scholarly journals Hereditary nonspherocytic hemolytic anemia due to a new hexokinase variant with reduced stability

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 690-697 ◽  
Author(s):  
M Magnani ◽  
V Stocchi ◽  
L Cucchiarini ◽  
G Novelli ◽  
S Lodi ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Specific Activity ◽  
Blood Platelets ◽  
Phenotypic Variability ◽  
Ph Dependence ◽  
Glucose Consumption ◽  
Heat Stability ◽  
Normal Activity ◽  
Red Cell ◽  
Hexokinase Activity

A 27-year-old woman with severe chronic hemolytic anemia was found to have reduced red cell hexokinase activity when the degree of reticulocytosis was considered. This enzyme had normal pH-dependent activity, normal Km for glucose, fructose, and mannose, normal Km for Mg adenosine triphosphate (ATP)2- and Ki for glucose-1,6-diphosphate. Furthermore, the pH-dependence and orthophosphate dependence of Ki for glucose-1,6-diphosphate were normal. However, this hexokinase was inactivated rapidly at 44 degrees C. No abnormalities were found in the red cell hexokinase isozymic pattern when it was compared with the profile obtained from cells of similar age. The hexokinase specific activity was reduced in all the red blood cell fractions obtained by density gradient ultracentrifugation; a marked difference in the distribution of cells through the gradient was evident. Among the glycolytic intermediates, a significant decrease of 2,3- diphosphoglycerate was evident. ATP and glucose 6-phosphate were also reduced when compared with cells of similar. Glucose consumption of the hexokinase-deficient cells decreased, but the rate of glucose metabolized through the hexose monophosphate shunt was unchanged. Although the total hexokinase activity in lymphocytes was only reduced by 37%, a marked hexokinase deficiency was detected in blood platelets (20% to 25% of normal activity). The parents and one of two siblings of the patient were heterozygous for the defect, with 66% to 74% of normal erythrocyte hexokinase activity and reduced heat stability of the enzyme. These results, when compared with those obtained in previously reported cases of hexokinase deficiency, provide further evidence of the broad phenotypic variability that characterizes this disorder. Furthermore, it is suggested that failure of energy generation is probably the primary cause of hemolytic anemia in hexokinase deficiency.

scholarly journals Hereditary nonspherocytic hemolytic anemia due to a new hexokinase variant with reduced stability

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 690-697 ◽  
Author(s):  
M Magnani ◽  
V Stocchi ◽  
L Cucchiarini ◽  
G Novelli ◽  
S Lodi ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Specific Activity ◽  
Blood Platelets ◽  
Phenotypic Variability ◽  
Ph Dependence ◽  
Glucose Consumption ◽  
Heat Stability ◽  
Normal Activity ◽  
Red Cell ◽  
Hexokinase Activity

Abstract A 27-year-old woman with severe chronic hemolytic anemia was found to have reduced red cell hexokinase activity when the degree of reticulocytosis was considered. This enzyme had normal pH-dependent activity, normal Km for glucose, fructose, and mannose, normal Km for Mg adenosine triphosphate (ATP)2- and Ki for glucose-1,6-diphosphate. Furthermore, the pH-dependence and orthophosphate dependence of Ki for glucose-1,6-diphosphate were normal. However, this hexokinase was inactivated rapidly at 44 degrees C. No abnormalities were found in the red cell hexokinase isozymic pattern when it was compared with the profile obtained from cells of similar age. The hexokinase specific activity was reduced in all the red blood cell fractions obtained by density gradient ultracentrifugation; a marked difference in the distribution of cells through the gradient was evident. Among the glycolytic intermediates, a significant decrease of 2,3- diphosphoglycerate was evident. ATP and glucose 6-phosphate were also reduced when compared with cells of similar. Glucose consumption of the hexokinase-deficient cells decreased, but the rate of glucose metabolized through the hexose monophosphate shunt was unchanged. Although the total hexokinase activity in lymphocytes was only reduced by 37%, a marked hexokinase deficiency was detected in blood platelets (20% to 25% of normal activity). The parents and one of two siblings of the patient were heterozygous for the defect, with 66% to 74% of normal erythrocyte hexokinase activity and reduced heat stability of the enzyme. These results, when compared with those obtained in previously reported cases of hexokinase deficiency, provide further evidence of the broad phenotypic variability that characterizes this disorder. Furthermore, it is suggested that failure of energy generation is probably the primary cause of hemolytic anemia in hexokinase deficiency.

scholarly journals Hereditary nonspherocytic hemolytic anemia and hexokinase deficiency

Blood ◽  
1978 ◽  
Vol 51 (5) ◽  
pp. 935-940 ◽  
Author(s):  
E Beutler ◽  
PG Dyment ◽  
F Matsumoto
Keyword(s):  
Thermal Stability ◽  
Hemolytic Anemia ◽  
Potent Inhibitor ◽  
Glucose Consumption ◽  
Kinetic Properties ◽  
Red Cell ◽  
Hexokinase Activity ◽  
Thermal Stability Of ◽  
Chronic Hemolytic Anemia ◽  
2,3 Dpg

An 11-yr-old child with mild chronic hemolytic anemia was found to have decreased red cell hexokinase activity in spite of the reduced mean age of her red cell population. Similar decreases in red cell hexokinase activity were documented in the patient's parents and in one sib. The red cells were morphologically normal. Red cell 2,3-DPG levels were normal and ATP and glucose-6-phosphate levels were diminished. The kinetic properties, electrophoretic mobility, and thermal stability of the residual red cell hexokinase were normal or nearly so. Glucose consumption of the hexokinase-deficient cells was not appreciably decreased, probably because less of the potent inhibitor glucose-6- phosphate was present in the erythrocytes. It is likely, although not certain, that in this patient nonspherocytic hemolytic anemia resulted from hexokinase deficiency.

scholarly journals Hereditary nonspherocytic hemolytic anemia and hexokinase deficiency

Blood ◽  
1978 ◽  
Vol 51 (5) ◽  
pp. 935-940 ◽  
Author(s):  
E Beutler ◽  
PG Dyment ◽  
F Matsumoto
Keyword(s):  
Thermal Stability ◽  
Hemolytic Anemia ◽  
Potent Inhibitor ◽  
Glucose Consumption ◽  
Kinetic Properties ◽  
Red Cell ◽  
Hexokinase Activity ◽  
Thermal Stability Of ◽  
Chronic Hemolytic Anemia ◽  
2,3 Dpg

Abstract An 11-yr-old child with mild chronic hemolytic anemia was found to have decreased red cell hexokinase activity in spite of the reduced mean age of her red cell population. Similar decreases in red cell hexokinase activity were documented in the patient's parents and in one sib. The red cells were morphologically normal. Red cell 2,3-DPG levels were normal and ATP and glucose-6-phosphate levels were diminished. The kinetic properties, electrophoretic mobility, and thermal stability of the residual red cell hexokinase were normal or nearly so. Glucose consumption of the hexokinase-deficient cells was not appreciably decreased, probably because less of the potent inhibitor glucose-6- phosphate was present in the erythrocytes. It is likely, although not certain, that in this patient nonspherocytic hemolytic anemia resulted from hexokinase deficiency.

Severe-glucose-6-phosphate dehydrogenase (G6PD) deficiency associated with chronic hemolytic anemia, granulocyte dysfunction, and increased susceptibility to infections: description of a new molecular variant (G6PD Barcelona)

Blood ◽  
1982 ◽  
Vol 59 (2) ◽  
pp. 428-434 ◽  
Author(s):  
JL Vives Corrons ◽  
E Feliu ◽  
MA Pujades ◽  
F Cardellach ◽  
C Rozman ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
G6pd Deficiency ◽  
Specific Activity ◽  
Heat Stability ◽  
Molecular Characteristics ◽  
Functional Studies ◽  
Leukocyte Alkaline Phosphatase ◽  
Severe Deficiency ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Increased Susceptibility

Abstract Molecular, kinetic, and functional studies were carried out on erythrocytes and leukocytes in a Spanish male with G6PD deficiency, congenital nonspherocytic hemolytic anemia (CNSHA), and increased susceptibility to infections. G6PD activity was absent in patient's red cells and was about 2% of normal in leukocytes. Molecular studies using standard methods (WHO, 1967) showed G6PD in the patient to have a slightly fast electrophoretic mobility at pH 8.0 with otherwise normal properties (heat stability at 46 degrees C, apparent affinity for substrates, optimum pH, and utilization of substrate analogues). Other tests showed the patient's granulocytes to engulf latex particles normally, but to have impaired reduction of nitroblue tetrazolium and ferricytochrome-c as well as reduced iodination. Chemotaxis and random migration of the patient's granulocytes were normal as were myeloperoxidase, leukocyte alkaline phosphatase (LAP), and ultrastructural features. The molecular characteristics of G6PD in the patient differed from those of all previously reported variants associated with CNSHA, so the present variant was provisionally called G6PD Barcelona to distinguish it from other G6PD variants previously described. Possible mechanisms for the severe deficiency of G6PD in erythrocytes and granulocytes was investigated by studies on the immunologic specific activity of the mutant enzyme.

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 Severe-glucose-6-phosphate dehydrogenase (G6PD) deficiency associated with chronic hemolytic anemia, granulocyte dysfunction, and increased susceptibility to infections: description of a new molecular variant (G6PD Barcelona)

Blood ◽  
1982 ◽  
Vol 59 (2) ◽  
pp. 428-434
Author(s):  
JL Vives Corrons ◽  
E Feliu ◽  
MA Pujades ◽  
F Cardellach ◽  
C Rozman ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
G6pd Deficiency ◽  
Specific Activity ◽  
Heat Stability ◽  
Molecular Characteristics ◽  
Functional Studies ◽  
Leukocyte Alkaline Phosphatase ◽  
Severe Deficiency ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Increased Susceptibility

Molecular, kinetic, and functional studies were carried out on erythrocytes and leukocytes in a Spanish male with G6PD deficiency, congenital nonspherocytic hemolytic anemia (CNSHA), and increased susceptibility to infections. G6PD activity was absent in patient's red cells and was about 2% of normal in leukocytes. Molecular studies using standard methods (WHO, 1967) showed G6PD in the patient to have a slightly fast electrophoretic mobility at pH 8.0 with otherwise normal properties (heat stability at 46 degrees C, apparent affinity for substrates, optimum pH, and utilization of substrate analogues). Other tests showed the patient's granulocytes to engulf latex particles normally, but to have impaired reduction of nitroblue tetrazolium and ferricytochrome-c as well as reduced iodination. Chemotaxis and random migration of the patient's granulocytes were normal as were myeloperoxidase, leukocyte alkaline phosphatase (LAP), and ultrastructural features. The molecular characteristics of G6PD in the patient differed from those of all previously reported variants associated with CNSHA, so the present variant was provisionally called G6PD Barcelona to distinguish it from other G6PD variants previously described. Possible mechanisms for the severe deficiency of G6PD in erythrocytes and granulocytes was investigated by studies on the immunologic specific activity of the mutant enzyme.

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.

Newborn Platelet Dysfunction: a Storage Pool and Release Defect

1976 ◽  
Vol 36 (01) ◽  
pp. 200-207 ◽  
Author(s):  
Donald G. Corby ◽  
Thomas F. Zuck
Keyword(s):  
Specific Activity ◽  
Platelet Rich Plasma ◽  
Adenine Nucleotides ◽  
Blood Platelets ◽  
Newborn Infants ◽  
Specific Radioactivity ◽  
High Dose ◽  
Platelet Dysfunction ◽  
Paired Samples ◽  
Normal Adults

SummaryPer cent aggregation, release and content of adenine nucleotides, and specific radioactivity were evaluated in citrated platelet-rich plasma (PRP) prepared from paired samples of maternal and cord blood. Platelets of newborn infants aggregated normally in response to high dose ADP (20 μM), strong collagen suspensions, and thrombin; however, when compared with PRP from the mothers or from normal adults, per cent aggregation in response to lower concentrations of ADP (2 μM), weak collagen, and part particularly epinephrine was markedly reduced. Nucleotide release after stimulation of the newborns’ PRP with the latter two inducers was also impaired. ATP and ADP content of the newborns’ platelets was also significantly less than that of their mothers or of normal adults, but specific activity was normal. The data suggest that the impairment of ADP release in the platelets of newborn infants is due to decreased sensitivity to external stimuli. Since metabolic ATP is necessary for the platelet release reaction, it is postulated that the platelet dysfunction results from a lack of metabolic ATP.

scholarly journals HEMOLYTIC SYSTEMS CONTAINING ANIONIC DETERGENTS

1946 ◽  
Vol 30 (1) ◽  
pp. 15-23 ◽  
Author(s):  
Eric Ponder
Keyword(s):  
Shape Change ◽  
Ph Dependence ◽  
Cell Ultrastructure ◽  
Effect Of Temperature ◽  
Red Cell ◽  
Inhibitory Effects ◽  
Serum Globulin ◽  
Hemolytic Action ◽  
Anionic Detergents ◽  
Protein Components

1. The members of the homologous series of anionic detergents, the sodium salts of the sulfated straight chain alcohols with the general formula CnH2n+1·SO3·Na, are hemolytic, the lytic activity being at a maximum when the compound contains 14 carbon atoms in the chain. In systems in which lysis is comparatively rapid, the hemolytic effect increases with increasing pH, but in systems containing quantities of lysin near the asymptotic concentrations the pH dependence of the activity is reversed. The effect of temperature is principally one on the velocity constant of the lytic reaction, with smaller effects on the position of the asymptotes of the time-dilution curves and on their shape. 2. The quantities of the detergents which produce disk-sphere transformations are approximately one-tenth of those required to produce complete hemolysis. In most cases, the shape change occurs when there are too few detergent molecules present to cover the red cell surfaces with a monolayer. 3. Plasma inhibits the hemolytic action of these detergents, and, in the quantities in which they occur in plasma, lecithin, serum globulin, cholesterol, and serum albumin, produce inhibitory effects which increase in that order in systems containing the C-14 sulfate. It can be inferred from these inhibitory effects that the anionic detergents can form compounds or complexes with lipid, lipoprotein, and protein components of the red cell ultrastructure.

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