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J Grozdea

Publications and source records attributed to J Grozdea.

At least 37 records · Page 2Linked to original sources

Isoelectric focusing of human neutrophil alkaline phosphatase isoenzymes in agarose gel.

An isoelectric focusing technique for human neutrophil alkaline phosphatase isoenzymes is described. After sonication with Zwittergent 3-12, butanol extraction and ultracentrifugation, dialysis of cytosols precedes focusing. Focusing patterns show a heterogeneity with two enzymatic activity areas: a main component at pI 6.7-6.8 with a minor component at pI 4.8-5.0 which is difficult to visualize due to its sensitivity to experimental conditions. The addition of 3 mmol/l ZnCl2 to the agarose gel improved the staining of focused bands and in particular the anodic component.

Alkaline Phosphatase↗

Thin-layer agarose isoelectric focusing of alkaline phosphatase isoenzymes from human neutrophils.

An improved method for thin-layer agarose isoelectric focusing of alkaline phosphatases (AP) from human neutrophils is described. The solubilization of AP isoenzymes was studied with four detergents. The best results were obtained after sonication with Zwittergent 3-12 (1% final concentration), followed by butanol extraction and ultracentrifugation 105,000 x g for 1 h. The cytosol can be stored at 0 degrees C or -80 degrees C, but not at -20 degrees C. Dialysis of the cytosol against a Tris buffer, pH 7.5, was imperative prior to focusing for removal of the detergent. Enzyme visualization is enhanced by incorporation of ZnCl2 (3 mM) into the agarose gel. The focusing patterns consist of two sets of isoenzymes: two main zones with pI 6.4 and 6.8 and minor components with pI 4.2, 4.8 and 5.2.

Alkaline Phosphatase↗

Heat resistance, immunological and quantitative changes of neutrophil alkaline phosphatase in trisomy 21 pregnancies.

Neutrophil alkaline phosphatase (NAP) was analysed in 25 pregnant women with trisomy 21 foetuses whose chromosomal aberration was recognized by cytogenetic study after amniocentesis. Enzyme investigation was performed at 20-22 weeks of gestation using cytochemical and biochemical techniques. Twenty-nine women at the same stage of normal pregnancies were selected as controls. In parallel, each mother was karyotyped. Ten subjects from each series underwent biochemical and immunological investigation: measurement of enzyme levels, thermostability study and immunological tests with alkaline phosphatase isoenzyme antibodies. NAP from pregnant women with trisomy 21 foetuses was characterized by: (1) a lower rate of enzyme activity, (2) a large amount of heat-stable enzyme (T = 56 degrees C for biochemical assays, T = 85 degrees C for cytochemical tests), and (3) a marked loss of liver antigenicity. These findings suggest the presence in trisomy 21 pregnancies of a non-specific alkaline phosphatase isoenzyme which appears as an "enzyme marker" in maternal circulating neutrophils.

Adult↗

An enzymatic marker in mothers of trisomy 21 children: neutrophil alkaline phosphatase.

Neutrophil alkaline phosphatase (NAP) from 12 mothers of normal children was investigated and the results compared to those of 7 mothers with trisomy 21 offsprings, in an attempt to determine a parental molecular change in this chromosomal abnormality. The biochemical properties of the enzyme were analyzed by the procedures of isoenzyme characterization, i.e. enzyme assays, thermostability, inhibition patterns and slab gel electrophoresis. Immunological properties were determined on 5 samples from normal mothers and on the same sample number of mothers with affected children. In these latter NAP showed characteristics that were to some extent different from the ones of normal controls. The following changes were observed: highly significant loading of membrane and nucleus pellets in NAP activity, poor effect of inhibitors on thermostable component and immunodepletion measured by a significant decrease of the normal affinity for antiliver and antiplacental alkaline phosphatase antisera. These findings are discussed in the light of our knowledge of alkaline phosphatase isoenzymes.

Adult↗

Cytochemical and biochemical studies on neutrophil alkaline phosphatase in parents of trisomy 21 children.

A study of karyotypes and neutrophil alkaline phosphatase (NAP) was carried out for 66 parents (33 couples) of trisomy 21 children and for 60 control parents (30 couples). Enzyme activity was determined simultaneously by biochemical and cytochemical techniques. In the mothers of trisomy 21 children we found: (a) by biochemical techniques, a significant increase of NAP activity in polymorphonuclear leukocyte (PMN) homogenates (P less than 0.01) and a lower supernatant/pellet ratio (P less than 0.01); (b) by cytochemical techniques (with or without thermal and urea treatments) NAP activity was significantly higher (P less than 0.001) than in control mothers; in the fathers, the two techniques gave normal NAP activity results.

Alkaline Phosphatase↗

Liver- and placenta-like alkaline phosphatases (EC 3.1.3.1) isoenzymes in normal human neutrophils.

Alkaline phosphatase (AP) has been characterized in the neutrophil polymorphonuclear leukocytes (PMN) from 14 healthy adults. The biochemical properties of the isoenzymes were analyzed by the following methods: enzyme assays, slab gel electrophoresis, thermostability, inhibition pattern using four chemical inhibitors, and immunological specificity. In normal PMNs, two types of AP have been found: a thermolabile isoenzyme identical to the liver/bone/kidney type and a thermostable isoenzyme with respect to its electrophoretic properties, its level of thermostability, its immunological specificity, the thermostable isoenzyme resembles the placental form more closely than the intestinal form. These results are consistent with the proposal that granulocyte AP is a heterogeneous enzyme. Two enzyme forms appear to be expressed in this blood cell.

Alkaline Phosphatase↗

Cytochemical profile of peripheric blood lymphocytes in 7 to 27 weeks' pregnancy.

A comparative study of alpha-naphthyl (nonspecific) esterase (ANAE), acid phosphatase (AcP), and beta-glucuronidase (beta-GLUS) in the blood lymphocytes was carried out on 26 healthy pregnant women (7 to 27 weeks' amenorrhea) and on 32 normal nonpregnant women, all 58 of whom were 19 to 34 years old. In pregnant women, we found a significantly lower percentage of ANAE-positive lymphocytes (after 11 weeks' amenorrhea), a strong reduction in AcP, and a slight decrease in beta-GLUS. The modified enzyme levels of circulating lymphocytes in the early stages of pregnancy seem to be related to the depressed cell-mediated immunity.

Acid Phosphatase↗

Neutrophil alkaline phosphatase in children with trisomy 21 and their parents.

A comparative study of karyotypes and neutrophil alkaline phosphatase (NAP) was carried out for 70 parents (35 couples) of trisomy 21 children and their 35 trisomy 21 children and for 110 control parents (55 couples) and their normal children. In the trisomy 21 families we found a significant increase in NAP: mother P less than 10-4; father P less than 10-4; children P less than 10-9; the NAP level in affected child is approximately equal to the sum of the NAP levels of the two parents (P = 0.80; sigma2 = 5%). In one parent of a trisomy 21 child, a karyotype anomaly was present.

Adolescent↗

Increase of neutrophil alkaline phosphatase in the parents of trisomy 21 children. Hematological and cytogenetic studies.

A comparative study of karyotypes, hematological variables and neutrophil alkaline phosphatase (NAP) was performed in 106 parents (53 couples) of children with free, homogeneous trisomy 21 and of 220 parents (110 couples) of normal children. In the parents of trisomy 21 children we found a significant increase of the NAP (p <0.05) and a nonsignificant difference in the number of white blood cells. In 4 cases, we also found an anomaly in the karyotype. A modification of the metabolism of the neutrophils seems possible.

Adolescent↗

Increase of neutrophil alkaline phosphatase in the mothers of leukemic children.

A comparative study of the polymorphonuclear neutrophiles of 62 parents of children suffering from acute leukemia and of 110 couples with normal children revealed a marked increased of Neutrophil Alkaline Phosphatase (p less than 0.05) in the mothers of the leukemic children. The other parameters (periodic acid Schiff and myeloperoxidase) were unchanged.

Acute Disease↗