Cow red blood cells. I. Effect of purines, pyrimidines, and nucleosides in bovine red cell glycolysis

1979 ◽  
Vol 236 (5) ◽  
pp. C255-C261 ◽  
Author(s):  
M. J. Seider ◽  
H. D. Kim
Keyword(s):  
Pyruvate Kinase ◽  
Enzyme Activities ◽  
Blood Cells ◽  
Glycolytic Enzyme ◽  
Red Cells ◽  
The Other ◽  
Red Cell ◽  
Hexokinase Activity ◽  
Glycolytic Rate

Cow red cells, under in vitro incubation conditions, exhibit a comparatively low glycolytic rate of 0.56 +/- 0.05 micromol/(ml cells.h), with a ratio of lactate formed to glucose consumed of 1.58. It has been found that this low glycolytic rate can be stimulated 50--60% above the basal level in the presence of a variety of purine and pyrimidine compounds including adenosine, inosine, adenine, hypoxanthine, xanthine, and uracil. In contrast, calf red cells, which have a much higher glycolytic rate, display no discernible response to these agents. In attempts to elucidate the mechanism by which this stimulation takes place, both glucose transport and glycolytic enzyme activities were determined in the presence of these stimulators. Glucose influx in cow red cells, measured using the glucose analog 3-O-methyl-glucose, exhibits both a low Km of 117 microM and a Vmax of 0.38 micromol/(ml cells.min), and is unaltered in the presence of adenosine. On the other hand, hexokinase, which in normal hemolysates of cow red cells has an activity of 0.49 +/- 0.03 micromol/(g Hb.min). was found to be stimulated to 0.73 micromol/(g Hb.min) in the presence of adenine. Both pyruvate kinase and phosphofructokinase were unaffected by this compound. These data suggest that certain purines and pyrimidine compounds may exert their stimulatory effect on hexokinase activity, resulting in an augmentation of cow red cell glycolysis.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393 ◽  
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

Abstract The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals Metabolic-membrane coupling in red blood cells of trout: the effects of anoxia and adrenergic stimulation

1989 ◽  
Vol 143 (1) ◽  
pp. 149-164 ◽  
Author(s):  
R. A. Ferguson ◽  
R. G. Boutilier
Keyword(s):  
Blood Cells ◽  
Energy Production ◽  
Red Cells ◽  
Salmo Gairdneri ◽  
Red Cell ◽  
Adrenergic Stimulation ◽  
Tight Coupling ◽  
Metabolic Coupling ◽  
Stimulation Of

Under oxygenated conditions, in vitro, the highly aerobic red cells of the rainbow trout (Salmo gairdneri) exhibit tight coupling between energy (i.e. nucleotide triphosphate, NTP)-consuming and NTP-producing metabolic activity, as shown by strict maintenance of red cell NTP:haemoglobin ratios. This coupling is maintained following adrenergic stimulation of oxygenated red cells when the increased NTP demands of ion transporting systems are met by enhanced energy production via aerobic metabolism. In unstimulated anoxic red cells, membrane-metabolic coupling is preserved via the arrest of NTP-consuming processes. Adrenergic stimulation of anoxic red cells, however, leads to a functional uncoupling of membrane metabolism with the result that NTP levels decline rapidly. At this time, cellular [NTP] is negatively correlated with [Na+]i and [Cl-]i and positively correlated with [K+]i. This, in addition to the fact that the pH of the intracellular compartment is also highly dependent on cellular NTP levels, provides evidence for the integration of energy and membrane metabolisms.

Allosteric Hemoglobin Effector Loaded Erythrocytes Improve the Efficiency of Transfusion to Prevent Sickling.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1418-1418
Author(s):  
Vanessa Bourgeaux ◽  
Olivier Hecquet ◽  
Dominique Rigal ◽  
Alain Francina ◽  
Yann Godfrin
Keyword(s):  
Red Blood Cells ◽  
Experimental Model ◽  
Blood Cells ◽  
Red Cells ◽  
Red Blood Cell Transfusion ◽  
Dependent Manner ◽  
Red Cell ◽  
Transfusion Therapy ◽  
Blood Samples

Abstract Sickle cell disease (SCD) is characterised by abnormal haemoglobin S (HbS). Under hypoxic conditions, HbS crystallizes, inducing sickling of red blood cells. Consequently, patients have a high risk of vaso-occlusive painful crisis. Red cell exchange transfusions remain an effective therapy in the acute and chronic treatment of SCD: the patient’s red blood cells (RBC) are removed and replaced by homologous normal red cells. Red cell exchange can provide needed oxygen carrying capacity while reducing the overall viscosity of blood (P.S. Swerdlow, 2006). We propose a novel preventive and therapeutic approach for SCD based on red blood cell transfusion. We hypothesise that loading RBC with an allosteric effector of hemoglobin can reduce RBC sickling. Indeed, the entrapment of Inositol Hexaphosphate (IHP) inside RBC reduces the oxygen-hemoglobin affinity, which is measured by a right shift of the oxygen dissociation curve. Thus, RBC-IHP have an increased capacity to deliver oxygen to tissues. It is also expected that the deoxygenation of SS RBC is reduced and sickling is avoided. IHP was entrapped into human RBC by hypotonic reversible lysis followed by a resealing step. RBC-IHP were characterised by the amount of IHP entrapped into RBCs and the P50 measurement. Unprocessed human RBC were used as control. The potential anti-sickling effect of RBC-IHP was investigated using an in vitro model. Firstly, an experimental model to observe the relationship between sickling and oxygen concentration was set up : patients cells were submitted to deoxygenation by nitrogen bubbling for 30 min, and then re-oxygenated with different concentrations of oxygen (2, 5, 8, 15, 22%) for 30 min. The percentage of sickled cells was assessed by microscopy (about 500 cells checked). We observed that sickled cells recovered a normal shape upon reoxygenation (>15%O2), and a steady state between 5 and 8 % of oxygen, allowing the development of a reliable experimental model. Next, patient blood samples (n=6), harvested just prior to red cell exchange, were studied. RBC were washed 3 times with phoshate buffer before use. Different proportions of RBC-IHP (10%, 30% or 50%) were mixed with patients red cells and submitted to deoxygenation (0% O2) for 30 min and reoxygenation (5% O2) for 30 min. The final hematocrit of the suspensions was approximately 15%. The percentage of sickled cells in the suspensions was evaluated by microscopy and corrected according to the appropriate dilution factor. After full deoxygenation, 10% to 50% of cells were sickled, which appeared to be dependent on the HbS level in the blood samples. For all patients, RBC-IHP exhibited an enhanced anti-sickling effect: sickling was reduced by 19, 34, and 67% according to the RBC-IHP proportions 10%, 30% and 50%, respectively. Indeed, for equivalent RBC proportions RBC-IHP (50%) was 1.4 to 9 times more efficient compared to the unprocessed control RBC. Thus, RBC-IHP has the capacity to prevent sickling in a dose-dependent manner and is efficient at low proportions (10%). Consequently, RBC-IHP can improve classical transfusion therapy in terms of transfused volume, frequency and preventive sickling effect.

Red Cell Enzyme Activities and Properties of Mutant Pyruvate Kinase after Long-Term Storage of Red Cells in Liquid Nitrogen

Vox Sanguinis ◽  
1979 ◽  
Vol 36 (6) ◽  
pp. 362-366
Author(s):  
S. Miwa ◽  
H. Fujii ◽  
Y. Ishida ◽  
E. Oda ◽  
T. Nakatsuji ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Pyruvate Kinase ◽  
Enzyme Activities ◽  
Red Cells ◽  
Red Cell ◽  
Long Term Storage ◽  
Cell Enzyme ◽  
Term Storage

A COMPARISON OF THE ACTION OF B.WELCHII TOXIN WITH OTHER HAEMOTOXINS ON HUMAN AND RABBIT RED CELLS IN VITRO

1930 ◽  
Vol 2 (1) ◽  
pp. 91-100
Author(s):  
J. H. Orr ◽  
W. A. Campbell ◽  
G. B. Reed
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Average Diameter ◽  
Regular Sequence ◽  
Red Cells ◽  
The Other ◽  
Staph Aureus

The effect of various representative hæmotoxins on human and rabbit red blood cells in vitro was studied. It was found that as a result of the action of B. welchii toxins produced from a variety of strains of the organism a definite anisocytosis was produced and that the change in size of the cells followed a regular sequence. The first change to be noted was a development of cells having an average diameter less than the normal (microcyte stage). Further action of the toxin resulted in the replacement of these microcytes by cells having an average diameter greater than the normal (macrocyte stage). Following this macrocyte stage it was found that there was a return of the cells to a diameter very closely approximating the normal. This change in the size of the cells did not appear as a result of the action of any of the other hæmotoxins worked with viz., B. tetani, V. septique, Strepto. scarlatinœ, Staph. aureus.

scholarly journals AG-348 enhances pyruvate kinase activity in red blood cells from patients with pyruvate kinase deficiency

Blood ◽  
2017 ◽  
Vol 130 (11) ◽  
pp. 1347-1356 ◽  
Author(s):  
Charles Kung ◽  
Jeff Hixon ◽  
Penelope A. Kosinski ◽  
Giovanni Cianchetta ◽  
Gavin Histen ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Pyruvate Kinase ◽  
Blood Cells ◽  
Kinase Activity ◽  
Red Cell ◽  
Pyruvate Kinase Activity ◽  
Pyruvate Kinase Deficiency ◽  
Key Points ◽  
Allosteric Activator

Key Points AG-348 is a small-molecule allosteric activator of WT red cell pyruvate kinase as well as mutant enzymes associated with hemolytic anemia. Activity in vitro, in mice, and in red blood cells suggests it may address the underlying molecular pathology in PK deficiency patients.

Cow red blood cells. II. Stimulation of bovine red cell glycolysis by plasma

1979 ◽  
Vol 236 (5) ◽  
pp. C262-C267 ◽  
Author(s):  
M. J. Seider ◽  
H. D. Kim
Keyword(s):  
Guinea Pig ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Red Cells ◽  
Red Cell ◽  
Cellular Components ◽  
Glycolytic Rate ◽  
Purines And Pyrimidines ◽  
Stimulation Of

Cow red cell glycolysis, which can be stimulated by a variety of purines and pyrimidines, was also found to be elevated by its own plasma. Dialyzed or charcoal-treated plasma could no longer stimulate glycolysis, suggesting that the stimulating factors may be purines or pyrimidines. Determination of purines or pyrimidines in plasma revealed the presence of xanthine (0.31 muM), hypoxanthine (0.60 muM), and adenosine (0.05 muM), as well as unknown compounds. A physiologic level of hypoxanthine, with or without xanthine and adenosine approximating their concentrations in plasma, resulted in the stimulation of cow red cell glycolytic rate by 16% (P less than 0.01). These findings suggest that plasma-borne purines may act on cow red cells in concert with as yet unidentified factors. Moreover, exchanging calf and cow plasmas produced no stimulatory effect on either calf or cow red cell glycolysis, suggesting that a) calf red cells lack some of the cellular components that respond to this stimulator and, b) only cow plasma contains this specific stimulator. In other species, including dog, cat, rabbit, rat, guinea pig, and human, stimulation of glycolysis by plasma was not observed.

Red Cell Enzyme Activities and Properties of Mutant Pyruvate Kinase after Long-Term Storage of Red Cells in Liquid Nitrogen

Vox Sanguinis ◽  
1979 ◽  
Vol 36 (6) ◽  
pp. 362-366 ◽  
Author(s):  
H. Fujii ◽  
S. Miwa ◽  
Y. Ishida ◽  
E. Oda ◽  
T. Nakatsuji ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Pyruvate Kinase ◽  
Enzyme Activities ◽  
Red Cells ◽  
Red Cell ◽  
Long Term Storage ◽  
Cell Enzyme ◽  
Term Storage

VERY LARGE AND COMPACT AGGREGATES OF RED CELLS IN HEART DISEASE AND CANCER: POSSIBLY AN ANALOGOUS ROLE IN THE MICROCIRCULATION AS PLATELET AGGREGATES

1987 ◽  
Author(s):  
L Dintenfass
Keyword(s):  
Blood Cells ◽  
Red Cells ◽  
Cell Aggregates ◽  
Red Cell ◽  
Compact Type ◽  
Platelet Aggregates ◽  
The One ◽  
Native Plasma

Very large aggregates of red blood cells, showing compact morphology, are easily observed in vitro, and might be of importance in the in vivo microcirculation. Blood from patients with myocardial infarction, WaldenstrBm1s macroglobulinaemia, or varoous carcinomas. etc., was anticoagulated with EDTA, and adjusted to haematocrit of 0.30, using native plasma. All tests were carried out in the slit-capillary photoviscometer, at temp, of 22°C. Micro and macrophotographs were taken during flow and stasis, using slits of 12.5 and 50 micron gaps. Studies showed that very large (two-dimensional) red cell aggregates are formed, such aggregates (clumps) containing up to 50,000 red cells in a single clump. The architecture of such aggregates differed according to the origin of blood: both rouleaux type and random / compact type of aggregates were observed; in principle, a spectrum of morphologies can be seen. These observations form a link between the earlier work of FAHRAEUS, on the one hand, and that of KNISELY, on the other; whic works appeared at the time to be contradictory and irreconcilable.

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