Blood Cell Enzyme Research Articles

Clustering patients with late-life depression by blood glutathione-dependent enzymatic activities for stratification of a heterogeneous group

IntroductionWe have previously found significant alterations in activities of glutathione dependent enzymes in blood cells of patients with late-life depression (LLD) compared with age-matched controls.ObjectivesThe revealing subgroups of LLD patients by glutathione-metabolism enzymes’ activities in blood cells using cluster analysis.Methods LLD patients (n=101) of 60-86 age (69 patients with recurrent depression (RD), 23 with bipolar disorder (BD) and 9 patients with a single depressive episode (DE)) were assessed by Hamilton depression rating scale (HAMD-17), and Hamilton Anxiety Rating Scale (HARS). Activity levels of glutathione reductase (GR) and glutathione S-transferase (GST) were determined in patients’ platelets (-pl) and erythrocytes (-er). The control group consisted of 51 peoples 55-84 years old without mental pathology. Cluster analysis module of the STATISTICA software was used for clustering the patients by baseline blood parameters.Results Three clusters of patients were obtained: C1, n=39, C2, n=31, C3, n=31, differing significantly in all biochemical parameters (Kruskal-Wallis test, p<0.001), except GST. When compared with control group by Mann-Whitney test, GST-pl, GST-er, and GR-er were significantly decreased in C1; GST-er was significantly increased in C2; GST-pl, GR-pl, and GR-er were significantly decreased in C3. Several significant correlations were found between the measured parameters and scores by HDRS or HAMD-17. In C1, baseline activity of GST-er correlated with total scores by HAMD-17 (R=0.335, p=0.043) after treatment. In C2, baseline activity of GR-er correlated with total scores by HARS (R=-0.376, p=0,037) after treatment and GR-pl correlated with delta scores by HAMD-17 under the treatment (R=0.484, p=0.006). No significant correlations were found in C3. Patients with BD distributed significantly unevenly between C1, C2, and C3, with significantly more BD patients clustering in C1 (61%) compared with C2 and C3 (Yetes-corrected Chi-square =7.73, p=0.0054), whereas patients with RD and DE distributed evenly.Conclusions Patterns of activity levels for glutathione-dependent enzymes in patients with BD differ from those in patients with RD and DE. Significant correlations of the measured biochemical parameters with scores by HDRS or HAMD-17 assessed after the treatment and evidenced for the treatment efficacy seem to be promising biomarkers for further evaluation of the treatment efficacy in heterogeneous group of LLD patients using the proposed approach to their stratification into subgroups.Disclosure of InterestNone Declared

Activity of energy, glutamate, and glutathione metabolism enzymes in blood cells of elderly patients with depression

Heterogeneity of depression in older adults is a challenge for the development of person-centered treatment. To address this, we studied glutamate and glutathione metabolism enzymes in blood cells in 53 older adult patients with depression and 20 controls. Patients with depression had decreased platelet glutathione-S-transferase and erythrocyte glutathione reductase. The biochemical and clinical data contributed to three clinical clusters that are not linked to the onset of depression or its duration but were related to anxiety, cerebrovascular and cardiovascular comorbidities, including parkinsonian features such as tremor, akathisia, and rigidity. These findings could aid person-centered diagnosis and outcomes and timely successful treatment(s) of depression in older adults.

Blood cells enzymes as an indicator of adaptive processes in newborns delivered off by mothers with iron deficiency anemia

Aim. To explore the enzymes in blood cells of newborns from mothers with iron deficiency anemia.Methods. During the study, 150 infants and their mothers underwent clinical and laboratory examination, of whom 130 infants had antenatal iron deficiency of different severity (main group), 20 children were included in the control group. Cytochemical analysis of peripheral blood was performed on 1st, 3rd, 7th day after birth by smear staining and determination of succinate dehydrogenase in lymphocytes, myeloperoxidase in neutrophils, acid and alkaline phosphatase in neutrophils and lymphocytes by microscopic analysis.Results.Succinate dehydrogenase activity was reduced on the 3rd day of life in lymphocytes of the newborns of the first group who were delivered off by mothers with latent iron deficiency. Average level of acid phosphatase in lymphocytes and neutrophils was higher compared to controls throughout the study (pConclusion. Cytochemical studies of peripheral blood neutrophils and lymphocytes may be used as a screening test to assess the immunity in newborns who were delivered off by mothers with iron deficiency, and for personalized management of patients with comorbidities.

Non-enzymatic antioxidant capacity assays: Limitations of use in biomedicine

The ‘Total antioxidant capacity’ (TAC) is a parameter frequently used for characterization of food products and of the antioxidant status of the body. This mini-review shows shortcomings of TAC assays and points of concern that should be considered when performing and interpreting results of such assays. The term TAC is not optimal since the assay measures only part of antioxidant capacity, usually excluding enzymatic activities. Antioxidant and oxidant-regenerating enzymes in blood cells and the blood vessel wall have a profound impact on the antioxidant properties of blood plasma, which is not reflected in the in vitro assays of isolated plasma. The term ‘Non-enzymatic antioxidant capacity’ (NEAC) is suggested as more relevant than TAC. NEAC is estimated by various methods, which yield different values and results obtained using different methods do not always show satisfactory correlation. One reason for the discrepancy of results is the use of different oxidants in NEAC assays. The use of hydroxyl radical as the oxidant is not recommended in view of the high and non-specific reactivity of this species.

Oxidative stress induces GSTP1 and CYP3A4 expression in the human erythroleukemia cell line, K562.

To explore the effects of oxidative stress on metabolizing enzymes in blood cells, we examined the effect of hydrogen peroxide (H(2)O(2)) on glutathione S-transferase (GST) and cytochrome P450 (CYP) expression in a human erythroleukemic cell line, K562. After adding H(2)O(2) (up to 100 microM) to the culture medium of K562 cells, the expression levels of GST and CYP enzymes were monitored by RT-PCR. The expression of GSTP1 and CYP3A4 was induced by oxidative stress. Quantitative PCR and immunoblot analysis revealed a 3- to 4-fold increase in GSTP1 and CYP3A4 mRNA, and a 2- to 3-fold increase in GSTP1 and CYP3A4 protein levels, 3 d after the addition of 100 microM H(2)O(2). Induction was H(2)O(2) dose-dependent and also depended on the length of culture. Our results suggest that oxidative stress may affect GST and/or CYP expression in human blood cells, which may alter the metabolism of drugs and xenobiotics and thus, the toxicity of these compounds to the blood cells.

Copper intake and assessment of copper status

The diagnosis of marginal copper deficiency has not been perfected despite an increased understanding of the physiologic roles of copper. The use of nonstandardized procedures and the effects of factors other than copper nutriture have impeded identification of an ideal indicator of copper nutritional status in humans. A review of studies of experimental copper deprivation conducted in adult humans over the past 12 y indicated that between 1.0 and 1.25 mg Cu/d is needed by adults for copper maintenance for periods of up to 6 mo and that < or = 2.6 mg Cu/d for periods of up to 42 d is not sufficient for recovery from copper deprivation. Copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet cytochrome-c oxidase, may be better indicators of metabolically active copper and copper stores than plasma concentrations of copper or ceruloplasmin because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture.

Effect of topically applied sulphur mustard on antioxidant enzymes in blood cells and body tissues of rats.

The effect of sulphur mustard (0.5 LD50, percutaneous) on antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px)) in blood cells (erythrocytes (RBC), leucocytes (WBC) and platelets) and body tissues (liver, kidney, spleen and brain) of rats has been investigated 24 h post exposure. The SOD activity was significantly decreased in WBC, platelets, spleen and brain as compared to control. The CAT activity was significantly inhibited in RBC, WBC and spleen as compared to control. The GSH-Px activity was signficantly depressed in WBC, spleen and liver as compared to control. It is concluded that sulphur mustard at a sublethal dose inhibited antioxidant enzyme activities in WBC and spleen. Thus, antioxidant enzymes in lymphatic tissues may be used as suitable models for assessing mustard toxicity. The study suggests the formation of reactive oxygen species in sulphur mustard intoxication.

Assessment of copper nutritional status

Despite increased interest in the role of copper deficiency in clinical problems and an increased understanding of the physiological roles of copper, the diagnosis of a marginal deficiency has not been perfected. The use of non-standardized procedures and the effects of factors other than copper nutriture have impeded identification of the "ideal" indicator of copper nutritional status in adult humans. The specific activity of copper enzymes, or of copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet or leukocyte cytochrome c oxidase, may be a better indicator of metabolically active copper stores than the serum concentration of copper or ceruloplasmin, because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture. A single index, such as serum copper concentration, is inadequate for assessing the total body copper nutriture of an individual and must be supported by corroborating evidence.

HLA gene and haplotype frequencies in galicia (NW Spain)

The genetic structure of six populations of Iran (Turks, Kurds, Lurs, Zabolis, Baluchis and Zoroastrians) was examined using data on blood groups, serum proteins and cell enzymes. Our results show conclusively that there are genetic differences among the six populations and the analysis of superimposed R and S matrices defined by Harpending & Jenkins (1973) show that the dispersion of some of the alleles correspond to the dispersion of the populations. The FST estimates are not large enough to favour selection on any of the loci studied. The FIT and FIS estimates are positive and moderately high suggesting that the genetic differentiation to some extent is influenced by inbreeding.

Population genetics of the people of Iran II. Genetic differentiation and population structure

The genetic structure of six populations of Iran (Turks, Kurds, Lurs, Zabolis, Baluchis and Zoroastrians) was examined using data on blood groups, serum proteins and cell enzymes. Our results show conclusively that there are genetic differences among the six populations and the analysis of superimposed R and S matrices defined Harpending & Jenkins (1973) show by that the dispersion of some of the alleles correspond to the dispersion of the populations. The FST estimates are not large enough to favour selection on any of the loci studied. The FIT and FIS estimates are positive and moderately high suggesting that the genetic differentiation to some extent is influenced by inbreeding.

Gametic equilibrium between 24 polymorphic markers

Pairs of unlinked genetic markers (blood cell antigens and enzymes and serum proteins) were tested for gametic equilibrium (GE) in data from 508 unrelated individuals from different regions of North America. A total of 24 markers were considered in the study. Only 14 of 262 pairs (5.3%) of markers deviated from GE at the 5% significance level and only 6 pairs (2.3%) remained in significant disequilibrium after accounting for genetic heterogeneity among regional groups. The particular combinations of alleles or one-locus genotypes that showed significant association were identified in case they might be replicated in future studies. On the basis of this study alone, however, there is no reason to suspect disequilibrium between the pairs of markers studied.

Parentage testing of dogs using variants of blood proteins: description of five new plasma protein polymorphisms

Plasma samples of 1126 dogs belonging to 21 different European breeds were analysed by two-dimensional agarose gel (pH 5.4 or pH 8.6)--horizontal polyacrylamide gel (pH 9.0) electrophoresis, followed by general-protein staining of gels. Genetic polymorphism was detected for five as yet unidentified proteins designated pretransferrin-1 and -2 (Prt1 and Prt2) and postalbumin-1, -2 and -3 (Pa1, Pa2 and Pa3). Three alleles are reported in the Prt1 and Prt2 systems and two alleles in the Prt2, Pa1 and Pa3 systems. While Prt2 variation was observed only in the cocker spaniel breed, each of the other four proteins showed a high degree of polymorphism in most of the breeds studied. Pa3 fractions were clearly observed only in samples stored at -20 degrees C for more than 2 years. Prt1, Pa1 and Pa2 proteins are additional useful markers for parentage control in dogs. This study corroborated previously published results that dog plasma proteins, in general, show considerably more polymorphism than that reported for haemoglobin and for several blood cell enzymes in this species.

Absence of Processing Enzymes for Inactive Renin in Mouse Blood but Polymorphonuclear Neutrophils Contain Active Renin

The possible role of enzymes in plasma and blood cells for the conversion of inactive renin was investigated in mice of the Balb C strain. The concentrations of both inactive and active renin were unchanged in unfractionated plasma and whole blood during incubation at 37 degrees C for 5 h, indicating, that inactive renin had not been activated. Purified platelets, mononuclear cells (monocytes and lymphocytes), and polymorphonuclear leukocytes were incubated separately with partially purified inactive renin at 37 degrees C for 1 h. Inactive renin was neither activated nor degraded by whole blood cells. Polymorphonuclear neutrophils were also obtained in mice after i.p. injection of fluid thioglycollate medium. Extracts of the polymorphonuclear neutrophils were incubated with partially purified, inactive renin. During incubation at pH 7.5 for 1 h at 37 degrees C, inactive renin was stable and no activation was registered, whereas degradation, without a concomitant increase in active renin, occurred at pH 4.5. Extracts of polymorphonuclear neutrophils were found to contain active, but no inactive renin. The mean amount of renin per cell was 0.5 X 10(-11) Goldblatt units (GU) (range 0.3 X 10(-11) GU - 0.9 X 10(-11) GU; n = 6). These findings argue against a role for the blood cells and plasma in a physiological activation of inactive renin, and do not support the concept that activation of inactive renin in the circulation may participate in homeostasis.

Red cell enzyme levels in the Australian aborigines

Ethnic differences are known to occur in the red cell levels of ATP1, reduced glutathione2 and the activity of pyridoxal kinase3. We have now measured the activities of 12 different enzymes in the red blood cells of full blood Aborigines representing 3 populations and compared them with Caucasian subjects. Some of the enzyme activities were similar in the 3 Australian Aboriginal populations, as well as the Caucasians, while others showed significant differences. However, these differences were present both between Aborigines and Caucasians and among the Aboriginal populations. It appears that there are no ethnic differences in the levels of red cell enzymes measured in the present study.

Trimethoprim-induced elevation of dihydrofolate reductase activity in human leukocytes.

The administration of trimethoprim (TMP)--a diamino benzylpyrimidine compound which binds very tightly the bacterial dihydrofolate reductase--was accompanied by the appearance of measurable levels of dihydrofolate reductase in peripheral leukocytes from patients with nonhematological diseases. In all instances, enzyme activity rose rapidly between the fourth and eighth day after TMP. The time course of the rise and fall of dihydrofolate activity approaches cellular life span and is similar to that obtained after methotrexate or triamterene administration. Dihydrofolate reductases, partially purified from leukocytes of patients treated with TMP, bone marrow and leukemic leukocytes, had simila molecular weights, pH optima, Ki of inhibitor (methotrexate); they were stimulated to the same degree by KCl and urea. Electrophoresis of the enzyme on cellulose acetate strip resulted in the separation of two enzymatically active protein components. No differences in the electrophoretic behavior of the three blood cell enzymes were noted. The findings noted above are consistent with the suggestion that the observed rise in dihydrofolate reductase activity is a quantitative one. Moreover, the effect of TMP in vivo is discussed in comparison with the currently held hypothesis for methotrexate action (stabilization by the drug of a previously synthetized enzyme).

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.

Activities of haem synthetic enzymes in blood cells of pre-natal flexed-tailed (f/f) anaemic mice

Levels of activity of beta-aminolaevulinate synthetase, delta-aminolaevulinate dehydratase, porphyrinogen synthetase and haem synthetase in circulating reticulocytes of pre-natal FL4/Re +/+ and congenitally anaemic FL1/Re f/f mice have been determined. The activities of delta-aminolaevulinate synthetase and delta-aminolaevulinate dehydratase were found to be decreased in proportion to the hypochromia observed in mature liver-derived erythrocytes in neonatal f/f mice, but activities of enzymes later in the haem synthetic pathway were relatively undisturbed. No significant differences were found in levels of haem synthetic enzymes in foetal livers of +/+ and f/f mice. These results indicate that the severe anaemia of late prenatal and neonatal f/f mice is due both to reduction in haem synthesis expressed at the reticulocyte stage of erythroid differentiation and to restricted production of erythroid progenitor cells. Retarded foetal growth and skeletal abnormalities, both characteristic of the pre-natal expression of the f/f gene complement, can also be related to reduced levels of haem synthesis, but the abnormal distribution of pigment cells seen in f/f animals appears to be a secondary effect of reduced tissue oxygenation resulting from pre-natal anaemia.

Inherited Hyperammonemic Syndromes

The term congenital hyperammonemia is generic, and the condition can be caused by inborn deficiencies of urea cycle enzymes, by some disorders of ornithine and lysine metabolism, and by some inborn errors of branched-chain amino acid catabolism. Ammonia toxicity produces similar clinical manifestations in all disorders causing hyperammonemia. The toxicity is primarily neurological, other metabolic consequences of hyperammonemia being less serious. Electroencephalographic changes, metabolic disturbances, disruption of neuronal microtubules, and histopathological appearance of Alzheimer type II astrocyte nuclei all occur with experimental as well as clinical hyperammonemia. When blood ammonia levels are excessive, particularly with alkalosis, fever, hypoxia, or hypoglycemia, there is irrationality, irritability, gastrointestinal upset, alternating hypotonia and hypertonia, convulsions, coma, and death. The early effects of hyperammonemia are reversible, but recurrent or persistent hyperammonemia can result in irreversible brain damage in those who survive. Characteristic aggravation or precipitation of symptoms by increased dietary protein or constipation, or a history of protein aversion, should raise the suspicion of hyperammonemia. After the confirmation of hyperammonemia, the precise biochemical diagnosis will require testing for abnormalities of blood and urine amino acids, and may require assays for specific enzymes in blood cells, cultured cells, or even liver biopsies. Treatment is by restriction of protein intake and prevention of constipation. Moderately severe hyperammonemia can be successfully treated with a low protein diet, perhaps supplemented by lactulose or other cathartic therapy. Severe hyperammonia is lethal in the newborn period, and attempts at treatment have often been unsuccessful. Prevention of recurrence is possible by genetic counseling and by antenatal diagnosis of some of these disorders.

Blood cell enzymes in trisomy E (18) syndrome.

Blood cell enzymes in trisomy E (18) syndrome.