Up to 21-day banked red blood cells collected by apheresis and stored for 14 days after automated wash at different times of storage

BACKGROUND AND OBJECTIVES: A closed-system technology (ACP-215, Haemonetics, Braintree, MA) enables automated washing and extended storage of frozen red blood cells (RBC). This technology was applied to wash banked RBC for removal of undesirable protein and metabolites before transfusion. We studied protein and metabolite depletion as well as RBC metabolism and viability up to 14 days postwash with regard to various pre-storage times. MATERIALS AND METHODS: Thirty RBC units were collected by means of apheresis and subdivided into three arms based on prewash storage time period (6 days/group 1, 14 days/group 2, 21 days/group 3). Wash efficacy (protein depletion, IgA), RBC metabolism (pH, lactate, potassium, haemolysis) and cell viability (ATP) were analysed immediately and 14 days after washing. RESULTS: Total protein and IgA postwash were lowered by automated wash in all groups and uniformly met EC guidelines. Potassium (mmol/l) was below 1.2 mmol/l postwash and significantly below prewash values in all groups, even after 14 days of storage (prewash vs. postwash; P < 0.05). RBCs washed after 14 and 21 days, respectively, showed significantly lower pH values and lower ATP content than RBCs washed after only 6 days of storage. Haemolysis rate remained significantly below 0.8%, the maximum level recommended by the EC guidelines, immediately and 14 days after washing in all units. CONCLUSION: Our data confirm that RBC units banked up to 21 days can be effectively protein- and potassium-depleted with the ACP-215 independent from prewash storage time. With respect to high ATP levels and pH, postwash storage of 2 weeks should be limited to units not older than 7 days before wash. This new washing technology ensures better standardization in washed RBC and provides blood centres with a logistical alternative to 24-h washed RBC products.     

Extended storage of whole blood before the preparation of blood components: in vitro effects on red blood cells in Erythro-Sol environment

Background and Objectives  Routine procedures for extended storage of whole blood (WB) before the preparation of blood components are of interest primarily for logistical reasons. We stored red cell units in either Erythro-Sol 2 (E-Sol 2, test units, 150 ml added) or in saline-adenine-glucose–mannitol (SAG-M) (reference units, 100 ml added) that were prepared after storage of WB at room temperature for 8, 12, 16 or 19 h after blood collection.
Study Design and Methods  Red blood cells were stored for 42 days. We measured pH, glucose, lactate, haemolysis, red blood cell adenosine triphosphate and 2,3-diphosphoglycerate on days 1, 7, 14, 21, 28, 35 and 42.
Results  Haematocrits were significantly lower in E-Sol 2 than in SAG-M due to the higher volume of E-Sol 2 added compared to SAG-M. Significantly reduced levels were found in E-Sol 2 of extracellular pH (throughout storage after 8-h hold and initially after 12-, 16- or 19-h hold), of lactate (initially after 8-h hold and throughout storage after 12-, 16- or 19-h hold), and of haemolysis from day 35 in the 8-h and on day 42 in the 12-h hold group. Sigificantly increased levels of adenosine triphosphate were seen in E-Sol 2 after 8-h hold (from day 14) and after 12-h hold (at days 21, 35 and 42) compared to SAG-M. Significantly higher concentrations of 2,3-diphosphoglycerate were noticed primarily after 8-h hold of WB.
Conclusion  The use of E-Sol 2 as a replacement for SAG-M does not significantly improve in vitro data after extended storage of WB at room temperature before preparation of blood components. However, after 8-h hold in vitro characteristics similar to or better than in fresh blood will be maintained for several weeks in E-Sol 2, a situation that makes E-Sol 2 superior to SAG-M when storage of WB is limited to 8 h. Some improvement was noted after 12-h hold as well.     

Modified formulation of CPDA for storage of whole blood, and of SAGM for storage of red blood cells, to maintain the concentration of 2,3-diphosphoglycerate

BACKGROUND AND OBJECTIVES: A dramatic decrease in the level of 2,3-diphosphoglycerate (2,3-DPG) takes place during the storage of whole blood (WB) in CPDA (citrate-phosphate-dextrose-adenine) and a similar decrease occurs during the storage of red blood cells (RBCs) in SAGM (saline-adenine-glucose-mannitol). The aim of the present study was to prevent this decrease by modifying CPDA and SAGM. MATERIALS AND METHODS: The pH of WB anticoagulant or RBC preservative solution was maintained at 7.6 by autoclaving the dextrose solution separately, by incorporating ascorbic acid and nicotinic acid into both CPDA and SAGM (to produce modified CPDA and SAGM solutions), and by reducing the concentration of adenine and adding citrate to the modified SAGM solution. The concentration of 2,3-DPG in WB after 28 days of storage in modified CPDA, and in RBCs stored in modified SAGM, was compared with that in WB or RBCs stored in unmodified solutions. RESULTS: The initial 2,3-DPG levels were maintained after 28 days in the modified formulations [10.63 +/- 2.58 microM/g of haemoglobin (Hb) in the case of modified CPDA and 12.07 +/- 1.47 microM/g of Hb in the case of modified SAGM], whereas in standard CPDA and SAGM solutions, the concentration of 2,3-DPG decreased to very low levels (0.86 +/- 0.97 microM/g Hb for CPDA and 0.12 +/- 0.008 for SAGM). CONCLUSIONS: Our modification in the formulation of CPDA or SAGM is effective in arresting the dramatic decrease in the level of 2,3-DPG that occurs during storage of WB and RBCs in unmodified solutions.     

Evaluation of a new, integral, whole blood filter (RS2000) system for prestorage leucodepletion of SAG-M red cells

Residual donor leucocytes are responsible for many adverse transfusion reactions. Prestorage leucodepletion may ameliorate these effects and enhance product quality. We studied a bottom and top (BAT) system incorporating an integral filter for whole blood leucodepletion. Our evaluation assessed leucodepletion efficiency as well as in vitro SAG-M red cell quality and storage characteristics.   Sixty-six units of blood were collected; test units into the Optipac^®- p L u S system and controls into the standard triple pack configuration. Test units were held for 4–6 h at room temperature (rt) or 12–18 h at 4°C. The mean leucocyte counts for the SAG-M red cells in the quality and storage trial were 0.6×10⁶ (rt hold), 0.05×10⁶ (4°C hold) and 2500×10⁶ (controls). We observed no significant differences between the groups for Na⁺, ATP, 2,3-DPG, glucose, lactate and pH during the 49 d storage. The control group, however, showed a greater increase in haemolysis and K⁺ with time. Autologous in vivo 24 h red cell recovery, after 42 d storage, was >75%. Adjustment of processing parameters in subsequent studies gave leucodepleted SAG-M red cells with minimal cell loss (9–19%) plus acceptable haemoglobin content (46–76 g/U) and haematocrit (54–62%). This system achieved >3.5 log leucodepletion with all but one unit containing <1×10⁶ leucocytes. The product quality is good and the system suitable for routine use in blood centres.     

Biochemistry of packed red blood cell concentrates stored in PAGGS-sorbitol solution for 42 days

We investigated metabolic, blood gas and acid-base balance modifications of erythrocyte concentrates resuspended in PAGGS-sorbitol solution during 6 weeks of storage. Glucose utilization was impaired during the last 2 weeks, while the intraerythrocytic ATP level decreased only 50% from the 1st to the 6th week. The K+ and Hb concentration in the medium showed a progressive increase which was more pronounced during the last 2 weeks. The decrease in pH progressed to reach 6.27 at the 6th week of storage. Intraerythrocytic 2,3-DPG dropped rapidly in the course of the 1st week and the percentage of oxyhemoglobin showed a remarkable increase after the 1st week (90%). Our data suggest that erythrocyte concentrates in PAGGS-sorbitol should be transfused, for optimal efficacy, within 4 weeks.     

Biochemistry of whole blood in poly(ethylene-co-ethylacrylate) experimental blood containers

The biochemical status of whole blood stored in containers fabricated of ethylene ethylacrylate (EEA) film was monitored at several times during 4 weeks of storage at 4 degrees C. Fifteen biochemical indicators were studied to reflect on erythrocyte integrity, cellular metabolism, plasma protein stability, and microaggregate formation. Comparison to storage in polyvinyl chloride (PVC) containers was made by distributing aliquots from each unit of blood among the containers being compared. Whole blood in EEA developed significantly higher levels of plasma hemoglobin, erythrocyte osmotic fragility, and D-glycerate-2,3-diphosphate (2,3-DPG), and somewhat greater glucose utilization, lactate production, and pH. These biochemical differences were not of great magnitude and the data suggest that EEA containers are compatible with the storage of whole blood.     

Improved Blood Preservation with 0.5CPD Erythro-Sol. Coagulation Factor VIII Activity and Erythrocyte Quality after Delayed Separation of Blood

Background and objectives: Delay between blood collection and the separation of its components may result in lowered yield of factor VIII (FVIII) and loss of 2,3-biphosphoglycerate (2,3-BPG). This study was to see whether the use of 0.5 CPD resulted in better preservation of FVIII and maintenance of 2,3-BPG. Materials and methods: 55 units of blood were collected in 0.5CPD and 48 in CPD SAG-M. Ten of the collections were paired, so that the same donors were bled a single session partly in an 0.5 CPD system and partly in CPD SAG-M. After collection, the blood was promptly cooled to 20°C and stored at that temperature for up to 24 h. Results: Preservation of FVIII activity was significantly better in 0.5CPD compared with CPD. The content of von Willebrand factor was stable in the anticoagulant solutions for 24 h at that temperature. Plasma separated from both media had low levels of prothrombin fragment 1 + 2 and complement activation. Paired collections substantiated previous reports that red cell storage is significantly improved in 0.5CPD compared with CPD SAG-M with respect to 2,3-BPG and haemolysis. Conclusions: Red cell metabolism and oxygen-releasing capacity are kept at acceptable levels in 0.5CPD blood for 24 h at 20°C before component separation. The concentration of red cell 2,3-BPG remained at normal or slightly subnormal levels during further storage in 0.5CPD at 4°C for 2–4 weeks before gradual decay to an average of 39% at 48 days.     

Clinical and Laboratory Experience with Erythrocyte and Platelet Preparations from a 0.5CPD Erythro-Sol Opti System

Background: Red blood cells stored as concentrates or suspensions in additive solutions change rapidly their oxygen affinity mainly due to the loss of 2,3-diphosphoglycerate (2,3-DPG). When collected in CPD with half of the normal concentration of citrate and citric acid (0.5CPD) and stored in a new additive solution (Erythro-Sol), 2,3-DPG is better maintained. No studies of the oxygen affinity of red cells stored under these conditions have been published. In Erythro-Sol, red cells have a satisfactory in vivo recovery for 49 days but the conditions after 28 days, within which time most red cell units are transfused, have not been investigated. Of importance is also to be able to make platelet concentrates (PCs) from 0.5CPD blood. Little data are available concerning the clinical usefulness of platelets prepared from 0.5CPD buffy coats (BCs). Methods: Blood was collected in 0.5CPD, held at 20°C for 3–4 h, then separated with the bottom-and-top technique into red cells, plasma and BC. In a storage experiment with 6 U the 2,3-DPG and P₅₀ values were determined weekly and a number of in vitro parameters were tested on day 28. In 6 donors the in vivo recovery and survival of red cells were determined using a single-chromium technique. Transfusions of 212 0.5CPD-Erythro-Sol red cell units were given to hematological patients under supervision. PCs derived from pools of 0.5CPD BCs suspended in PAS2 (T-Sol) were transfused to 20 thrombocytopenic patients and compared with CPD-BC-PCs suspended in PAS1. Corrected count increments (CCI) were determined. Results: The erythrocyte 2,3-DPG and P₅₀ values were normal or slightly subnormal initially but increased to supernormal levels during the 1 week, and remained at these levels for a further 1–3 weeks; the 2,3-DPG was two thirds of normal after 28 days, the P₅₀ was 3.72±0.28 kPa after 14 days and 2.84±0.41 after 28 days (mean ± SD). The P₅₀ values corresponded closely (r²= 0.903) to 2,3-DPG. The in vivo recovery of 4-week-stored red cells was 89.6±5.5% and the T₅₀ was 32.2±2.0 days. No adverse effects were observed in the transfusions. The CCI values did not differ between test and control groups; in both, 3- to 5-day-stored PCs gave lower CCI than fresh (0–2 days) PCs. Patients with acute myeloid leukemia AML (n = 11) had significantly lower CCI values than patients with myelodysplastic syndrome, myeloma and lymphoma (n = 9; CCI_{1 h}: p = 0.001; CCI_{24 h}: p = 0.006). Conclusions: Red cells stored in Erythro-Sol sustain a normal or slightly lowered oxygen affinity for 2–4 weeks, their viability is excellent, and they are well tolerated in clinical transfusions. Platelets prepared from 0.5CPD-BCs cause CCl_s of the same magnitude as CPD-BCs.     

In vitro quality of platelets during prolonged storage after washing with three platelet additive solutions

Background and Objectives Patients with anaphylactic transfusion reactions require washed platelet concentrates (PCs) for subsequent platelet (PLT) transfusions. New PLT additive solutions (PASs) contain substances that might be beneficial for the preservation of PLT function during storage. This study compares the quality of PLTs washed and stored with T‐Sol, Composol or SSP+. Study Design and Methods Fifteen buffy coats were pooled and divided into three parts. PCs with 30% plasma and 70% PAS (T‐Sol, Composol or SSP+) were prepared. Washing was performed on day 5 of storage. Ten PCs were prepared and washed with each PAS. In vitro variables including haemostatic function (clotting time and clot retraction) were analysed on day 5 before, directly after and up to 2 days after washing. Results Swirling was well preserved, and pH was within acceptable limits (6·4–7·4) during storage for all PASs. The PLT number was reduced by washing for all PASs, and T‐Sol PCs had a further decrease during storage. PLTs in T‐Sol were spontaneously more activated and had lower capacity to respond to an agonist than Composol or SSP+ PLTs. The haemostatic function was only slightly changed by washing and during postwashing storage. Conclusion PLTs washed with T‐Sol, Composol or SSP+ had good in vitro quality for two days after washing despite absence of glucose. PLTs in T‐Sol were more affected by the washing procedure and subsequent storage than Composol or SSP+ PLTs as judged by higher spontaneous activation.     

Red Cell ATP and 2,3-Diphosphoglycerate Concentrations as a Function of Dihydroxyacetone Supplementation of CPD Adenine

Abstract. Units of CPDA-1 whole blood were subdivided and each treated with additions of dihydroxyacetone (DHA) to give final concentrations from 0 to 80m M . The 'optimum' concentration of DHA to maintain 2,3-diphosphoglycerate (2,3-DPG) with minimal loss of ATP during 42 days of storage appeared to be 30m M of DHA. With this formulation, red cell 2, 3-DPG concentrations rose to 130–140% of normal by 14 days and then decreased in a near-linear manner to 50–60% normal by 42 days, while maintaining adequate ATP levels. In addition, packed red cells were prepared from CPD fresh blood and treated with adenine, glucose, and various concentrations (0-80m M of DHA. The cells also responded most favorably to 30m M DHA, although the response was not as positive as whole blood. This concentration of DHA produced nearly 100% maintenance of 2,3-DPG at 14 days with subsequent fall to 30% of normal by 42 days.     

Effects of 2,3-diphosphoglycerate on the mechanical properties of erythrocyte membrane

Investigation by Schindler et al and Sheetz and Casaly have indicated that high (approximately 10 mmol/L) concentrations of 2,3- diphosphoglycerate (2,3-DPG) have a destabilizing effect on erythrocyte membrane and the membrane skeleton. We have investigated changes in the membrane mechanical properties that occur at elevated 2,3-DPG levels in both intact cells and ghosts. The membrane shear modulus, viscoelastic recovery time constant, critical force, "plastic" viscosity, and material relaxation time constant were measured by standard micropipette and flow channel techniques. Intact cells showed no change in properties at physiologic ionic strength and 2,3-DPG concentrations of about 20 mmol/L, except for an increase in membrane viscosity resulting from an increased cellular hemoglobin concentration that occurs when the 2,3-DPG concentration is elevated. At ionic strengths 20% below physiologic and 2,3-DPG concentrations of approximately 20 mmol/L, decreases in membrane shear modulus and membrane viscosity were observed. In ghosts, no changes in these properties were observed at a 2,3-DPG concentration of 10 mmol/L and ionic strengths as low as 25% below physiologic, but a decrease in the force required to form tethers (critical force) was observed at physiologic ionic strength. The decrease in membrane shear modulus and viscosity of intact cells and the reduced critical force in ghosts are consistent with the results of other investigators. However, the difference in the effects of 2,3-DPG on ghosts and intact cells indicates that the effects of 2,3-DPG depend strongly on the conditions of the experiment. It appears unlikely that 2,3-DPG affects erythrocyte membrane material properties under physiologic conditions.     

Cytokine generation in whole blood, leukocyte-depleted and temporarily warmed red blood cell concentrates

BACKGROUND AND OBJECTIVES: It has been suggested that inflammatory cytokines such as Interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha (TNF-alpha) and IL-8 might be responsible for a large number of non-antibody-mediated adverse reactions to the transfusion of blood components, especially of platelet concentrates (PCs). The aim of this study was to compare the levels of proinflammatory cytokines in different blood components containing red cells such as buffy-coat-free packed red cells (RBCs), filtered RBCs and whole blood (WB) during storage under several conditions. MATERIALS AND METHODS: WB (CPD-A1, n = 16) was stored for 35 days at 2-6 degrees C; samples were taken on days 0, 21 and 35. Buffy-coat-poor RBCs in additive solution PAGGS-M (n = 16) were divided into halves, one half was leukocyte (WBC)-depleted by filtration on day 0, both halves were stored for 49 days at 2-6 degrees C (samples: days 0, 21, 49). Furthermore, buffy-coat-poor, unfiltered SAG-M RBCs (n = 16) were halved immediately after production and stored at 2-6 degrees C until day 42 (samples: days 0, 21, 42). One half remained at room temperature for 24 h on day 3. Cytokine levels were determined with commercial enzyme-linked immunosorbent assays. RESULTS: Levels of IL-1beta and TNF-alpha rose during storage of WB and RBCs. IL-6 could be detected markedly above the detection threshold in WB only. At the end of storage, we detected IL-8 in 1 of 16 units of WB tested, in 10 of 16 standard PAGGS-M RBCs and in 15 of 16 temporarily warmed SAG-M RBCs. Prestorage filtration of RBCs prevented the accumulation of IL-1beta and TNF-alpha. Temporarily warming of RBCs for 24 h did not cause any substantial increase in cytokine levels other than IL-8. RBCs stored in different additive solutions (PAGGS-M versus SAG-M) showed similar cytokine concentrations during storage. The cytokine content of WB was very similar to that of buffy-coat-poor RBCs. CONCLUSION: Cytokine levels measured in WB and buffy-coat-poor RBCs result in levels which are unlikely to cause febrile reactions even in the case of massive transfusion. We conclude that, according to present knowledge, there is no reason for prestorage filtration of buffy-coat-poor RBCs or WB to avoid febrile transfusion reactions due to cytokine accumulation during storage.     

Cellular properties of human erythrocytes preserved in saline-adenine-glucose-mannitol in the presence of L-carnitine

L-Carnitine (LC) in the preservation medium during storage of red blood cells (RBC) can improve the mean 24-hr percent recovery in vivo and increase RBC life-span after reinfusion. The purpose of the study was to investigate the differences in the biochemical properties of RBCs stored in the presence or absence of LC, and the cell-age related responses to storage conditions and to LC. RBC concentrates in saline-adenine-glucose-mannitol (SAG-M) were stored in the presence or absence of 5 mM LC at 4 degrees C for up to 8 weeks. RBC subpopulations of different densities were prepared by centrifugation on Stractan density gradient. Cells were sampled at 0, 3, 6, and 8 weeks, and hematological and cellular properties analyzed (MCV, MCHC, 4.1a/4.1b ratio as a cell age parameter, intracellular Na(+) and K(+)). After 6 weeks, MCV of RBC stored in the presence of LC was lower than that of controls (6 weeks MCV: controls 95.4 +/- 1.8 fl; LC 91.5 +/- 2.0 fl; n = 6; P < 0.005). This was due to swelling of control cells, and affected mainly older RBCs. LC appeared to reduce or retard cell swelling. Among the osmotically active substances whose changes during storage could contribute to cell swelling, only intracellular Na(+) and K(+) differed between stored control RBCs and LC-treated cells. LC reduces the swelling of older cells during storage at 4 degrees C in SAG-M, possibly by acting on the permeability of cell membrane to monovalent cations.     

Killing of fabric-associated bacteria in hospital laundry by low-temperature washing

Hospitals using 71.1 C water for laundering consume vast amounts of energy. We studied whether washing at 22 C would result in fabric-associated bacterial counts significantly different from those remaining after the high-temperature wash procedure in general use. Using a standard method to enumerate fabric-associated bacteria, we found that soiled sheets and terry cloth items were contaminated, respectively, with 10(6) and 10(8) cfu/100 cm2 of fabric area, predominantly gram-negative rods (especially Enterobacteriaceae and Pseudomonadaceae). Staphylococcus species were the most common gram-positive organisms. A standard low-temperature washing cycle without laundry chemicals removed 3 log10 of bacteria by agitation, dilution, and drainage. When low-temperature laundry chemicals were used, 3 log10 of bacteria were killed after the bleach was added, and sheets and terry cloth items had postwash colony counts of 10(1)-10(2) cfu/100 cm2. Drying removed an additional 1-2 log10 organisms. Bacterial counts and species from low- and high-temperature washed fabrics were comparable. Low-temperature washing is therefore as effective as high-temperature washing for eliminating pathogenic bacteria from hospital laundry.     

Evaluation of Adenosine Triphosphate and 2,3-Diphosphoglycerate after Twenty-Four Hours in Red Cells Washed for Neonatal Transfusion

Adenosine triphosphate and 2,3-diphosphoglycerate concentrations in 14 U of CPDA-1 stored red cells (PRBC) and washed red cells (WRBC) were measured to assess indirectly the quality of WRBC for neonatal transfusion after 24 h. The results indicate that there is no difference in red cell ATP and 2,3-DPG concentrations between PRBC and WRBC after a 24-hour period.     

Hemoglobin function in stored blood

Summary Inorganic phosphate which is known to stimulate red cell glycolysis is present in one of the preservatives for storing whole blood, citrate-phosphate-dextrose (CPD), but not the other, acid-citrate-dextrose (ACD). Both of these preservatives for liquid storage were developed before 2,3-diphosphoglycerate (2,3-DPG) was found to be necessary for normal hemoglobin function. In a recent study we have shown that very high concentrations of phosphate (10, 15, and 20 mM) were deleterious for maintaining 2,3-DPG. In the present study a lower range of phosphate concentrations (2, 4, 6, and 8 mM) was studied for maintenance of 2,3-DPG and ATP during storage under blood banking conditions. The lowest concentration, 2 mM, which corresponds to CPD was found to be the best concentration for maintaining 2,3-DPG and thus hemoglobin function. Four mM phosphate was not quite as good but better than no phosphate. Six and 8 mM phosphate were considerably worse.

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Zusammenfassung Im Anschluß an vorangegangene Arbeiten wurde die Anwendung von geringen Phosphatmengen (2–8 mM) zur Stabilisierung von 2,3-DPG und ATP während der Lagerung von Blut unter Blutbankbedingungen geprüft, um die Hämoglobinfunktion aufrecht zu erhalten. Am besten bewährte sich die 2mM-Konzentration des anorganischen Phosphats, während die 4mM-Konzentration weniger wirkungsvoll war. 6–8 mM-Konzentrationen wurden als ungeeignet befunden, die Stabilisierung von 2,3-DPG und ATP zu gewährleisten.

     

Erythrocyte 2,3-Diphosphoglycerate in Liver Diseases

The erythrocyte 2,3-diphosphoglycerate (2,3-DPG) was studied in patients with liver diseases, chronic obstructive pulmonary disease, and in normal subjects. The level of 2,3-DPG in liver diseases occurred in the following increasing order: chronic persistent hepatitis, chronic active hepatitis, liver cirrhosis, and cirrhosis with hepatocellular carcinoma. A significant negative correlation between the 2,3-DPG concentration and serum albumin concentration was found in the liver diseases. The 2,3-DPG level was correlated to the serum concentration of total bile acids and to the arterial blood pH. A negative correlation was found between the arterial blood pH and the serum albumin concentration. The level of 2,3-DPG in hepatocellular carcinoma and/or liver cirrhosis was higher than that in more hypoxic chronic obstructive pulmonary disease. And an increased level of 2,3-DPG was also shown in nonhypoxic patients with liver diseases. These results suggest that the level of erythrocyte 2,3-DPG increases according to the severity of the liver disease, and compared to the level in hypoxic chronic obstructive pulmonary disease, the level of erythrocyte 2,3-DPG is higher in both hepatocellular carcinoma and liver cirrhosis.     

The oxygen transport system of red blood cells during diabetic ketoacidosis and recovery

Daily evaluations of 8 newly detected ketoacidotic diabetics showed the Bohr-effect of haemoglobin to be decreased by 50% while erythrocyte 2,3-DPG was decreased below 10 mumoles/g Hb. 2,3-DPG correlated strongly with pH during acidosis and with plasma inorganic phosphate (Pi) subsequently to the first insulin administration. Oxygen affinity of haemoglobin, measured as P50 act pH, was unchanged in ketoacidosis compared to the time, however, P50 act pH fell striking (p less than 0.001) and remained decreased up to 7 days depending upon the resynthesis of 2,3-DPG in relation to Pi. The Hill-coefeficient in reflecting the slope of the oxygen dissociation curve was diminished in ketoacidosis (p less than 0.005), and decreased further after pH-normalization (p less than 0.005). There was a close association of n with 2,3-DPG (p less than 0.001) and additionally with Pi at 2,3-DPG-levels below 10 mumoles/g Hb. Based on these findings a decreased erythrocyte oxygen release of one fifth during acidosis and more than one third after pH-correction can be hypothesised. In view of the intimate relation of Pi to the oxygen transport system it is suggesed that treatment of ketoacidosis should include Pi-sugstitution.     

Erythrocyte glycolysis and its marked alterations by muscular exercise in type VII glycogenosis

Levels of erythrocyte glycolytic intermediates after the phosphofructokinase (PFK) step, including 2,3-bisphosphoglycerate (2,3- DPG), were decreased at rest in patients from separate families with type VII glycogenosis. The concentration of 2,3-DPG was about half of the normal control value during a period of unrestricted daily activity but was further decreased to one third of normal after a one-day bed rest. Mild ergometric exercise rapidly increased the levels of fructose- 1,6-bisphosphate, dihydroxyacetone phosphate plus glyceraldehyde-3- phosphate, and 2,3-DPG in patients' circulating erythrocytes but did not in those of normal subjects. This indicated that a crossover point at the PFK step in glycolysis disappeared after physical exercise and, consequently, the 2,3-DPG concentration, which had decreased because of blockage of the PFK step, was restored considerably. This apparently exercise-related alteration in intermediary metabolism at the beginning of glycolysis was reproduced in vitro by incubating normal erythrocytes in the presence of inosine or ammonia, both of which have increased levels in circulating blood during and after exercise in this disorder. We conclude that physical activity in addition to a genetic deficiency in erythrocyte PFK affects glycolysis in erythrocytes in type VII glycogenosis and that myogenic factors released from exercising muscles may be responsible for this change.     

Storage of Whole Blood for up to 24 Hours at Ambient Temperature prior to Component Preparation

The effect of rapid cooling to 20-24 degrees C of whole blood immediately after collection, using 'cooling units' with butane-1,4-diol and prolonged storage up to 24 h at ambient temperature was investigated in the whole blood and the subsequently prepared plasma, buffy coat and buffy-coat-poor red cell concentrate (BC-poor RCC) in saline-adenine-glucose-mannitol (SAG M) solution. Factor VIII:C content of the plasma (n = 10), after 24 h storage was 80 +/- 3% of the initial value. In routine procedures factor VIII:C content in the plasma (n = 129 pools of 20 donor units plasma) was 0.77 +/- 0.078 IU/ml, after storage of the whole blood for 16-20 h. In whole blood (n = 10), the 2,3-diphosphoglycerate (2,3-DPG) content of the red cells decreased from 4.36 +/- 0.55 to 1.47 +/- 0.6 mumol/ml red cells after 24 h storage at 20-24 degrees C. After storage of the BC-poor RCC (n = 10) at 2-6 degrees C for 1 week, the 2,3-DPG had dropped to 0.76 +/- 0.46 mumol/ml red cells. During the first 24 h of storage of whole blood, the adenine triphosphate (ATP) levels of the red cells remained stable. A mean increase of 20% of the initial value was observed after addition of SAG M solution. In the BC-poor RCC the ATP slowly decreased to 81 +/- 5% after 5 weeks and to 68 +/- 6.6% of the initial value after 6 weeks storage.(ABSTRACT TRUNCATED AT 250 WORDS)