scholarly journals Normal and Variant Isoenzymes of Human Blood Cell Hexokinase and the Isoenzyme Patterns in Hemolytic Anemia

Blood ◽  
1970 ◽  
Vol 36 (2) ◽  
pp. 219-227 ◽  
Author(s):  
CIGDEM ALTAY ◽  
CHESTER A. ALPER ◽  
DAVID G. NATHAN
Keyword(s):  
Hemolytic Anemia ◽  
Cell Metabolism ◽  
Electrophoretic Pattern ◽  
Red Cells ◽  
Red Cell ◽  
Hexokinase Activity ◽  
Individual Bands ◽  
Agarose Electrophoresis ◽  
Human Blood Cell ◽  
Isoenzyme Patterns

Abstract Electrophoresis of red cell hexokinase in agarose electrophoresis revealed two major (1 and 2) and two minor (3 and 4) bands. Platelet and leukocyte hexokinase patterns differed from those of red cells. There was a strong band 1, but considerably faster bands termed 5, 6 and 7 were also observed which were sensitive to changes in glucose concentration. The presence of contaminating leukocytes can significantly alter the electrophoretic pattern of "red cell" hexokinase activity. Bands 2, 3 and 4 of red cells appeared to be synthesized independently of band 1 and absence of band 1 did not effect either red cell metabolism or survival. Absence of bands 2, 3 and 4 may be associated with hemolytic anemia, decreased erythrocyte hexokinase activity and decreased erythrocyte glycolysis. Young red cells had increased activity of all bands, particularly band 2. No influence of hemoglobin type on hexokinase patterns was observed, nor was there any selective influence of cell storage, medium glucose, or 2-mercaptoethanol on individual bands. None of the various isoenzyme patterns were associated with abnormal hexokinase kinetics.

scholarly journals Generalized hexokinase deficiency in the blood cells of a patient with nonspherocytic hemolytic anemia

Blood ◽  
1983 ◽  
Vol 61 (1) ◽  
pp. 12-18 ◽  
Author(s):  
G Rijksen ◽  
JW Akkerman ◽  
AW van den Wall Bake ◽  
DP Hofstede ◽  
GE Staal
Keyword(s):  
Hemolytic Anemia ◽  
Blood Platelets ◽  
Clinical Signs ◽  
Electrophoretic Pattern ◽  
Residual Activity ◽  
Red Cells ◽  
Type I ◽  
Ph Optimum ◽  
Hexokinase Activity ◽  
Type Iii

In a patient with nonspherocytic hemolytic anemia, a hexokinase deficiency was detected in the red cells (residual activity about 25% of normal) and in blood platelets (20%-35% of normal activity). Although the total hexokinase activity in lymphocytes was normal, the amount of hexokinase type I was decreased to about 50% of normal. However, the deficiency was compensated for by the appearance of type III hexokinase. Compartmentation studies with controlled digitonin- induced cell lysis showed that this type III enzyme was localized in the cytosol, while almost all hexokinase activity in normal lymphocytes is particulate. No abnormal lymphocyte functions could be detected. The patient was homozygous for the defect. The parents and three of five sibs of the patient were apparently heterozygous with residual activities of 50%-67% of normal in their red cells, but did not show any clinical signs of hexokinase deficiency. The variant enzyme had a slightly decreased affinity for MgATP2- and a strongly increased inhibition constant for glucose-1,6-P2. Affinity for glucose, heat stability, and pH optimum were normal. In the electrophoretic pattern of red cell hexokinase, only one subtype of hexokinase I could be detected, while in normal red cells, at least three subtypes are present. In the heterozygous individuals, no enzymatic abnormalities could be detected, except for an aberration in the electropherogram of one sib.

scholarly journals Generalized hexokinase deficiency in the blood cells of a patient with nonspherocytic hemolytic anemia

Blood ◽  
1983 ◽  
Vol 61 (1) ◽  
pp. 12-18 ◽  
Author(s):  
G Rijksen ◽  
JW Akkerman ◽  
AW van den Wall Bake ◽  
DP Hofstede ◽  
GE Staal
Keyword(s):  
Hemolytic Anemia ◽  
Blood Platelets ◽  
Clinical Signs ◽  
Electrophoretic Pattern ◽  
Residual Activity ◽  
Red Cells ◽  
Type I ◽  
Ph Optimum ◽  
Hexokinase Activity ◽  
Type Iii

Abstract In a patient with nonspherocytic hemolytic anemia, a hexokinase deficiency was detected in the red cells (residual activity about 25% of normal) and in blood platelets (20%-35% of normal activity). Although the total hexokinase activity in lymphocytes was normal, the amount of hexokinase type I was decreased to about 50% of normal. However, the deficiency was compensated for by the appearance of type III hexokinase. Compartmentation studies with controlled digitonin- induced cell lysis showed that this type III enzyme was localized in the cytosol, while almost all hexokinase activity in normal lymphocytes is particulate. No abnormal lymphocyte functions could be detected. The patient was homozygous for the defect. The parents and three of five sibs of the patient were apparently heterozygous with residual activities of 50%-67% of normal in their red cells, but did not show any clinical signs of hexokinase deficiency. The variant enzyme had a slightly decreased affinity for MgATP2- and a strongly increased inhibition constant for glucose-1,6-P2. Affinity for glucose, heat stability, and pH optimum were normal. In the electrophoretic pattern of red cell hexokinase, only one subtype of hexokinase I could be detected, while in normal red cells, at least three subtypes are present. In the heterozygous individuals, no enzymatic abnormalities could be detected, except for an aberration in the electropherogram of one sib.

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 K-Na EXCHANGE ACCOMPANYING THE PROLYTIC LOSS OF K FROM HUMAN RED CELLS

1947 ◽  
Vol 30 (5) ◽  
pp. 379-387 ◽  
Author(s):  
Eric Ponder
Keyword(s):  
Steady State ◽  
Cell Metabolism ◽  
Ionic Composition ◽  
Surface Membrane ◽  
Sodium Taurocholate ◽  
Red Cells ◽  
Red Cell ◽  
Membrane Changes ◽  
Kinetics Of ◽  
Human Red Cells

In systems containing human red cells and sodium taurocholate as a lysin, or distearyl lecithin as a sphering agent, the prolytic loss of K at 25°C. is accompanied by a gain of Na by the cell, the gain being somewhat greater than the K loss. A small volume increase accompanies the exchange. The kinetics of the K loss and the Na gain are similar to those already described; i.e., the changes are rapid at first, and slow down so that after 12 to 20 hours it appears that a new steady state is being approached. Similar, but smaller, losses of K and gains of Na occur when the cells stand in isotonic NaCl at 25°C. without the addition of a lysin or sphering agent. On these and other experimental grounds, it is impossible to retain the idea that the mammalian red cell in general is impermeable to cations. The cells nevertheless seem to be in a steady state with respect to their environment, their ionic composition changing as the composition of the environment is changed. The possible processes by means of which one steady state can be exchanged for another—changes in the permeability of a surface membrane, changes in the velocity of an active ion transfer process dependent on red cell metabolism, and changes in the activity of the ions in the red cell interior as a result of changes in an orderly internal structure—are discussed.

scholarly journals Lipid analyses and fluidity studies by electron spin resonance of red cell membranes in hereditary high red cell membrane phosphatidylcholine hemolytic anemia

Blood ◽  
1984 ◽  
Vol 64 (5) ◽  
pp. 1129-1134 ◽  
Author(s):  
Y Yawata ◽  
T Sugihara ◽  
M Mori ◽  
S Nakashima ◽  
Y Nozawa
Keyword(s):  
Electron Spin Resonance ◽  
Cell Membrane ◽  
Membrane Fluidity ◽  
Hemolytic Anemia ◽  
Electron Spin ◽  
Order Parameter ◽  
Red Cells ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Spin Resonance

Abstract Membrane lipid analyses and electron spin resonance (ESR) studies of membrane fluidity were carried out on the red cells of a Japanese patient with hereditary high red cell membrane phosphatidylcholine hemolytic anemia (HPCHA). Increased amounts of phosphatidylcholine (PC) and cholesterol were found in the membrane lipids of the affected patient, despite normal plasma lipids. The order parameter of cholesterol-free pure phospholipid liposomes prepared from this patient's red cells was decreased, apparently because of the increased PC. In contrast, the order parameter of the total red cell lipid liposomes (containing free cholesterol) was essentially normal. The overall fluidity of the intact red cells was determined by ESR with a spin probe, 5-SAL. Again, the order parameters were normal in the intact red cells of the patient with HPCHA. This suggests that the concomitant increase of membrane cholesterol and phosphatidylcholine serves to maintain normal membrane fluidity in the HPCHA red cells.

RED CELL METABOLISM IN THE PREMATURE INFANT

PEDIATRICS ◽  
1965 ◽  
Vol 36 (1) ◽  
pp. 104-112
Author(s):  
Frank A. Oski ◽  
J. Lawrence Naiman
Keyword(s):  
Premature Infant ◽  
Premature Infants ◽  
Metabolic Profile ◽  
Cell Metabolism ◽  
Glucose Consumption ◽  
Red Cells ◽  
Red Cell ◽  
Term Infants ◽  
Atp Levels ◽  
Red Cell Metabolism

The erythrocytes of premature infants, term infants, and adults were studied with respect to ATP levels, ATP stability, glucose consumption, glutathione stability, and the tendency to develop morphologic abnormalities during short periods of incubation. The erythrocytes of both the premature and term infants had higher ATP levels and glucose consumption, greater ATP and glutathione instability, and more marked morphologic abnormalities than the adult red cells. The erythrocytes of the premature infants showed the greatest degrees of abnormality. It is suggested that the premature infant at birth possesses a metabolic profile in his erythrocytes which results in ATP instability.

RED CELL METABOLISM IN THE PREMATURE INFANT

PEDIATRICS ◽  
1967 ◽  
Vol 39 (5) ◽  
pp. 689-695
Author(s):  
Frank A. Oski
Keyword(s):  
Premature Infants ◽  
Pentose Phosphate Pathway ◽  
Cell Metabolism ◽  
Glucose Consumption ◽  
Pentose Phosphate ◽  
Red Cells ◽  
Red Cell ◽  
O2 Production ◽  
Cell Glucose ◽  
Normal Adults

Red cell glucose consumption and C14 O2 production from glucose-I-C14 and glucose-U-C14 were measured in 17 premature infants, 15 normal adults and 7 patients with reticulocytosis. The red cells from the premature infants consumed significantly more glucose and produced more C14 O2 than did the cells from normal adults but did not differ in this regard from patients with elevated reticulocyte counts. The percent of glucose metabolized by the pentose phosphate pathway in the red cells of the infants was as great, and often greater, than that observed in the other two groups. The red cells of the premature infants also demonstrated an ability to respond to methylene blue. The pentose phosphate pathway would appear to be intact in the red cells from these infants and inadequate generation of NADPH2 is not responsible for the ease with which these cells develop oxidative denaturation of hemoglobin.

scholarly journals The Effects of Concentrated Eluted Anti-Red Blood Cell Antibodies on the in Vivo Survival of Normal Red Blood Cells

Blood ◽  
1960 ◽  
Vol 15 (4) ◽  
pp. 525-533 ◽  
Author(s):  
NEIL W. CULP ◽  
HUGH CHAPLIN
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Hemolytic Anemia ◽  
Normal Control ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Red Cells ◽  
Significant Alteration ◽  
Red Cell

Abstract 1. A method has been described for the preparation and sterilization of a concentrated eluate from human red cell stroma. 2. Red cells sensitized by such an eluate prepared from normal control red cells showed entirely normal in vivo survival, as did cells sensitized by eluate from anti-H coated cells. 3. Sensitization of red cells by concentrated eluates from a patient with Coombs-negative acquired hemolytic anemia and from a patient with Coombs-positive acquired hemolytic anemia did not cause significant alteration in the in vivo survival of the red cells. 4. Red cells sensitized by the concentrated eluate from anti-D sensitized cells disappeared from the recipient’s circulation very rapidly and were sequestered in the spleen, indicating preservation of the physiologic properties of the antibody throughout the elution, concentration and sterilization procedures.

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