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An ultrastructural study of vascular proliferation and vascular alkaline phosphatase activity in the developing cerebral cortex of the rat.

Alkaline phosphatase cytochemistry was employed to study the distribution of this enzyme in blood vessels during vascular differentiation and maturation during the postnatal development of the rat cerebral cortex. Enzyme reaction product was present in early vascular sprouts, and also throughout the subsequent maturation and differentiation of capillaries and arterial vessels. Cerebral capillaries appeared to be patent soon after the fusion of a sprout tip with another vessel; no evidence for delayed or synchronous opening was obtained. The distribution of alkaline phosphatase reaction product in vessel walls changed during vascular maturation. In vascular sprouts, reaction product was found mainly in the narrow lumen. As vessels became patent, reaction product appeared also on abluminal surfaces, at first chiefly in the narrow spaces between overlapping vascular cells. As vessels matured, reaction product became more generally distributed around the abluminal surface. In relatively mature capillaries and arterial vessels, it was restricted largely to endothelial cell surfaces and the spaces between smooth muscle cells. The significance of this distribution is unknown. Some possible explanations, including the possibility of artifact, are discussed. No alkaline phosphatase reaction product was found in differentiated veins.

Alkaline Phosphatase↗

Alkaline phosphatase in auxiliary liver transplantation.

Auxiliary liver allografts were inserted into baboons and allowed to undergo rejection. The incorporation of (14)C leucine into alkaline phosphatase was measured in both host and donor livers with the aid of an anti-alkaline phosphatase antibody. The results indicate that the increased serum alkaline phosphatase levels found under these circumstances are due to increased synthesis of the enzyme by both livers. Furthermore, it was shown that in the absence of biliary obstruction, this enhanced production of alkaline phosphatase is accompanied by an increase in the excretion of the enzyme into bile. It is suggested that the stimulus for the enhanced production of alkaline phosphatase is a substance which is normally produced in the liver and excreted into the bile.

Alanine Transaminase↗

Effect of the antiallergic drug disodium cromoglycate and various derivatives on alkaline phosphatase.

Disodium cromoglycate (DSCG) inhibits alkaline phosphatase in a non-competitive manner, the enzyme undergoing a conformational change which is attenuated by the presence of calcium ions. The structurally related pyranone and benzoic acid are weak inhibitors of the enzyme and produce a similar conformational change. Coumarin does not induce any conformational change in the enzyme suggesting that the 4-oxo group in DSCG may be essential for its effect.

Alkaline Phosphatase↗

The solubilization of tetrameric alkaline phosphatase from human liver and its conversion into various forms by phosphatidylinositol phospholipase C or proteolysis.

When membrane-bound human liver alkaline phosphatase was treated with a phosphatidylinositol (PI) phospholipase C obtained from Bacillus cereus, or with the proteases ficin and bromelain, the enzyme released was dimeric. Butanol extraction of the plasma membranes at pH 7.6 yielded a water-soluble, aggregated form that PI phospholipase C could also convert to dimers. When the membrane-bound enzyme was solubilized with a non-ionic detergent (Nonidet P-40), it had the Mr of a tetramer; this, too, was convertible to dimers with PI phospholipase C or a protease. Butanol extraction of whole liver tissue at pH 6.6 and subsequent purification yielded a dimeric enzyme on electrophoresis under nondenaturing conditions, whereas butanol extraction at pH values of 7.6 or above and subsequent purification by immunoaffinity chromatography yielded an enzyme with a native Mr twice that of the dimeric form. This high molecular weight form showed a single Coomassie-stained band (Mr = 83,000) on electrophoresis under denaturing conditions in sodium dodecyl sulfate, as did its PI phospholipase C cleaved product; this Mr was the same as that obtained with the enzyme purified from whole liver using butanol extraction at pH 6.6. These results are highly suggestive of the presence of a butanol-activated endogenous enzyme activity (possibly a phospholipase) that is optimally active at an acidic pH. Inhibition of this activity by maintaining an alkaline pH during extraction and purification results in a tetrameric enzyme. Alkaline phosphatase, whether released by phosphatidylinositol (PI) phospholipase C or protease treatment of intact plasma membranes, or purified in a dimeric form, would not adsorb to a hydrophobic medium. PI phospholipase C treatment of alkaline phosphatase solubilized from plasma membranes by either detergent or butanol at pH 7.6 yielded a dimeric enzyme that did not absorb to the hydrophobic medium, whereas the untreated preparations did. This adsorbed activity was readily released by detergent. Likewise, alkaline phosphatase solubilized from plasma membranes by butanol extraction at pH 7.6 would incorporate into phosphatidylcholine liposomes, whereas the enzyme released from the membranes by PI phospholipase C would not incorporate. The dimeric enzyme purified from a butanol extract of whole liver tissue carried out at pH 6.6 did not incorporate. We conclude that PI phospholipase C converts a hydrophobic tetramer of alkaline phosphatase into hydrophilic dimers through removal of the 1,2-diacylglycerol moiety of phosphatidylinositol. Based on these and others' findings, we devised a model of alkaline phosphatase's conversion into its various forms.

Alkaline Phosphatase↗

Increased plasma pyridoxal-5'-phosphate when alkaline phosphatase activity is reduced in moderately zinc-deficient rats.

It is generally believed that the zinc metalloenzyme alkaline phosphatase is required to hydrolyze phosphorylated forms of vitamin B-6 prior to their use. To test this hypothesis, rats were fed a liquid diet containing either adequate or moderately low zinc during gestation and lactation. Zinc deficiency was produced in dams evidenced by significant reductions in zinc concentration of plasma (49%), liver (25%), and femur (24%), and plasma alkaline phosphatase activity (48%). Plasma pyridoxal-5'-phosphate (PLP), which significantly increased (61%) in these same rats, was negatively correlated (r = -0.74, P < 0.02) with plasma alkaline phosphatase activity. Maternal liver PLP concentration was unaffected by zinc status. The zinc and vitamin B-6 relationship seen in dams was less observable in offspring. Stimulation of erythrocyte alanine aminotransferase activity by exogenously added PLP in vitro tended to be higher in both moderately zinc-deficient mothers and their offspring, but the difference was not significant. Our results support the hypothesis that alkaline phosphatase activity is required for the hydrolysis of plasma PLP. Our results also suggest that zinc status as alkaline phosphatase activity should be defined in an individual if plasma PLP is to be used as an indicator of vitamin B-6 status.

Alkaline Phosphatase↗

Structural evidence that human liver and placental alkaline phosphatase isoenzymes are coded by different genes.

Human liver alkaline phosphatase [ortho-phosphoric monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] was purified, and some of its physical and chemical properties were examined and compared to those of human placental alkaline phosphatase. The results indicated a different peptide structure for each, based upon HB2-terminal residue sequence, two-dimensional tryptic peptide maps, and different amino acid compositions. These data are interpreted to indicate that the enzymes are synthesized by different structural genes. Other molecular properties differentiating the two enzymes were a higher apparent molecular weight for the liver enzyme from sodium dodecyl sulfate gel electrophoresis, a higher S20,w value, different carbohydrate content, and a different isoelectric point. The immunochemical specificity of each enzyme was not affected by removal of sialic acid groups. Both enzymes are similar in that they are dimers of equal molecular weight subunits, and are probably homodimers.

Alkaline Phosphatase↗

Heterogeneous nature of alkaline phosphatase from rat liver.

1. Alkaline phosphatase from rat liver was separated into tow components by gel filtration. 2. Both components had the same temperature and pH optima, were inhibited by high concentrations of urea and had the same activity towards hydrolysis of phenyl disodium phosphate and p-nitrophenyl phosphate. 3. They differed in electrophoretic mobility, sensitivity to inactivation by heat and digestion by neuraminidase, response to low concentrations of urea, and catalytic activity with sodium pyrophosphate and sodium phytate.

Alkaline Phosphatase↗

[Alkaline phosphatase activity in goblet cells in the small intestine of piglets experimentally infected with the coccidium Isospora suis].

Alkaline phosphatase activity (EC. 3.1.3.1.) in goblet cells was investigated in the small intestine of 16 gnotobiotic piglets infected one day after delivery (DAD) by different rates of oocysts of Isospora suis coccidia. At a high infection rate of I. suis (750,000) the goblet cells were found to be highly positive to alkaline phosphatase on day 3 to day 4 after infection (DAI). In piglets infected by a low infection rate of I. suis oocysts (100,000) the activity of alkaline phosphatase activity in goblet cells was proved on days 4 to 10 after infection. In the first group of piglets, the positive goblet cells prevailed in the middle region of jejunum, with the peak on 4th DAI. It the second group of piglets a marked increase in the alkaline phosphatase activity was recorded in the goblet cells in the posterior part of jejunum on days 4 to 5 after infection and on 10th DAI. No alkaline phosphatase activity in the goblet cells was demonstrated in the control gnotobiotic piglets at the age of two to seven days.

Alkaline Phosphatase↗

Inhibitors of green crab (Scylla serrata) alkaline phosphatase.

Green crab (Scylla serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme which catalyzes the nonspecific hydrolysis of phosphate monoesters. The effects of several inhibitors on its activity have been studied. The results show that Na2HPO4, Na2HAsO4 and Na2WO4 are competitive inhibitors, L-phenylalanine is an uncompetitive inhibitor, while L-cysteine is a mixed-type inhibitor. The equilibrium constants were determined for inhibitor binding. The order of potency was Na2HPO4 (KI = 0.95 mM) > Na2AsO4 (1.10 mM) > Na2WO4 (1.55 mM).

Alkaline Phosphatase↗

Serum alkaline phosphatase activity during pregnancy and the post-partum period in Ghanaian women.

Serum alkaline phosphatase activity was determined in 454 Ghanaian women at various stages of pregnancy and the eighteen months following delivery and compared with the levels found in 45 non-pregnant women who were not nursing babies. There was a statistically significant fall in mean enzyme activity in the second trimester, followed by a rise above normal in the third trimester. During the eighteen months following delivery, the mean serum alkaline phosphatase activities were significantly raised. Increase in plasma volume which occurs during pregnancy is suggested as the cause of the fall in serum enzyme activity in the second trimester whereas the rise in enzyme activity in the third trimester is attributed to the well-known increase in serum alkaline phosphatase of placental origin. The prolonged elevation of serum alkaline phosphatase activity in the post-partum months is most likely due to the traditional prolonged breast-feeding that practised in Ghana.

Alkaline Phosphatase↗

Mucosal alkaline phosphatase and bile lipids in the gallbladder in cholecystitis.

Histochemically, alkaline phosphatase was localised in epithelial brush-border and capillary endothelium of the lamina propria in 41 gallbladders studied. Three distinct patterns of the brush-border enzyme distribution were observed, namely, complete delineation, patchy localisation and a brush-border totally devoid of enzyme activity. Where no histochemical activity was found, biliary chenodeoxycholate was 50 per cent. higher than in cases of patchy distribution and twice as high as in cases with complete staining of the brush-border. Conversely, biliary cholate levels were significantly higher in completely stained cases. Biliary cholesterol was higher in the cases with no histochemical activity and this was reflected in a significantly higher lithogenic index. Significant quantities of cholesterol esters were found in bile from the focal activity group only. Total alkaline phosphatase activity assayed in gallbladders from 112 patients with cholecystitis and/or lithiasis showed that activity was significantly lower in patients with pigment stones than patients with cholecystitis and/or cholesterol stones. This activity was not related to biliary lipid composition in the 77 bile samples analysed. This evidence indicates that biliary lipid composition is associated with the histochemical distribution of alkaline phosphatase in gallbladder mucosa.

Alkaline Phosphatase↗

Pancreatic alkaline phosphatase and a tumour variant.

Alkaline phosphatase has been extracted from human pancreas with n-butanol and has been shown to have distinctive isoenzyme characteristics when compared with the enzymes of small intestine and of normal serum. The enzyme has been demonstrated histochemically in a carcinoma of the pancreas and also in an islet cell tumour and the tumour enzyme has been shown to be unique, not only electrophoretically, but in being remarkably heat sensitive and in being activated by low molarities of urea.

Adenocarcinoma↗

Monitoring of murine osteosarcoma by serial alkaline phosphatase determinations.

To establish that alkaline phosphatase (AP) was released by osteosarcoma cells, we measured this enzyme in C3H/HeJ mice with im-implanted osteosarcoma and in in vitro cultures of neoplastic cells subjected to short-term incubation. We found that 10(5) osteosarcoma cells synthesized a significant amount of AP in vitro in 30 minutes at 37 degrees C. A good correlation existed between pulmonary metastatic tumors and the AP values. Serum AP measurements indicated approximate sizes of disseminated and localized tumors, but could not monitor early localized tumors.

Alkaline Phosphatase↗

[Isolated elevation of serum alkaline phosphatase].

The isoenzymes of serum alkaline phosphatase (AP) were determined in 17 children and 3 adults who had transiently or persistently raised serum AP levels without clinical, laboratory or radiographic evidence of hepatobiliary or bone disease. Liver and bone AP were both raised in 8 of 11 children having transient hyperphosphatasia whereas a rise in only one of the two enzymes (liver- or bone-AP) was detected in the serum of children and adults having persistent hyperphosphatasia. Isolated hyperphosphatasaemia is an anomaly having no pathological significance which, when found, has importance since expensive and invasive investigation methods that come into question can be avoided.

Adult↗

Multiple forms of alkaline phosphatase isoenzymes of Serratia marcescens.

Alkaline phosphatase (APase) isoenzymes produced by different strains of Serratia marcescens were examined. Variation of isoenzyme patterns with respect to number and their mobilities in starch gels after electrophoresis were observed. Ten strains gave a 1-isoenzyme pattern with 5 different mobilities; 7 strains gave a 2-isoenzyme pattern with 3 different mobilities; 9 strains gave a 3-isoenzyme pattern with 5 different mobilities; and 3 strains gave a 4-isoenzyme pattern. Three strains synthesized two electrophoretically distinct APases in low phosphate medium. A high concentration of inorganic phosphate induced the synthesis of one of these APase isoenzymes.

Alkaline Phosphatase↗

Developmental changeover in rat duodenal alkaline phosphatase.

The specific activity of alkaline phosphatase in the rat duodenum increased just prior to birth, leveled off and then increased again at about 20 days after birth. During the postnatal rise in activity the electrophoretic pattern of the enzyme on sodium dodecyl sulphate (SDS) polyacrylamide gels changed from a single activity band to three bands. The changed pattern was identical with that of the adult duodenal enzyme. The fetal- and adult-type enzymes had different sensitivities to neuraminidase, suggesting that there is a significant difference in the sialic acid content of these two forms. This molecular conversion may be necessary for adaptation to eating a solid food diet, because it occurs just before the weaning period.

Aging↗

Leukocyte alkaline phosphatase in the newborn.

Leukocyte alkaline phosphatase (LAP) activity was studied in 50 women at delivery and in the newborn after delivery. The following findings were observed: (1) the LAP activity of newborns is lower than that of the mothers; (2) significant decrease in the LAP activity has been observed in newborns on the 4th day after delivery; (3) there are no significant differences between newborns from normal and prolonged labor; (4) immediately after delivery the LAP scores of male newborns are lower than those of females, but 4 days later these scores were reversed, and (5) 4 days after delivery, the LAP scores do correlate to the weight of the newborn.

Alkaline Phosphatase↗

Reactive thiol groups in calf-intestinal alkaline phosphatase.

Incubation of calf intestinal alkaline phosphatase with iodoacetamide results in time-dependent changes in catalytic activity. Above 2 mmol/l, inactivation occurs but at lower iodoacetamide concentrations there is a relatively slow increase in activity. The changes induced by the treatment, i.e. both activation and inactivation, are irreversible, affect Vmax only, and are not prevented by the presence of the substrate analogue Pi. Total inactivation is associated with the incorporation of 4 mol iodoacetamide/mol of enzyme but the majority of the enzymic activity is lost after incorporation of only 1 mol of iodoacetamide. Thus any structural alterations of one subunit consequent to modification may be transmitted to the partner subunit, i.e. activity loss seems to be a cooperative process.

Alkaline Phosphatase↗