Comparative study of the various methods for the ultrastructural localization of alkaline phosphatase activity

Summary Renal, intestinal and developing dental tissues were studied for alkaline phosphatase localization at the ultrastructural level using several metal salt modifications of the Gomori method and a new cadmium capture technic developed by Seligman. Except for particle size differences in the precipitates, localization both extra- and intracellularly were similar.This study was supported by USPHS Research Grant DE 2800-02, National Institutes of Health.     

Correlation of blood zinc and alkaline phosphatase

Out a group of 28 sheep the serum values of zinc and alkaline phosphatase were checked. A positive correlation between low values of zinc and alkaline phosphatase was ascertained. This could help in the diagnosis of zinc deficiency.     

Isolation and electrophoretic characteristics of 3 lymphocyte populations of normal human blood]

Cell electrophoresis allows separation of normal human blood lymphocytes into two main groups which are a function of their relative rates of migration, with regard to the reference speed (1 mum.sec.-1V-1.cm): the lymphocytes which have a greater mobility than this value seem to be T-lymphocytes (80,1 per cent for 42 healthy adults); on the contrary, B-lymphocytes have an inferior mobility (19,9 per cent). Two known methods are used for the selection of the lymphoid populations: spontaneous rosetting with sheep's red blood cells, which are characteristic of T lymphocytes, and adherence to nylon wool columns, which is dominant in the case of B-lymphocytes. This method confirms the fact that T-lymphocytes have a rapid migration and B-lymphocytes a slow migration. We have isolated a third population, having neither the T markers, nor the B markers. It has a very homogeneous migration, centered on the two classes 1,05 and 1,10 mum.sec.-1.V.-1.cm.     

Comparative effects of antiepileptics on the alkaline phosphatase and gamma glutamyltransferase activities in plasma and leukocytes]

Alkaline phosphatase and gamma glutamyltransferase activities have been compared in plasma and leukocytes from presumably healthy subjects and from epileptic patients under treatment with different antiepileptic drugs. Plasma enzyme activities are always increased by antiepileptic treatment, leukocytic enzyme activities are increased only for some patients. No relation has been observed between the variations in any of the two enzyme activities and plasma level of anticonvulsant drugs.     

Stability of isoenzymes of alkaline phosphatase in various buffer systems.

The stability of isoenzymes of alkaline phosphatase from liver, bones and small intestine was compared after addition to inactivated serum in the buffer systems: glycine, 2-amino-2-methyl-1-propanol, diethanolamine and 2-amino-2-methyl-1,3-propandiol at 37 degrees C. The mentioned isoenzymes were inactivated to different extents in glycine and 2-amino-2-methyl-1-propanol buffers. In diethanolamine and 2-amino-2-methyl-1,3-propandiol buffers sufficient stability of isoenzymes is obtained so that only these buffers are suitable for activity determinations of alkaline phosphatase at 37 degrees C.     

Developmental characteristics of the human lumbar spinal column at various periods of ontogeny

Regularities in development and differentiation of vertebrae have been investigated according to ageing. The data concerning the size of the vertebral canal at the level of the lumbar vertebral bodies and intervertebral discs, intensity of increase of these dimensions in various age periods, the character of spatial relations of sizes and form of the canal has been analysed. The transversal section area of the canal and the body in the corresponding segment varies between 1:3 and 1:5.5. By means of x-ray osteophotometry the mineralization degree of the lumbar vertebral bodies has been determined in men of various age. An important fact has been stated on a close connection between mineral saturation in juvenile and first mature ages with induces of general strength.     

Co-purification of type I alkaline phosphatase and type I phosphoprotein phosphatase from various animal tissues

A purification procedure, which included ethanol treatment as a step for dissociating the large molecular forms of type I phosphoprotein phosphatase, was employed for the studies of the alkaline phosphatase and phosphoprotein phosphatase activities in bovine brain, heart, spleen, kidney, and uterus, rabbit skeletal muscle and liver, and lobster tail muscle. The results indicate that the major phosphoprotein phosphatase (phosphorylase a as a substrate) and alkaline phosphatase (p-nitrophenyl phosphate as a substrate; Mg²⁺ and dithiothreitol as activators) activities in the extracts of all tissues studied were copurified as an entity of M_r = 35,000. The purified enzymes from different tissues exhibit similar physical and catalytic properties with respect to either the phosphoprotein phosphatase or the alkaline phosphatase activity. The present findings indicate that (a) the M_r = 35,000 species, which represents a catalytic entity of the large molecular forms of type I phosphoprotein phosphatase, is widespread in animal tissues, indicating that it is a multifunctional phosphatase; (b) the association of type I alkaline phosphatase activity with type I phosphoprotein phosphatase is a general phenomenon.     

Properties of placental alkaline phosphatase. I. Molecular size and electrical charge of the various electrophoretic components of the six common phenotypes

The properties of the six common phenotypes (F₁, S₁, I₁, F₁S₁, S₁I₁, F₁I₁) of placental alkaline phosphatase have been studied by DEAE-Sephadex chromatography, gel filtration, two-dimensional paper-starch gel and starch block electrophoresis. The rapidly migrating component (component I) had the same molecular weight (118,000) in all phenotypes studied but different negative charge (F₁>I₁>S₁). The fastest of the slow-moving components (component II) had a molecular weight of 130,000 and a higher absolute negative charge, but slightly lower net negative charge than I. The very slow-moving components (II) were usually composed of two major and several minor components. The major components had a higher absolute negative charge but a lower net negative charge than II. All II components had a molecular weight of over 200,000. It is suggested that II is an enzyme bound to a small molecule, while II components are aggregates of enzyme with various substances or with itself.This work was supported in part by U.S. Public Health Service Grant HD02552. K.H. is a Career Scientist of the Health Research Council of the City of New York (I-513).     

A rare electrophoretic enzyme variant of serum alkaline phosphatase in a group of Icelanders.

An electrophoretic isoenzyme variant of serum alkaline phosphatase was found in 10 out of 343 subjects belonging to an Icelandic population in Husavik and the Husavik region. 9 of the variant-positive subjects were women. The enzyme variant differs from normal isoenzymes in electrophoretic mobility, substrate specificity, and response to inhibitors. It could be demonstrated that nine of the subjects with the enzyme variant were related with each other.     

Induction of E. coli alkaline phosphatase synthesis in cells preincubated at low temperatures]

Cell preincubation at lowered t degrees was found to result in increased alcaline phosphatase synthesis. The ability of cells for increased alcaline phosphatase synthesis correlates with increased content of cis-vaccinic acid and higher liquidity of lipids. It has been ascertained that modifications caused by cell preincubation at lowered t degrees favour the greater stability of mRNA coding the alcaline phosphatase.     

Comparative characteristics of cyclodextrin glycosyltransferases from various groups of microorganisms]

Cyclodextrin glycosyltransferases (CGT-ase, 1.4-alpha-glucanotransferase, cyclizing, EC 2.4.1.19) produced by some thermophilic, alkalophilic and mesophilic bacterial strains, were isolated and characterized. It was shown that thermophilic and mesophilic CGT-ases represent a mixture of alpha-, beta- and gamma-cyclodextrins (CD), alpha-cyclodextrin being the predominant component. Alkalophilic enzymes produce only beta-CD and are able to produce CD not only from starch but also from maltose, melibiose, maltotriose, amylose and amylopectin. The optimal conditions for the catalytic activity of the enzymes were determined. It was found that calcium, magnesium and zinc ions have a beneficial effect on the specific activity of these enzymes. The amino acid composition of the enzymes was studied.