Showing papers on "Alkaline phosphatase published in 1973"
TL;DR: The results suggest that the more differentiated upper villus cells incorporate more labeled sugar precursors into membrane glycoproteins than the less differentiated lower villus or crypt cells and that these membrane gly coproteins are associated with membrane-bound enzymes.
1,059 citations
TL;DR: The embryonal carcinoma enzyme resembles the enzymes from kidney and placenta in kinetics of thermal inactivation and sensitivity to the inhibitor L-phenylalanine, but is distinguishable from the alkaline phosphatases of liver and intestine.
Abstract: In tumors and embryoid bodies of mouse teratoma a correlation has been established between specific activity of alkaline phosphatase (EC 3.1.3.1) and content of embryonal carcinoma, the stem cell of the tumor. A histochemical study of embryoid bodies has shown that high levels of the enzyme are confined to embryonal carcinoma. Fifteen tissue culture lines could be classified into three groups: (a) lines identifiable as pluripotential embryonal carcinoma by their morphology, tumorigenicity, and capacity to differentiate in vivo; (b) nullipotential embryonal carcinoma, resembling pluripotential embryonal carcinoma in morphology and malignancy but giving rise to undifferentiated tumors; and (c) lines of apparently nonmalignant somatic cells. Both types of embryonal carcinoma possess levels of alkaline phosphatase 5- to a 100-fold higher than the somatic cell lines. The embryonal carcinoma enzyme resembles the enzymes from kidney and placenta in kinetics of thermal inactivation and sensitivity to the inhibitor L-phenylalanine, but is distinguishable from the alkaline phosphatases of liver and intestine. These findings are discussed in relation to the use of teratoma for the study of cell differentiation.
576 citations
TL;DR: Observations indicate that the two types of PMN granules discharge in a sequential manner, specific granules fusing with the vacuole before azurophils, thereby providing optimal conditions for coordinated activity of granule contents.
Abstract: The sequential discharge of neutrophilic polymorphonuclear leukocyte (PMN) granules—azurophils and specifics—was investigated by electron microscopy and cytochemistry. Thus the enzyme content of PMN phagocytic vacuoles was determined at brief intervals after phagocytosis of bacteria, utilizing peroxidase as a marker enzyme for azurophil granules, and alkaline phosphatase for specifics. At 30 s, approximately half the phagocytic vacuoles were reactive for alkaline phosphatase, whereas none contained peroxidase. Peroxidase-containing vacuoles were rarely seen at 1 min, but by 3 min, vacuoles containing both enzymes were consistently present. Alkaline phosphatase was found in both small and large vacuoles, whereas peroxidase was visible only in large ones. By 10 min, very big phagocytic vacuoles containing considerable amounts of reaction product for both enzymes were evident. These observations indicate that the two types of PMN granules discharge in a sequential manner, specific granules fusing with the vacuole before azurophils. In an earlier paper, we reported that the pH of phagocytic vacuoles drops to 6.5 within 3 min and to ∼4 within 7–15 min. Substances known to be present in specific granules (alkaline phosphatase, lysozyme, and lactoferrin) function best at neutral or alkaline pH, whereas most of those contained in azurophil granules (i.e., peroxidase and the lysosomal enzymes) have pH optima in the acid range. Hence the sequence of granule discharge roughly parallels the change in pH, thereby providing optimal conditions for coordinated activity of granule contents.
259 citations
TL;DR: In all dosage groups the most marked relief of pain was at sites of Paget's involvement of long bones, and there was a dose-response relation of administered disodium etidronate to improvement of pain, with moderate or marked improvement ofPain in six of seven subjects on 20 mg per kilogram per day.
Abstract: A six-month double-blind controlled study was performed to determine effectiveness and dose response of disodium etidronate on Paget's disease (osteitis deformans). Forty-seven subjects were studied with pain related to radiologically documented Paget's bone sites as well as marked elevation of serum alkaline phosphatase and hydroxyproline excretion. There was significant (p<0.05) reduction of alkaline phosphatase, hydroxyproline and radiofluoride or radiostrontium uptake over Paget's bone lesions after orally administered disodium etidronate at doses of 5, 10 and 20 mg per kilogram per day for six months. There was a dose-response relation of administered disodium etidronate to improvement of pain, with moderate or marked improvement of pain in six of seven subjects on 20 mg per kilogram per day. In all dosage groups the most marked relief of pain was at sites of Paget's involvement of long bones. Apparent side effects were limited to occasional abdominal cramps. (N Engl J Med 289:1379–1384, 1973)
239 citations
TL;DR: The neutral peptidase and aminopeptidase M, both of which can attack insulin B chain, differed not only in response to inhibitors and activators but also in the inhibitory effect of a guinea-pig antiserum raised to rabbit aminopesptidases M.
Abstract: 1. A method for the preparation of brush border from rabbit kidneys is described. Contamination by other organelles was checked by electron microscopy and by the assay of marker enzymes and was low. 2. Seven enzymes, all hydrolases, were substantially enriched in the brush-border preparation and are considered to be primarily located in this structure. They are: alkaline phosphatase, maltase, trehalase, aminopeptidase A, aminopeptidase M, γ-glutamyl transpeptidase and a neutral peptidase assayed by its ability to hydrolyse [125I]iodoinsulin B chain. 3. Adenosine triphosphatases were also present in the preparation, but showed lower enrichments. 4. Alkaline phosphatase was the most active phosphatase present in the preparation. The weak hydrolysis of AMP may well have been due to this enzyme rather than a specific 5′-nucleotidase. 5. The two disaccharidases in brush border were distinguished by the relative heat-stability of trehalase compared with that of maltase. 6. The individuality of the four peptidases was established by several means. The neutral peptidase and aminopeptidase M, both of which can attack insulin B chain, differed not only in response to inhibitors and activators but also in the inhibitory effect of a guinea-pig antiserum raised to rabbit aminopeptidase M. This antiserum inhibited both the purified and the brush-border activities of aminopeptidase M. The neutral peptidase and γ-glutamyl transpeptidase were unaffected but aminopeptidase A was weakly inhibited. The characteristic responses to Ca2+ and serine with borate served to distinguish aminopeptidase A and γ-glutamyl transpeptidase from other peptidases. 7. No dipeptidases, tripeptidases or carboxypeptidases were identified as brush-border enzymes. 8. Incubation of brush border with papain released almost all the aminopeptidase M activity but only about half the activities of maltase, γ-glutamyl transpeptidase and aminopeptidase A. No release of alkaline phosphatase, trehalase or the neutral peptidase was observed.
195 citations
TL;DR: Entering into phophorus deficiency was accompanied by a loss of heterocysts, a decline in chlorophyll a, protein, RNA, and cellular phosphorus, and an increase in carbohydrate per unit dry weight.
Abstract: SUMMARY
Several aspects of the metabolism and composition of a strain of Anabaena have been studied during phosphorus deficiency. The effects of medium composition, substrate concentration, temperature, pH, and illumination on alkaline phosphatase activity and phosphate uptake have been examined. Of particular interest among these results was the dependence of maximum alkaline phosphatase activity on Ca and of phosphate uptake on Mg. Depletion of dissolved phosphate from the culture medium runs accompanied by a marked increase in alkaline phosphatase activity, initial rate of phosphate uptake, and total amount of phosphate taken up to satisfaction of the phosphorus debt. Readdition of phosphate to a phosphorus-deficient culture resulted in a rapid decline in the ability to take up phosphate but no loss of alkaline phosphatase beyond dilution of activity already present. Entry into phophorus deficiency was accompanied by a loss of heterocysts, a decline in chlorophyll a, protein, RNA, and cellular phosphorus, and an increase in carbohydrate per unit dry weight. The possible use of these changes as physiological indicators of phosphorus limitation in natural situations is discussed.
167 citations
TL;DR: It is suggested that the phoS and phoT genes are primarily involved in P(i) transport and, as a result of this function, play a role in the regulation of alkaline phosphatase synthesis.
Abstract: Two classes of alkaline phosphatase constitutive mutations which comprise the original phoS locus (genes phoS and phoT) on the Escherichia coli genome have been implicated in the regulation of alkaline phosphatase synthesis. When these mutations were introduced into a strain dependent on a single system, the pst system, for inorganic phosphate (Pi) transport, profound changes in Pi transport were observed. The phoT mutations led to a complete Pi− phenotype in this background, and no activity of the pst system could be detected. The introduction of the phoS mutations changed the specificity of the pst system so that arsenate became growth inhibitory. Changes in the phosphate source led to changes in the levels of constitutive alkaline phosphatase synthesis found in phoS and phoT mutants. When glucose-6-phosphate or l-α-glycerophosphate was supplied as the sole source of phosphate, phoT mutants showed a 3- to 15- fold reduction in constitutive alkaline phosphatase synthesis when compared to the maximal levels found in limiting Pi media. However, these levels were still 100 times greater than the basal level of alkaline phosphatase synthesized in wild-type strains under these conditions. The phoS mutants showed only a two- to threefold reduction when grown with organic phosphate sources. The properties of the phoT mutants selected on the basis of constitutive alkaline phosphatase synthesis were similar in many respects to those of pst mutants selected for resistance to growth inhibition caused by arsenate. It is suggested that the phoS and phoT genes are primarily involved in Pi transport and, as a result of this function, play a role in the regulation of alkaline phosphatase synthesis.
165 citations
TL;DR: Phosphorylated non-histone chromatin proteins are shown to stimulate the synthesis of RNA in a cell-free system using rat liver RNA polymerase and rat DNA as template.
138 citations
TL;DR: Although mitotic rate was subnormal, several other factors attest to the essential viability of the cultured intestine: L-leucine incorporation into protein, as well as the synthesis of a specific vitamin D3-induced calcium-binding protein (CaBP), increased over a 48-h culture period, and the electropotential gradient was maintained throughout the culture period as was a concentration gradient for calcium.
Abstract: Duodena from 20-day-old chick embryos can be maintained in large scale organ culture on specially designed stainless-steel grids in contact with serum-free medium for 48 h with excellent preservation of mucosal structure at both the light and electron microscope levels. Although mitotic rate was subnormal, several other factors attest to the essential viability of the cultured intestine: L-leucine incorporation into protein, as well as the synthesis of a specific vitamin D3-induced calcium-binding protein (CaBP), increased over a 48-h culture period, and the electropotential gradient across the intestine was maintained throughout the culture period as was a concentration gradient for calcium. The tissue responded to vitamin D3 in the medium by synthesizing the calcium-binding protein within 6 h and by exhibiting enhanced 45Ca uptake within 12–24 h. Concentrations of vitamin D3, or its 25-hydroxylated derivative, higher than necessary for CaBP induction, also increased the activity of alkaline phosphatase. The 1,25-dihydroxylated derivative of vitamin D3, at a level extremely potent in CaBP induction, did not stimulate alkaline phosphatase. Mucosal to serosal transport of 45Ca could also be measured in everted duodenal sacs, subsequent to culture under similar conditions, and was also increased by vitamin D3 in the medium. Other embryonic organs, esophagus, stomach, liver, pancreas, lung, skin, and muscle, did not produce CaBP in response to vitamin D3 in the culture medium. However, CaBP-synthesizing capacity was present in the entire intestinal tract, exclusive of the rectum. 59Fe and 32P uptake by cultured duodenum were also stimulated by vitamin D3. The system has proven quite useful in the study of the vitamin D-mediated calcium absorptive mechanism but should be applicable to the study of the absorption of other nutrients, drugs, hormones, etc., as well as other studies of intestinal function.
99 citations
Journal Article•
92 citations
TL;DR: Results of this study suggest that examination of the isoenzymes by gel electrophoresis is more sensitive than the biochemical methods in tracing the cause responsible for the enzyme changes in plasma.
TL;DR: Alkaline phosphatase purified from Escherichia coli contains up to 2 molecules of tightly bound inorganic phosphate per enzyme molecule, which suggests that the active sites on the alkalineosphatase molecule may be equivalent and independent, at least in the pre-steady state.
TL;DR: Acidic extracts of the largest granules contained lysozyme and cationic proteins; apparently one or more of these proteins may be connected with the antibacterial capacity of peroxidaseless PMNs from chickens.
Abstract: sucrose, were heterogeneous, and contained acid hydrolases. Alkaline phosphatase was absent. Acidic extracts of the largest granules contained lysozyme and cationic proteins; these extracts, at concentrations of 20-35 [tg/ml, inhibited Escherichia coli, Serratia marcescens, and Staphylococcus albus. Evidently one or more of these proteins may be connected with the antibacterial capacity of peroxidaseless PMNs from chickens.
TL;DR: A rough strain of Salmonella typhimurium, which has a defect in the "inner core" region of its lipopolysaccharide, was shown to release more alkaline phosphatase into the medium during growth than normal.
Abstract: A rough strain of Salmonella typhimurium, which has a defect in the "inner core" region of its lipopolysaccharide, was shown to release more alkaline phosphatase into the medium during growth than ...
TL;DR: Nickel had no effect on copper content and only a slight effect on zinc content, but significantly increased the iron concentrations of these tissues, and caused an accumulation of iron in all the cellular fractions of these two tissues.
TL;DR: Alterations in specific canalicular enzymes in two forms of cholestasis suggest that these changes may be involved in the pathogenesis of bile secretory failure, or may result from cholESTasis.
Abstract: Bile secretory failure (cholestasis) may result from several possible mechanisms involved in bile secretion. We have examined the possibility that abnormalities in enzyme content, composition, and turnover of liver plasma membrane constituents are altered in cholestasis. Severe and mild cholestasis were produced by 5 days of bile duct ligation and ethinyl estradiol administration, respectively. Bile duct ligation but not ethinyl estradiol treatments was associated with elevations of the serum bilirubin level and 5'-nucleotidase activity. However, basal bile flow and bilirubin transport maximum (T(m)) were significantly reduced after ethinyl estradiol treatment. Liver plasma membrane fractions rich in canalicular membranes were prepared from groups of rats in each of three categories; normal, after bile duct ligation, or ethinyl estradiol administration, and their respective controls. Electron microscopy and enzyme marker studies demonstrated plasma membrane fractions free of significant contamination. Plasma membrane fractions prepared from mild as well as severe cholestasis had increased alkaline phosphatase activity, and reduced 5'-nucleotidase and Mg(2+)-ATPase activities. Co(2+)-CMPase activity was unchanged. Kinetic analysis of 5'-nucleotidase and Mg(2+)-ATPase activities in plasma membrane fractions demonstrated reduced V(maz) (but unaltered K(m)). Reducted V(maz) was unrelated to addition in vitro of di-or trihydroxy bile salts or ethinyl estradiol and, therefore, suggests that reduced activities in cholestasis are due to decreased enzyme content. Cholestasis was not associated with changes in the synthesis or degradation rate of pulse-labeled plasma membrane proteins or alterations in the major protein bands separated on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Plasma membrane cholesterol, phospholipid, and neutral sugar content was unaltered, but sialic acid content was significantly increased in both forms of cholestasis. Alterations in specific canalicular enzymes in two forms of cholestasis suggest that these changes may be involved in the pathogenesis of bile secretory failure, or may result from cholestasis.
TL;DR: It is demonstrated that serum from patients with acute viral hepatitis, chronic Australia antigenemia, common bile duct obstruction, primary biliary cirrhosis, hepatic necrosis secondary to halothane, and alcoholic Cirrhosis suppresses DNA synthesis by stimulated normal lymphocytes.
TL;DR: Levels of alkaline phosphatase and inorganic phosphorus varied with both age and sex, and the magnitude of these variations in normal ranges is of clear importance in assessing data from individual paediatric or adolescent patients.
Abstract: In a cross-sectional survey 624 schoolchildren were screened for plasma calcium, inorganic phosphate, and alkaline phosphatase levels. Plasma magnesium and alkaline phosphatase isoenzymes were also estimated in some cases.No significant difference was found between adult and childhood values for calcium and magnesium. Levels of alkaline phosphatase and inorganic phosphorus varied with both age and sex. The magnitude of these variations in normal ranges is of clear importance in assessing data from individual paediatric or adolescent patients.
TL;DR: Results confirm that enterokinase is attached to the external side of the duodenal brush border membrane.
TL;DR: It is revealed that about one third of cystic fibrosis patients have serologic evidence of hepatobiliary disease before clinical signs are evident and that γ-GTP, the transaminases, and the liver isoenzyme of alkaline phosphatase are the most reliable indicators of this complication.
TL;DR: It is shown that bile duct ligation causes an early increase in synthesis of hepatic alkaline phosphatase whereas the incresase in 5'nucleotidase and γ-glutamyl transpeptidase is delayed until the phase of biles duct proliferation.
TL;DR: Parathyroid hormone is important in maintaining the concentration of plasma ionized calcium in normal children, but the regulation of total serum calcium is different from that in adults, and the factors regulating it may have a direct or indirect influence on the growth process.
Abstract: Extract: Simultaneous measurements of immunoreactive parathyroid hormone (iPTH), calcium, magnesium, phosphorus, and alkaline phosphatase in serum were performed in 120 normal subjects who ranged from 6 months to 20 years of age. When plotted as a function of age, these extracellular indices of calcium homeostasis showed changes throughout the growth period and differences in their interrelationships in early childhood, middle childhood, and adolescence. Mean serum concentration of minerals was highest during early childhood and decreased parallel to decreases in iPTH and alkaline phosphatase. Between the ages of 6 and 12 years, calcium, magnesium, and phosphorus tended to plateau, iPTH was lower, and alkaline phosphatase increased. During adolescence, calcium, phosphorus, and alkaline phosphatase decreased to adult values, magnesium showed no change, and iPTH increased to adult values. Plasma ionized calcium, which was higher in middle childhood (mean, 4.59 mg/100 ml) than at ages 12 to 20 years (mean, 4.33 mg/100 ml), showed the expected negative correlation with iPTH (P < 0.005). Total serum calcium did not correlate with iPTH; however, the pattern of decrease of serum calcium with age was similar to growth velocity curves, especially in girls, in whom mean serum calcium was higher (9.93 mg/100 ml) between 8 and 16 years than in boys (9.75 mg/100 ml). Speculation: Parathyroid hormone is important in maintaining the concentration of plasma ionized calcium in normal children, but the regulation of total serum calcium is different from that in adults. The total serum calcium or factors regulating it may have a direct or indirect influence on the growth process.
TL;DR: The results suggest that the activation by Mg2+ proceeds through a binding of the metal with free enzyme which might be followed by a conformational change, rather than by an action on the substrate.
TL;DR: It was concluded that the alkaline phosphatase of B. ruminicola is firmly bound to a structural component within the periplasmic area of the cell wall and that the enzyme is released in large amounts only when the cells break down.
Abstract: Of the three species (Bacteroides ruminicola, B succinogenes, and Megasphaera elsdenii) of anaerobic gram-negative rumen bacteria studied, only B ruminicola produced significant amounts of alkaline phosphatase This enzyme, which is constitutive, showed a greater affinity for p-nitrophenylphosphate than for sodium-beta-glycerophosphate and was shown to be located exclusively in the periplasmic space of log-phase cells Small amounts of this enzyme were released from these cells in stationary-phase cultures, but washing in 001 M MgCl(2) and the production of spheroplasts by using lysozyme in 001 M MgCl(2) did not release significant amounts of the enzyme Exposure to 02 M MgCl(2) did not release significant amounts of the periplasmic alkaline phosphatase of the cell, and when these cells were spheroplasted with lysozyme in 02 M MgCl(2) only 25% of the enzyme was released Spheroplasts were formed spontaneously in aging cultures of B ruminicola, but even these cells retained most of their periplasmic alkaline phosphatase It was concluded that the alkaline phosphatase of B ruminicola is firmly bound to a structural component within the periplasmic area of the cell wall and that the enzyme is released in large amounts only when the cells break down The behavior of alkaline phosphatase in this bacterium contrasts with that of conventional periplasmic enzymes of aerobic bacteria, which are released upon conversion into spheroplasts by lysozyme and ethylenediaminetetraacetic acid and by other types of cell wall damage All three species of bacteria studied here, as well as bacteria found in mixed populations in the rumen, have thick, complex layers external to the double-track layer of their cell walls In addition, B ruminicola produces a loose extracellular material
TL;DR: Serum enzymes that show changed activities in diabetes mellitus can be divided into four groups: Group I includes some lysosomal enzymes—β-glucuronidase N-acetyl-β- glucosaminidase, acid phosphatase, and amylase—that show increased activity correlated with blood sugar concentration, and others that may be more active in diabetics with complications such as hepatic and renal involvement and obesity.
Abstract: Serum enzymes that show changed activities in diabetes mellitus can be divided into four groups: Group I includes some lysosomal enzymes—β-glucuronidase N-acetyl-β-glucosaminidase, acid phosphatase, and amylase—that show increased activity correlated with blood sugar concentration. Because lysosomal enzymes as well as liver amylase show latency and may be "activated" by several agents, their increased activity in the serum of diabetics might be a manifestation of an activation occurring in tissues. Group II includes alkaline phosphatase and trehalase, which are increased but not correlated with blood sugar concentration. Their enhanced activity may reflect tissue metabolic disorders. Group III includes enzymes that increase in the postketotic period almost regularly—phosphohexose isomerase —or in only the most severe cases—aminotransferases and several dehydrogenases—because of tissue damage caused by metabolic and circulatory alterations. Cholinesterase, on the other hand, is decreased. Group IV includes any of the above-mentioned enzymes, and still others, that may be more active in diabetics with complications such as hepatic and renal involvement and obesity.
TL;DR: The hypothesis that the tumor and the placental alkaline phosphatases are products of the same gene is strongly supported.
Abstract: An alkaline phosphatase immunochemically similar to placental alkaline phosphatase (EC 3.1.3.1) was purified from liver metastases of a giant-cell carcinoma of the lung. Some properties of its physical and chemical structure were determined and compared to those of purified placental alkaline phosphatase. The purified tumor phosphatase and the placental phosphatase were similar with regard to the following properties: (1) NH2-terminal sequence, (2) peptide map, (3) subunit molecular weight, and (4) isoelectric point. The physical properties and NH2-terminal sequence of alkaline phosphatase isoenzyme of liver differed from the placental and the tumor enzyme. The data from the present study strongly support the hypothesis that the tumor and the placental alkaline phosphatases are products of the same gene.
TL;DR: The distribution of myoepithelial cells in the rat parotid is unusual and the cells run longitudinally around intercalary ducts and send processes on to the bases of adjoining acini but do not embrace the acini.
Abstract: Alkaline phosphatase activity has been studied in the parotid glands of rats at the light and electron microscopical levels. Reaction product was found to outline the plasma membranes of myoepithelial cells. It was also found in the walls of many capillaries and on the luminal surface and between apposing cells in some intercallary ducts.
TL;DR: Adult male rats injected with methylmercuric chloride or mercuric bichloride in a daily dosage of 1·0 mg Hg/kg body weight died after 4 wk, and Histochemical studies of the enzymes glucose 6-phosphatase, alkaline phosphat enzyme, acid phosphatases, adenosine triph phosphatase (ATPase) and succinic dehydrogenase performed on liver, kidney and brain revealed general decreases and moderate increases.
Journal Article•
TL;DR: An enzymatic, immunochemical test system has been developed which allows quantitation of minute amounts of the Regan isoenzyme of alkaline phosphatase, which was found in 89 of 91 normal healthy adults and in 106 of 112 patients with neoplastic discase.
Abstract: An enzymatic, immunochemical test system has been developed which allows quantitation of minute amounts of the Regan isoenzyme of alkaline phosphatase The basis of the method is the concentration (tenfold) of the Regan isoenzyme by reaction with a monospecific antiserum to placental alkaline phosphatase, insolubilized by polymerization with ethyl chloroformate This concentration is achieved by incubating heat-inactivated serum with the antibody and by subsequent centrifugation The pellet containing the active enzyme-antibody complex is immediately assayed for enzyme activity
We studied sera from 91 normal healthy adults and 112 cancer patients to determine the presence of the Regan isoenzyme Detectable Regan isoenzyme activity was found in 89 of 91 normal healthy adults Three of the normal sera contained a level of the isoenzyme that fell above 2 SD from the mean In the case of patients with neoplastic discase, 106 of 112 sera had detectable Regan isoenzyme activity and only 11 sera showed elevated activities Cancer sera with abnormal Regan levels contained 3 to 300 times the average normal value