Dehydrogenase Activity In Erythrocytes Research Articles

INFLUENCE OF MELATONIN ON AGE-RELATED CHANGES IN CARBOHYDRATE METABOLISM AND ANTIOXIDANT CAPACITY IN THE BLOOD OF ALLOXAN DIABETIC RATS

Melatonin and its metabolites have potent antioxidant/anti-inflammatory properties, and they have proven to be highly effective in a variety of disorders linked to inflammation and oxidative stress. The object of this experimental research was to ascertain the influence of aging on the level of basal glycemia and activities of glucose-6-phosphate dehydrogenase [EC1.1.1.49], pyruvate kinase [EC 2.7.1.40] and glutathione reductase [EC1.6.4.2] in erythrocytes of alloxan diabetic rats on the background of melatonin injections. Methods: We used 100 male Wistar rats, two age groups: the - 2-month (adult), and II - 4-month (old). Alloxan diabetes was evoked via injecting the rats with a 5% solution of alloxan monohydrate intraperitoneally in a dose of 170 mg/kg. Four days after diabetes induction, rats were divided into diabetic (untreated) and melatonin-diabetic group (10 mg/kg, daily and intraperitoneally for six weeks). Blood was taken from the tail vein evaluate the basal glycemia on 5-th and 47-th day after the injection of alloxan. Rats were sacrificed at the 47-th day of the experiment accordance with the ethical treatment of animals. Determinations of the enzymes activities were by standard methods. Statistical analysis was performed using Statistica 10 StatSoft Inc. Results. The level of basal glycemia on the fifth day of the experiment in animals of both groups increased on average by 115% from baseline values. We founded that on 47-th day this index was higher in group of old rats on 20% more than in adult rats. Pyruvate kinase activity in erythrocytes of adult and old animals with diabetes decreased by 34% and 51% respectively compared with the control. glucose-6-phosphate dehydrogenase activity in erythrocytes of adult and old animals with diabetes decreased by 25% and 44% respectively compared with the control on 47-th day. The changes may be the result of age-related disorders of glucose metabolism due to disturbances in free radical mechanisms. Glutathione reductase activity in erythrocytes of adult and old animals with diabetes decreased by 30% and 36% respectively compared with the control on 47-th day. A 42-days injection of melatonin to the alloxan diabetic rats of both groups contributed to a normalization of the level of basal glycemia, the activities of pyruvate kinase and glutathione reductase in the rat blood, as well as to a considerable increase of the activity of glucose-6-phosphate dehydrogenase, whose level exceeded by average 9% this particular index in the control group of animals. Under the influence of melatonin increase activity of glucose-6-phosphate dehydrogenase in the blood of rats may be due to the increasing number of substrate for glucose-6-phosphate dehydrogenase (stimulating the flow of glucose into cells and its phosphorylation) and direct action. Conclusion. In this case melatonin probably increases use of glucose for regeneration of NADPH2 and aerobic oxidation of glucose that indicate an acceleration of antioxidative protection and energy production in blood of adult and old diabetic rats.

AGE-RELATED CHANGES OF GLYCOLYTIC ACTIVITY AND ANTIOXIDANT CAPACITY IN THE BLOOD OF ALLOXAN DIABETIC RATS

The increasing incidence of type 1 diabetes coupled with advances in treatment of type 1 diabetes has resulted in an unprecedented number of older adults living with and controllable type 1 diabetes. The objective of this experimental study was to assess the impact of aging on the level of basal glycaemia and activities of glucose-6-phosphate dehydrogenase [EC1.1.1.49], pyruvate kinase [EC 2.7.1.40] and glutathione reductase [EC1.6.4.2] in erythrocytes of alloxan-diabetic rats. Methods: We used 100 male Wistar rats, divided into two age groups: I group included- 2-month (adult) animals, and II group was made up of 4-month (old) animals. Diabetes was modelled by injecting the rats with 5% solution of alloxan monohydrate intraperitoneally in a dose of 170 mg/kg. Blood was taken from the tail vein to evaluate the basal glycaemia on 5-th and 47-th day after the alloxan injection. Rats were sacrificed on the 47-th day of the experiment in accordance with the regulations on ethical treatment of vertebrates. The assessment of the activity of the enzymes was carried out by standard methods. Statistical analysis was performed by using Statistica 10 StatSoft Inc. Results. The level of basal glycaemia on the fifth day of the experiment in the animals of both groups went up on average by 115% from baseline values. We founded that on 47-th day this index was higher in group of old rats by 20% than in adult rats. Pyruvate kinase activity in erythrocytes of adult and old animals with diabetes decreased by 35% and 50% respectively compared with the control. Glucose-6-phosphate dehydrogenase activity in erythrocytes of adult and old animals with diabetes decreased by 27% and 45% respectively compared with the control on 47-th day. The changes may be considered as the result of age-related disorders of glucose metabolism due to disturbances in free radical mechanisms. Glutathione reductase activity in erythrocytes of adult and old animals with diabetes decreased by 29% and 35% respectively compared with the control on 47-th day. Conclusion. We have determined when getting aged, the alloxan-diabetic rats demonstrate changes in the sensitivity of pyruvate kinase, glucose-6-phosphate dehydrogenase and glutathione reductase activities in erythrocytes resulted from the effect of diabetes mellitus factors (hyperglycaemia). We can suggest that glycaemic control is key purpose for older patients with type 1 diabetes in order to prevent of complication, which can be aggravated with age.

Modification of the Catalytic Properties of Erythrocyte Aldehyde Dehydrogenase in Rats after Nitric Oxide Inhalation.

We studied aldehyde dehydrogenase activity in erythrocytes from healthy rats and animals with thermal trauma after NO inhalation. NO had an activating effect on catalytic properties of aldehyde dehydrogenase in healthy rats and burned animals. The effect of NO was more pronounced during burn disease.

WITHDRAWN: Modified high-performance liquid chromatographic conditions for determination of inosine 5′-monophosphate dehydrogenase activity inerythrocytes

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RETRACTED: Validation and application of a high-performance liquid chromatographic-based assay for determination of the inosine 5′-monophosphate dehydrogenase activity in erythrocytes

Thiopurine drug monitoring has become an important issue in treating children with acute lymphoblastic leukaemia (ALL). In this population, a genetic polymorphism causes wide differences in the activity of thiopurine S-methyletransferase (TPMT)--the rate-limiting enzyme of the thiopurine degradation metabolism--leading to the necessity of drug dose adjustments. It is not yet known if similar differences exist in the inosine 5'-monophosphate dehydrogenase (IMPDH; EC 1.1.1.205), the rate-limiting enzyme of the thiopurine synthesis. To test this, we established and validated a high-performance liquid chromatographic (HPLC)-based assay to determine the IMPDH enzyme activity in erythrocytes. The remarkable features of this assay are its simple erythrocyte separation/haemolysis and assay conditions and a distinct segregation of xanthosine 5'-monophosphate (XMP) from the clear supernatant after precipitation. The probes were processed without a time-consuming extraction and heating procedure and the assay demonstrated a good intra- and interday stability as well as a recovery rate of approximately 100%. The IMPDH enzyme activity was measured in erythrocytes of 75 children with diagnosis of ALL before starting antileukaemic therapy and their activity compared to those of 35 healthy adult controls. The measured enzyme activity was wide ranging in both groups. The individual enzyme activity differences observed in children with ALL might led to differences in the thionucleotide levels in those undergoing the standard thiopurine dose regimen.

Glucose-6-phosphate dehydrogenase activity in erythrocytes from chronically copper-poisoned sheep

The present study was undertaken to investigate the capacity to generate reducing equivalents in erythrocytes from experimentally copper-poisoned sheep. Ten ewes were dosed orally with CuSO 4 to induce the Cu toxicity. Copper dosing was stopped at the first day of hemolysis. The activity of glucose-6-phosphate dehydrogenase (G6PD) in the erythrocytes, the levels of reduced glutathione (GSH) and glucose (in serum and erythrocytes) was examined at frequent intervals. The copper-poisoned sheep had reduced levels (25–35% less) of glucose in serum and erythrocytes than controls. The activity of G6PD in erythrocytes from sheep was 50–60% of typical levels found in human erythrocytes. Immediately before the hemolytic period, the copper-poisoned sheep showed decreased activity of G6PD, declining to 65% of the initial activity. In addition, we found decreased blood levels of reduced GSH in copper-poisoned sheep. There appears to be a relationship between decreased capacity to generate reducing equivalents and the overload of copper in sheep erythrocytes.

Effect of ageing on erythrocyte aldose reductase and sorbitol dehydrogenase activity

We measured erythrocyte aldose reductase and sorbitol dehydrogenase activity in erythrocytes in healthy individuals aged from 16 to 91 years to determine the mechanism of age-dependent sorbitol accumulation. Erythrocyte aldose reductase activity increased significantly with age but ageing had no effect on sorbitol dehydrogenase activity. Age and the aldose reductase/sorbitol dehydrogenase ratio were positively correlated. These findings suggest that an increase in the ratio of aldose reductase to sorbitol dehydrogenase may contribute to the tissue accumulation of sorbitol in the elderly and may be a mechanism of a disease that is common in elderly individuals.

Effect of haemodialysate and its peptide fractions on lactate dehydrogenase activity in erythrocytes from healthy subjects and patients with end-stage renal disease.

The effect of haemodialysate and its 3 peptide fractions on lactate dehydrogenase activity in erythrocytes was studied in patients treated by repeated haemodialysis for end-stage renal disease and in healthy subjects. Erythrocytes from dialyzed patients showed significantly lower LDH activity than those from healthy subjects. After a 3-h incubation with haemodialysate (675 and 450 micrograms protein/ml), or its peptide fraction III (270 and 190 micrograms protein/ml), a significant inhibition of LDH activity was observed. On the other hand, neither haemodialysate nor its peptide fractions inhibited LDH activity in red blood cells from patients with end-stage renal disease treated by repeated haemodialysis.

Optimized Spectrophotometric Determination of Aldehyde Dehydrogenase Activity in Erythrocytes

We describe a reliable and sensitive semiautomated spectrophotometric assay of aldehyde dehydrogenase (ALDH; EC 1.2.1.3) activity in erythrocytes. The hemolysate can be stabilized with sucrose, and the technique involves only microliters of hemolysate on a centrifugal analyzer. The use of microcolumns to remove interfering hemoglobin is avoided, and reproducibility of the assay has been improved by manipulating the inherent lactate dehydrogenase activity of erythrocytes by adding lactate and oxalate to the reaction mixture. These modifications have decreased the analytical imprecision of the assay, allowing a better appraisal of aldehyde dehydrogenase activity in erythrocytes as a biological marker of excess alcohol consumption. Erythrocytic ALDH activity was significantly less in 40 alcoholics than in 145 teetotallers (median activity 128 vs 219 mU/g of hemoglobin, respectively; P = 0.0001), indicating the potential of this assay as a useful marker of excess alcohol consumption.

SORBITOL DEHYDROGENASE IN HUMAN LENS: STUDY OF THE ENZYME IN VARIOUS HUMAN TISSUES

Several patients with congenital cataracts were described to have a deficient sorbitol dehydrogenase activity in erythrocytes (Human Genet. 61:338, 1982; J. Inner. Metab. Dis. 7 Suppl.2:151, 1984). Since there were obscure findings concerning the erythrocyte enzyme activity and the inheritance, we have studied the enzyme in lens and compared its property with that in other tissues such as liver and blood. The specific activity of sorbitol dehydrogenase in lens was higher than in liver (4.9-5.4 nmol/min/mg protein vs. 2.9-4.2 n=8 each). The Michaelis constant for sorbitol was low in lens compared to that in other tissues (0.9 nmol/L in lens, 2.6 mmol/L in liver). Polyacrylamide gel iso-electrofocusing of the enzyme in lens showed multiple bands between pH 4.4-8.1, while in liver 2-4 bands at pH 7.0-8.6 and in erythrocytes 3-4 bands at pH 7.6-8.4. Xylitol inhibited the enzyme in all samples studied by 30-50 %, whereas xylulose activated the enzyme by 200% in liver, in lens and erythrocytes by 40-60 %. These results indicate that sorbitol dehydrogenase in lens is unique compared with that in liver or erythrocytes.

Sorbitol dehydrogenase deficiency in a family with congenital cataracts.

Cataract is one of the important clinical manifestations of galactosaemia due to galactokinase or galactose-1-phosphate uridyltransferase deficiency. It has been known that the accumulation of polyols, sorbitol or galactitol in a lens plays a key role in the formation of diabetic and galactosaemic cataracts (van Heyningen, 1976). It is also well established that the sorbitol pathway involving aldolase reductase (EC 1.1.1.21) and sorbitol dehydrogenase (EC 1.1.1.14) is important in cataract formation (Varma et al., 1977). Recently Vaca et al. (1982) reported the first case of red cell sorbitol dehydrogenase deficiency in a family with cataracts. Therefore we have decided to determine the sorbitol dehydrogenase activity in erythrocytes in addition to galactokinase, galactose-1-phosphate uridyltransferase and UDP-galactose epimerase activities in a selected population with congenital cataracts.KeywordsGlycogen Storage DiseaseCongenital CataractMaple Syrup Urine DiseaseSorbitol DehydrogenaseDiabetic RabbitThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

The Interference of Leukocytes and Platelets With Measurement of Glucose-6-Phosphate Dehydrogenase Activity of Erythrocytes With Low Activity Variants of the Enzyme

AbstractComplete removal of leukocytes and platelets from whole blood showed that the glucose-6-phosphate dehydrogenase (G6PD) activity in “pure” erythrocytes from G6PD-deficient hemizygous Sardinian subjects is consistently lower than reported in the literature. Thus, although none of 27 hemizygous subjects showed undetectable erythrocyte G6PD activity, their levels ranged between 0.0015 and 0.008 IU/g Hb, as compared with a mean value of 4.5 IU/g Hb in normal subjects. Most of the biochemical properties that were formerly ascribed to erythrocyte G6PD appear to be those of the enzyme from contaminating leukocytes and (or) platelets.

The interference of leukocytes and platelets with measurement of clucose-6-phosphate dehydrogenase activity of erythrocytes with low activity variants of the enzyme

Complete removal of leukocytes and platelets from whole blood showed that the glucose-6-phosphage dehydrogenase (G6PD) activity in “pure” erythrocytes from G6PD deficient hemizygous Sardinian subjects is consistently lower than reported in the literature. Thus, although non of 27 hemizygous subjects showed undetectable erythrocyte G6PD activity, their levels ranged between 0.0015 and 0.008 IU/g Hb, as compared with a mean value of 4.5 IU/g Hb in normal subjects. Most of the biochemical peoperties that were formerly ascribed to erythrocyte G6PD appear to be those of the enzyme from contaminating leukocytes and (or) platelets.

Activities of catalase in leucocytes and glucose-6-phosphate dehydrogenase in erythrocytes of hypocatalasemia and acatalasemia.

Catalase activity in leucocytes was found to be half the normal value in hypocatalasemia and extremely low in acatalasemia. Glucose-6-phosphate dehydrogenase activity in erythrocytes was not significantly different between normal, hypocatalasemia and acatalasemia in three families of acatalasemia, but in one family lower activities than normal were found in hypocatalasemia and actalasemia erythrocytes. Other enzyme activities in blood, such as alkaline phosphatase, lactate dehydrogenase, glutamic oxaloacetic and glutamic pyruvic transaminases were not significantly different between normal subjects, hypocatalasemia and acatalasemia.

Metabolism of [4-14C]estrone by sheep erythrocytes around the time of parturition

The metabolism of [4-14C]estrone in vitro by red blood cells of sheep in late pregnancy and after partuirition has been studied. [14C]estrone (600 ng) was incubated with 0.5 ml erythrocytes plus 0.5 ml of Krebs-Ringer phosphate buffer, pH 7.4, for 2 h at 37 degrees C in an atmosphere of air. After incubation, [3H]estrogens were added to the incubation medium as internal standards for identification and for correction for procedural losses. Metabolites were isolated and purified by chromatography, acetate derivative formation, and recrystallization to a constant 3H/14C ratio. Approximately 20% and 2% of added estrone were converted to 17beta-estradiol and 17 alpha-estradiol, respectively. The remainder was recovered unchanged. Daily measurements of 17 beta-hydroxysteroid dehydrogenase activity in erythrocytes of five ewes, over the period 8 days prepartum to 4 days postpartum, showed no significant change in activity.

A technique for the cytochemical demonstration in the electron microscope of glucose-6-phosphate dehydrogenase activity in erythrocytes of malaria-infected animals.

SUMMARYA technique for the cytochemical demonstration in the electron microscope of glucose‐6‐phosphate dehydrogenase activity in malaria‐infected erythrocytes is described. Reaction product occurred as diffuse electron‐dense deposits of the formazan of MTT in the red cells. No activity was observed within the malaria parasites themselves, except in the contents of food vacuoles. The results of a series of control experiments are also described.

An electron cytochemical study of glucose-6-phosphate dehydrogenase activity in erythrocytes of malaria-infected mice, monkeys and chickens

An electron cytochemical study of glucose-6-phosphate dehydrogenase activity in erythrocytes of malaria-infected mice, monkeys and chickens

Electron cytochemical study on glucose-6-phosphate dehydrogenase activity in erythrocytes of Aotus monkeys infected with Plasmodium falciparum

Journal Article Electron cytochemical study on glucose-6-phosphate dehydrogenase activity in erythrocytes of Aotus monkeys infected with Plasmodium falciparum Get access R.D.G. Theakston, R.D.G. Theakston Search for other works by this author on: Oxford Academic PubMed Google Scholar K.A. Fletcher K.A. Fletcher Search for other works by this author on: Oxford Academic PubMed Google Scholar Transactions of The Royal Society of Tropical Medicine and Hygiene, Volume 65, Issue 4, 1971, Page 417, https://doi.org/10.1016/0035-9203(71)90113-1 Published: 01 January 1971

Glucose 6-phosphate dehydrogenase activity in erythrocytes. Absence of diurnal variation.

Glucose 6-phosphate dehydrogenase activity in erythrocytes. Absence of diurnal variation.

G-6-PD deficiency gene dynamics in a Brazilian population.

SummaryStudies on the glucose-6-phosphate dehydrogenase activity of erythrocytes from 416 males and 820 females are reported. The total number of deficient males (intermediate + complete) varied from 3.9% among the whites (102 persons) to 15.1% among Dark Mulattoes (99 individuals); the incidence in the Negroid sample in general (316 persons) was 12.4%. The data obtained among the women were utilized to study problems of penetrance of the G-6-PDD genes. Results obtained in both sexes were employed to verify, unsuccessfully, possible selection differentials through age analysis and the comparison of the gene incidences of people affected with minor ailments and controls. Problems of gene flow analysis in populations like this one in which the “Negro” and “White” forms of the deficiency are present were stressed.