Serum Lactic Dehydrogenase,Leucine Aminopeptidase and 5-Nucleotidase Activity: Observations in Patients with Carcinoma of the Pancreas and Hepatobiliary Disease

Serum lactic dehydrogenase, leucine aminopeptidase, 5-nucleotidase and alkaline phosphatase activities were investigated in a number of diseases involving the hepatobiliary system.Leucine aminopeptidase was found to be a sensitive indicator of biliary obstruction, serum 5-nucleotidase slightly less sensitive, and alkaline phosphatase appreciably less sensitive. Leucine aminopeptidase and 5-nucleotidase activities were often increased by malignant infiltration of the liver and primary hepatic disease even in the absence of jaundice.Serum lactic dehydrogenase was frequently increased in primary hepatic disease and malignant disorders but was not apparently affected by bile duct obstruction per se. Thirty-five of 45 patients with proved malignancy had increased lactic dehydrogenase levels.The highest leucine aminopeptidase levels were encountered in carcinoma of the head of the pancreas. The frequent increase in both serum lactic dehydrogenase and leucine aminopeptidase activities in patients with carcinoma of the head of the pancreas suggests that these combined estimations are useful laboratory procedures in the diagnosis of malignant extrahepatic obstruction.     

Antipili antibody affords protection against ascending pyelonephritis in rat: evaluated by renal brush border membrane enzymes

Kinetic parameters (Km and Vmax) of renal brush border membrane (BBM) enzymes alkaline phosphatase, maltase, leucine aminopeptidase and gamma-glutamyltranspeptidase were worked out in control, infected and immunized-infected rats. There was no significant change in the Km of all the enzymes studied in three groups. The Vmax of all the enzymes studied decreased significantly (p less than 0.05) 3 or 4 days postinfection and onwards in the left obstructed kidney of infected and immunised-infected animals. However, in the right unobstructed kidney the Vmax of alkaline phosphatase and leucine aminopeptidase increased significantly (p less than 0.05) in the early stages and decreased (p less than 0.05) in later stages in both the experimental groups. The significant difference (p less than 0.05) in the Vmax of infected and immunized-infected groups at various stages of infection revealed the partial protective role of antipili antibody against ascending pyelonephritis.     

Ectoenzymes in Prorocentrum minimum

Ectoenzymes, or enzymes associated with the cell-surface or periplasmic space, play an important role in organic matter cycling by rendering certain forms of dissolved organic matter bioavailable. Ectoenzyme activities may thereby help meet the nutritional demands of harmful algae such as Prorocentrum minimum. The activities of two ectoenzymes; leucine aminopeptidase and alkaline phosphatase, have been studied in axenic cultures of P. minimum. Leucine aminopeptidase releases non-polar amino acids such as leucine from the N-terminus of polypeptides, whereas alkaline phosphatase is an enzyme that is able to hydrolyze phosphate from phosphomonoesters. P. minimum alkaline phosphatase is the better studied of the two ectoenzymes and its characteristics are reviewed herein. Future research on P. minimum physiology will benefit from a growing suite of tools available for assessing the activity of alkaline phosphatase and other ectoenzymes in field populations and ultimately the work done with P. minimum will be useful for studies of other harmful species.     

Is there a plasma membrane-located anion-sensitive ATPase? IV. Distribution of the enzyme in rat pancreas.

The intracellular localization of anion-sensitive Mg2+-ATPase in rat pancreas was studied by differential centrifugation, density gradient centrifugation and by the use of inhibitors of mitochondrial Mg2+-ATPase. The anion-sensitive MG2+-ATPase appears to be localized almost exclusively in a mitochondrial (15 min, 15 000 times g) fraction which shows two peaks after density gradient centrifugation. Both peaks coincide with the highest levels of cytochrome c oxidase activity, but not with alkaline phosphatase, (Na+ plus K+)-ATPase and leucine aminopeptidase activities or RNA. They appear to be equal sensitive to inhibition by oligomycin, aurovertin D and the rat liver mitochondrial inhibitor protein, at least when 1 mM EDTA is present in the isolation media. We conclude that no significant plasma membrane-located anion-sensitive Mg2+-ATPase activity is present in rat pancreas.     

Effect of antiulcerogenic drug, ranitidine, on intestinal absorption and digestive functions in mice

Oral administration of antiulcerogenic drug ranitidine significantly inhibits glucose and amino acid uptake in small intestinal segments. It also inhibits activities of brush border membrane disaccharidases and alkaline phosphatase but increases the activity of leucine aminopeptidase. Kinetic analysis reveals noncompetitive and mixed type of inhibition for disaccharidases and alkaline phosphatase, respectively. In vitro addition of the drug to membrane preparation shows similar type of results as seen in vivo with the inhibition constant (ki) for sucrase, lactase, maltase and alkaline phosphatase as 12.5, 5, 11.5 and 19.5 mM, respectively.     

Intestinal brush border hydrolase topography. Effects of vitamin D-3 and filipin

Intestinal brush borders were isolated from vitamin D-3-treated and vitamin D-deficient chicks, and protein topography in the paired preparations assessed by the enzymatic release of four marker hydrolases. Exposure of the brush borders to the protease bromelain resulted in soluble levels of alkaline phosphatase, leucine aminopeptidase, maltase, and sucrase activities from preparations of vitamin D-3-treated birds that were 42%, 75%, 64%, and 56%, respectively, of corresponding activities released in preparations from rachitic chicks. Analyses for recovery of enzyme activity revealed that bromelain treatment selectively inactivated 43% of the alkaline phosphatase activity of brush borders obtained from vitamin D-3-replete birds, and preferentially diminished recovered sucrase activity in preparations from vitamin D-deficient chicks. In additional experiments, brush borders isolated from rachitic birds were treated in vitro with the polyene antibiotic filipin or an equivalent volume of vehicle. Subsequent exposure of such preparations to bromelain resulted in little or no differences in levels of marker hydrolase specific activities released from filipin- or vehicle-treated brush borders. However, analyses of membrane-bound specific activities after treatment of brush border preparations with a range of filipin concentrations, revealed a biphasic inhibition of approx. 30% for both maltase and sucrase, relative to vehicle controls, and a smaller effect on alkaline phosphatase and leucine aminopeptidase.     

Potentials of enzyme tests and radioisotope hepatography in detecting early functional changes in the liver

The enzymic tests and radionuclide hepatography were used to study and compare liver function after rabbits were exposed to tetrachloromethane poisoning. The activity of serum enzymes of cholinesterase, alkaline phosphatase, aldolase and leucine aminopeptidase was determined. Hepatography was made with the use of 198Au-colloid with an activity 0.74 MBC. The enzymic tests were demonstrated to be more sensitive than radionuclide hepatography in detecting the earliest parenchymatous lesions in the liver. The data obtained correlate with the data of the pathohistological examinations, which demonstrated the presence of marked vacuole parenchymatous fatty dystrophy. The authors recommend that the enzymic tests should be used for detecting early hepatic lesions induced by tetrachloromethane.     

Subcellular localization and properties of lipase activities in human polymorphonuclear leukocytes

A fluorimetric assay for lipase activity has been optimized for measurement of the enzyme in human neutrophils. Activity was maximal at acid (4.5) and alkaline (9.5) pH, although there was also a neutral peak of activity at pH 6.5. Neutrophils were homogenised in isotonic sucrose and subjected to analytical subcellular fractionation by sucrose density gradient centrifugation. The gradient fractions were assayed for acid, neutral and alkaline lipase activity and for the principal organelle marker enzymes. Neutral lipase showed a unimodal distribution with an equilibrium density of 1.19 g . cm-3, corresponding to the distribution of particulate leucine aminopeptidase. Acid and alkaline lipase activities showed very similar distribution profiles to each other with both soluble components and a broad peak of particulate activity. The broad modal density of 1.19-1.22 g . cm-3 suggests that acid and alkaline lipase activities could be localised to more than one population of cytoplasmic granule. Fractionation experiments with neutrophils homogenised in sucrose medium containing digitonin confirmed the localisation of neutral lipase and leucine aminopeptidase to the same cytoplasmic granule, and suggested that at least part of the acid lipase activity was localised to the specific granule. No lipase activity could be attributed to the alkaline phosphatase-containing granule. Neutrophils were isolated from control subjects, patients with chronic granulocytic leukaemia and women in the third trimester of pregnancy. The specific activity of acid, neutral and alkaline lipase, and leucine aminopeptidase, in contrast to that of alkaline phosphatase, were similar in the three patient groups.     

The enzyme histochemistry of the pineal gland in rats

Synopsis The enzyme histochemistry of the adult rat pineal is reviewed with particular reference to the probable endocrine activity of this organ. The parenchymal cells contain large amounts of oxidative enzymes and non-specific esterase, rather less leucine amino peptidase and acid phosphatase, and only small amounts of phosphorylase and alkaline phosphatase. In addition, high concentrations of alkaline phosphatase are present in the walls of capillary vessels. Leucine aminopeptidase is also seen in the connective tissue around blood vessels.     

EXTRACELLULAR ENZYMES OF THE MICROCYSTIS AERUGINOSA PCC 7813 STRAIN ARE INHIBITED IN THE PRESENCE OF HYDROQUINONE AND PYROGALLOL,ALLELOCHEMICALS PRODUCED BY AQUATIC PLANTS1

Several cyanobacterial species have a high potential to dominate in marine environments and freshwater reservoirs, and the ecological and physiological reasons for this phenomenon are not understood comprehensively. In this study, the ability of a Microcystis aeruginosa Kütz. strain to produce free dissolved enzymes was documented. We have observed that this highly toxic strain releases alkaline phosphatase, leucine aminopeptidase, and β‐glucosidase into the ambient environment. Additionally, the inhibitory activity of selected phenols produced by aquatic plants on the activity of these enzymes was analyzed. The investigated compounds, pyrogallol and, to a lesser degree, hydroquinone, decreased the activity of extracellular enzymes produced by M. aeruginosa, with leucine aminopeptidase being the most sensitive to the inhibitors. The noncompetitive character of enzymatic inhibition suggests that the polyphenols produced by aquatic plants are able to influence the activity of different extracellular or membrane‐bound enzymes.     

Localization of leucine aminopeptidase and vitamin B-12 binding protein in rabbit peripheral blood polymorphonuclear leukocytes

Polymorphonuclear leukocytes were isolated from the peripheral blood of rabbits by Ficoll-Hypaque centrifugation followed by dextran sedimentation. The granulocytes were homogenized in isotonic sucrose and subjected to analytical subcellular fractionation by sucrose density gradient centrifugation. Leucine aminopeptidase, when assayed with L-leucine-7-amido-4-methyl-coumarin as substrate, showed a similar distribution to N-acetyl-ß-glucosaminidase and thus is localized to the tertiary granules. There was no leucine aminopeptidase associated with the plasma membrane of these cells. Further experiments with purified plasma membranes and inhibitor studies using diazotized sulphanilic acid further confirmed that leucine aminopeptidase had a purely intracellular localization. Vitamin B-12 binding protein showed a similar localization to alkaline phosphatase indicating that, as in human polymorphonuclear leukocytes, vitamin B-12 binding protein is located to the specific granules.     

A high yield preparation for rat kidney brush border membranes. Different behaviour of lysosomal markers

Rat kidney cortex slices were homogenized with a polytron in a isoosmotic medium containing 5 mmol/l EGTA. By two precipitations with MgCl2 (12 mmol/l) and differential centrifugation, brush border membranes were purified. The brush border marker enzymes alkaline phosphatase and aminopeptidase M were found to be enriched 17.0 +/- 5.3-fold and 16.7 +/- 3.7-fold, respectively. By this method, a high yield of brush border membranes was obtained (48.3 +/- 7.9% for alkaline phosphatase; 47.0 +/- 9.5% for aminopeptidase M). The acid phosphatase was enriched 5-fold, whereas other lysosomal enzymes (glucosaminidase, glucuronidase, cathepsin D) were enriched only 0.2-fold. Acid phosphatase activity could not be washed out, but could be separated from alkaline phosphatase and leucine aminopeptidase by means of free flow electrophoresis and sucrose density gradient centrifugation. Vesicles prepared by the presently described Mg/EGTA-method show better transport properties, compared to vesicles prepared by the calcium method of Evers et al. (Evers, C., Haase, W., Murer, H. and Kinne, R. (1978) Membrane Biochem. 1, 203-219), whereas by SDS-polyacrylamide gel electrophoresis, no differences in the protein patterns were observed.     

Brush border enzymes as markers for ascending pyelonephritis: active immunization with pili

Activities of renal brush border membrane (BBM) enzymes, alkaline phosphatase, maltase and leucine aminopeptidase, were determined in control, pyelonephritic and immunized-infected rats. The activities of all enzymes decreased significantly (P < 0.05) in obstructed kidney while activities of alkaline phosphatase and leucine aminopeptidase increased significantly (P < 0.05) in unobstructed kidney in early stages of infection. The affinity constant (K_m) of all enzymes remained unaltered in control and experimental groups. Significant differences (P < 0.05) in the activities of BBM enzymes of infected and immunized-infected animals suggested a protective role of active immunization with pili.     

Alterations in the activities of intestinal enzymes in vitamin-C-deficient guinea pigs

The effect of vitamin C deficiency on various enzymes of the intestinal epithelium has been studied in guinea pigs. Brush border sucrase and alkaline phosphatase activities were considerably enhanced (p less than 0.001), but leucine aminopeptidase levels were reduced in scorbutic animals compared to the control group. There was essentially no change in the activity of maltase under these conditions. Kinetic studies with sucrase and alkaline phosphatase in control and scorbutic animals revealed that augmentation of the enzyme activities in scurvy is due to enhanced enzyme contents. Lactate dehydrogenase, succinate dehydrogenase, glucose-6-phosphatase and Mg+2 ATPase also exhibited reduced activities in the intestine of vitamin-C-deficient animals. Observed alterations in the activities of intestinal enzymes in scurvy were restored to control levels upon feeding of vitamin C to scorbutic guinea pigs.     

Histochemical studies on the adrenal glands of the marmosets (Callithrix jacchus and Callithrix penicillata).

The histrochemistry of the adrenal glands was studied in four adult male marmosets (two Callithrix jacchus and two Callithrix penicillata). It was impossible to demonstrate any reactivity to UDPG-GT, ADH, alanyl aminopeptidase, leucine aminopeptidase, xilitol (NAD-dependent) dehydrogenase, beta-glucuronidase and aryl-sulfatase in these glands. Total phosphorylase was found in scattered cells of the glomerulosa and adjacent outer fasciculata of one C. penicillata. The dehydrogenases (LDH, G-6-PDH,6-PGDH, NADPH2-TR,ICDH,SDH,NADH2-TR, alpha-GPDH, beta-OHBDH) as well as the hydrolases (except alkaline phosphatase, ATPase, and acetylcholinesterase) showed a stonger reactivity in the cortical part. Some hydrolases (naphthol acetate esterase, acid phosphatase) and cytochrome oxidase were less reactive in the zona glomerulosa, where the dehydrogenases were more abundant. The outer fasciculata and the reticularis also showed a strong dehydrogenase reactivity.     

Enzymatic histochemistry of mouse kidney in plastic

Two-micrometer sections of methacrylate-embedded kidney were used to investigate the enzymatic activities of mouse kidney where the proximal tubule and Bowman's capsule from the same corpuscle were viewed in the same section. Alkaline phosphatase, acid phosphatase, 5'-nucleotidase, gamma-glutamyl transpeptidase, N-acetyl-beta-glucosaminidase, leucine aminopeptidase, alpha-naphthyl butyrate esterase, and adenosine triphosphatase activities were observed in the proximal tubule, but only 5'-nucleotidase, alpha-naphthyl butyrate esterase, and alkaline phosphatase were observed in the squamous portion of the parietal epithelium of Bowman's capsule. The use of methacrylate-embedded tissue allowed more precise localization of enzymatic activity than is possible with most frozen sections. This may provide interesting applications not only for characterization of kidney diseases but also for characterization of other normal and abnormal tissues.     

Urinary enzyme release following aminoglycoside administration in single low dose

Urinary enzyme excretion was investigated in healthy volunteers before and after infusion of single low doses of aminoglycoside (gentamicin, tobramycin and amikacin) in the same subjects. Significant increases were detected in urinary leucine aminopeptidase, lactate dehydrogenase and alkaline phosphatase following gentamicin infusion; after amikacin administration only urinary release of leucine aminopeptidase was found to be increased. No difference was detected compared to basal values in enzyme excretion after tobramycin infusion. Urinary enzyme release in our conditions suggests low tubular damage after single doses of gentamicin and amikacin.     

Enzyme levels in bronchoalveolar lavage fluid and serum of active pulmonary tuberculosis patients

Bronchoalveolar lavage (BALF) was found to be a useful index of cell damage. It has been observed that the enzymes in BALF could give an idea of cell damage in a pulmonary disease. Acid phosphatase, alkaline phosphatase, leucine aminopeptidase and gamma-glutamyl-transpeptidase were assessed in mild, moderate, and severe pulmonary tuberculosis patients and Mantoux-negative normal controls. Activities of these enzymes were found to be higher in patients and were increasing with the severity of the disease. Increase in these enzymes in pulmonary tuberculosis patients could be attributed to lung tissue damage.     

Prokaryotic extracellular enzymatic activity in relation to biomass production and respiration in the meso- and bathypelagic waters of the (sub)tropical Atlantic

Prokaryotic extracellular enzymatic activity, abundance, heterotrophic production and respiration were determined in the meso- and bathypelagic (sub)tropical North Atlantic. While prokaryotic heterotrophic production (PHP) decreased from the lower euphotic layer to the bathypelagic waters by two orders of magnitude, prokaryotic abundance and cell-specific PHP decreased only by one order of magnitude. In contrast to cell-specific PHP, cell-specific extracellular enzymatic activity (alpha- and beta-glucosidase, leucine aminopeptidase, alkaline phosphatase) increased with depth as did cell-specific respiration rates. Cell-specific alkaline phosphatase activity increased from the intermediate water masses to the deep waters up to fivefold. Phosphate concentrations, however, varied only by a factor of two between the different water masses, indicating that phosphatase activity is not related to phosphate availability in the deep waters. Generally, cell-specific extracellular enzymatic activities were inversely related to cell-specific prokaryotic leucine incorporation. Thus, it is apparent that the utilization of deep ocean organic matter is linked to higher cell-specific extracellular enzymatic activity and respiration and lower cell-specific PHP than in surface waters.     

Subacute thyroiditis associated with systemic multi-organ disorders

Subacute thyroiditis is generally thought to be a self-limited inflammatory disease of the thyroid gland. This paper describes serial observations on the clinical course of a typical patient with subacute thyroiditis. This patient showed specific features of destructive thyrotoxicosis with increases in the serum levels of acute phase reactants and in the erythrocyte sedimentation rate. She also showed signs of liver dysfunction [slightly increased alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (gamma-GTP), and leucine aminopeptidase (LAP)], slight anemia, glucose intolerance, increased pancreatic enzymes, splenomegaly, and an increase in peripheral Leu 7 positive (NK/K) cells. These abnormalities all improved with recovery from disease. These findings indicate that in this patient with subacute thyroiditis inflammation is not limited to the thyroid gland but also involves the liver, pancreas and spleen. Thus the subacute thyroiditis in this patient may be a systemic multi-organ disease.