Red Blood Cell Enzyme Research Articles

The effects of splenectomy on the hematology and on the activity of delta-aminolevulinic acid dehydratase (ALA-D) in hematopoietic tissues of the european eel, Anguilla anguilla L.

1. 1. The effects of splenectomy on hematological parameters and on the activity of ALA-D in some tissues of the European eel, Anguilla anguilla L., were studied after 1, 2, 4, 6 and 13 weeks. 2. 2. No marked differences in red blood cell variables (hematocrit, hemoglobin and RBC count) or enzyme activity were observed between sham-operated and splenectomized eels. 3. 3. An initial lymphopenia followed later by neutrophilia were observed in both operated groups. 4. 4. Abnormal erythrocytes were present in the blood from splenectomized eels. 5. 5. The spleen seems to function primarily as an erythrocyte reservoir and destruction organ for erythrocytes and has only a subsidiary role as an erythrocyte-producing organ. Lymphocytopoiesis seems to occur partly in the spleen, while granulocytopoiesis probably occurs elsewhere.

Red blood cell fatty acid synthetase nonidentity with the enzyme from liver

1. 1. Fatty acid synthetase has been purified 680-fold from rat erythrocytes. 2. 2. The enzyme obtained gave one major staining band on disc gel electrophoresis. 3. 3. The molecular weight of the enzyme was estimated to be 310,000 and the enzyme is composed of two half-molecular weight subunits. 4. 4. The red blood cell enzyme synthesizes almost equal amounts of C 14 and C 16 fatty acids (39 and 43%, respectively). 5. 5. The enzyme isolated from rat red blood cells differs from the fatty acid synthetases isolated from the rat adipose tissue, mammary gland, liver and lung in its lack of cross-reactivity with antibody to rat liver fatty acid synthetase. 6. 6. This evidence suggests that the red blood cell fatty acid synthetase is structurally different from that of rat liver.

The diagnosis of zygosity in twins.

Uniovular and diovular twin pairs can be classified with at least 90% accuracy by careful questioning or by ratings by observers of similarity in appearance, eye color, ear conformation, and the like. This article outlines the methods required for highly accurate zygosity diagnosis using blood typing, fingerprints, and anthropometry. The logic behind the use of discrete Mendelian phenotypes is explained and tables are provided which give the odds against a dizygotic twin pair showing concordance on eight blood group systems, four serum proteins, and six red blood cell enzymes. These tables are suitable for use with American twins of European ancestry; the method of generating such tables is explained so that similar tables based on different gene frequency estimates or for additional genetic markers can be easily constructed. The logic behind the use of metric, polygenic traits in zygosity diagnosis is also presented, together with tables giving the odds against a dizygotic twin pair showing various degrees of similarity in ponderal index, in cephalic index, and in Slater's Z score, a measure of fingerprint similarity. Taken together, the 18 serological markers should misclassify dizygotic twins as monozygotic fewer than two times in 1000. With the three anthropometric variables, this average probability of misclassification drops about 1 order of magnitude.

Biochemical response of squirrel monkeys to ozone.

Biochemical studies were performed on blood and lung tissue of squirrel monkeys (Saimiri sciureus) following acute exposure to 0.75 ppm ozone (O3) for 4 h/d for 4 consecutive days. One group of animals was sacrificed at the end of the last exposure day and another group was sacrificed 4 d later after the last exposure. Evidence was sought for oxidation-induced changes known to occur in rodents when high levels of O3 are inhaled. A significant increase in red blood cell membrane fragility was observed, as well as significant decreases in red blood cell glutathione and erythrocyte acetylcholinesterase; however, the red blood cell enzymes, lactic acid dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G6PDH) were not changed significantly. Lung tissue analysis showed that lipid peroxidation was markedly increased and tissue vitamin E levels were significantly decreased. The tissue enzymes G6PDH, glutathione reductase, and LDH significantly increased in activity. No significant changes were seen in either superoxide dismutase or malic acid dehydrogenase. The results of this experiment indicate that O3, or reaction products resulting from O3-tissue interaction in the lung, pass the air-blood barrier and are capable of producing biochemical changes in blood as well as in lung tissue.

Detection and incidence of B and C vitamin deficiency in alcohol-related illness.

The activity of the red blood cell enzymes transketolase, glutathione reductase, and aspartate transaminase, and their activation by the coenzymes thiamine, riboflavin, and pyridoxine, the pyruvate tolerance test, the leucocyte vitamin C concentration, and the activity in serum of gamma-glutamyl transferase were measured in a series of 35 patients with alcohol-related illness. The incidence of thiamine deficiency was 31% as assessed by the activation of transketolase, and 55% as assessed by the pyruvate tolerance test. The incidence of riboflavin deficiency was 23% and of ascorbic acid deficiency 91%. No cases of pyridoxine deficiency were detected. The pyruvate tolerance test was found to be a more sensitive test of thiamine deficiency than the transketolase activation, and the activation of red blood cell aspartate transaminase was found to be a poor indicator of pyridoxine deficiency. There was a poor correlation of the gamma-glutamyl transferase activity with the degree of vitamin deficiency, suggesting that alcohol exposure is only partly responsible for the observed vitamin deficiency.

Studies of red blood cell and serum polymorphisms among the Matagi.

Serum samples from 159 Matagi were tested for nine blood group systems, haptoglobins, transferrins, Gc, 10 red blood cell enzyme polymorphisms and for the Gm and Inv allotypes to obtain evidence concerning their ethnic origin. The data exclude all claims except that they are of Japanese origin.

METABOLISM OF CATECHOL ESTROGEN BY HUMAN ERYTHROCYTES

In a study designed to evaluate the kinetics of catechol estrogen formation from plasma estrone in vivo, we obtained evidence that the red blood cell (RBC) enzyme, catechol-O-methyl transferase, catalyzes the transformation of 2-hydroxyestrone to 2-methoxyestrone. Under in vitro conditions, the rate of conversion of 2-hydroxyestrone to 2-methoxyestrone by human RBC's was such that 7 nmol of 2-methoxyestrone were formed per h per ml of RBC.

Gene Markers in Human Bone Marrow Transplantation1

One or more differences between donors and recipients were found in polymorphic red blood cell antigens and enzymes in each of 56 bone marrow transplant sibling pairs. These results identify those polymorphic traits which are potentially most informative for detection of donor cells in a recipient following transplantation.

Simultaneous determination of superoxide dismutase and catalase in biological materials by polarography

The polarographic method of catalytic currents applied to a wave of oxygen permits the simultaneous assay of superoxide dismutase and catalase in biological materials with high speed and reproducibility and minimal manipulation of tissues. Washed red blood cells and tissue homogenates give rise to a strong polarographic maximum, apparently due to heme proteins, which interferes with the measurement. This maximum is suppressed by addition of approximately 0.2% plasma. Therefore, the determination of the two enzymes in red blood cells can be carried out by direct addition of whole blood to the polarographic solution. Thirty microliters of blood are enough for optimal determination of both enzymes. The method can determine superoxide dismutase and catalase at concentrations as low as 2 × 10 −11 m and 5 × 10 −10 m, respectively, and shows a linear correlation between measured activity and enzyme levels. The average values of the two enzymes in human red blood cells was found by this method to be 2.6 × 10 −6 m for catalase and 1.8 × 10 −6 m for superoxide dismutase, which agree with previously reported values.

Gene Markers in Human Bone Marrow Transplantation

One or more differences between donors and recipients were found in polymorphic red blood cell antigens and enzymes in each of 56 bone marrow transplant sibling pairs. These results identify those polymorphic traits which are potentially most informative for detection of donor cells in a recipient following transplantation.

Action of Growth Hormone on Erythropoiesis: Changes in Red Blood Cell Enzyme Activities in Growth-retarded Patients with and without Growth Hormone Deficiency

Fifteen red cell enzyme activities of growth-retarded patients with and without growth hormone (GH) deficiency were investigated before and after GH administration. The 15 enzymes were Hexokinase, phosphoglucomutase, glucose phosphate, isomerase, phosphofructokinase, fructose diphosphate aldolase, glyceraldehyde-3-phosphae dehydrogenase, triosephosphate isomerase, 2,3-diphosphoglycerate mutase, 3-phosphoglycerate kinase, 3-phosphoglycerate mutase, enolase, pyruvate kinase, glycose-6-phosphate dehydrogenase, 6-phosphogluconic dehydrogenase, glutathione reducase. Sixty-six subjects were studied: 30 normal control subjects (group N) and 36 patients (aged 5-23 years) with short stature. Complete endocrine evaluation showed 21 (group I) to have GH deficiency (10 patients with isolated GH deficiency) and 15 (group II) to have normal hypothalamic and pituitary function except for two patients with a moderate hypothyroidism. Both had been receiving thyroid hormone treatment for a long time before our studies. All 36 patients were treated with 2 mg human growth hormone intramuscularly for 7 days. Before GH treatment no significant difference was observed between hematologic data in group I (GH deficiency) and group II (no GH deficiency). After GH therapy there was a significant increase in reticulocyte count in both groups of patients with short stature. The mean pretreatment value in group I was 1.294% +/- 0.084 (SEM); the mean post-treatment value was 2.081% +/- 0.287 (SEM)< P less than 0.005. The mean pretreatment value in group II was 1.0% 0.184 (SEM); the mean post-treatment value was 1.407% +/- 0.193 (SEM), P less than 0.01. In group II (no GH deficiency) mean pretreatment erythrocyte enzyme activities were not significantly different from those activities observed in normal control subjects (group N). However, in patients who lacked GH, the pretreatment activities of five red cell enzymes (glucose phosphate isomerase, triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, 2,3-diphosphoglycerate mutase, 3-phosphoglycerate kinase) were significantly decreased before GH administration compared with the values in normal control subjects...

Protein-calorie malnutrition: A study of red blood cell and serum enzymes during and after crisis

This paper reports a study of changes in red blood cell enzymes and some serum parameters during and after treatment of protein-calorie malnutrition. The red cell GSH levels were low during the crisis, together with the levels of GSSG:NADPH reductase, GSH:H 2O 2 peroxidase, aspartate aminotransferase and alanine aminotransferase. After treatment the levels of all these enzymes increased significantly to normal values. Of the serum parameters investigated, significant reduction in the activity of the enzymes cholinesterase, catecholamine oxidase, total proteins, albumin, urea and electrolytes were obvious, and returned to normal values after treatment. Ceruloplasmin activity remained low even after three weeks' treatment and could not be related to copper levels. The results are discussed in relation to anemia and liver damage that may accompany the syndrome.

Glucose-6-phosphate dehydrogenase Ferrara. A new variant of g-6-PD identified in Northern Italy.

A new variant of glucose-6-phosphate dehydrogenase (G-6-PD) has been discovered in Northern Italy, in the district of Ferrara. This variant is characterized by high decrease of red blood cell enzyme activity (less than 5% of normal), high affinity for G-6-P and NADP, increased utilization of deamino-NADP and 2-deoxy-G-6-P, and faster electrophoretic mobility in the buffer systems commonly used for the classification of the G-6-PD variants. The new G-6-PD type was never associated with clinical manifestations in any cases except neonatal jaundice in some of the newborns with this enzyme deficiency. The frequency of the new variant in the Ferrara district indicates that it has probably appeared in this area by mutation some centuries ago. It is suggested that this variant should be named G-6-PD Ferrara.

Metabolism of Human Erythrocytes

Deficiency states have been well established in nine enzymes of anaerobic glycolysis and suggested, but inadequantely confirmed, in others. In additon, hemolytic anemia has been defined in glucose-6-phosphate dehydrogenase deficiency, in certain enzymes glutathione metabolism of importance to the function of the oxidative hexsemonophosphate shunt, and in certain enzymes of nucleotide metabolism. In heterogeneous dyserythropoietic states with ineffective erythropoiesis, enzyme ratios are grossly distorted and differ greatly from those of either normal or reticullocyte-rich blood. Red blood cell (RBC) enzyme deficiencies show noticeable genetic polymorphism, most apparently recessively transmitted anemias actually resulting from inheritance of two different mutant, allelic genes. In addition to causing hemolytic syndromes in selected instances, deficiencies of enzymes more important to other tissues than to the RBC itself are detectable by assay of hemolystates.

Red cell enzymes—I. Glycolytic enzymes and reduction of glutathione in bovine red blood cells

1. 1. Spectrophotometric assays of twelve different enzymes of red blood cells were conducted in six breeds of cattle. 2. 2. Activity of fructose diphosphate aldolase was significantly higher in Sahiwal and crossbred [( Bos indicus) Sahiwal x ( Bos taurus) Red Polls] than in the other breeds. 3. 3. The rates of regeneration of reduced glutathione (GSH) and the activity of the enzymes associated with the regeneration of GSH (hexokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase) were not significantly different between the breeds studied.

Aktivitätsbestimmung und Normalwerte von L-Glutamat-L-Cystein-γ-Ligase (EC 6.3.2.2) in menschlichen Erythrocyten; Glutathionbiosynthese V.

For the determination of the activity of glutamate: cysteine-gamma-ligase one method was optimized and a new micromethod was developed: a) Utilizing [14C] glutamate and cysteine as substrates, the product [14C] gamma-glutamylcysteine was isolated as its cadmium mercaptide and determined by liquid scintillation counting gamma-glutamyl-[14C] aminobutyrate, which was synthesized from glutamate and [14C] alpha-aminobutyrate was separated from remaining radioactive substrate by paper electrophoresis and counted. Both methods were used to determine the activity of the enzyme in human red blood cells. For 20 parallel determinations a standard deviation of +/- 9% and +/- 5% was obtained for method a and b, respectively. The specific activity of the enzyme in erythrocytes of adults was found to be 0.40 +/- 0.05 U/g hemoglobin with method a, and 0.50 +/- 0.05 U/g hemoglobin with method b.

Effect of malathion on oxygen consumption and blood cholinesterase of the hen.

The effect of malathion on oxygen consumption by liver and kidney slices, and on cholinesterase enzyme (ChE) of plasma and red blood cells (RBC) has been investigated in hens. Single oral doses of 262.40 and 393.60 mg/kg (equivalent to 50% and 75% of the LD50 respectively), resulted in an initial rise of plasma cholinesterase followed by a fall. The RBC cholinesterase on the other hand was significantly inhibited right from the beginning. The plasma enzyme activity recovered earlier returning to almost normal values in six to eight days while the RBC enzyme continued to be significantly inhibited even after ten days. Six hours after a single oral dose of malathion, the oxygen consumption by liver slices was significantly inhibited and inhibition persisted up to 36 hours. Thereafter, it gradually returned to normal values in the next 36 hours. No change in oxygen consumption by kidney slices was observed.

Hemolysis in the newborn infant resulting from deficiencies of red blood cell enzymes: Diagnosis and management

Inherited deficiencies of red cell enzymes are recognized causes of hemolysis in newborn infants. Neonatal manifestations include erythroblastosis fetalis, anemia, hyperbilirubinemia, or a combination of these. Some infants with enzyme deficiencies demonstrate no neonatal manifestations; in others death may result from hydrops fetalis or kernicterus. In cases of suspected hemolysis, simple diagnostic measures will identify the most common causes of the anemia and/or the hyperbilirubinemia; these are not inherited deficiencies of enzymes but are red cell sensitization and congenital and acquired infections. Persistent hemolysis and abnormalities in the G-6-PD screening test, in the Heinz body preparation, or in an autohemolysis test support a tentative diagnosis of enzyme deficiency. The diagnosis is confirmed by specific enzyme assay. Appropriate therapy depends on the severity of anemia and/or hyperbilirubinemia and may include transfusion of packed red cells or an exchange transfusion. Careful follow-up is essential to determine the chronicity and severity of hemolysis.

Genetic variation of red blood cell enzymes in the dog. Use of soluble glutamic oxaloacetic transaminase as proof of chimerism.

One hundred ten dogs, of whom at least 63 were unrelated, were examined for electrophoretically distinguishable, genetic variants of 22 different red blood cell enzymes and of hemoglobin. A variant of soluble glutamic oxalaocetic transaminase (S-GOT) was found to occur with high frequency in dogs of the Basenji breed, and infrequent variants of indophenol oxidase and acid phosphatase were found in other breeds. The S-GOT variant was utilized to establish the presence of donor- and the absence of recipient-type cells following marrow transplantation.

Purification of human erythrocyte phosphofructokinase

Phosphofmctokinase* has been isolated from several mammalian tissues in highly purified or even crystallized forms [l-6] . Human PFK received greater attention when a new human disease was detected, which could be characterized by a deficiency of muscle PFK and a decrease of the enzyme in red blood cells to about 50% of normal. These findings led to comparative studies of various properties of both enzymes, which revealed characteristic differences between them [7-91 . In these studies only partially purified erythrocyte PFK with a specific activity of 12wasused [7]. Recently, we described the occurrence of various oligomeric forms of human erythrocyte PFK, and determined the molecular weights of their SDS cleavage products [lo] . In this paper the procedure for getting a highly purified, stable and homogeneous human erythrocyte PFK preparation with a specific activity of about 130 is described. Some special properties of the enzyme are also presented.