylene blue.

In a pilot study the optimal concentration of inosine for 2,3-DPG maintenance in a CPD-adenine (0.25 mM) preservative was confirmed to be at lease 10 mM. In these experiments, 2,3-DPG maintenance was nearly normal for six weeks of storage in CPD-adenine-inosine (10 mM) preservative with or without methylene blue. The control preservative lacking inosine showed a statistically significant decrease in 2,3-DPG concentrations after the 3rd week. Finally, 2,3-DPG levels were significantly better maintained in CPD-adenine preservatives that contained 15 mM concentrations of inosine, whether methylene blue was present or not (10(-6)M), compared to CPD-adenine-inosine preservatives that contained 5 mM inosine, with or without methylene blue. The methylene blue effect, while it can be demonstrated in most experiments to help the red blood cell maintain 2,3-DPG during prolonged blood storage, is judges to be a slight value. However, inosine is of great value in maintaining 2,3-DPG for prolonged (five to six weeks) liquid storage.

Adenine↗
PubMedDate not supplied

Blood preservation XXVI, CPD-adenine packed cells: benefits of increasing the glucose.

In searching for the optimal glucose concentration, this lab has monitored ATP, 2,3-DPG, pH, and glucose levels of samples taken from full blood units stored for 6 weeks at 4 C. The blood was collected into CPD-adenine containing 100, 125, 150, 175, and 200 per cent of the glucose present in CPD. The units were stored as whole blood, soft packed (50 to 70% Hct), or hard packed units (80 to 95% Hct). ATP values in general did not decrease very greatly in whole blood units and only moderately in soft packed units. However, in hard packed units a steady progressive decrease in the ATP values was seen to begin at day 14. In these hard-packed units the only improvement with extra glucose was seen beginning at day 14 when ATP maintenance was better with 200 per cent glucose, but the improvement was not significant until day 42. However, at 35 days the ATP values for 200 and 175 per cent glucose were noticeably better than for the other preservatives. Therefore, it appears from this study that the glucose concentration in CPD-adenine for hard-packed cells should be at least 175 per cent of that in regularly formulated CPD. Also, there would appear to be an advantage of having 200 per cent glucose in those units of blood that may be stored beyond 35 days for emergency blood shortage times.

Acid-Base Equilibrium↗
PubMedDate not supplied

Preservation of neutrophils in CPD-adenine.

Human neutrophils were stored in vitro in CPD or CPD-adenine plasma under conditions designed to simulate those of leukapheresis followed by short-term preservation. Viability, morphology by light microscopy, and phagocytic ability of cells stored in either anticoagulant were similar when studied sequentially during 72 hours of storage. Thus, CPD-adenine offered no advantage, under the conditions employed, over standard CPD without adenine.

Adenine↗
PubMed1993

Half-strength citrate CPD and new additive solutions for improved blood preservation. I. Studies of six experimental solutions.

Poor stability of plasma factor VIII in whole blood and loss of erythrocyte 2,3-bis-phosphoglycerate (BPG) during red cell storage are limitations with systems for blood component preparation in current use. This study presents attempts to improve post-collection storage conditions in both these respects using half-strength citrate CPD solution (0.5CPD) for blood collection, which has been shown by others to improve the stability of factor VIII, and some compositions of hypotonic additive solutions for red cell storage containing citrate, adenine, mannitol, and phosphate. Guanosine was also included in some of the media. The erythrocyte BPG concentration was maintained at a normal level for 3-4 weeks with the best of the tested compositions. Total adenine nucleotide concentration was maintained at the original level for 49 days and adenosine triphosphate for 28 days. Spontaneous storage haemolysis was low, 0.31% (mean) +/- 0.08-0.10% (SD) after 49 days in the two best compositions. The intracellular pH was 0.2-0.3 pH units higher than the extracellular pH at the beginning of storage, but this difference gradually diminished and disappeared after 4-5 weeks. We suggest two likely explanations of the effects: the maintenance of intracellular pH at a level sufficiently high not to impair BPG synthesis until after several weeks of storage, and a sufficient supply of phosphate needed in the synthesis of organic phosphate compounds. The content of citrate was selected such that the total amount supplied to a patient in a massive transfusion, when using a combination of 0.5CPD plasma and red cell suspension, would be smaller than that provided by a transfusion of CPD whole blood.

2,3-Diphosphoglycerate↗
PubMed1975

Studies on citrate-phosphate-dextrose (CPD) blood supplemented with adenine.

The effect of varying adenine concentrations in citrate-phosphate-dextrose (CPD) blood was studied in an attempt to optimize the storage conditions for human erythrocytes with regard to posttransfusion viability and oxygen release function. The maintenance of diphosphoglycerate (DPG) was impaired by adenine supplementation; this effect was closely related to the adenine concentration. A 0.25 mM adenine concentration in CPD blood improved the adenosine triphosphate (ATP) levels and the posttransfusion viability markedly, without appreciably impairing the DPG maintenance. The results suggest that CPD solution supplemented with adenine to give a 0.25 mM concentration in the blood is a better preservative for human erythrocytes than the commonly used acid-citrate-dextrose (ACD), CPD, and ACD-adenine solutions with regard to posttransfusion viability and oxygen release function. Adenine addition to this low concentration is not expected to cause renal damage even after massive transfusion.

Adenine↗
PubMed2004

CPD-photolyase adenovirus-mediated gene transfer in normal and DNA-repair-deficient human cells.

Cyclobutane pyrimidine dimers (CPDs) are the most frequent and deleterious lesions generated in the mammalian genome after UV-C irradiation. The persistence of these lesions in DNA can be toxic and mutagenic, and also represents a specific signal to apoptosis. To investigate the CPDs repair in situ and consequent UV-induced apoptosis in human cells, we generated a recombinant adenovirus vector containing the gene encoding a CPD-photolyase-EGFP fusion protein (Adphr-EGFP). Adphr-EGFP-infected cells are proficient in photorepair, which prevents apoptotic cell death in comparison with samples kept in the dark, indicating that the fusion protein is functional in CPD recognition and removal. By using local UV irradiation, foci of the photolyase fusion protein were observed in UV-damaged areas of the nuclei in colocalization with NER enzymes. Phr-EGFP migration to CPD sites and redistribution after photorepair was followed, and shown to present similar kinetics in normal or DNA-repair-deficient cells. To our knowledge, this is the first report of an investigation of CPDs repair in situ employing a CPD-photolyase-EGFP enzyme. The Adphr-EGFP vector can be an informative tool to investigate the repair and cellular consequences of UV-induced lesions in primary human cells.

Adenoviridae↗
PubMed2005

CPD photolyase gene from Spinacia oleracea: repair of UV-damaged DNA and expression in plant organs.

The UV-B radiation contained in solar radiation has deleterious effects on plant growth, development and physiology. Specific damage to DNA caused by UV radiation involves the cyclobutyl pyrimidine dimers (CPD) and the pyrimidine (6-4) pyrimidone photoproducts. CPDs are repaired by CPD photolyase via a UV-A/blue light-dependent mechanism. The gene for the class II CPD photolyase has been cloned from higher plants such as Arabidopsis, cucumbers and rice. We isolated and characterized the cDNA and a genomic clone encoding the spinach class II CPD photolyase. The gene consisted of 3777 bases and 9 exons. The sequence of amino acids predicted from the nucleotide sequence of the cDNA of the gene was highly homologous to that of the higher plants listed above. When a photolyase-deficient Escherichia coli strain was transformed with the cDNA, photoreactivation activity was partially restored, by the illumination with photoreactivating light, resulting in an increased survival and decreased content of CPDs in the Escherichia coli genome. In both the male and female plants, the gene was highly expressed in leaves and flowers under the condition of 14-h light and 10-h dark cycle. The expression in the roots was quite low compared with the other organs.

Amino Acid Sequence↗
PubMed1984

High yield of platelet-rich plasma from CPD blood compared to ACD blood.

The yields of platelet-rich plasma (PRP) obtained by centrifugation of CPD (citrate-phosphate-dextrose) blood and ACD (acid-citrate-phosphate) blood were compared. The volumes of PRP from 5 ml blood in test tubes and 200 ml blood in bags were larger by 4% and 4.5%, respectively, when CPD was used as an anticoagulant. In addition, the number of platelets in PRP from CPD blood was higher than that from ACD blood. These data suggest that the optimal centrifugal condition for CPD blood should be different from that for ACD blood.

Blood Platelets↗
PubMed1984

High yield of platelet-rich plasma from CPD blood compared to ACD blood.

The yields of platelet-rich plasma (PRP) obtained by centrifugation of CPD (citrate-phosphate-dextrose) blood and ACD (acid-citrate-phosphate) blood were compared. The volumes of PRP from 5 ml blood in test tubes and 200 ml blood in bags were larger by 4% and 4.5%, respectively, when CPD was used as an anticoagulant. In addition, the number of platelets in PRP from CPD blood was higher than that from ACD blood. These data suggest that the optimal centrifugal condition for CPD blood should be different from that for ACD blood.

Blood Platelets↗
PubMed1990

Anti-retroviral effect of chlorophyll derivatives (CpD-D) by photosensitization.

A new photosensitizer, CpD(chlorophyll derivatives), previously reported as a promising agent for tumor therapy, was studied to determine its inhibitory effects on Gross leukemia virus(GLV), a mouse retrovirus isolated from the GLV-producing TGV cell line, and the cytocidal effect on the GLV infected cells in vitro, following photodynamic treatment with CpD-D and red light, the viral inactivation and infectivity were examined by measuring the reverse transcriptase(RT) activity of the virus itself and that in cell-free culture supernatant of freshly GLV-infected secondary mouse embryo cells respectively. The cytocidal activity was measured by trypan blue exclusion test. Inhibition of GLV associated RT activity resulted from CpD-D and red light treatment. The RT inhibition effect was immediate and the infectivity of these photodynamically treated GLV to mouse embryo cells was also inhibited. However, specific cytotoxicity of GLV infected cells was not found. Thus, it is concluded that CpD-D may be used as an effective antiviral agent.

AKR murine leukemia virus↗
PubMed1979

[Changes in ionized calcium and citrate levels in dogs during mechanical autotransfusion with heparin, ACD and CPD].

The control of ionized calcium (Ca++), total calcium, and citrate levels in serum were determined in dogs during autotransfusion (AT) of blood stabilized with heparin, ACD (formula B) and CPD. Blood samples were taken according to the changes of aortic pressure (AOP), which was continuously monitored. Taking the values during the stable phase of AOP preceding the AT as baseline, Ca++ dropped by 27% with ACD and by 34% with CPD at the maximum decrease of AOP immediately after the AT. The corresponding increase of citrate was 174% with ACD and 521% with CPD, while total calcium remained stable. Thus cardiac depression after AT of citrated blood seems to be mainly caused by the drop of Ca++, which is significantly more pronounced with CPD, corresponding to the higher content of citrate.

Animals↗
PubMed1995

Cross-sectional analysis of erythropoietin use in CPD: its relation to azotemic index clearances.

The association between the use of erythropoietin and urea or creatinine clearance was studied in two populations on continuous peritoneal dialysis (CPD) residing either at an altitude of 1600 m (n = 194) or at sea level (n = 108). Among peritoneal and total KT/V urea and creatinine clearance (CCr) indices, only total CCr was lower in the high altitude group receiving erythropoietin than in the corresponding group not receiving erythropoietin (68.0 +/- 34.9 vs 82.9 +/- 40.9 L/1.73 m2 weekly, p < 0.01). However, 24-hour urine volume and urinary KT/V urea and CCr were consistently lower in the groups receiving erythropoietin than in those not receiving erythropoietin. Total weekly KT/V urea < or = 1.70 and CCr < or = 52 L/1.73 m2 were considered indicators of inadequate CPD. Although the percent of patients receiving erythropoietin did not differ overall between groups with adequate and those with inadequate CPD, a trend towards more frequent use of erythropoietin was found in the sea level group with inadequate CCr versus the group with adequate CCr (28.2% vs 16.9%, p = 0.084). In CPD decreased renal function is associated with more frequent use of erythropoietin. Whether inadequate total urea or creatinine clearance is also associated with more frequent erythropoietin use requires further study.

Altitude↗