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Sample records for high alkaline phosphatase

  1. Alkaline phosphatase of Physarum polycephalum is insoluble.

    Science.gov (United States)

    Furuhashi, Kiyoshi

    2008-02-01

    The plasmodia of Physarum polycephalum grow as multinucleated cells in the presence of sufficient humidity and nutriment. Under non-illuminating conditions, stresses such as low temperature or high concentrations of salts transform the plasmodia into spherules whereas dehydration induces sclerotization. Some phosphatases including protein phosphatase and acid phosphatase have been purified from the plasmodia, but alkaline phosphatase remains to be elucidated. Phosphatase of the plasmodia, spherules and sclerotia was visualized by electrophoresis gel-staining assay using 5-bromo-4-chloro-3-indolyl phosphate. Insoluble fractions of the sclerotia were abundant in phosphatase activity. The phosphatase which was extracted by nonionic detergent was subjected to column chromatography and preparative electrophoresis. Purified phosphatase showed the highest activity at pH 8.8, indicating that this enzyme belongs to alkaline phosphatase. The apparent molecular mass from sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-reducing condition was estimated to be 100 kDa whereas that under reducing was 105 kDa. An amount of 1% sodium dodecyl sulfate or 0.5 M NaCl had no effects on the activity although the phosphatase showed heat instability, Mg(2+)-dependency and sensitivity to 2-glycerophosphate or NaF. The extracting conditions and enzymatic properties suggest that this alkaline phosphatase which is in a membrane-bound form plays important roles in phosphate metabolism.

  2. Association of alkaline phosphatase phenotypes with arthritides

    Directory of Open Access Journals (Sweden)

    Padmini A

    2004-01-01

    Full Text Available Arthritides, a symmetrical polyarticular disease of the bone are a heterogenous group of disorders in which hereditary and environmental factors in combination with an altered immune response appear to play a causative and pathogenic role in its occurrence. Alkaline phosphatase (ALP is an enzyme found in all tissues, with particularly high concentrations of ALP observed in the liver, bile ducts, placenta, and bone.Alkaline phosphatase is an orthophosphoric monoester phosphohydrolase catalyzing the hydrolysis of organic esters at alkaline pH, indicating that alkaline phosphatase is involved in fundamental biological processes.1 The present study envisages on identifying the specific electromorphic association of alkaline phosphatase with arthritides. Phenotyping of serum samples was carried out by PAGE (Polyacrylamide gel electrophoresis following Davies (19642 protocol on 41 juvenile arthritis, 150 rheumatoid arthritis and 100 osteo arthritis apart from, 25 normal children and 100 adult healthy subjects. Phenotyping of alkaline phosphatase revealed an increase in preponderance of p+ and p++ phenotypes in juvenile, rheumatoid and osteo arthritic patients. However a significant association of these phenotypes was observed only with rheumatoid arthritis condition (c2:17.46. Similarly, a significant increase of p+ phenotypes in female rheumatoid arthritis patients was observed (c2:14.973, suggesting that the decrease in p° (tissue non specific synthesis/secretion of alkaline phosphatase could be associated with decreased mineralization and ossification process in arthritis condition.

  3. High Serum Alkaline Phosphatase, Hypercalcaemia, Race, and Mortality in South African Maintenance Haemodialysis Patients

    Science.gov (United States)

    Duarte, Raquel; Naicker, Saraladevi

    2017-01-01

    Objective. To determine the association between serum total alkaline phosphatase (TAP) and mortality in African maintenance haemodialysis patients (MHD). Patients and Methods. The study enrolled a total of 213 patients on MHD from two dialysis centers in Johannesburg between January 2009 and March 2016. Patients were categorized into a low TAP group (≤112 U/L) versus a high TAP group (>112 U/L) based on a median TAP of 112 U/L. Results. During the follow-up period of 7 years, there were 55 (25.8%) deaths. After adjusting for cofounders such as age, other markers of bone disorder, and comorbidity (diabetes mellitus), patients in the high TAP group had significantly higher risk of death compared to patients in the low TAP group (hazard ratio, 2.50; 95% CI 1.24–5.01, P = 0.01). Similarly, serum calcium >2.75 mmol/L was associated with increased risk of death compared to patients within levels of 2.10–2.37 mmol/L (HR 6.34, 95% CI 1.40–28.76; P = 0.02). The HR for death in white patients compared to black patients was 6.88; 95% CI 1.82–25.88; P = 0.004. Conclusion. High levels of serum alkaline phosphatase, hypercalcaemia, and white race are associated with increased risk of death in MHD patients. PMID:28168054

  4. Identification of novel chromone based sulfonamides as highly potent and selective inhibitors of alkaline phosphatases.

    Science.gov (United States)

    al-Rashida, Mariya; Raza, Rabia; Abbas, Ghulam; Shah, Muhammad Shakil; Kostakis, George E; Lecka, Joanna; Sévigny, Jean; Muddassar, Muhammad; Papatriantafyllopoulou, Constantina; Iqbal, Jamshed

    2013-08-01

    A new series of structurally diverse chromone containing sulfonamides has been developed. Crystal structures of three representative compounds (2a, 3a and 4a) in the series are reported. All compounds were screened for their inhibitory potential against alkaline phosphatases (ALPs). Two main classes of ALP isozymes were selected for this study, the tissue non-specific alkaline phosphatase (TNALP) from bovine and porcine source and the tissue-specific intestinal alkaline phosphatases (IALPs) from bovine source. All sulfonamide compounds had a marked preference for IALP (K(i), up to 0.01 ± 0.001 μM) over TNALPs. Kinetics studies of the compounds showed competitive mode of inhibition. Molecular docking studies were carried out in order to characterize the selective inhibition of the compounds. An additional interesting aspect of these chromone sulfonamides is their inhibitory activity against ecto-5'-nucleotidase enzyme. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  5. [Alkaline phosphatase in Amoeba proteus].

    Science.gov (United States)

    Sopina, V A

    2005-01-01

    In free-living Amoeba proteus (strain B), 3 phosphatase were found after disc-electrophoresis of 10 microg of protein in PAGE and using 1-naphthyl phosphate as a substrate a pH 9.0. These phosphatases differed in their electrophoretic mobilities - "slow" (1-3 bands), "middle" (one band) and "fast" (one band). In addition to 1-naphthyl phosphate, "slow" phosphatases were able to hydrolyse 2-naphthyl phosphate and p-nitrophenyl phosphate. They were slightly activated by Mg2+, completely inhibited by 3 chelators (EDTA, EGTA and 1,10-phenanthroline), L-cysteine, sodium dodecyl sulfate and Fe2+, Zn2+ and Mn2+ (50 mM), considerably inactivated by orthovanadate, molybdate, phosphatase inhibitor cocktail 1, p-nitrophenyl phosphate, Na2HPO4, DL-dithiothreitol and urea and partly inhibited by H2O2, DL-phenylalanine, 2-mercaptoethanol, phosphatase inhibitor cocktail 2 and Ca2+. Imidazole, L-(+)-tartrate, okadaic acid, NaF and sulfhydryl reagents -p-(hydroxy-mercuri)benzoate and N-ethylmaleimide - had no influence on the activity of "slow" phosphatases. "Middle" and "fast" phosphatases, in contrast to "slow" ones, were not inactivated by 3 chelators. The "middle" phosphatase differed from the "fast" one by smaller resistance to urea, Ca2+, Mn2+, phosphates and H2O2 and greater resistance to dithiothreitol and L-(+)-tartrate. In addition, the "fast" phosphatase was inhibited by L-cysteine but the "middle" one was activated by it. Of 5 tested ions (Mg2+, Cu2+, Mn2+, Ca2+ and Zn2+), only Zn2+ reactivated "slow" phosphatases after their inactivation by EDTA treatment. The reactivation of apoenzyme was only partial (about 35 %). Thus, among phosphatases found in amoebae at pH 9.0, only "slow" ones are Zn-metalloenzymes and may be considered as alkaline phosphatases (EC 3.1.3.1). It still remains uncertain, to which particular phosphatase class "middle" and "fast" phosphatases (pH 9.0) may belong.

  6. Multisystemic functions of alkaline phosphatases.

    Science.gov (United States)

    Buchet, René; Millán, José Luis; Magne, David

    2013-01-01

    Human and mouse alkaline phosphatases (AP) are encoded by a multigene family expressed ubiquitously in multiple tissues. Gene knockout (KO) findings have helped define some of the precise exocytic functions of individual isozymes in bone, teeth, the central nervous system, and in the gut. For instance, deficiency in tissue-nonspecific alkaline phosphatase (TNAP) in mice (Alpl (-/-) mice) and humans leads to hypophosphatasia (HPP), an inborn error of metabolism characterized by epileptic seizures in the most severe cases, caused by abnormal metabolism of pyridoxal-5'-phosphate (the predominant form of vitamin B6) and by hypomineralization of the skeleton and teeth featuring rickets and early loss of teeth in children or osteomalacia and dental problems in adults caused by accumulation of inorganic pyrophosphate (PPi). Enzyme replacement therapy with mineral-targeting TNAP prevented all the manifestations of HPP in mice, and clinical trials with this protein therapeutic are showing promising results in rescuing life-threatening HPP in infants. Conversely, TNAP induction in the vasculature during generalized arterial calcification of infancy (GACI), type II diabetes, obesity, and aging can cause medial vascular calcification. TNAP inhibitors, discussed extensively in this book, are in development to prevent pathological arterial calcification. The brush border enzyme intestinal alkaline phosphatase (IAP) plays an important role in fatty acid (FA) absorption, in protecting gut barrier function, and in determining the composition of the gut microbiota via its ability to dephosphorylate lipopolysaccharide (LPS). Knockout mice (Akp3 (-/-)) deficient in duodenal-specific IAP (dIAP) become obese, and develop hyperlipidemia and hepatic steatosis when fed a high-fat diet (HFD). These changes are accompanied by upregulation in the jejunal-ileal expression of the Akp6 IAP isozyme (global IAP, or gIAP) and concomitant upregulation of FAT/CD36, a phosphorylated fatty acid

  7. Low serum alkaline phosphatase activity in Wilson's disease.

    Science.gov (United States)

    Shaver, W A; Bhatt, H; Combes, B

    1986-01-01

    Low values for serum alkaline phosphatase activity were observed early in the course of two patients with Wilson's disease presenting with the combination of severe liver disease and Coombs' negative acute hemolytic anemia. A review of other cases of Wilson's disease revealed that 11 of 12 patients presenting with hemolytic anemia had values for serum alkaline phosphatase less than their respective sex- and age-adjusted mean values; in eight, serum alkaline phosphatase activity was less than the lower value for the normal range of the test. Low values for serum alkaline phosphatase were much less common in Wilson's disease patients with more chronic forms of presentation. Copper added in high concentration to serum in vitro did not have an important effect on serum alkaline phosphatase activity. The mechanism responsible for the decrease in serum alkaline phosphatase activity in patients is uncertain.

  8. Prophylactic treatment with alkaline phosphatase in cardiac surgery induces endogenous alkaline phosphatase release

    NARCIS (Netherlands)

    Kats, Suzanne; Brands, Ruud; Hamad, Mohamed A. Soliman; Seinen, Willem; Schamhorst, Volkher; Wulkan, Raymond W.; Schoenberger, Jacques P.; van Oeveren, Wim

    2012-01-01

    Introduction: Laboratory and clinical data have implicated endotoxin as an important factor in the inflammatory response to cardiopulmonary bypass. We assessed the effects of the administration of bovine intestinal alkaline phosphatase (bIAP), an endotoxin detoxifier, on alkaline phosphatase levels

  9. Persistently increased intestinal fraction of alkaline phosphatase

    DEFF Research Database (Denmark)

    Nathan, E; Baatrup, G; Berg, H

    1984-01-01

    Persistent elevation of the intestinal fraction of the alkaline phosphatase (API) as an isolated finding has to our knowledge not been reported previously. It was found in a boy followed during a period of 5.5 years. The only symptom was transient periodic fatigue observed at home, but not appare...

  10. [Leucocyte alkaline phosphatase in normal and pathological pregnancy (author's transl)].

    Science.gov (United States)

    Stark, K H; Zaki, I; Sobolewski, K

    1981-01-01

    The activities of leucocyte alkaline phosphatase were determined in 511 patients with normal and pathological pregnancy. Mean values were compared and the enzyme followed up, and the conclusion was drawn that leucocyte alkaline phosphatase was no safe indicator of foetal condition. No direct relationship were found to exist between leucocyte alkaline phosphatase, total oestrogens, HSAP, HLAP, HPL, and oxytocinase.

  11. A High Level of Intestinal Alkaline Phosphatase Is Protective Against Type 2 Diabetes Mellitus Irrespective of Obesity.

    Science.gov (United States)

    Malo, Madhu S

    2015-12-01

    Mice deficient in intestinal alkaline phosphatase (IAP) develop type 2 diabetes mellitus (T2DM). We hypothesized that a high level of IAP might be protective against T2DM in humans. We determined IAP levels in the stools of 202 diabetic patients and 445 healthy non-diabetic control people. We found that compared to controls, T2DM patients have approx. 50% less IAP (mean +/- SEM: 67.4 +/- 3.2 vs 35.3 +/- 2.5 U/g stool, respectively; p diabetes status. With each 25 U/g decrease in stool IAP, there is a 35% increased risk of diabetes. The study revealed that obese people with high IAP (approx. 65 U/g stool) do not develop T2DM. Approx. 65% of the healthy population have diabetes', and might develop T2DM and other metabolic disorders in the near future. In conclusion, high IAP levels appear to be protective against diabetes irrespective of obesity, and a 'temporal IAP profile' might be a valuable tool for predicting 'the incipient metabolic syndrome', including 'incipient diabetes'.

  12. Tissue non-specific alkaline phosphatase production by human dental pulp stromal cells is enhanced by high density cell culture.

    Science.gov (United States)

    Tomlinson, Matthew J; Dennis, Caitriona; Yang, Xuebin B; Kirkham, Jennifer

    2015-08-01

    The cell surface hydrolase tissue non-specific alkaline phosphatase (TNAP) (also known as MSCA-1) is used to identify a sub-population of bone marrow stromal cells (BMSCs) with high mineralising potential and is found on subsets of cells within the dental pulp. We aim to determine whether TNAP is co-expressed by human dental pulp stromal cells (hDPSCs) alongside a range of BMSC markers, whether this is an active form of the enzyme and the effects of culture duration and cell density on its expression. Cells from primary dental pulp and culture expanded hDPSCs expressed TNAP. Subsequent analyses revealed persistent TNAP expression and co-expression with BMSC markers such as CD73 and CD90. Flow cytometry and biochemical assays showed that increased culture durations and cell densities enhanced TNAP expression by hDPSCs. Arresting the hDPSC cell cycle also increased TNAP expression. These data confirm that TNAP is co-expressed by hDPSCs together with other BMSC markers and show that cell density affects TNAP expression levels. We conclude that TNAP is a potentially useful marker for hDPSC selection especially for uses in mineralised tissue regenerative therapies.

  13. Dephosphorylation of endotoxin by alkaline phosphatase in vivo

    NARCIS (Netherlands)

    Poelstra, Klaas; Bakker, W.W; Klok, P.A; Kamps, J.AAM; Hardonk, M.J; Meijer, D.K F

    1997-01-01

    Natural substrates for alkaline phosphatase (AP) are at present not identified despite extensive investigations. Difficulties in imagining a possible physiological function involve its extremely high pH optimum for the usual exogenous substrates and its localization as an ecto-enzyme. As endotoxin i

  14. Effect of carbon source on alkaline phosphatase production and excretion in Aspergillus caespitosus.

    Science.gov (United States)

    Guimarães, Luis Henrique Souza; Jorge, João Atilio; Terenzi, Héctor Francisco; Jamur, Maria Célia; Oliver, Constance; De Lourdes Teixeira De Moraes Polizeli, Maria

    2003-01-01

    The effect of several carbon sources on the production of alkaline phosphatase by the thermotolerant Aspergillus caespitosus was analysed. The fungus released high levels of alkaline phosphatases into the medium after being cultured for long periods with xylan or industrial residues such as wheat raw and sugar cane bagasse in the culture media. In contrast, the alkaline phosphatase activities were found only intracellulary when the fungus was cultured in glucose-supplemented media. The pH of the medium likely affects the process of enzyme secretion according to the carbon source used. Addition of xylan or industrial residues in the culture medium stimulated the secretion of phosphatases. In contrast, media supplemented with glucose or disaccharides promoted retention of these enzymes into the cells. The subcellular location activities of alkaline phosphatases were studied using histochemical and immunochemical methods and showed that alkaline phosphatases were present in the mycelial walls and septa.

  15. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Jemmerson, R.; Low, M.G.

    1987-09-08

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  16. Alkaline Phosphatase, an Unconventional Immune Protein.

    Science.gov (United States)

    Rader, Bethany A

    2017-01-01

    Recent years have seen an increase in the number of studies focusing on alkaline phosphatases (APs), revealing an expanding complexity of function of these enzymes. Of the four human AP (hAP) proteins, most is known about tissue non-specific AP (TNAP) and intestinal AP (IAP). This review highlights current understanding of TNAP and IAP in relation to human health and disease. TNAP plays a role in multiple processes, including bone mineralization, vitamin B6 metabolism, and neurogenesis, is the genetic cause of hypophosphatasia, influences inflammation through regulation of purinergic signaling, and has been implicated in Alzheimer's disease. IAP regulates fatty acid absorption and has been implicated in the regulation of diet-induced obesity and metabolic syndrome. IAP and TNAP can dephosphorylate bacterial-derived lipopolysaccharide, and IAP has been identified as a potential regulator of the composition of the intestinal microbiome, an evolutionarily conserved function. Endogenous and recombinant bovine APs and recombinant hAPs are currently being explored for their potential as pharmacological agents to treat AP-associated diseases and mitigate multiple sources of inflammation. Continued research on these versatile proteins will undoubtedly provide insight into human pathophysiology, biochemistry, and the human holobiont.

  17. Alkaline Phosphatase, an Unconventional Immune Protein

    Science.gov (United States)

    Rader, Bethany A.

    2017-01-01

    Recent years have seen an increase in the number of studies focusing on alkaline phosphatases (APs), revealing an expanding complexity of function of these enzymes. Of the four human AP (hAP) proteins, most is known about tissue non-specific AP (TNAP) and intestinal AP (IAP). This review highlights current understanding of TNAP and IAP in relation to human health and disease. TNAP plays a role in multiple processes, including bone mineralization, vitamin B6 metabolism, and neurogenesis, is the genetic cause of hypophosphatasia, influences inflammation through regulation of purinergic signaling, and has been implicated in Alzheimer’s disease. IAP regulates fatty acid absorption and has been implicated in the regulation of diet-induced obesity and metabolic syndrome. IAP and TNAP can dephosphorylate bacterial-derived lipopolysaccharide, and IAP has been identified as a potential regulator of the composition of the intestinal microbiome, an evolutionarily conserved function. Endogenous and recombinant bovine APs and recombinant hAPs are currently being explored for their potential as pharmacological agents to treat AP-associated diseases and mitigate multiple sources of inflammation. Continued research on these versatile proteins will undoubtedly provide insight into human pathophysiology, biochemistry, and the human holobiont. PMID:28824625

  18. Alkaline phosphatase as a periodontal disease marker

    Directory of Open Access Journals (Sweden)

    Malhotra Ranjan

    2010-01-01

    Full Text Available Background: The potential of alkaline phosphatase (ALP as an important diagnostic marker of gingival crevicular fluid (GCF has been the subject to investigation since 1970. ALP is stored in specific granules and secretory vesicles of the neutrophils and is mainly released during their migration to the site of infection. It is also present in bacteria within dental plaque, osteoblasts and fibroblasts. It has, thus, become important to elucidate whether GCF levels of ALP are potential measures of the inflammatory activity occurring in the adjacent periodontal tissues. Objective: The aim of this study was to assess the total activity of ALP in the GCF collected from healthy sites, sites with gingivitis and with chronic adult periodontitis. An attempt was also made to establish the correlation of ALP activity with plaque index, gingival index, bleeding index and probing depth. Materials and Methods: A total of 18 patients were divided into three groups: viz., healthy sites, Group I; gingivitis, Group II; chronic periodontitis, Group III. Clinical parameters like plaque index, bleeding index, gingival index and probing depth were recorded. The ALP level in GCF of all three groups was determined by spectrophotometric analysis. Results: Total enzyme activity of ALP was significantly higher in periodontitis as compared with that in healthy and gingivitis sites, and was significantly and positively correlated with probing depth. Conclusion: ALP can be considered as a periodontal disease marker as it can distinguish between healthy and inflamed sites. However, to better define its capacity for periodontal diagnosis, additional longitudinal studies are required.

  19. Alkaline Phosphatase, an Unconventional Immune Protein

    Directory of Open Access Journals (Sweden)

    Bethany A. Rader

    2017-08-01

    Full Text Available Recent years have seen an increase in the number of studies focusing on alkaline phosphatases (APs, revealing an expanding complexity of function of these enzymes. Of the four human AP (hAP proteins, most is known about tissue non-specific AP (TNAP and intestinal AP (IAP. This review highlights current understanding of TNAP and IAP in relation to human health and disease. TNAP plays a role in multiple processes, including bone mineralization, vitamin B6 metabolism, and neurogenesis, is the genetic cause of hypophosphatasia, influences inflammation through regulation of purinergic signaling, and has been implicated in Alzheimer’s disease. IAP regulates fatty acid absorption and has been implicated in the regulation of diet-induced obesity and metabolic syndrome. IAP and TNAP can dephosphorylate bacterial-derived lipopolysaccharide, and IAP has been identified as a potential regulator of the composition of the intestinal microbiome, an evolutionarily conserved function. Endogenous and recombinant bovine APs and recombinant hAPs are currently being explored for their potential as pharmacological agents to treat AP-associated diseases and mitigate multiple sources of inflammation. Continued research on these versatile proteins will undoubtedly provide insight into human pathophysiology, biochemistry, and the human holobiont.

  20. Combination of alkaline phosphatase anti-alkaline phosphatase (APAAP)- and avidin-biotin-alkaline phosphatase complex (ABAP)-techniques for amplification of immunocytochemical staining of human testicular tissue.

    Science.gov (United States)

    Davidoff, M S; Schulze, W; Holstein, A F

    1991-01-01

    An amplification procedure was developed for the visualization of antigens in human testis using monoclonal antibodies against desmin and vimentin. The technique combines the high sensitive and specific APAAP- and ABAP-methods. Depending on the quality of the antibodies used and the processing of the material prior to the immunocytochemical staining the amplification technique may be applied either as a single APAAP and ABAP- or as a double APAAP and ABAP-combination. Especially after the double amplification reaction a distinct increase of the staining intensity of the vimentin- (in Sertoli cells, myofibroblasts of the lamina propria, and fibroblasts of the interstitium) and desmin- (in myofibroblasts of the lamina propria and smooth muscle cells of the blood vessels) like immunoreactivity was observed. If different diazonium salts were used for the visualization of the alkaline phosphatase activity (e.g. Fast Red TR Salt, Fast Blue BB Salt) desmin- and vimentin-like immunoreactivity can be demonstrated in the same tissue section in a double sequential staining approach. For double staining, the alkaline phosphatase technique may be combined successfully with a technique or a combination that uses peroxidase as a marker.

  1. Overexpression of Human Bone Alkaline Phosphatase in Pichia Pastoris

    Science.gov (United States)

    Karr, Laurel; Malone, Christine, C.; Rose, M. Franklin (Technical Monitor)

    2000-01-01

    The Pichiapastoris expression system was utilized to produce functionally active human bone alkaline phosphatase in gram quantities. Bone alkaline phosphatase is a key enzyme in bone formation and biomineralization, yet important questions about its structural chemistry and interactions with other cellular enzymes in mineralizing tissues remain unanswered. A soluble form of human bone alkaline phosphatase was constructed by deletion of the 25 amino acid hydrophobic C-terminal region of the encoding cDNA and inserted into the X-33 Pichiapastoris strain. An overexpression system was developed in shake flasks and converted to large-scale fermentation. Alkaline phosphatase was secreted into the medium to a level of 32mgAL when cultured in shake flasks. Enzyme activity was 12U/mg measured by a spectrophotometric assay. Fermentation yielded 880mgAL with enzymatic activity of 968U/mg. Gel electrophoresis analysis indicates that greater than 50% of the total protein in the fermentation is alkaline phosphatase. A purification scheme has been developed using ammonium sulfate precipitation followed by hydrophobic interaction chromatography. We are currently screening crystallization conditions of the purified recombinant protein for subsequent X-ray diffraction analyses. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

  2. Dairy products and the French paradox: Could alkaline phosphatases play a role?

    Science.gov (United States)

    Lallès, Jean-Paul

    2016-07-01

    The French paradox - high saturated fat consumption but low incidence of cardiovascular disease (CVD) and mortality - is still unresolved and continues to be a matter of debate and controversy. Recently, it was hypothesised that the high consumption of dairy products, and especially cheese by the French population might contribute to the explanation of the French paradox, in addition to the "(red) wine" hypothesis. Most notably this would involve milk bioactive peptides and biomolecules from cheese moulds. Here, we support the "dairy products" hypothesis further by proposing the "alkaline phosphatase" hypothesis. First, intestinal alkaline phosphatase (IAP), a potent endogenous anti-inflammatory enzyme, is directly stimulated by various components of milk (e.g. casein, calcium, lactose and even fat). This enzyme dephosphorylates and thus detoxifies pro-inflammatory microbial components like lipopolysaccharide, making them unable to trigger inflammatory responses and generate chronic low-grade inflammation leading to insulin resistance, glucose intolerance, type-2 diabetes, metabolic syndrome and obesity, known risk factors for CVD. Various vitamins present in high amounts in dairy products (e.g. vitamins A and D; methyl-donors: folate and vitamin B12), and also fermentation products such as butyrate and propionate found e.g. in cheese, all stimulate intestinal alkaline phosphatase. Second, moulded cheeses like Roquefort contain fungi producing an alkaline phosphatase. Third, milk itself contains a tissue nonspecific isoform of alkaline phosphatase that may function as IAP. Milk alkaline phosphatase is present in raw milk and dairy products increasingly consumed in France. It is deactivated by pasteurization but it can partially reactivate after thermal treatment. Experimental consolidation of the "alkaline phosphatase" hypothesis will require further work including: systematic alkaline phosphatase activity measurements in dairy products, live dairy ferments and

  3. Expression and Characterization of Recombinant Thermostable Alkaline Phosphatase from a Novel Thermophilic Bacterium Thermus thermophilus XM

    Institute of Scientific and Technical Information of China (English)

    Jianbo LI; Limei XU; Feng YANG

    2007-01-01

    A gene (tap) encoding a thermostable alkaline phosphatase from the thermophilic bacterium Thermus thermophilus XM was cloned and sequenced. It is 1506 bp long and encodes a protein of 501 amino acid residues with a calculated molecular mass of 54.7 kDa. Comparison of the deduced amino acid sequence with other alkaline phosphatases showed that the regions in the vicinity of the phosphorylation site and metal binding sites are highly conserved. The recombinant thermostable alkaline phosphatase was expressed as a His6-tagged fusion protein in Escherichia coli and its enzymatic properties were characterized after purification. The pH and temperature optima for the recombinant thermostable alkaline phosphatases activity were pH 12 and 75 ℃. As expected, the enzyme displayed high thermostability, retaining more than 50% activity after incubating for 6 h at 80 ℃. Its catalytic function was accelerated in the presence of 0.1 mM Co2+, Fe2+, Mg2+, or Mn2+ but was strongly inhibited by 2.0 mM Fe2+. Under optimal conditions, the Michaelis constant (Km) for cleavage of p-nitrophenyl-phosphate was 0.034 mM. Although it has much in common with other alkaline phosphatases, the recombinant thermostable alkaline phosphatase possesses some unique features, such as high optimal pH and good thermostability.

  4. Characterization of Human Bone Alkaline Phosphatase in Pichia Pastoris

    Science.gov (United States)

    Malone, Christine C.; Ciszak, Eva; Karr, Laurel J.

    1999-01-01

    A soluble form of human bone alkaline phosphatase has been expressed in a recombinant strain of the methylotrophic yeast Pichia pastoris. We constructed a plasmid containing cDNA encoding for human bone alkaline phosphatase, with the hydrophobic carboxyl terminal portion deleted. Alkaline phosphatase was secreted into the medium to a level of 32mg/L when cultured in shake flasks, and enzyme activity was 12U/mg, as measured by a spectrophotometric assay. By conversion to a fermentation system, a yield of 880mg/L has been achieved with an enzyme activity of 968U/mg. By gel electrophoresis analysis, it appears that greater than 50% of the total protein in the fermentation media is alkaline phosphatase. Although purification procedures are not yet completely optimized, they are expected to include filtration, ion exchange and affinity chromatography. Our presentation will focus on the purification and crystallization results up to the time of the conference. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

  5. The tillage effect on the soil acid and alkaline phosphatase activity

    Directory of Open Access Journals (Sweden)

    Lacramioara Oprica

    2011-12-01

    Full Text Available Phosphatases (acid and alkaline are important in soils because these extracellular enzymes catalyze the hydrolysis of organic phosphate esters to orthophosphate; thus they form an important link between biologically unavailable and mineral phosphorous. Phosphatase activity is sensitive to environmental perturbations such as organic amendments, tillage, waterlogging, compaction, fertilizer additions and thus it is often used as an environmental indicator of soil quality in riparian ecosystems. The aim of the study was to assess the effect of tillage systems on phosphatases activity in a field experiment carried out in Ezăreni farm. The phosphatase activitiy were determined at two depths (7-10 cm and 15-25cm layers of a chernozem soil submitted to conventional tillage (CT in a fertilised and unfertilised experiment. Monitoring soil alkaline phosphatase activity showed, generally, the same in fertilized soil profiles collected from both depths; the values being extremely close. In unfertilized soils, alkaline phosphatase activity is different only in soils that were exposed to unconventional work using disc harrows and 30cm tillage. Both works type (no tillage and conventional tillage cause an intense alkaline phosphatase activity in 7-10 cm soil profile. Acid phosphatase activity is highly fluctuating in both fertilized as well unfertilized soil, this enzyme being influenced by the performed works.

  6. The catalytic properties of alkaline phosphatases under various conditions

    Science.gov (United States)

    Atyaksheva, L. F.; Chukhrai, E. S.; Poltorak, O. M.

    2008-11-01

    A comparative study was performed to examine the catalytic properties of alkaline phosphatases from bacteria Escherichia coli and bovine and chicken intestines. The activity of enzyme dimers and tetramers was determined. The activity of the dimer was three or four times higher than that of the tetramer. The maximum activity and affinity for 4-nitrophenylphosphate was observed for the bacterial alkaline phosphatase ( K M = 1.7 × 10-5 M, V max = 1800 μmol/(min mg of protein) for dimers and V max = 420 μmol/(min mg of protein) for tetramers). The Michaelis constants were equal for two animal phosphatases in various buffer media (pH 8.5) ((3.5 ± 0.2) × 10-4 M). Five buffer systems were investigated: tris, carbonate, hepes, borate, and glycine buffers, and the lowest catalytic activity of alkaline phosphatases at equal pH was observed in the borate buffer (for enzyme from bovine intestine, V max = 80 μmol/(min mg of protein)). Cu2+ cations formed a complex with tris-(oxymethyl)-aminomethane ( tris-HCl buffer) and inhibited the intestine alkaline phosphatases by a noncompetitive mechanism.

  7. Direct Promotion of Collagen Calcification by Alkaline Phosphatase

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Alkaline phosphatase promotes hydrolysis of phosphate containing substrates, causes a rise in inorganic phosphate and, therefore, enhances calcification of biological tissues. In this work, the calcification of collagen in a model serum was used as a model of collagenous tissue biomaterials to study the possible calcification promotion mechanism of alkaline phosphatase. In the enzyme concentration range of 0.10.5mg/mL, the enzyme shows a direct calcification promoting effect which is independent of the hydrolysis of its phosphate containing substrates but proportional to the enzyme concentration. Potassium pyrophosphate somewhat inhibits the calcification promotion.

  8. Human placental alkaline phosphatase electrophoretic alleles: Quantitative studies

    Science.gov (United States)

    Lucarelli, Paola; Scacchi, Renato; Corbo, Rosa Maria; Benincasa, Alberto; Palmarino, Ricciotti

    1982-01-01

    Human placental alkaline phosphatase (ALP) activity has been determined in specimens obtained from 562 Italian subjects. The mean activities of the three common homozygotes (Pl 2 = 4.70 ± 0.24, Pl 1 = 4.09 ± 0.08, and Pl 3 = 2.15 ± 0.71 μmol of p-nitrophenol produced) were significantly different. The differences among the various allelic forms account for 10% of the total quantitative variation of the human placental alkaline phosphatase. PMID:7072721

  9. Intestinal alkaline phosphatase prevents metabolic syndrome in mice.

    Science.gov (United States)

    Kaliannan, Kanakaraju; Hamarneh, Sulaiman R; Economopoulos, Konstantinos P; Nasrin Alam, Sayeda; Moaven, Omeed; Patel, Palak; Malo, Nondita S; Ray, Madhury; Abtahi, Seyed M; Muhammad, Nur; Raychowdhury, Atri; Teshager, Abeba; Mohamed, Mussa M Rafat; Moss, Angela K; Ahmed, Rizwan; Hakimian, Shahrad; Narisawa, Sonoko; Millán, José Luis; Hohmann, Elizabeth; Warren, H Shaw; Bhan, Atul K; Malo, Madhu S; Hodin, Richard A

    2013-04-23

    Metabolic syndrome comprises a cluster of related disorders that includes obesity, glucose intolerance, insulin resistance, dyslipidemia, and fatty liver. Recently, gut-derived chronic endotoxemia has been identified as a primary mediator for triggering the low-grade inflammation responsible for the development of metabolic syndrome. In the present study we examined the role of the small intestinal brush-border enzyme, intestinal alkaline phosphatase (IAP), in preventing a high-fat-diet-induced metabolic syndrome in mice. We found that both endogenous and orally supplemented IAP inhibits absorption of endotoxin (lipopolysaccharides) that occurs with dietary fat, and oral IAP supplementation prevents as well as reverses metabolic syndrome. Furthermore, IAP supplementation improves the lipid profile in mice fed a standard, low-fat chow diet. These results point to a potentially unique therapy against metabolic syndrome in at-risk humans.

  10. Endotoxin detoxification by alkaline phosphatase in cholestatic livers

    NARCIS (Netherlands)

    Poelstra, K; Bakker, WW; Hardonk, MJ; Meijer, DKF; Wisse, E; Knook, DL; Balabaud, C

    1997-01-01

    Increased expression of alkaline phosphatase (AP) in the liver is a hallmark of cholestasis but the pathophysiological role of this is not clear. We argue that deprotonation of carboxyl groups at the active site of the enzyme may be a prerequisite for optimal AP activity. Such a creation of negative

  11. Induction of glomerular alkaline phosphatase after challenge with lipopolysaccharide

    NARCIS (Netherlands)

    Kapojos, Jola Jovita; Poelstra, Klaas; Borghuis, Theo; van den Berg, Anke; Baelde, Hans J.; Klok, P.A; Bakker, W.W

    2003-01-01

    Alkaline phosphatase (AP) can be considered as a host defence molecule since this enzyme is able to detoxify bacterial endotoxin at physiological pH. The question emerged whether this anti-endotoxin principle is inducible in the glomerulus and if so, which glomerular cells might be involved in the e

  12. Bone alkaline phosphatase and mortality in dialysis patients

    NARCIS (Netherlands)

    C. Drechsler; M. Verduijn; S. Pilz; R.T. Krediet; F.W. Dekker; C. Wanner; M. Ketteler; E.W. Boeschoten; V. Brandenburg

    2011-01-01

    Serum alkaline phosphatase (AP) is associated with vascular calcification and mortality in hemodialysis patients, but AP derives from various tissues of origin. The aim of this study was to assess the effect of bone-specific AP (BAP) on morbidity and mortality in dialysis patients. From a prospectiv

  13. Involvement of CD36 and intestinal alkaline phosphatases in fatty acid transport in enterocytes, and the response to a high-fat diet.

    Science.gov (United States)

    Lynes, Matthew D; Widmaier, Eric P

    2011-02-28

    The vertebrate intestine is notable for its plasticity in response to environmental, pathologic, reproductive, and dietary challenges. The molecular mechanisms of intestinal adaptations typically involve both morphologic and functional changes. In response to chronic ingestion of a high-fat diet, for example, the mammalian small intestine quickly adapts to efficiently accommodate increased transport of long-chain fatty acids across the mucosa. Whereas this may be adaptive in the short term, in the long term it may contribute to the pathologies associated with chronic high-fat diets in humans and other mammals. This review focuses on some of the known and putative mechanisms by which fatty acids are transported across the intestinal epithelium in addition to simple diffusion, and how these mechanisms may be regulated in part by a high-fat diet. A model is proposed in which two key proteins, CD36 and the enzyme intestinal alkaline phosphatase, work in a coordinated manner to optimize fatty acid transport across enterocytes in mice. Copyright © 2010 Elsevier Inc. All rights reserved.

  14. High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China

    Directory of Open Access Journals (Sweden)

    Han Ju

    2012-02-01

    Full Text Available Abstract Background Osteosarcoma is a malignant tumor with high ability to form invasion and metastasis. Identifying prognostic factor in osteosarcoma is helpful to select those patients for more aggressive management. Our study evaluated serum alkaline phosphatase (ALP cooperating with matrix metalloproteinase-9 (MMP-9 as an important prognostic predictor for local recurrence and distant metastasis of osteosarcoma. Methods 177 cases were included from the osteosarcoma patients treated at 1st Affiliated Hospital of Sun Yat-sen University (1999-2008. Pre-chemotherapy serum ALP (pre-ALP were studied and correlated with tumor recurrence, lung metastasis and patient survival. MMP-9 protein in tumor tissues was detected by immunohistochemistry and correlated with pre-ALP level. Results Pre-ALP were partitioned into normal, high, and very high groups, in each group the incidence of metastases was 12.2%, 21.2% and 34.6%, respectively (p = 0.007. In the three groups the mean disease-free survival (DFS was 57 ± 3.15, 28 ± 3.57 and 14 ± 3.35 months, respectively (p Conclusions Pre-ALP was an independent prognostic factor for the survival of osteosarcoma patients in south China, and correlated with MMP-9 expression and lung metastasis. ALP can also serve as a prognostic marker for treatment, and merit large-scale validation studies.

  15. High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China.

    Science.gov (United States)

    Han, Ju; Yong, Bicheng; Luo, Canqiao; Tan, Pingxian; Peng, Tingsheng; Shen, Jingnan

    2012-02-15

    Osteosarcoma is a malignant tumor with high ability to form invasion and metastasis. Identifying prognostic factor in osteosarcoma is helpful to select those patients for more aggressive management. Our study evaluated serum alkaline phosphatase (ALP) cooperating with matrix metalloproteinase-9 (MMP-9) as an important prognostic predictor for local recurrence and distant metastasis of osteosarcoma. 177 cases were included from the osteosarcoma patients treated at 1st Affiliated Hospital of Sun Yat-sen University (1999-2008). Pre-chemotherapy serum ALP (pre-ALP) were studied and correlated with tumor recurrence, lung metastasis and patient survival. MMP-9 protein in tumor tissues was detected by immunohistochemistry and correlated with pre-ALP level. Pre-ALP were partitioned into normal, high, and very high groups, in each group the incidence of metastases was 12.2%, 21.2% and 34.6%, respectively (p = 0.007). In the three groups the mean disease-free survival (DFS) was 57 ± 3.15, 28 ± 3.57 and 14 ± 3.35 months, respectively (p lung metastasis rate decreased (p = 0.028); DFS and OS were both prolonged (p osteosarcoma patients in south China, and correlated with MMP-9 expression and lung metastasis. ALP can also serve as a prognostic marker for treatment, and merit large-scale validation studies.

  16. Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase.

    Science.gov (United States)

    Kaku, Yoshihiro; Noguchi, Akira; Marsh, Glenn A; Barr, Jennifer A; Okutani, Akiko; Hotta, Kozue; Bazartseren, Boldbaatar; Fukushi, Shuetsu; Broder, Christopher C; Yamada, Akio; Inoue, Satoshi; Wang, Lin-Fa

    2012-01-01

    Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and HeV-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2μl) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity.

  17. Fever of unknown origin (FUO) due to large B-cell lymphoma: the diagnostic significance of highly elevated alkaline phosphatase and serum ferritin levels.

    Science.gov (United States)

    Cunha, Burke A; Petelin, Andrew

    2013-01-01

    Determining the cause of fever of unknown origin (FUO) is often a vexing and difficult diagnostic process. In most cases, the signs and symptoms in adult FUOs suggest a malignant, infectious, or rheumatic/inflammatory etiology. The diagnosis of FUO may be narrowed if specific findings are present (eg, hepatosplenomegaly) that limit the diagnostic possibilities. Infectious causes of FUO with hepatosplenomegaly include miliary tuberculosis, typhoid fever, and visceral leishmanosis (kala-azar). However, FUOs with hepatosplenomegaly are most often attributable to malignant neoplasms, ie, Hodgkin lymphoma, non-Hodgkin lymphoma, hepatoma, hypernephroma (renal-cell carcinoma), or preleukemia. We present a middle-aged woman with FUO and hepatosplenomegaly. Inpatient nonspecific laboratory findings included a highly elevated erythrocyte sedimentation rate, and elevated levels of vitamin B12, lactate dehydrogenase, angiotensin-converting enzyme, ferritin, and alkaline phosphatase. These individual findings are nonspecific, but together point to a lymphoma. An important test in differentiating malignant from infectious FUOs is the Naprosyn test, and her Naprosyn test was positive, indicating malignancy. A gallium scan suggested a uterine lymphoma. A computed tomography scan revealed hepatosplenomegaly, but the gallium uptake was not increased in her liver and spleen. Uterine and bone marrow biopsies were negative for lymphoma. We present a case of FUO with hepatosplenomegaly attributable to large B-cell lymphoma as diagnosed via liver biopsy. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. High levels of both serum gamma-glutamyl transferase and alkaline phosphatase are independent preictors of mortality in patients with stage 4-5 chronic kidney disease.

    Science.gov (United States)

    Caravaca-Fontán, Fernando; Azevedo, Lilia; Bayo, Miguel Ángel; Gonzales-Candia, Boris; Luna, Enrique; Caravaca, Francisco

    High serum gamma-glutamyl transferase (GGT) levels are associated with increased mortality in the general population. However, this association has scarcely been investigated in patients with chronic kidney disease (CKD). This study aims to investigate the clinical characteristics of CKD patients with abnormally elevated serum GGT, and its value for predicting mortality. Retrospective observational study in a population cohort of adults with stage 4-5 CKD not yet on dialysis. Demographic, clinical, and biochemical parameters of prognostic interest were recorded and used to characterise CKD patients with high levels of GGT (>36 IU/l). Cox proportional hazard regression models were used to analyse the influence of baseline serum GGT and alkaline phosphatase (ALP) levels on mortality for whatever reason. The study group consisted of 909 patients (mean age 65±15 years). Abnormally elevated GGT or ALP levels at baseline were observed in 209 (23%) and 172 (19%) patients, respectively, and concomitant elevations of GGT and ALP in 68 (7%). High GGT levels were associated with higher comorbidity burden, and a biochemical profile characterised by higher serum concentration of uric acid, triglycerides, alanine aminotransferase, ferritin, and C-reactive. During the study period, 365 patients (40%) died (median survival time=74 months). In adjusted Cox regression models, high levels of GGT (hazard ratio [HR]=1.39;CI 95%: 1.09-1.78, P=.009) and ALP (HR=1.31; CI95%: 1.02-1.68, P=.038) were independently associated with mortality. High serum levels of GGT are independent predictors of mortality in CKD patients. Copyright © 2016 Sociedad Española de Nefrología. Published by Elsevier España, S.L.U. All rights reserved.

  19. Acid- and alkaline phosphatase in amniotic fluid in normal and complicated pregnancy.

    Science.gov (United States)

    Beckman, G; Beckman, L; Löfstrand, T

    1978-01-01

    171 samples of amniotic fluid were obtained by abdominal amniocentesis from 67 women with complicated pregnancies (isoimmunization, diabetes mellitus or toxaemia). The levels of heat-labile alkaline phosphatase (HLAP), heat-stable alkaline phosphatase (HSAP) and acid phosphatase (AcP) were determined and compared to the enzyme levels in 179 samples from women with normal pregnancies of corresponding gestational ages. HLAP showed two "peaks" of activity, one in the 5th-22nd week and the other at term. HSAP and AcP showed increased activity at term. HSAP was decreased (p less than 0.01) in isoimmunization between the 36th and 40th week. 11 cases of toxaemia with placental insufficiency showed no differences in the levels of HLAP and HSAP compared with normal pregnancy. AcP showed no differences between normal and complicated pregnancy. Samples contaminated by blood showed no significant increase in the acid- and alkaline phosphatase levels. Samples contaminated by meconium showed a complex pattern. Some samples had normal enzyme levels, some had high levels of HLAP only and some had high levels of HSAP and AcP. The origin of the enzymes is not known with certainty. HSAP in amniotic fluid is most likely not of placental but intestinal origin. Determinations of acid- and alkaline phosphatase in amniotic fluid seem to be of little values in the clinical management of complicated pregnancy.

  20. Activation of Calf Intestinal Alkaline Phosphatase by Trifluoroethanol

    Institute of Scientific and Technical Information of China (English)

    曹志方; 徐真; 朴龙斗; 周海梦

    2001-01-01

    Alkaline phosphatase is a stable enzyme which is strongly resistant to urea, guanidine hydrochloride, acid pH, and heat. But there have been few studies on the effect of organic cosolvents on the activity and structure of alkaline phosphatase. The activity of calf intestinal alkaline phosphatase (CIAP) is markedly increased when incubated in solutions with elevated trifluoroethanol (TFE) concentrations. The activation is a time dependent course. There is a very fast phase in the activation kinetics in the mixing dead time (30 s) using convential methods. Further activation after the very fast phase follows biphasic kinetics. The structural basis of the activation has been monitored by intrinsic fluorescence and far ultraviolet circular dichroism. TFE (0 - 60%) did not lead to any significant change in the intrinsic fluorescence emission maximum, indicating no significant change in the tertiary structure of CIAP. But TFE did significantly change the secondary structure of CIAP, especially increasing α-helix content. We conclude that the activation of ClAP is due to its secondary structural change. The time for the secondary structure change induced by TFE preceds that of the activity increase. These results suggest that a rapid conformational change of ClAP induced by TFE results in the enhancement of ClAP activity, followed by further increase of this activity because of the further slightly slower rearrangements of the activated conformation. It is concluded that the higher catalytic activity of ClAP can be attained with various secondary structures.

  1. High frequencies of elevated alkaline phosphatase activity and rickets exist in extremely low birth weight infants despite current nutritional support

    Directory of Open Access Journals (Sweden)

    Parker Bruce R

    2009-07-01

    Full Text Available Abstract Background Osteopenia and rickets are common among extremely low birth weight infants (ELBW, Methods We evaluated all ELBW infants admitted to Texas Children's Hospital NICU in 2006 and 2007. Of 211 admissions, we excluded 98 patients who were admitted at >30 days of age or did not survive/stay for >6 weeks. Bone radiographs obtained in 32 infants were reviewed by a radiologist masked to laboratory values. Results In this cohort of 113 infants, P-APA was found to have a significant inverse relationship with BW, gestational age and serum phosphorus. In paired comparisons, P-APA of infants Conclusion Elevation of P-APA >600 IU/L was very common in ELBW infants. BW was significantly inversely related to both P-APA and radiologic rickets. No single value of P-APA was related to radiological findings of rickets. Given the very high risk of osteopenia and rickets among ELBW infants, we recommend consideration of early screening and early mineral supplementation, especially among infants

  2. phoD Alkaline Phosphatase Gene Diversity in Soil.

    Science.gov (United States)

    Ragot, Sabine A; Kertesz, Michael A; Bünemann, Else K

    2015-10-01

    Phosphatase enzymes are responsible for much of the recycling of organic phosphorus in soils. The PhoD alkaline phosphatase takes part in this process by hydrolyzing a range of organic phosphoesters. We analyzed the taxonomic and environmental distribution of phoD genes using whole-genome and metagenome databases. phoD alkaline phosphatase was found to be spread across 20 bacterial phyla and was ubiquitous in the environment, with the greatest abundance in soil. To study the great diversity of phoD, we developed a new set of primers which targets phoD genes in soil. The primer set was validated by 454 sequencing of six soils collected from two continents with different climates and soil properties and was compared to previously published primers. Up to 685 different phoD operational taxonomic units were found in each soil, which was 7 times higher than with previously published primers. The new primers amplified sequences belonging to 13 phyla, including 71 families. The most prevalent phoD genes identified in these soils were affiliated with the orders Actinomycetales (13 to 35%), Bacillales (1 to 29%), Gloeobacterales (1 to 18%), Rhizobiales (18 to 27%), and Pseudomonadales (0 to 22%). The primers also amplified phoD genes from additional orders, including Burkholderiales, Caulobacterales, Deinococcales, Planctomycetales, and Xanthomonadales, which represented the major differences in phoD composition between samples, highlighting the singularity of each community. Additionally, the phoD bacterial community structure was strongly related to soil pH, which varied between 4.2 and 6.8. These primers reveal the diversity of phoD in soil and represent a valuable tool for the study of phoD alkaline phosphatase in environmental samples.

  3. Alkaline phosphatase in stallion semen: characterization and clinical applications.

    Science.gov (United States)

    Turner, R M O; McDonnell, S M

    2003-06-01

    Significant amounts of alkaline phosphatase (AP) activity have been found in semen plasma from numerous species. In species in which the majority of semen plasma AP (SPAP) activity originates from the epididymis and testicle, SPAP activity can be used clinically as a marker to differentiate testicular origin azoospermia or oligospermia from ejaculatory failure. Information on SPAP activity in stallions to date has been limited. In this study, a standard clinical chemistry analyzer was used to determine AP activity in pre-ejaculatory fluid and ejaculates from groups of normal stallions. Additionally, accessory glands, epididymides, testicles and other components of the urogenital tract of normal stallions were assayed to determine which tissues contain SPAP activity. The results indicated that levels of AP activity are low in pre-ejaculatory fluid, but significantly higher in ejaculatory fluid from normal stallions. Spermatozoa were not a significant source of SPAP activity. High levels of SPAP activity were found in the testes and epididymides. These findings suggest that SPAP activity is a candidate for a sperm-independent marker for ejaculation in the stallion. Finally, AP activity was determined in ejaculatory fluid from a stallion with bilaterally blocked ampullae, both before and after relief of the blockage. While the blockage was present, AP activity in ejaculatory fluid was low. However, following relief of the blockage, AP activity in ejaculatory fluid rose dramatically, thus suggesting that AP activity will be useful as an inexpensive, simple clinical assay for differentiating ejaculatory failure or excurrent duct blockages from testicular origin azoospermia and oligospermia.

  4. Gallium nitrate inhibits alkaline phosphatase activity in a differentiating mesenchymal cell culture.

    Science.gov (United States)

    Boskey, A L; Ziecheck, W; Guidon, P; Doty, S B

    1993-02-01

    The effect of gallium nitrate on alkaline phosphatase activity in a differentiating chick limb-bud mesenchymal cell culture was monitored in order to gain insight into the observation that rachitic rats treated with gallium nitrate failed to show the expected increase in serum alkaline phosphatase activity. Cultures maintained in media containing 15 microM gallium nitrate showed drastically decreased alkaline phosphatase activities in the absence of significant alterations in total protein synthesis and DNA content. However, addition of 15 microM gallium nitrate to cultures 18 h before assay for alkaline phosphatase activity had little effect. At the light microscopic and electron microscopic level, gallium-treated cultures differed morphologically from gallium-free cultures: with gallium present, there were fewer hypertrophic chondrocytes and cartilage nodules were flatter and further apart. Because of altered morphology, staining with an antibody against chick cartilage alkaline phosphatase appeared less extensive; however, all nodules stained equivalently relative to gallium-free controls. Histochemical staining for alkaline phosphatase activity was negative in gallium-treated cultures, demonstrating that the alkaline phosphatase protein present was not active. The defective alkaline phosphatase activity in cultures maintained in the presence of gallium was also evidenced when cultures were supplemented with the alkaline phosphatase substrate, beta-glycerophosphate (beta GP). The data presented suggest that gallium inhibits alkaline phosphatase activity in this culture system and that gallium causes alterations in the differentiation of mesenchymal cells into hypertrophic chondrocytes.

  5. Chromatographic separation of alkaline phosphatase from dental enamel

    DEFF Research Database (Denmark)

    Moe, D; Kirkeby, S; Salling, E

    1989-01-01

    Alkaline phosphatase (AP) was prepared from partly mineralized bovine enamel by extraction in phosphate buffer, centrifugation and various chromatographic techniques. Chromatofocusing showed that the enamel enzyme possessed five isoelectric points at the acid pH level ranging from pH 5.7 to pH 4.......4. Three enzyme peaks were eluted using low pressure chromatography with a Bio-gel column. With a HPLC gel filtration column the separation of the enamel extract resulted in only one peak with AP activity. The fractions of this peak were used to produce an antibody against bovine AP....

  6. ALKALINE PHOSPHATASE ACTIVITY AS A MARKER OF DOG SEMEN FREEZABILITY

    Directory of Open Access Journals (Sweden)

    KOSINIAK-KAMYSZ K.

    2007-01-01

    Full Text Available The investigation was performed to evaluate the dog semen freezability and itsquality after thawing allowing its use for artificial insemination (AI. On the basis ofsperm motility, concentration and alkaline phosphatase (AP activity in semenplasma it was possible to establish that AP activity corresponds with the basic factorof semen examination. Significant statistical differences occurred between thequality of ejaculates which were qualified or disqualified to deep freezing and AI.These results show that AP activity in raw dog semen plasma can be used as amarker for the dog semen qualification for deep freezing and AI with 95%probability of the prognosis of the results.

  7. Plasma calcium, magnesium, phosphorus, and alkaline phosphatase levels in normal British schoolchildren.

    Science.gov (United States)

    Round, J M

    1973-07-21

    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.

  8. Isozymes of bovine intestinal alkaline phosphatase. Characterization and functional studies

    Energy Technology Data Exchange (ETDEWEB)

    Besman, M.J.A.

    1986-01-01

    The membrane-associated alkaline phosphatases of calf and adult bovine small intestines have been isolated to homogeneity by a novel method developed to purify large quantities of enzyme. Chromatofocusing revealed the existence of two isozymes in calf tissue while only one form was present in the adult. The three amphiphilic metallo protein dimers were characterized as to total amino acid and carbohydrate content, zinc stoichiometries and mode of carbohydrate linkage. The molecular relationship between the three enzymes was defined by tryptic peptide HPLC-mapping and N-terminal sequencing, and demonstrated the existence of two calf isozymes of unique primary sequence, only one of which is expressed in the adult animal. In the presence of protease inhibitors, two new, higher M/sub r/ species (66,000 and 62,000 daltons vs 60,000 daltons) of adult bovine alkaline phosphatase were demonstrated by electrophoresis of /sup 32/P/sub i/-labeled tissue, probing gels by autoradiography and Western blotting. The in vivo enzyme was isolated using a modified, rapid procedure; the two higher M/sub r/ species copurified.

  9. Kinetic aspects of human placental alkaline phosphatase enzyme membrane.

    Science.gov (United States)

    Roig, M G; Serrano, M A; Bello, J F; Cachaza, J M; Kennedy, J F

    1991-01-01

    The crosslinking of alkaline phosphatase of human placenta with human serum albumin has been optimized. During the physico-chemical characterization of this immobilized biocatalyst, special attention was paid to attributes such as the irreversibility of the enzyme support bonding, the stability of the catalytic activity, and the effects of pH and temperature on this activity. Regarding stability, patterns of denaturation are proposed, to account for inactivation curves over time and under storage/operation conditions. These patterns, in some cases, indicate the existence of different populations of immobilized enzyme molecules, with a different degree of sensitivity to denaturation. The activity vs pH profiles are clearly modified by the immobilization process. This is because the pH of the free homogeneous solution, measurable with a pH-meter, differs from the real pH of the immediate microenvironment of the immobilized enzyme molecules due to the effects of proton accumulation in the microenvironment (in the reaction catalysed by alkaline phosphatase, protons are produced), to limitations to the free diffusion of H+ and to the possible partition effects of H+ due to polar interactions with residues or molecules of the enzyme membrane. In the experimental working conditions, the apparent optimum temperatures are centered at 40 degrees C, inactivation (thermal denaturation) occurring above this temperature. In the temperature range 10-40 degrees C, the kinetic control over the overall activity of the immobilized enzyme was observed, causing the Arrhenius profiles to be linear.

  10. Establishing Quantitative Standards for Residual Alkaline Phosphatase in Pasteurized Milk

    Science.gov (United States)

    Chon, Jung-Whan; Kim, Hyunsook; Kim, Kwang-Yup

    2016-01-01

    The alkaline phosphatase (ALP) assay is a rapid and convenient method for verifying milk pasteurization. Since colorimetric ALP assays rely on subjective visual assessments, their results are especially unreliable near the detection limits. In this study, we attempted to establish quantitative criteria for residual ALP in milk by using a more objective method based on spectrophotometric measurements. Raw milk was heat-treated for 0, 10, 20, 30, and 40 min and then subjected to ALP assays. The quantitative criteria for residual ALP in the milk was determined as 2 μg phenol/mL of milk, which is just above the ALP value of milk samples heat-treated for 30 min. These newly proposed methodology and criteria could facilitate the microbiological quality control of milk. PMID:27194927

  11. Measurement of bone alkaline phosphatase and relative study with osteosarcoma

    Institute of Scientific and Technical Information of China (English)

    YANG Zhiping; HUO Yanqing; SUN Guangzhi; LI Jianmin; LI Xin

    2007-01-01

    The objective of this paper is to explore the value of bone alkaline phosphatase (BALP) for diagnosing osteosarcoma,evaluating the effect of the chemotherapy,judging the prognosis and supervising the relapse and metastasis.The immunoassay was used to check the BALP of the blood serum that was from 42 primary osteosarcoma patients.Alkaline phosphatase (ALP) in blood serum was checked with auto biochemistry equipment.The biopsy tissue and the lesion resected in operation were treated with pathology and histological response was counted.The patients were followed up from five months to 49 months with an average of 24.3 months.Eighteen cases relapsed and transferred,among which,16 of them were dead,and others were survival to the end of the follow-up.BALP was more sensitive than ALP in diagnosing osteosarcoma (P = 0.015).Fifteen cases decreased to normal value in ALP after preoperative chemotherapy,and 34 cases decreased in BALP.Both ALP and BALP in all cases decreased to normal value in postoperative.There was significant difference in positive correlation between the decrease of BALP and the increase of histological response (P = 0.001,r = 0.642).In the followup,there was significant difference in BALP between the group of relapse and transfer and the group of free disease survival (P=0.000).As a check marker in blood serum,BALP,reflecting the process of ossification,has a higher sensitivity than ALP.It has applied value in the diagnosis of osteosarcoma,reflection of the effect of chemotherapy and forecast the prognosis.

  12. Alkaline phosphatase protects against renal inflammation through dephosphorylation of lipopolysaccharide and adenosine triphosphate

    NARCIS (Netherlands)

    Peters, E; Geraci, S; Heemskerk, S; Wilmer, M J; Bilos, A; Kraenzlin, B; Gretz, N; Pickkers, P; Masereeuw, R

    2015-01-01

    BACKGROUND AND PURPOSE: Recently, two phase-II trials demonstrated improved renal function in critically ill patients with sepsis-associated acute kidney injury treated with the enzyme alkaline phosphatase. Here, we elucidated the dual active effect on renal protection by alkaline phosphatase presum

  13. Activity of alkaline phosphatase adsorbed and grafted on "polydopamine" films.

    Science.gov (United States)

    Ball, Vincent

    2014-09-01

    The oxidation of dopamine in slightly basic solutions and in the presence of oxygen as an oxidant allows for the deposition of dopamine-eumelanin ("polydopamine") films on almost all kinds of materials allowing for an easy secondary functionalization. Molecules carrying nucleophilic groups like thiols and amines can be easily grafted on those films. Herein we show that alkaline phosphatase (ALP), as a model enzyme, adsorbs to "polydopamine" films and part of the adsorbed enzyme is rapidly desorbed in contact with Tris buffer. However a significant part of the enzyme remains irreversibly adsorbed and keeps some enzymatic activity for at least 2 weeks whereas ALP adsorbed on quartz slides is rapidly and quantitatively deactivated. In addition we estimated the Michaelis constant Km of the enzyme irreversibly bound to the "polydopamine" film. The Michaelis constant, and hence the affinity constant between paranitrophenol phosphate and ALP are almost identical between the enzyme bound on the film and the free enzyme in solution. Complementarily, it was found that "polydopamine" films display some phosphatase like catalytic activity.

  14. Alkaline phosphatase levels in patients with coronary heart disease saliva and its relation with periodontal status

    Science.gov (United States)

    Yunita, Dina Suci; Masulili, Sri Lelyati C.; Tadjoedin, Fatimah M.; Radi, Basuni

    2017-02-01

    Coronary heart disease (CHD) is a disease that causes narrowing of the coronary arteries. Currently, there is a hypothesis regarding periodontal infection that increases risk for heart disease. Alkaline phosphatase (ALP) as a marker of inflammation will increase in atherosclerosis and periodontal disease. The objective of this research is analyzing the relationship between the levels of alkaline phosphatase in saliva with periodontal status in patients with CHD and non CHD. Here, saliva of 104 subjects were taken, each 1 ml, and levels of Alkaline Phosphatase was analyzed using Abbott ci4100 architect. We found that no significant difference of Alkaline Phosphatase levels in saliva between CHD patients and non CHD. Therefore, it can be concluded that Alkaline Phosphatase levels in patients with CHD saliva was higher than non CHD and no association between ALP levels with periodontal status.

  15. Comparative evaluation of Schistosoma mansoni, Schistosoma intercalatum, and Schistosoma haematobium alkaline phosphatase antigenicity by the alkaline phosphatase immunoassay (APIA).

    Science.gov (United States)

    Cesari, I M; Ballén, D E; Mendoza, L; Ferrer, A; Pointier, J-P; Kombila, M; Richard-Lenoble, D; Théron, A

    2014-04-01

    To know if alkaline phosphatase (AP) from schistosomes other than Schistosoma mansoni can be used as diagnostic marker for schistosomiasis in alkaline phosphatase immunocapture assay (APIA), we comparatively tested n-butanol extracts of adult worm membranes from a Venezuelan (JL) strain of S. mansoni (Ven/AWBE/Sm); a Cameroonian (EDEN) strain of Schistosoma intercalatum (Cam/AWBE/Si) and a Yemeni strain of Schistosoma haematobium (Yem/AWBE/Sh). APIA was evaluated with sera of patients from Venezuela, Senegal, and Gabon infected with S. mansoni, from Gabon infected with S. intercalatum or S. haematobium, from Chine infected with Schistosoma japonicum and from Cambodian patients infected with Schistosoma mekongi. Results indicate that 92.5% (37/40) of Venezuela sera, 75% (15/20) of Senegal sera, 39.5% (17/43) of S. haematobium sera, and 19.2% (5/26) S. intercalatum sera were APIA-positive with the Ven/AWBE/Sm preparation. APIA with the Cam/AWBE/Si preparation showed that 53.8% of S. intercalatum-positive sera had anti-AP antibodies, and 51.2% S. haematobium-positive sera cross-immunocapturing the S. intercalatum AP. APIA performed with Yem/AWBE/Sh showed that 55.8% S. haematobium sera were positive. Only two out of nine S. japonicum sera were APIA-positive with the Ven/AWBE/Sm and Cam/AWBE/Si, and no reaction was observed with Cambodian S. mekongi-positive sera. AP activity was shown to be present in all the schistosome species/strains studied. The use of APIA as a tool to explore the APs antigenicity and the presence of Schistosoma sp. infections through the detection of anti-Schistosoma sp. AP antibodies in a host, allowed us to demonstrate the antigenicity of APs of S. mansoni, S. intercalatum, and S. haematobium.

  16. Bone mineralisation in premature infants cannot be predicted from serum alkaline phosphatase or serum phosphate

    DEFF Research Database (Denmark)

    Faerk, J; Peitersen, Birgit; Petersen, S

    2002-01-01

    BACKGROUND: The bone mineral content of premature infants at term is lower than in mature infants at the same postconceptional age. Serum alkaline phosphatase and serum phosphate are often used as indicators of bone mineralisation. OBJECTIVE: To analyse the association between bone mineral content...... and serum alkaline phosphatase and serum phosphate. METHODS: Serum alkaline phosphatase and phosphate were measured at weekly intervals during admission in 108 premature infants of gestational age below 32 weeks (mean (SD) gestational age 29 (2) weeks; mean (SD) birth weight 1129 (279) g). Bone mineral...... content was measured at term (mean gestational age 41 weeks) by dual energy x ray absorptiometry and corrected for body size. RESULTS: Serum alkaline phosphatase was significantly negatively associated with serum phosphate (p serum alkaline...

  17. Alkaline Phosphatase Assay for Freshwater Sediments: Application to Perturbed Sediment Systems

    Science.gov (United States)

    Sayler, Gary S.; Puziss, Marla; Silver, Martin

    1979-01-01

    The p-nitrophenyl phosphate hydrolysis-phosphatase assay was modified for use in freshwater sediment. Laboratory studies indicated that the recovery of purified alkaline phosphatase activity was 100% efficient in sterile freshwater sediments when optimized incubation and sonication conditions were used. Field studies of diverse freshwater sediments demonstrated the potential use of this assay for determining stream perturbation. Significant correlations between phosphatase and total viable cell counts, as well as adenosine triphosphate biomass, suggested that alkaline phosphatase activity has utility as an indicator of microbial population density and biomass in freshwater sediments. PMID:16345464

  18. SERUM VALUES OF ALKALINE PHOSPHATASE AND LACTATE DEHYDROGENASE IN OSTEOSARCOMA

    Science.gov (United States)

    ZUMÁRRAGA, JUAN PABLO; BAPTISTA, ANDRÉ MATHIAS; ROSA, LUIS PABLO DE LA; CAIERO, MARCELO TADEU; CAMARGO, OLAVO PIRES DE

    2016-01-01

    ABSTRACT Objective: To study the relationship between the pre and post chemotherapy (CT) serum levels of alkaline phosphatase (AP) and lactate dehydrogenase (LDH), and the percentage of tumor necrosis (TN) found in specimens after the pre surgical CT in patients with osteosarcoma. Methods: Series of cases with retrospective evaluation of patients diagnosed with osteosarcoma. Participants were divided into two groups according to serum values of both enzymes. The values of AP and LDH were obtained before and after preoperative CT. The percentage of tumor necrosis (TN) of surgical specimens of each patient was also included. Results: One hundred and thirty seven medical records were included from 1990 to 2013. Both the AP as LDH decreased in the patients studied, being the higher in pre CT than post CT. The average LHD decrease was 795.12U/L and AP decrease was 437.40 U/L. The average TN was 34.10 %. There was no statistically significant correlation between the serums values and the percentage of tumoral necrosis. Conclusion: The serum levels values of AP and LDH are not good predictors for the chemotherapy-induced necrosis in patients with osteosarcoma. Level of Evidence IV, Case Series. PMID:27217815

  19. Interplay between intestinal alkaline phosphatase, diet, gut microbes and immunity.

    Science.gov (United States)

    Estaki, Mehrbod; DeCoffe, Daniella; Gibson, Deanna L

    2014-11-14

    Intestinal alkaline phosphatase (IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP's role in the intestine is to dephosphorylate toxic microbial ligands such as lipopolysaccharides, unmethylated cytosine-guanosine dinucleotides and flagellin as well as extracellular nucleotides such as uridine diphosphate. IAP's ability to detoxify these ligands is essential in protecting the host from sepsis during acute inflammation and chronic inflammatory conditions such as inflammatory bowel disease. Also important in these complications is IAP's ability to regulate the microbial ecosystem by forming a complex relationship between microbiota, diet and the intestinal mucosal surface. Evidence reveals that diet alters IAP expression and activity and this in turn can influence the gut microbiota and homeostasis. IAP's ability to maintain a healthy gastrointestinal tract has accelerated research on its potential use as a therapeutic agent against a multitude of diseases. Exogenous IAP has been shown to have beneficial effects when administered during ulcerative colitis, coronary bypass surgery and sepsis. There are currently a handful of human clinical trials underway investigating the effects of exogenous IAP during sepsis, rheumatoid arthritis and heart surgery. In light of these findings IAP has been marked as a novel agent to help treat a variety of other inflammatory and infectious diseases. The purpose of this review is to highlight the essential characteristics of IAP in protection and maintenance of intestinal homeostasis while addressing the intricate interplay between IAP, diet, microbiota and the intestinal epithelium.

  20. Phosphate solubilizing bacteria and alkaline phosphatase activity in coastal waters off Trivandrum

    Digital Repository Service at National Institute of Oceanography (India)

    Mamatha, S.S.; Gobika, A.; Janani, P.

    Phosphorus is a key nutrient in marine environment. Phosphate solubilising bacteria (PSB) have the ability to solubilise ionic forms of orthophosphoric acid to free form of phosphrous in the water column. Both PSB and alkaline phosphatase activity...

  1. Barley seed coating with free and immobilized alkaline phosphatase to improve P uptake and plant growth

    OpenAIRE

    Pilar Izquierdo, María Concepción; Ortega Santamaría, Natividad; Pérez Mateos, Manuel; Busto Núñez, Mª Dolores

    2012-01-01

    Coating barley seeds with free and immobilized alkaline phosphatase was investigated as a potential means to enhance plant utilization of accumulated soil phosphorus (P). Two coating techniques were studied: film-coating and pelleting. The highest phosphatase activity retention in the coating layer, ranging from 0·48 to 0·67, was observed when seeds were film-coated with phosphatase–polyresorcinol complex (PPC). The germination of seeds film-coated or pelleted with alkaline phosph...

  2. Kidney alkaline phosphatase in mercuric chloride injected chicks resistant and susceptible to leukosis

    Energy Technology Data Exchange (ETDEWEB)

    Miller, V.L.; McIntyre, J.A.; Bearse, G.E.

    1969-01-01

    Two strains of chickens were selected for resistance and susceptibility to avian leukosis. Researchers found that the resistant chicks retained two to four times as much mercury in the liver and kidneys as did the susceptible chicks following injection of mercuric chloride or phenylmercuric acetate. Differences in alkaline phosphatase in the kidneys of the resistant and susceptible chicks, and the effect of the mercuric chloride injection on the alkaline phosphatase activity were reported in this paper. 19 references, 2 tables.

  3. A study on the occurrence of alkaline phosphatase in the sutura interfrontalis of Wistar rats.

    Science.gov (United States)

    Markens, I S; Oudhof, H A

    1979-01-01

    The aim of the present study was to determine the presence of alkaline phosphatase during various stages in development and closure of the sutura interfrontalis. The histological sections reveal that this enzyme could primarily be demonstrated in the dura mater of this suture. In further developmental stages, alkaline phosphatase could be observed within the intermediate zone as well as the pericranium. These findings are brought in relation with the occurrence of synostosis which can be induced under experimental conditions.

  4. Imaging of alkaline phosphatase activity in bone tissue.

    Directory of Open Access Journals (Sweden)

    Terence P Gade

    Full Text Available The purpose of this study was to develop a paradigm for quantitative molecular imaging of bone cell activity. We hypothesized the feasibility of non-invasive imaging of the osteoblast enzyme alkaline phosphatase (ALP using a small imaging molecule in combination with (19Flourine magnetic resonance spectroscopic imaging ((19FMRSI. 6, 8-difluoro-4-methylumbelliferyl phosphate (DiFMUP, a fluorinated ALP substrate that is activatable to a fluorescent hydrolysis product was utilized as a prototype small imaging molecule. The molecular structure of DiFMUP includes two Fluorine atoms adjacent to a phosphate group allowing it and its hydrolysis product to be distinguished using (19Fluorine magnetic resonance spectroscopy ((19FMRS and (19FMRSI. ALP-mediated hydrolysis of DiFMUP was tested on osteoblastic cells and bone tissue, using serial measurements of fluorescence activity. Extracellular activation of DiFMUP on ALP-positive mouse bone precursor cells was observed. Concurringly, DiFMUP was also activated on bone derived from rat tibia. Marked inhibition of the cell and tissue activation of DiFMUP was detected after the addition of the ALP inhibitor levamisole. (19FMRS and (19FMRSI were applied for the non-invasive measurement of DiFMUP hydrolysis. (19FMRS revealed a two-peak spectrum representing DiFMUP with an associated chemical shift for the hydrolysis product. Activation of DiFMUP by ALP yielded a characteristic pharmacokinetic profile, which was quantifiable using non-localized (19FMRS and enabled the development of a pharmacokinetic model of ALP activity. Application of (19FMRSI facilitated anatomically accurate, non-invasive imaging of ALP concentration and activity in rat bone. Thus, (19FMRSI represents a promising approach for the quantitative imaging of bone cell activity during bone formation with potential for both preclinical and clinical applications.

  5. Modeling catalytic promiscuity in the alkaline phosphatase superfamily.

    Science.gov (United States)

    Duarte, Fernanda; Amrein, Beat Anton; Kamerlin, Shina Caroline Lynn

    2013-07-21

    In recent years, it has become increasingly clear that promiscuity plays a key role in the evolution of new enzyme function. This finding has helped to elucidate fundamental aspects of molecular evolution. While there has been extensive experimental work on enzyme promiscuity, computational modeling of the chemical details of such promiscuity has traditionally fallen behind the advances in experimental studies, not least due to the nearly prohibitive computational cost involved in examining multiple substrates with multiple potential mechanisms and binding modes in atomic detail with a reasonable degree of accuracy. However, recent advances in both computational methodologies and power have allowed us to reach a stage in the field where we can start to overcome this problem, and molecular simulations can now provide accurate and efficient descriptions of complex biological systems with substantially less computational cost. This has led to significant advances in our understanding of enzyme function and evolution in a broader sense. Here, we will discuss currently available computational approaches that can allow us to probe the underlying molecular basis for enzyme specificity and selectivity, discussing the inherent strengths and weaknesses of each approach. As a case study, we will discuss recent computational work on different members of the alkaline phosphatase superfamily (AP) using a range of different approaches, showing the complementary insights they have provided. We have selected this particular superfamily, as it poses a number of significant challenges for theory, ranging from the complexity of the actual reaction mechanisms involved to the reliable modeling of the catalytic metal centers, as well as the very large system sizes. We will demonstrate that, through current advances in methodologies, computational tools can provide significant insight into the molecular basis for catalytic promiscuity, and, therefore, in turn, the mechanisms of protein

  6. Cinacalcet Lowers Serum Alkaline Phosphatase in Maintenance Hemodialysis Patients

    Science.gov (United States)

    Belozeroff, Vasily; Goodman, William G.; Ren, Lulu; Kalantar-Zadeh, Kamyar

    2009-01-01

    Background and objectives: Studies suggest an association between elevated serum alkaline phosphatase (AP) and increased mortality in hemodialysis patients, but the effect of existing therapies on AP is not fully understood. We assessed the effects of cinacalcet on AP in a secondary analysis of controlled trial data. Design, setting, participants, & measurements: This was a post hoc analysis of data from three 26-wk randomized, double-blind, placebo-controlled, phase 3 trials and a 26-wk double-blind, placebo-controlled extension trial that investigated cinacalcet in secondary hyperparathyroidism treatment in dialysis patients. Hemodialysis patients (n = 890) with intact parathyroid hormone ≥300 pg/ml and serum calcium ≥8.4 mg/dl received cinacalcet plus standard therapy or standard therapy alone for up to 52 wk. Total, not bone-specific, AP was assessed (proportion of cinacalcet/control subjects achieving a ≥20% or any AP reduction from baseline; the proportion of subjects with AP ≥120 U/L) at baseline; the end of titration; and study weeks 26, 42, and 52. Results: At 52 wk, a greater proportion of cinacalcet-treated patients had either a ≥20% (39 versus 18%) or any (58 versus 36%) AP reduction compared with control subjects, respectively. The likelihood of achieving either a ≥20% or any AP reduction (determined by relative proportion) was 2.33 (95% confidence interval 1.50 to 3.61) and 1.74 (95% confidence interval 1.31 to 2.31), respectively, at week 52. Cinacalcet treatment tended toward a decreased percentage of patients with AP ≥120 U/L (baseline, 42.6%; week 52, 30.6%) compared with control (35.0 to 48.6%, respectively). Conclusions: In this combined analysis of controlled trials of patients who were receiving hemodialysis, cinacalcet lowered total serum AP. PMID:19261825

  7. Observations on the alkaline phosphatase isoenzyme distribution in maternal and amniotic fluid compartments in Nigerian parturients.

    Science.gov (United States)

    Okpere, E; Okorodudu, A; Gbinigie, O

    1988-01-01

    Estimation of the alkaline phosphates isoenzymes in paired maternal serum and amniotic fluids in term uncomplicated pregnancies and in patients with pre-eclampsia, showed poor correlation coefficients between the levels of both heat stable and heat labile isoenzymes. There was a statistically significant fall in AF (P less than .05) HSAP in pre-eclampsia and a highly significant rise of HLAP in meconial liquor. It is concluded that the poor correlation between the levels of HSAP in maternal serum and amniotic fluid (despite their common source of origin), the normal levels of HLAP in maternal serum in the presence of significantly high levels of HSAP in maternal serum in the presence of significantly diminished levels in amniotic fluid point to a state of relatively diminished permeability of the chorioamniotic membranes to the alkaline phosphatase isoenzymes in Nigerians.

  8. Alkaline phosphatase expression during relapse after orthodontic tooth movement

    Directory of Open Access Journals (Sweden)

    Pinandi Sri Pudyani

    2014-03-01

    Full Text Available Background: The increasing of osteoblast activities during bone formation will be accompanied with the increasing expression of alkaline phosphatase enzyme (ALP. ALP can be obtained from clear fluid excreted by gingival crevicular fluid (GCF. Bone turnover, especially bone formation process, can be monitored through the expression of ALP secreted by GCF during orthodontic treatment. Thus, retention period is an important period that can be monitored through the level of bone metabolism around teeth. Purpose: This research were aimed to determine the relation of distance change caused by tooth relapse and ALP activities in gingival crevicular fluid after orthodontic; and to determine ALP as a potential biomarker of bone formation during retention period. Methods: Lower incisors of 25 guinea pigs were moved 3 mm to the distally by using open coil spring. Those relapse distance were measured and the gingival crevicular fluid was taken by using paper points to evaluate ALP levels on days 0, 3, 7, 14 and 21 respectivelly by using a spectrophotometer (405 nm. t-test and ANOVA test were conducted to determine the difference of ALP activities among the time intervals. The correlation regression analysis was conducted to determine the relation of distance change caused by the relapse tooth movement and ALP activities. Results: The greatest relapse movement was occurred on day 3 after open coil spring was removed. There was significant difference of the average of distance decrease among groups A1-A5 (p<0.05. It was also known that ALP level was increased on day 3, but there was no significant difference of the average level of ALP among groups A1-A5 (p>0.05. Finally, based on the results of correlation analysis between the ALP level decreasing and the relapse distance on both right and left of mesial and distal sides, it is known that there was no relation between those two variables (p>0.05. Conclusion: It can be concluded that relapse after orthodontic

  9. Associations between renal hyperfiltration and serum alkaline phosphatase.

    Directory of Open Access Journals (Sweden)

    Se Won Oh

    Full Text Available Renal hyperfiltration, which is associated with renal injury, occurs in diabetic or obese individuals. Serum alkaline phosphatase (ALP level is also elevated in patients with diabetes (DM or metabolic syndrome (MS, and increased urinary excretion of ALP has been demonstrated in patients who have hyperfiltration and tubular damage. However, little was investigated about the association between hyperfiltration and serum ALP level. A retrospective observational study of the 21,308 adults in the Korea National Health and Nutrition Examination Survey IV-V databases (2008-2011 was performed. Renal hyperfiltration was defined as exceeding the age- and sex-specific 97.5th percentile. We divided participants into 4 groups according to their estimated glomerular filtration rate (eGFR: >120, 90-119, 60-89, and 120 mL/min/1.73 m2 showed the highest risk for MS, in the highest ALP quartiles (3.848, 95% CI, 1.876-7.892, compared to the lowest quartile. Similarly, the highest risk for DM, in the highest ALP quartiles, was observed in participants with eGFR >120 ml/min/1.73 m2 (2.166, 95% CI, 1.084-4.329. ALP quartiles were significantly associated with albuminuria in participants with eGFR ≥ 60 ml/min/1.73m2. The highest ALP quartile had a 1.631-fold risk elevation for albuminuria with adjustment of age and sex. (95% CI, 1.158-2.297, P = 0.005. After adjustment, the highest ALP quartile had a 1.624-fold risk elevation, for renal hyperfiltration (95% CI, 1.204-2.192, P = 0.002. In addition, hyperfiltration was significantly associated with hemoglobin, triglyceride, white blood cell count, DM, smoking, and alcohol consumption (P<0.05. The relationship between serum ALP and metabolic disorders is stronger in participants with an upper-normal range of eGFR. Higher ALP levels are significantly associated with renal hyperfiltration in Korean general population.

  10. Immobilization of Penaeus merguiensis alkaline phosphatase on gold nanorods for heavy metal detection.

    Science.gov (United States)

    Homaei, Ahmad

    2017-02-01

    Biotechnology of enzyme has gained popularity due to the growing need for novel environmental technologies and the development of innovative mass-production. The work describes the original application of biosensors based on Penaeus merguiensis alkaline phosphatase (PM ALP) immobilized on gold nanorods (GNRs) to heavy metal determination. Penaeus merguiensis alkaline phosphatase (PM ALP) was immobilized on gold nanorods (GNRs) by ionic exchange and hydrophobic interactions. The optimum pH and temperature for maximum enzyme activity for the immobilized PM ALP are identified to be 11.0 and 60°C, respectively, for the hydrolysis of para-Nitrophenylphosphate (p-NPP). The kinetic studies confirm the Michaelis-Menten behavior and suggests overall slightly decrease in the performance of the immobilized enzyme with reference to the free enzyme. Km and Vmax values were 0.32µm and 54µm. min(-1) for free and 0.39µm and 48µmmin(-1) for immobilized enzymes, respectively. Similarly, the thermal stability, storage stability and stability at extreme pH of the enzyme is found to increase after the immobilization. The inhibitory effect heavy metal ions was studied on free and immobilized PM ALP. The bi-enzymatic biosensor were tested to study the influence of heavy metal ions and pesticides on the corresponding enzyme. The obtained high stability and lower decrease in catalytic efficiency suggested the great potential and feasibility of immobilized PM ALP nanobiocatalyst in efficient and apply the biosensor in total toxic metal content determination.

  11. Efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels in distinguishing exudates from transudates

    OpenAIRE

    Gupta K; Ghalaut Veena; Gupta Prem; Arora Puneet; Tandon S

    2004-01-01

    The objective of present study was to evaluate the efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels to classify pleural fluids. A total of 80 patients were divided in transudates and exudates on the basis of extensive clinical, radiological and biochemical evaluation. The efficacy of pleural fluid alkaline phosphatase (P ALP) and pleural fluid / serum alkaline phosphatase ratio (P/S ALP) assessment along with that of Light′s criteria to accurately classify t...

  12. Relation of salivary calcium, phosphorus and alkaline phosphatase with the incidence of dental caries in children

    Directory of Open Access Journals (Sweden)

    Vijayaprasad K

    2010-09-01

    Full Text Available Aim: The purpose of this study was to assess possible relationship of Calcium, Phosphorus and Alkaline-phophatase levels in saliva with incidence of caries in child patients. Settings and Design: Children (n=75 attending Department of Pedodontics, St. Joseph Dental college, Eluru, with and without caries were categorized in to Group I: Consisting of 25 children with non-rampant caries, Group II: Consisting of 25 children with rampant caries, Group III: Consisting of 25 children without caries. (Control group. Materials and Methods: The samples of saliva were collected one week after oral prophylaxis. Unstimulated directly expectorated whole saliva samples were collected in clean, dry, sterilized glass bottles and fitted with proper rubber stoppers immediately. The samples were subjected to biochemical assay for estimation of calcium, phosphorus and alkaline phosphatase levels. Statistical analysis used: ANOVA. Results: The alkaline Phosphatase activity for rampant caries group was 18.66 K.A, and control group was 4.68 K.A. The values of alkaline phosphatase activity for minimal caries group was 6.16 KA. Conclusion: Saliva could reflect a caries risk situation was supported by the fact that alkaline phosphatase activity was very much significantly higher in caries prone groups.

  13. Relationship between serum heat—stable alkaline phosphatase level and p[regnancy

    Institute of Scientific and Technical Information of China (English)

    CaoGuo-Xian; DingMei-Juan; 等

    1998-01-01

    Serum heat-stable alkaline phosphatase(HSAP) level in 649 cases of normal pregnancy and 164 cases of high-risk pregnancy is measured by raioimmunoassay (RIA).The results indicate that the HSAP level in normal prenancy increased proportionally with gestation weeks(r=0.9843).In 33 cases of pregnancy induced hypertension and 21 cases of intrauterine fetal growth retardation,the HSAP level is significantly low.In 7 cases of neonatal asphyxia and 26 cases of ftal distress,the HSAP level in the mother's serum is also low.In 53 cases of intrahepatic cholestasis of pregnancy,the HSAP level in similar to those of normal pregnancy,This study illustrates that HSAP RIA can play an important role in the evaluation of placental function and fetal prognosis for cases of high-risk pregancy.

  14. Alkaline phosphatase immobilization onto Bio-Gide(R) and Bio-Oss(R) for periodontal and bone regeneration.

    NARCIS (Netherlands)

    Oortgiesen, D.A.W.; Plachokova, A.S.; Geenen, C.; Meijer, G.J.; Walboomers, X.F.; Beucken, J.J.J.P van den; Jansen, J.B.M.J.

    2012-01-01

    AIM: To evaluate the effect of alkaline phosphatase (ALP) immobilization onto Bio-Gide((R)) in vitro, and to study the in vivo performance of ALP-enriched Bio-Gide((R)) and/or Bio-Oss((R)) with the purpose to enhance periodontal regeneration. MATERIALS AND METHODS: Alkaline phosphatase ALP was

  15. Alkaline phosphatase immobilization onto Bio-Gide(R) and Bio-Oss(R) for periodontal and bone regeneration.

    NARCIS (Netherlands)

    Oortgiesen, D.A.W.; Plachokova, A.S.; Geenen, C.; Meijer, G.J.; Walboomers, X.F.; Beucken, J.J.J.P van den; Jansen, J.B.M.J.

    2012-01-01

    AIM: To evaluate the effect of alkaline phosphatase (ALP) immobilization onto Bio-Gide((R)) in vitro, and to study the in vivo performance of ALP-enriched Bio-Gide((R)) and/or Bio-Oss((R)) with the purpose to enhance periodontal regeneration. MATERIALS AND METHODS: Alkaline phosphatase ALP was immob

  16. 1-Step Versus 2-Step Immobilization of Alkaline Phosphatase and Bone Morphogenetic Protein-2 onto Implant Surfaces Using Polydopamine

    OpenAIRE

    Nijhuis, Arnold W.G.; van den Beucken, Jeroen J.J.P.; Boerman, Otto C.; Jansen, John A.; Leeuwenburgh, Sander C.G.

    2013-01-01

    Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the current study, a 1-step polydopamine-based approach was applied for alkaline phosphatase (ALP) and bone morphogenetic protein-2 (BMP-2) immobilization, and compared to the conventional 2-step polydop...

  17. Relationship between Salivary Alkaline Phosphatase Enzyme Activity and The Concentrations of Salivary Calcium and Phosphate Ions

    Directory of Open Access Journals (Sweden)

    Mina Jazaeri

    2015-04-01

    Full Text Available Although salivary alkaline phosphatase (ALP can balance de- and remineralization processes of enamel, there is no evidence regarding its effects on the concentrations of calcium and phosphate in saliva. The present study aims to determine the relationship between salivary ALP activity and the concentrations of calcium and phosphate in saliva. In this cross-sectional study, we evaluated salivary markers in 120 males, ages 19 to 44 years. All participants provided 5 mL of unstimulated whole saliva and the level of enzyme activity as well as calcium and phosphate concentrations were measured using a colorimetric method. Data were gathered and analyzed by statistical package for social sciences (SPSS 13.00 using Pearson correlation test. A p value of 0.05. According to the results of the present study, there was no significant relation between salivary ALP activity and calcium and phosphate concentrations in saliva. However, further research is highly recommended.

  18. Differentiating intracellular from extracellular alkaline phosphatase activity in soil by sonication.

    Directory of Open Access Journals (Sweden)

    Shuping Qin

    Full Text Available Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio  =  1/8 (w/v and power density  =  15 watt ml(-1], the activity of alkaline phosphomonoesterase (phosphatase in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first plateau of enzyme activity was reached between 60 and 100 s, and a second higher plateau after 300 s. We also found that sonication for 100 s under optimal conditions activated most (about 80% of the alkaline phosphatase that was added to an autoclaved soil, while total bacteria number was not affected. Sonication for 300 s reduced the total bacteria number by three orders of magnitude but had no further effects on enzyme activity. Our results indicate that the first plateau of alkaline phosphatase activity was derived from extracellular enzymes attached to soil particles, and the second plateau to the combination of extracellular and intracellular enzymes after cell lysis. We conclude that our adjusted sonication method may be an alternative to the currently used physiological and chloroform-fumigation methods for differentiating intracellular from extracellular phosphatase activity in soil. Further testing is needed to find out whether this holds for other soil types.

  19. Differentiating intracellular from extracellular alkaline phosphatase activity in soil by sonication.

    Science.gov (United States)

    Qin, Shuping; Hu, Chunsheng; Oenema, Oene

    2013-01-01

    Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio  =  1/8 (w/v) and power density  =  15 watt ml(-1)], the activity of alkaline phosphomonoesterase (phosphatase) in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first plateau of enzyme activity was reached between 60 and 100 s, and a second higher plateau after 300 s. We also found that sonication for 100 s under optimal conditions activated most (about 80%) of the alkaline phosphatase that was added to an autoclaved soil, while total bacteria number was not affected. Sonication for 300 s reduced the total bacteria number by three orders of magnitude but had no further effects on enzyme activity. Our results indicate that the first plateau of alkaline phosphatase activity was derived from extracellular enzymes attached to soil particles, and the second plateau to the combination of extracellular and intracellular enzymes after cell lysis. We conclude that our adjusted sonication method may be an alternative to the currently used physiological and chloroform-fumigation methods for differentiating intracellular from extracellular phosphatase activity in soil. Further testing is needed to find out whether this holds for other soil types.

  20. A role for intestinal alkaline phosphatase in the maintenance of local gut immunity

    NARCIS (Netherlands)

    Chen, K.T.; Malo, M.; Beasley-Topliffe, L.K.; Poelstra, K.; Millan, J.; Mostafa, G.; Alam, S.; Ramasamy, S.; Warren, H.; Hohmann, E.; Hodin, R.A.

    2010-01-01

    Background and Aims: Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor known to dephosphorylate lipopolysaccharide (LPS); however, the role of IAP in the gut response to luminal bacteria remains undefined. We investigated immune responses of wildtype (WT) and IAP-knockout (IAP-KO

  1. The effect of alkaline phosphatase coated onto titanium alloys on bone responses in rats.

    NARCIS (Netherlands)

    Schouten, C.; Beucken, J.J.J.P. van den; Jonge, L.T. de; Bronkhorst, E.M.; Meijer, G.J.; Spauwen, P.H.M.; Jansen, J.A.

    2009-01-01

    The enzyme alkaline phosphatase (ALP) was recently proposed as an implant coating material in order to improve the biological performance of orthopedic and dental implants. The present study evaluated the in vivo bone response to electrosprayed coatings, consisting of ALP, calcium phosphate (CaP) or

  2. Intestinal alkaline phosphatase contributes to the reduction of severe intestinal epithelial damage.

    NARCIS (Netherlands)

    Bol-Schoenmakers, M.; Fiechter, D.; Raaben, W.; Hassing, I.; Bleumink, R.; Kruijswijk, D.; Maijoor, K.; Tersteeg-Zijderveld, M.; Brands, R.; Pieters, R.

    2010-01-01

    Inflammatory bowel disease is characterized by chronic inflammation of the intestine and is accompanied by damage of the epithelial lining and by undesired immune responses towards enteric bacteria. It has been demonstrated that intestinal alkaline phosphatase (iAP) protects against the induction of

  3. PLACENTAL-LIKE ALKALINE-PHOSPHATASE AND DNA FLOW-CYTOMETRY IN SPERMATOCYTIC SEMINOMA

    NARCIS (Netherlands)

    DEKKER, [No Value; ROZEBOOM, T; DELEMARRE, J; OOSTERHUIS, JW; Dam, A.

    1992-01-01

    Immunohistochemical analysis was done on 7 testicular tumors classified as spermatocytic seminoma (SS) and 25 classic seminomas. Except for a few scattered cells, the spermatocytic seminomas were negative for placental-like alkaline phosphatase (PLAP); the classic seminomas were all positive for

  4. [Serum calcium and phosphorus concentration and alkaline phosphatase activity in healthy children during growth and development].

    Science.gov (United States)

    Savić, Ljiljana; Savić, Dejan

    2008-01-01

    Many changes happen during growth and development in an organism as a result of important hormon changes, especially biohumoral ones. These changes make a problem when interpreting biochemical results in pediatric population. The most important changes are intensive calcium and phosphorus metabolic turnover in bone tissue with changes in alkaline phosphatase activity as a result of osteoblast activity. The aim of this study was to follow the serum calcium and phosphorus concentration and alkaline phosphatase activity in children 1-15 years old in different growth and development period and of different sexes and to fortify the influence of growth and development dynamics on biohumoral status in healthy male and female children. We evaluated 117 healthy children of both sexes from 1-15 years of age and divided them into three age groups: 1-5, 6-10 and 11-15 years. We followed the serum calcium and phosphorus concentration and alkaline phosphatase activity in different groups and in different sexes. Our investigation found significantly higher values of serum calcium in boys than in girls with no important changes between the age groups and significantly higher values of serum phosphorus in the youngest age group in all children and in different sexes with no important sex differences. Alkaline phosphatase activity followed the growth spurt and was the biggest in 6-10 years group in girls and in 11-15 years group in boys.

  5. Alkaline phosphatase-polyresorcinol complex: characterization and application to seed coating.

    Science.gov (United States)

    Pilar, María C; Ortega, Natividad; Perez-Mateos, Manuel; Busto, María D

    2009-03-11

    An alkaline phosphatase (EC 3.1.3.1) from Escherichia coli ATCC27257 was immobilized by copolymerization with resorcinol. The phosphatase-polyresorcinol complex synthesized retained about 74% of the original enzymatic activity. The pH and temperature profile of the immobilized and free enzyme revealed a similar behavior. Kinetic parameters were determined: K(m) and K(i) values were 2.44 and 0.423 mM, respectively, for the phosphatase-polyresorcinol complex and 1.07 and 0.069 mM, respectively, for free phosphatase. The thermal and storage stabilities of the immobilized phosphatase were higher than those of the native one. On addition to soil, free enzyme was completely inactivated in 4 days, whereas the phosphatase-polyresorcinol complex was comparatively stable. Barley seed coated with the immobilized enzyme exhibited higher rhizosphere phosphatase activity. Under pot culture conditions, an increase in the soil inorganic phosphorus was detected when the seed was encapsulated with the phosphatase-polyresorcinol complex, and a positive influence on biomass and inorganic phosphorus concentration of shoot was observed.

  6. Somatic cell count and alkaline phosphatase activity in milk for evaluation of mastitis in buffalo.

    Science.gov (United States)

    Patil, M P; Nagvekar, A S; Ingole, S D; Bharucha, S V; Palve, V T

    2015-03-01

    Mastitis is a serious disease of dairy animals causing great economic losses due to a reduction in milk yield as well as lowering its nutritive value. The application of somatic cell count (SCC) and alkaline phosphatase activity in the milk for diagnosis of mastitis in buffalo is not well documented. Therefore, the present study was conducted to observe the SCC and alkaline phosphatase activity for evaluation of mastitis in buffalo. Milk samples of forty apparently healthy lactating buffaloes were selected and categorized into five different groups viz. normal buffaloes, buffaloes with subclinical mastitis with CMT positive milk samples (+1 Grade), (+2 Grade), (+3 Grade), and buffaloes with clinical mastitis with 8 animals in each group. The milk samples were analyzed for SCC and alkaline phosphatase activity. The levels of SCC (×10(5) cells/ml) and alkaline phosphatase (U/L) in different groups were viz. normal (3.21±0.179, 16.48±1.432), subclinical mastitis with CMT positive milk samples with +1 Grade (4.21±0.138, 28.11±1.013), with +2 Grade (6.34±0.183, 34.50±1.034), with +3 Grade (7.96±0.213, 37.73±0.737) and buffaloes with clinical mastitis (10.21±0.220, 42.37±0.907) respectively, indicating an increasing trend in the values and the difference observed among various group was statistically significant. In conclusion, the results of the present study indicate that the concentration of milk SCC and alkaline phosphatase activity was higher in the milk of buffaloes with mastitis than in the milk of normal buffaloes.

  7. Membrane-bound alkaline phosphatase from ectopic mineralization and rat bone marrow cell culture.

    Science.gov (United States)

    Simão, Ana Maria S; Beloti, Márcio M; Cezarino, Rodrigo M; Rosa, Adalberto Luiz; Pizauro, João M; Ciancaglini, Pietro

    2007-04-01

    Cells from rat bone marrow exhibit the proliferation-differentiation sequence of osteoblasts, form mineralized extracellular matrix in vitro and release alkaline phosphatase into the medium. Membrane-bound alkaline phosphatase was obtained by method that is easy to reproduce, simpler and fast when compared with the method used to obtain the enzyme from rat osseous plate. The membrane-bound alkaline phosphatase from cultures of rat bone marrow cells has a MW(r) of about 120 kDa and specific PNPP activity of 1200 U/mg. The ecto-enzyme is anchored to the plasma membrane by the GPI anchor and can be released by PIPLC (selective treatment) or polidocanol (0.2 mg/mL protein and 1% (w/v) detergent). The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10. This fraction hydrolyzes ATP (240 U/mg), ADP (350 U/mg), glucose 1-phosphate (1100 U/mg), glucose 6-phosphate (340 U/mg), fructose 6-phosphate (460 U/mg), pyrophosphate (330 U/mg) and beta-glycerophosphate (600 U/mg). Cooperative effects were observed for the hydrolysis of PPi and beta-glycerophosphate. PNPPase activity was inhibited by 0.1 mM vanadate (46%), 0.1 mM ZnCl2 (68%), 1 mM levamisole (66%), 1 mM arsenate (44%), 10 mM phosphate (21%) and 1 mM theophylline (72%). We report the biochemical characterization of membrane-bound alkaline phosphatase obtained from rat bone marrow cells cultures, using a method that is simple, rapid and easy to reproduce. Its properties are compared with those of rat osseous plate enzyme and revealed that the alkaline phosphatase obtained has some kinetics and structural behaviors with higher levels of enzymatic activity, facilitating the comprehension of the mineralization process and its function.

  8. Efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels in distinguishing exudates from transudates

    Directory of Open Access Journals (Sweden)

    Gupta K

    2004-01-01

    Full Text Available The objective of present study was to evaluate the efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels to classify pleural fluids. A total of 80 patients were divided in transudates and exudates on the basis of extensive clinical, radiological and biochemical evaluation. The efficacy of pleural fluid alkaline phosphatase (P ALP and pleural fluid / serum alkaline phosphatase ratio (P/S ALP assessment along with that of Light′s criteria to accurately classify transudates and exudates were analyzed. Up to 89% transudates misclassified by Light′s criteria were correctly classified by pleural fluid alkaline phosphatase (P ALP and pleural fluid/serum alkaline phosphatase ratio (P/S ALP evaluation. Similarly 92% exudates misclassified by Light′s criteria were correctly classified by pleural fluid alkaline phosphatase (P ALP and pleural fluid/serum alkaline phosphatase ratio (P/S ALP evaluation. By applying a cut off value of 40.0 IU for P ALP, a sensitivity of 85% and specificity of 75% was found. For P/S ALP, applying a cut off value of 0.25 a sensitivity of 85% and specificity of 80% was found. Both P ALP and P/S ALP had a PPV of 92%. However, their respective NPV were 63% and 70%.

  9. Development of conductometric biosensors based on alkaline phosphatases for the water quality control

    CERN Document Server

    Berezhetskyy, A

    2008-01-01

    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devoted to creation and optimization of conductometric biosensor based on alkaline phosphatase active microalgae and sol gel technology, the last chapter described application of the proposed algal biosensor for measurements of heavy metal ions toxicity of waste water, general conclusions stating the progresses achieved in the field of environmental monitoring

  10. Development of conductometric biosensors based on alkaline phosphatases for the water quality control

    Science.gov (United States)

    Berezhetskyy, A.

    2008-09-01

    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devoted to creation and optimization of conductometric biosensor based on alkaline phosphatase active microalgae and sol gel technology, the last chapter described application of the proposed algal biosensor for measurements of heavy metal ions toxicity of waste water, general conclusions stating the progresses achieved in the field of environmental monitoring

  11. Significance of bone specific alkaline phosphatase as a tumor marker in malignant bone tumor

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sug Jun; Jeon, Dae Geun; Huh, Kwang [Korea Cancer Center Hospital, Seoul (Korea, Republic of)

    1998-01-01

    The relationship between total alkaline phosphatase activity and bone forming lesion is a well known fact. But alkaline phosphatase consist mainly of two portion (liver, bone). To clarify the exact activity of bone forming tissue, quantitative measurement of BALP is essential. Two finds of tests were performed for their feasibility as a laboratory test (wheat germ lectin vs electrophoresis). We analyzed 40 bony lesion and got 58 samples. Lectin method was simple, economic, with reliable resproducability. Owing to the small number of test sample, we could not identify the relationship between the disease activity and measured BALP level. Further collection of clinical sample and analysis the pattern of BALP on each clinical settings. (author). 8 refs.

  12. The Effect of Titanium Dioxide Nanoparticles on Salivary Alkaline Phosphatase Activity

    Directory of Open Access Journals (Sweden)

    Eaman A. S. AL-Rubaee

    2015-12-01

    Full Text Available The structural and optical properties of the titanium dioxide nanoparticles [TiO2 NPs] have been investigated using [UV-Vis] spectrophotometer and SEM. The produced nanoparticles show small and about sharp round peaks around 220 nm. The produced nanoparticles have a spherical shape with an average particle size ˂50 nm. The effect of titanium dioxide NPs was studied on the activity of Alkaline Phosphatase [ALP] in the saliva of 25 patients with gingivitis in comparison to 20 healthy subjects with the average age about 22–23 years for both groups. The results correlated with the observation that salivary alkaline phosphatase activity increase in patient with gingivitis in comparison to control group and salivary ALP activity inhibited by titanium dioxide nanoparticles.

  13. [Development of conductometric biosensor based on alkaline phosphatase for determining concentration of cadmium ions].

    Science.gov (United States)

    Sosovs'ka, O F; Berezhets'kyĭ, A L

    2007-01-01

    The paper describes a novel conductometric biosensor sensitive to cadmium ions based on alkaline phosphatase immobilized on gold planar microelectrodes used as transducers. Assays have been carried out with paranitrophenyl phosphate as substrate for the immobilized enzyme. Various parameters such as reticulation time, along with pH, ionic strength and buffer concentration of the measuring solution were studied. The optimized biosensor was stable, reproducible and it exhibited a detection limit of 4.45 microM for cadmium ions.

  14. Development of conductometric biosensors based on alkaline phosphatases for the water quality control

    OpenAIRE

    2008-01-01

    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devote...

  15. Biochemical and functional properties of mammalian bone alkaline phosphatase isoforms during osteogenesis

    OpenAIRE

    Halling Linder, Cecilia

    2016-01-01

    The human skeleton is a living and dynamic tissue that constantly is being renewed in a process called bone remodeling. Old bone is resorbed by osteoclasts and new bone is formed by osteoblasts. Bone is a composite material made up by mineral crystals in the form of hydroxyapatite (calcium and phosphate) that provides the hardness of bone, and collagen fibrils that provides elasticity and flexibility. Alkaline phosphatase (ALP) is a family of enzymes that is present in most species and cataly...

  16. Synthesis, alkaline phosphatase inhibition studies and molecular docking of novel derivatives of 4-quinolones.

    Science.gov (United States)

    Miliutina, Mariia; Ejaz, Syeda Abida; Khan, Shafi Ullah; Iaroshenko, Viktor O; Villinger, Alexander; Iqbal, Jamshed; Langer, Peter

    2017-01-27

    New and convenient methods for the functionalization of the 4-quinolone scaffold at positions C-1, C-3 and C-6 were developed. The 4-quinolone derivatives were evaluated for their inhibitory potential on alkaline phosphatase isozymes. Most of the compounds exhibit excellent inhibitory activity and moderate selectivity. The IC50 values on tissue non-specific alkaline phosphatase (TNAP) were in the range of 1.34 ± 0.11 to 44.80 ± 2.34 μM, while the values on intestinal alkaline phosphatase (IAP) were in the range of 1.06 ± 0.32 to 192.10 ± 3.78 μM. The most active derivative exhibits a potent inhibition on IAP with a ≈14 fold higher selectivity as compared to TNAP. Furthermore, molecular docking calculations were performed for the most potent inhibitors to show their binding interactions within the active site of the respective enzymes.

  17. Intraspecific variation in alkaline phosphatase activity in Phaeodactylum tricornutum (Bacillariophyceae, Bohlin

    Directory of Open Access Journals (Sweden)

    Domênica Teixeira de Lima

    2016-01-01

    Full Text Available ABSTRACT To describe potential intraspecific variation in phosphorus incorporation in two strains of Phaeodactylum tricornutum (Bohlin, Ub3 and Ub7, alkaline phosphatase (AP activity was evaluated via enzyme-labeled fluorescence assay. Analysis using the probe ELF-97(r provides individual evaluation, and therefore can determine the nutritional status of inorganic phosphorus in phytoplanktonic cells. Bioassays compared the control treatment to both phosphate-enriched and phosphate-depleted treatments by varying only the phosphate concentration in the media. The P. tricornutum strains exhibited differences in their development when incubated in the phosphate-enriched media. The development of the Ub7 strain differed by exhibiting "luxury uptake" and utilization of organic phosphorus, and the alkaline phosphatase analysis indicated limitations of this clone under such conditions. The Ub7 strain showed higher AP activity, when compared to Ub3, in the P-enriched condition. P. tricornutum presented increases in AP activity and low variation in Surface/Volume ratio, by increasing biovolume and its maximum linear dimension, as strategies for phosphate incorporation. Our results highlight intraspecific differences in alkaline phosphatase activity, and hence differences in the incorporation of organic phosphorus, as the tested species regulated enzymatic activity under different external phosphate concentrations.

  18. Alkaline phosphatase activity at the southwest coast of India: A comparison of locations differently affected by upwelling..

    Digital Repository Service at National Institute of Oceanography (India)

    Mamatha, S.S.; Malik, A.; Varik, S.; Parvathi, V.; Jineesh, V.K.; Gauns, M.; LokaBharathi, P.A.

    coastal waters. As alkaline phosphatase activity (APA) indicates the status of P for primary production in aquatic environments, we asked the following question: is the level of APA indicative of P sufficiency or deficiency in coastal waters, especially...

  19. [Recovery of the sensitivity of J-41 cells to Coxsackie B3 virus by treatment with exogenous alkaline phosphatase].

    Science.gov (United States)

    Voronina, F V; Gulevich, N E; Khesin, Ia E

    1980-12-01

    It has been shown that injection of G-41 cell cultures, deficient as regards alkaline phosphatase and resistant to Coxsackie B3 virus, in conjunction with exposure to an alkaline phosphatase preparation from the calf intestine results in virus reproduction. Depending on the dose administered and multiplicity of infection there occur either complete destruction of the monolayer or death of some cells with the development of cytopathic changes specific for Coxackie virus.

  20. Structural characteristics of alkaline phosphatase from the moderately halophilic bacterium Halomonas sp. 593

    Energy Technology Data Exchange (ETDEWEB)

    Arai, Shigeki; Yonezawa, Yasushi [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan); Ishibashi, Matsujiro [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Matsumoto, Fumiko; Adachi, Motoyasu; Tamada, Taro [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan); Tokunaga, Hiroko [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Blaber, Michael [Florida State University, 1115 West Call Street, Tallahassee, FL 32306-4300 (United States); Tokunaga, Masao [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Kuroki, Ryota, E-mail: kuroki.ryota@jaea.go.jp [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan)

    2014-03-01

    In order to clarify the structural basis of the halophilic characteristics of an alkaline phosphatase derived from the moderate halophile Halomonas sp. 593 (HaAP), the tertiary structure of HaAP was determined to 2.1 Å resolution by X-ray crystallography. The structural properties of surface negative charge and core hydrophobicity were shown to be intermediate between those characteristic of halophiles and non-halophiles, and may explain the unique functional adaptation to a wide range of salt concentrations. Alkaline phosphatase (AP) from the moderate halophilic bacterium Halomonas sp. 593 (HaAP) catalyzes the hydrolysis of phosphomonoesters over a wide salt-concentration range (1–4 M NaCl). In order to clarify the structural basis of its halophilic characteristics and its wide-range adaptation to salt concentration, the tertiary structure of HaAP was determined by X-ray crystallography to 2.1 Å resolution. The unit cell of HaAP contained one dimer unit corresponding to the biological unit. The monomer structure of HaAP contains a domain comprised of an 11-stranded β-sheet core with 19 surrounding α-helices similar to those of APs from other species, and a unique ‘crown’ domain containing an extended ‘arm’ structure that participates in formation of a hydrophobic cluster at the entrance to the substrate-binding site. The HaAP structure also displays a unique distribution of negatively charged residues and hydrophobic residues in comparison to other known AP structures. AP from Vibrio sp. G15-21 (VAP; a slight halophile) has the highest similarity in sequence (70.0% identity) and structure (C{sup α} r.m.s.d. of 0.82 Å for the monomer) to HaAP. The surface of the HaAP dimer is substantially more acidic than that of the VAP dimer (144 exposed Asp/Glu residues versus 114, respectively), and thus may enable the solubility of HaAP under high-salt conditions. Conversely, the monomer unit of HaAP formed a substantially larger hydrophobic interior

  1. Enslavement in the water body by toxic Aphanizomenon ovalisporum, inducing alkaline phosphatase in phytoplanktons.

    Science.gov (United States)

    Bar-Yosef, Yehonathan; Sukenik, Assaf; Hadas, Ora; Viner-Mozzini, Yehudit; Kaplan, Aaron

    2010-09-14

    The hepatotoxin cylindrospermopsin (CYN) produced by certain cyanobacteria, including Aphanizomenon ovalisporum (hereafter Aphanizomenon) [1], seriously affects lake water quality [2], but its biological role is not known. Strong correlation between Aphanizomenon abundance in Lake Kinneret, Israel, and alkaline phosphatase (APase) activity suggests that inorganic phosphate (Pi) limitation induces the PHO regulon and APase secretion [3]. Staining lake samples with DAPI [4] revealed a high level of polyphosphate bodies (PPB) in Aphanizomenon. Application of enzyme-labeled fluorescence (ELF-APase) [5] showed APase in various organisms, but not in Aphanizomenon. ELF-APase signals and extracellular APase activity in Aphanizomenon were detected only after exploiting PPB under prolonged Pi deprivation in cultures or toward the end of its autumn bloom. Pi deprivation of Aphanizomenon induces CYN production, high-affinity Pi uptake, and an internal, not external, APase. Addition of Aphanizomenon spent media or CYN to various phytoplanktons, including Chlamydomonas reinhardtii, induced genes typically upregulated under Pi limitation and a rise in extracellular APase activity, despite ample surrounding Pi. Coculturing Aphanizomenon with Chlamydomonas or with Debarya sp. showed positive ELF-APase signals, but not in Aphanizomenon. CYN producers promote Pi supply by inducing APase secretion by other phytoplanktons, possibly explaining their increased abundance despite reduced Pi supply from watersheds.

  2. Surface alkaline phosphatase activities of macroalgae on coral reefs of the central Great Barrier Reef, Australia

    Science.gov (United States)

    Schaffelke, B.

    2001-05-01

    Inshore reefs of the Great Barrier Reef (GBR) are subject to episodic nutrient supply, mainly by flood events, whereas midshelf reefs have a more consistent low nutrient availability. Alkaline phosphatase activity (APA) enables macroalgae to increase their phosphorus (P) supply by using organic P. APA was high (~4.0 to 15.5 µmol PO4 3- g DW-1 h-1) in species colonising predominantly inshore reefs and low (study were much lower than data reported from other coral reef systems. In experiments with two Sargassum species tissue P levels were correlated negatively, and N:P ratios were positively correlated with APA. High APA can compensate for a relative P-limitation of macroalgae in coral reef systems that are subject to significant N-inputs, such as the GBR inshore reefs. APA and other mechanisms to acquire a range of nutrient species allow inshore species to thrive in habitats with episodic nutrient supply. These species also are likely to benefit from an increased nutrient supply caused by human activity, which currently is a global problem.

  3. Evidence of associations between feto-maternal vitamin D status, cord parathyroid hormone and bone-specific alkaline phosphatase, and newborn whole body bone mineral content

    Science.gov (United States)

    In spite of a high prevalence of vitamin D inadequacy in pregnant women and neonates, relationships among vitamin D status [25(OH)D], parathyroid hormone (PTH), bone specific alkaline phosphatase (BALP), and whole body bone mineral content (WBBMC) in the newborn are poorly characterized. The purpose...

  4. Intestinal alkaline phosphatase: multiple biological roles in maintenance of intestinal homeostasis and modulation by diet.

    Science.gov (United States)

    Lallès, Jean-Paul

    2010-06-01

    The diverse nature of intestinal alkaline phosphatase (IAP) functions has remained elusive, and it is only recently that four additional major functions of IAP have been revealed. The present review analyzes the earlier literature on the dietary factors modulating IAP activity in light of these new findings. IAP regulates lipid absorption across the apical membrane of enterocytes, participates in the regulation of bicarbonate secretion and of duodenal surface pH, limits bacterial transepithelial passage, and finally controls bacterial endotoxin-induced inflammation by dephosphorylation, thus detoxifying intestinal lipopolysaccharide. Many dietary components, including fat, protein, and carbohydrate, modulate IAP expression or activity and may be combined to sustain a high level of IAP activity. In conclusion, IAP has a pivotal role in intestinal homeostasis and its activity could be increased through the diet. This is especially true in pathological situations (e.g., inflammatory bowel diseases) in which the involvement of commensal bacteria is suspected and when intestinal AP is too low to detoxify a sufficient amount of bacterial lipopolysaccharide.

  5. Evaluation of biodegradation and biocompatibility of collagen/chitosan/alkaline phosphatase biopolymeric membranes

    Indian Academy of Sciences (India)

    E BERTEANU; D IONITA; M SIMOIU; M PARASCHIV; R TATIA; A APATEAN; M SIDOROFF; L TCACENCO

    2016-04-01

    The aim of this study was to develop a new variant of membranes based on collagen (COL), chitosan (CHI) and alkaline phosphatase (ALP) immobilized and cross-linking with glutaraldehyde (GA) at different concentrations. The biodegradation in the presence of collagenase was investigated. Biocompatibility was evaluated by MTT assay using a mouse fibroblast cell culture type NCTC (clone 929). Non-cross-linked samples were biocompatible and membranes cross-linked with low concentrations of GA (0.04, 0.08%) were also iocompatible. However, high concentrations of GA lead to a decreased biocompatibility. The adsorption behaviour of Ca$^{2+}$ ions to all membraneswere evaluated using the Freundlich isotherms. Haemolytic studies were performed in order to consider their applications in biomineralization process. By the addition of collagen and ALP to chitosan, the haemolytic indexdecreases, the COL–CHI–ALP membrane being in the non-haemolytic domain, while the COL–CHI–ALP–GA membrane has a haemolytic index greater than 2, and is slightly haemolytic.

  6. Responses of phosphate transporter gene and alkaline phosphatase in Thalassiosira pseudonana to phosphine.

    Science.gov (United States)

    Fu, Mei; Song, Xiuxian; Yu, Zhiming; Liu, Yun

    2013-01-01

    Phosphine, which is released continuously from sediment, can affect the eco-physiological strategies and molecular responses of phytoplankton. To examine the effects of phosphine on phosphorus uptake and utilization in Thalassiosira pseudonana, we examined the transcriptional level of the phosphate transporter gene (TpPHO) and the activity of alkaline phosphatase (AKP) in relation to supplement of various concentrations of phosphine. TpPHO expression was markedly promoted by phosphine in both the phosphate-deficient and phosphate-4 µM culture. However, high phosphine concentrations can inhibit TpPHO transcription in the declining growth phase. AKP activity was also higher in the phosphine treatment groups than that of the control. It increased with increasing phosphine concentration in the range of 0 to 0.056 µM but was inhibited by higher levels of phosphine. These responses revealed that phosphine can affect phosphate uptake and utilization in T. pseudonana. This result was consistent with the effect of phosphine on algal growth, while TpPHO expression and AKP were even more sensitive to phosphine than algal growth. This work provides a basic understanding for further research about how phosphine affects phytoplankton.

  7. Prognostic Significance of Serum Alkaline Phosphatase Level in Osteosarcoma: A Meta-Analysis of Published Data

    Directory of Open Access Journals (Sweden)

    Hai-Yong Ren

    2015-01-01

    Full Text Available Background. Serum alkaline phosphatase (SALP is commonly elevated in osteosarcoma patients. A number of studies have investigated the prognostic role of SALP level in patients with osteosarcoma but yielded inconsistent results. Method. Systematic computerized searches were performed in PubMed, Embase, and Web of Science databases for relevant original articles. The pooled hazard ratios (HRs and relative risks (RRs with corresponding confidence intervals (CIs were calculated to assess the prognostic value of SALP level. Results. Finally, 21 studies comprising 3228 patients were included. Overall, the pooled HRs of SALP suggested that elevated level had an unfavorable impact on osteosarcoma patients’ overall survival (OS (HR = 1.82; 95% CI: 1.61–2.06; p<0.001 and event-free survival (EFS (HR = 1.97; 95% CI: 1.61–2.42; p<0.001. Combined RRs of SALP indicated that elevated level was associated with presence of metastasis at diagnosis (RR = 5.55; 95% CI: 1.61–9.49; p=0.006. No significantly different results were obtained after stratified by variables of age range, cancer stage, sample size, and geographic region. Conclusion. This meta-analysis demonstrated that high SALP level is significantly associated with poor OS or EFS rate and presence of metastasis at diagnosis. SALP level is a convenient and effective biomarker of prognosis for osteosarcoma.

  8. Albumin-to-Alkaline Phosphatase Ratio: A Novel Prognostic Index for Hepatocellular Carcinoma

    Science.gov (United States)

    Chan, Anthony W. H.; Mo, Frankie K. F.; Wong, Grace L. H.; Wong, Vincent W. S.; Cheung, Yue-Sun; Chan, Henry L. Y.; Yeo, Winnie; Lai, Paul B. S.; To, Ka-Fai

    2015-01-01

    Prognosis of patients with hepatocellular carcinoma (HCC) depends on both tumour extent and hepatic function reserve. Liver function test (LFT) is a basic routine blood test to evaluate hepatic function. We first analysed LFT components and their associated scores in a training cohort of 217 patients who underwent curative surgery to identify LFT parameters with high performance (discriminatory capacity, homogeneity, and monotonicity of gradient). We derived a novel index, albumin-to-alkaline phosphatase ratio (AAPR), which had the highest c-index (0.646) and χ2 (24.774) among other liver biochemical parameters. The AAPR was an independent prognostic factor for overall and disease-free survival. The adjusted hazard ratio of death and tumour relapse was 2.36 (P = 0.002) and 1.85 (P = 0.010), respectively. The independent prognostic significance of AAPR on top of 5 commonly used and well established staging systems was further confirmed in 2 independent cohorts of patients receiving surgical resection (n = 256) and palliative therapy (n = 425). In summary, the AAPR is a novel index readily derived from a simple low-cost routine blood test and is an independent prognostic indicator for patients with HCC regardless of treatment options. PMID:25737613

  9. Albumin-to-Alkaline Phosphatase Ratio: A Novel Prognostic Index for Hepatocellular Carcinoma

    Directory of Open Access Journals (Sweden)

    Anthony W. H. Chan

    2015-01-01

    Full Text Available Prognosis of patients with hepatocellular carcinoma (HCC depends on both tumour extent and hepatic function reserve. Liver function test (LFT is a basic routine blood test to evaluate hepatic function. We first analysed LFT components and their associated scores in a training cohort of 217 patients who underwent curative surgery to identify LFT parameters with high performance (discriminatory capacity, homogeneity, and monotonicity of gradient. We derived a novel index, albumin-to-alkaline phosphatase ratio (AAPR, which had the highest c-index (0.646 and χ2 (24.774 among other liver biochemical parameters. The AAPR was an independent prognostic factor for overall and disease-free survival. The adjusted hazard ratio of death and tumour relapse was 2.36 (P=0.002 and 1.85 (P=0.010, respectively. The independent prognostic significance of AAPR on top of 5 commonly used and well established staging systems was further confirmed in 2 independent cohorts of patients receiving surgical resection (n=256 and palliative therapy (n=425. In summary, the AAPR is a novel index readily derived from a simple low-cost routine blood test and is an independent prognostic indicator for patients with HCC regardless of treatment options.

  10. Substrate and Transition State Binding in Alkaline Phosphatase Analyzed by Computation of Oxygen Isotope Effects.

    Science.gov (United States)

    Roston, Daniel; Cui, Qiang

    2016-09-14

    Enzymes are powerful catalysts, and a thorough understanding of the sources of their catalytic power will facilitate many medical and industrial applications. Here we have studied the catalytic mechanism of alkaline phosphatase (AP), which is one of the most catalytically proficient enzymes known. We have used quantum mechanics calculations and hybrid quantum mechanics/molecular mechanics (QM/MM) simulations to model a variety of isotope effects relevant to the reaction of AP. We have calculated equilibrium isotope effects (EIEs), binding isotope effects (BIEs), and kinetic isotope effects (KIEs) for a range of phosphate mono- and diester substrates. The results agree well with experimental values, but the model for the reaction's transition state (TS) differs from the original interpretation of those experiments. Our model indicates that isotope effects on binding make important contributions to measured KIEs on V/K, which complicated interpretation of the measured values. Our results provide a detailed interpretation of the measured isotope effects and make predictions that can test the proposed model. The model indicates that the substrate is deformed in the ground state (GS) of the reaction and partially resembles the TS. The highly preorganized active site preferentially binds conformations that resemble the TS and not the GS, which induces the substrate to adapt to the enzyme, rather than the other way around-as with classic "induced fit" models. The preferential stabilization of the TS over the GS is what lowers the barrier to the chemical step.

  11. New, improved lanthanide-based methods for the ultrastructural localization of acid and alkaline phosphatase activity.

    Science.gov (United States)

    Halbhuber, K J; Zimmermann, N; Linss, W

    1988-01-01

    New, improved techniques for the ultrastructural localization of acid and alkaline phosphatase activity using lanthanide cations as the trapping agent were developed. Delayed penetration of the capture ions and the incubation constituents into cellular compartments was prevented by pretreating specimens with borohydride/saponin. Both the concentration of the capture agent in the incubation medium and the incubation time of the tissue specimens were optimized to achieve a satisfactory cytochemical reaction and to avoid precipitation artefacts caused by local matrix effects. The conversion of cerium phosphate into the almost insoluble cerium fluoride minimized losses of the reaction product during postincubation processing. Moreover, lanthanum itself as well as lanthanides other than cerium, e.g., gadolinium and didymium (praseodymium, neodymium), were successfully applied and can be recommended as capture agents for phosphatase cytochemistry.

  12. Host plant effects on alkaline phosphatase activity in the whiteflies, Bemisia tabaci Biotype B and Trialeurodes vaporariorum.

    Science.gov (United States)

    Yan, Ying; Peng, Lu; Liu, Wan-Xue; Wan, Fang-Hao; Harris, Marvin K

    2011-01-01

    Bemisia tabaci (Gennadius) B-biotype and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) often coexist on greenhouse-grown vegetable crops in northern China. The recent spread of B. tabaci B-biotype has largely replaced T. vaporariorum, and B-biotype now overlaps with T. vaporariorum where common hosts occur in most invaded areas. The impact of the B-biotype on the agro eco system appears to be widespread, and involves the ability to compete with and perhaps replace other phytophages like T. vaporariorum. An emerging hypothesis is that the B-biotype is physiologically superior due at least in part to an improved ability to metabolically utilize the alkaline phosphatase pathway. To test this hypothesis, alkaline phosphatase activity was studied in the B-biotype and T. vaporariorum after feeding on a number of different hosts for a range of durations, with and without host switching. Alkaline phosphatase activity in T. vaporariorum was 1.45 to 2.53-fold higher than that of the B-biotype when fed on tomato for 4 and 24 h, or switched from tomato to cotton and cabbage for the same durations. However, alkaline phosphatase activity in the B-biotype was 1.40 to 3.35-fold higher than that of T. vaporariorum when the host switching time was ∼72 and ∼120 h on the same plant. Both short-term (4 h) and long-term (72 h) switching of plant hosts can significantly affect the alkaline phosphatase activity in the two species. After ∼120 h, feeding on tomato and cotton alkaline phosphatase activity in the B-biotype was significantly higher than that of T. vaporariorum. It was shown that alkaline phosphatase aids the species feeding on different plant species, and that the B-biotype is physiologically superior to T. vaporariorum in utilizing the enzyme compared to T. vaporariorum over longer periods of feeding.

  13. Alkaline phosphatase in nasal secretion of cattle: biochemical and molecular characterisation.

    Science.gov (United States)

    Ghazali, M Faizal; Koh-Tan, H H Caline; McLaughlin, Mark; Montague, Paul; Jonsson, Nicholas N; Eckersall, P David

    2014-09-05

    Nasal secretion (NS) was investigated as a source of information regarding the mucosal and systemic immune status of cattle challenged by respiratory disease. A method for the collection of substantial volumes (~12 ml) of NS from cattle was developed to establish a reference range of analytes that are present in the NS of healthy cattle. Biochemical profiles of NS from a group of 38 healthy Holstein-Friesian cows revealed high alkaline phosphatase (AP) activity of up to 2392 IU/L. The character and source of the high activity of AP in bovine NS was investigated. Histochemical analysis confirmed the localization of the AP enzyme activity to epithelial cells and serous glands of the nasal respiratory mucosa. Analysis of mRNA levels from nasal mucosa by end point RT-PCR and PCR product sequencing confirmed that the AP was locally produced and is identical at the nucleotide level to the non-specific AP splice variant found in bovine liver, bone and kidney. Analysis by isoelectric focussing confirmed that AP was produced locally at a high level in nasal epithelium demonstrating that AP from nasal secretion and nasal mucosa had similar pI bands, though differing from those of the liver, kidney, bone and intestine, suggesting different post-translational modification (PTM) of AP in these tissues. A nasal isozyme of AP has been identified that is present at a high activity in NS, resulting from local production and showing distinctive PTM and may be active in NS as an anti-endotoxin mediator.

  14. Intestinal alkaline phosphatase: a summary of its role in clinical disease.

    Science.gov (United States)

    Fawley, Jason; Gourlay, David M

    2016-05-01

    Over the past few years, there is increasing evidence implicating a novel role for Intestinal Alkaline Phosphatase (IAP) in mitigating inflammatory mediated disorders. IAP is an endogenous protein expressed by the intestinal epithelium that is believed to play a vital role in maintaining gut homeostasis. Loss of IAP expression or function is associated with increased intestinal inflammation, dysbiosis, bacterial translocation and subsequently systemic inflammation. As these events are a cornerstone of the pathophysiology of many diseases relevant to surgeons, we sought to review recent research in both animal and humans on IAP's physiologic function, mechanisms of action and current research in specific surgical diseases.

  15. Comparison of Inactivation and Unfolding of Calf Intestinal Alkaline Phosphatase in Guanidinium Chloride Solution

    Institute of Scientific and Technical Information of China (English)

    张英侠; 闫淑莲; 刘永利; 席宏伟; 周海梦

    2002-01-01

    The changes in activity and unfolding of calf intestinal alkaline phosphatase (CIP) during denaturation in guanidinium chloride solutions of different concentrations were investigated using ultraviolet difference absorption spectra and fluorescence emission spectra. Unfolding and inactivation rate constants were measured and compared. The inactivation course is much faster than that of unfolding, which suggests that the active site of CIP containing two zinc ions and one magnesium ion is situated in a limited and flexible region of the enzyme molecule, which is more fragile to the denaturant than the protein as a whole.

  16. [Effect of dental alloys on salivary alkaline and acid phosphatase, alpha amylase K+, Na+, and Cl-].

    Science.gov (United States)

    Todorov, I; Saprjanova, M

    1977-04-01

    Comparative studied were performed in healthy subjects without metals in their oral cavities and in individuals having different metal alloys (gold, steel, amalgam) in their mouths and presenting with various complaints such as xerostomia, burning mucosa, etc. It was found that the contents of alkaline and acid phosphatases, alpha-amylase, K+, Na+ and Cl- in saliva increased significantly with the increase in total corrosion potential when non-precious metal alloys, especially different types of alloys, were present. Parallel to this, the frequency and the intensity of the complaints increased.

  17. Effect of Diazinon on Acid and Alkaline Phosphatase Activities in Plasma and Organs of Clarias gariepinus

    OpenAIRE

    I.R. Inyang; E.R. Daka and E.N. Ogamba

    2011-01-01

    The aim of this study was to determine the effect of the pesticide, diazinon, on phosphatases in the plasma and organs on Clarias gariepinus. Adult Clarias gariepinus were exposed in four replicates to varying sublethal concentrations diazinon (ranging from 1.00 to 10.0 mg/L) in 30-day semi-static bioassays. Alkaline phoshatase (ALP) and acid phosphate (ACP) were determined in plasma and other organs (gastrointestinal tract - GIT, kidney, muscle, gill and liver) of the fish after the experime...

  18. A New Voltammetric Enzyme Immunoassay System for the Detection of Alkaline Phosphatase

    Institute of Scientific and Technical Information of China (English)

    KuiJIAO; WeiSUN; 等

    2002-01-01

    A new voltammetric enzyme immunoassay system was invesigated based on p-nitrophenyl phosphate (PNPP) as the subsrate for alkaline phosphatase (ALP). PNPP is enzymatically hydrolyzed and the product p-nitrophenol (PNP) is detected by differential pulse voltammetry (DPV), which can be oxidized at +1.02 V(vs.Ag/AgCl) on bare glass carbon electrode (GCE). The conditions for enzymatic reaction and electrochemical detection were studied. According to this method,ALP can be detected with a detection limit of 2.8×102 mU/L and a linear range of 4.0×102-1.0×106mU/L.

  19. A New Voltammetric Enzyme Immunoassay System for the Detection of Alkaline Phosphatase

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A new voltammetric enzyme immunoassay system was investigated based on p-nitrophenyl phosphate (PNPP) as the substrate for alkaline phosphatase (ALP). PNPP is enzymatically hydrolyzed and the product p-nitrophenol (PNP) is detected by differential pulse voltammetry (DPV), which can be oxidized at +1.02 V (vs. Ag/AgCl) on bare glass carbon electrode (GCE). The conditions for enzymatic reaction and electrochemical detection were studied. According to this method, ALP can be detected with a detection limit of 2.8′102 mU/L and a linear range of 4.0′102 ~ 1.0′106 mU/L.

  20. Distribution of alkaline and acid phosphatases in the duodenal wall of native blackgoats by using different fixatives

    Directory of Open Access Journals (Sweden)

    N. S. Ahmed

    2010-01-01

    Full Text Available Ten duodeni of adult goat were fixed in chilled acetone, 80% ethyl alcohol, alcohol-formalin solution, alcohol bouinssolution and buffered neutral formalin solution. The distribution of alkaline and acid phosphatases noticed in absorptive andgoblet cells that lining the duodenal mucosa of black goat, but different in their intensity and distribution according to differentfixatives. The distribution of alkaline phosphatase in absorptive columnar cells that lining intestinal glands was more intensethan other cells, whereas the concentration of acid phosphatase was more intense in goblet cells than other cells in the mucosaof goat duodenum specially in samples fixed in chilled acetone and ethyl alcohol 80%. The study revealed that the sampleswere fixed with chilled acetone gave highest reaction for alkaline and acid phosphatases than other fixative samples. Noreaction for alkaline and acid phosphatases included some absorptive cells lining villi, all cells lining the lower parts ofintestinal glands, paneth cells and submucosal glands in different fixatives, except submucosal glands revealed positivereaction for acid phosphatase in samples fixed in chilled acetone and 80% ethyl alcohol, paneth cells reveal positive reaction for the same enzyme in samples fixed in 80% ethyl alcohol in all examined areas of the duodenum wall of the native blackgoat.

  1. Salivary Alkaline Phosphatase as a Noninvasive Marker for Periodontal Disease in Children with Uncontrolled Type 1 Diabetes Mellitus.

    Science.gov (United States)

    Sridharan, Srirangarajan; Sravani, Paruchuri; Satyanarayan, Aparna; Kiran, K; Shetty, Varun

    The aim of this pilot study was to determine whether salivary alkaline phosphatase levels can be a non invasive marker for early inflammatory periodontal disease in children with uncontrolled type 1 diabetes mellitus. 10 healthy children (group 1), 10 children with recently diagnosed type 1 diabetes mellitus (group 2) and 10 children with type 1 diabetes mellitus for more than 4 years (group 3) were recruited for the study. All three groups were matched for age, gender and socioeconomic status. Periodontal health was assessed by plaque index, gingival index and probing pocket depth. Metabolic status was assessed by glycosylated hemoglobin levels, salivary alkaline phosphatase levels were determined by spectrophotometer. Data was analyzed by Kruskal Wallis ANOVA, Mann-Whitney U test and Spearman's rank correlation method. Salivary alkaline phosphatase levels correlated significantly with the periodontal parameters in the diabetic group. An increase in salivary alkaline phosphatase levels increased with increased values of gingival index and probing pocket depth. Group 3 showed greater correlation than group 2 and group 1. At p value pperiodontal disease parameters. Salivary alkaline phosphatase levels could be a useful tool in analyzing periodontal status of children with uncontrolled type I diabetes mellitus.

  2. Alkaline phosphatase activity and the phosphorus mineralization rate of Lake Taihu

    Institute of Scientific and Technical Information of China (English)

    GAO; Guang; ZHU; Guangwei; QIN; Boqiang; CHEN; Jun

    2006-01-01

    The phosphorus fractions, the alkaline phosphatase activity (APA) and other water chemical parameters were concomitantly monitored from April 2003 to October 2004 in different ecotype sites of Lake Taihu. During the stages of algae growth, the phosphorus fractions and their relationships with APA in different ecotype sites were discussed and the phosphorus mineralization rate was calculated. In the water of Lake Taihu, most of the phosphorus (70.2%) could be attributed to the suspended particulate phosphorus, while the dissolved reactive phosphorus (DRP) seems to contribute less than 7%. About 58% of the total phosphorus, however, can be hydrolyzed as inorganic phosphate to compensate for phosphorus deficiency of algae and bacteria growth. During the different algae growth stages, the APA and its Kinetic parameters were varied significantly between different ecotype sites of Lake Taihu. This trend is also visible by comparing the phosphorus mineralization rate,and the most rapidly phosphorus turnover time is only several minutes. The fast recycle of phosphorus can, to some extent, be explained that the phosphorus source of algal blooms. The phytoplankton seems to compensate for phosphorus deficiency by using the alkaline phosphatase to hydrolyze phosphomonoesters.

  3. Changes in plasma inorganic phosphorus and alkaline phosphatase activity during the adolescent growth spurt.

    Science.gov (United States)

    Round, J M; Butcher, S; Steele, R

    1979-01-01

    Longitudinal data on changes in the concentrations of plasma inorganic phosphorus and plasma alkaline phosphatase activity in 23 girls and 44 boys during the adolescent growth spurt are reported together with the height velocities, ages and sexual maturity ratings. The average age at the peak of the growth spurt was 12.5 years in the girls and 14.1 years in the boys with mean annual height gains of 7.0 and 9.7 cm/year respectively. In both sexes, plasma alkaline phosphatase activity rose and fell with the growth velocity during the growth spurt. Plasma inorganic phosphorus rose to reach a peak in the 4 months before the peak of the growth spurt in height; this rise was statisically significant in the boys but not in the girls. Values subsequently fell rapidly towards the normal adult concentrations. Plasma calcium, total protein, and albumin concentrations were also followed during this time, but were not at any point significantly different from normal adult values. These findings provide a guide for the interpretation of plasma biochemistry in adolescent patients.

  4. Ultrastructural localization of alkaline phosphatase in the eggs of Hydatigera taeniaeformis (Taenia taeniaeformis).

    Science.gov (United States)

    Ajayi, S T; Smith, B F; LeFlore, W B

    1985-01-01

    Freshly shed gravid proglottids from a three-month-old infection of Hydatigera taeniaeformis collected from the faecal droppings of infected cats were used for this study. They were treated for transmission electron microscopy (TEM) followed by incubation using the lead precipitate method. Control sections were incubated in a substrate-free medium, a substrate medium containing 1.0 mM sodium fluoride (NaF) (an inhibitor), and the last sections were denatured at 90 degrees C for 1 min prior to incubation. Intensive alkaline phosphatase activity in the embryophoric blocks and the outer embryophoric membrane was revealed. The reaction products were also indicated in the oncospheral membrane. However, no enzyme activity was seen in any other part of the egg. The enzyme was also absent in the control sections. The presence of alkaline phosphatase activity in the outer embryophoric and oncospheral membranes suggested that this enzyme may be involved in carbohydrate metabolism and nutritional absorption, and also may play a role in the transport of nutrients and other substances from the adult to the developing embryo, respectively.

  5. Prevention of antibiotic-associated metabolic syndrome in mice by intestinal alkaline phosphatase.

    Science.gov (United States)

    Economopoulos, K P; Ward, N L; Phillips, C D; Teshager, A; Patel, P; Mohamed, M M; Hakimian, S; Cox, S B; Ahmed, R; Moaven, O; Kaliannan, K; Alam, S N; Haller, J F; Goldstein, A M; Bhan, A K; Malo, M S; Hodin, R A

    2016-05-01

    To examine whether co-administration of intestinal alkaline phosphatase (IAP) with antibiotics early in life may have a preventive role against metabolic syndrome (MetS) in mice. A total of 50 mice were allocated to four treatment groups after weaning. Mice were treated with azithromycin (AZT) ± IAP, or with no AZT ± IAP, for three intermittent 7-day cycles. After the last treatment course, the mice were administered a regular chow diet for 5 weeks and subsequently a high-fat diet for 5 weeks. Body weight, food intake, water intake, serum lipids, glucose levels and liver lipids were compared. 16S rRNA gene pyrosequencing was used to determine the differences in microbiome composition. Exposure to AZT early in life rendered mice susceptible to MetS in adulthood. Co-administration of IAP with AZT completely prevented this susceptibility by decreasing total body weight, serum lipids, glucose levels and liver lipids to the levels of control mice. These effects of IAP probably occur as a result of changes in the composition of specific bacterial taxa at the genus and species levels (e.g. members of Anaeroplasma and Parabacteroides). Co-administration of IAP with AZT early in life prevents mice from susceptibility to the later development of MetS. This effect is associated with alterations in the composition of the gut microbiota. IAP may represent a novel treatment against MetS in humans. © 2016 John Wiley & Sons Ltd.

  6. Role of lysophosphatidylcholine in brush-border intestinal alkaline phosphatase release and restoration.

    Science.gov (United States)

    Nakano, Takanari; Inoue, Ikuo; Alpers, David H; Akiba, Yasutada; Katayama, Shigehiro; Shinozaki, Rina; Kaunitz, Jonathan D; Ohshima, Susumu; Akita, Masumi; Takahashi, Seiichiro; Koyama, Iwao; Matsushita, Makoto; Komoda, Tsugikazu

    2009-07-01

    Intestinal alkaline phosphatase (IAP) is a brush-border membrane ectoenzyme (BBM-IAP) that is released into the lumen (L-IAP) after a high-fat diet. We examined the effects of oil feeding and the addition of mixed-lipid micelles on the formation of L-IAP in oil-fed rat intestine, Caco-2 cell monolayers, and mouse intestinal loops. We localized IAP in the duodenum of rats fed corn oil using fluorescence microscopy with enzyme-labeled fluorescence-97 as substrate. Four hours after oil feeding, L-IAP increased approximately 10-fold accompanied by the loss of BBM-IAP, consistent with BBM-IAP release. Rat IAP isozyme mRNAs progressively increased 4-6 h after oil feeding, followed by the increase of IAP activity in the subapical location at 6 h, consistent with the restoration of IAP protein. Postprandial lipid-micelle components, sodium taurocholate with or without oleic acid, mono-oleylglycerol, cholesterol, or lysophosphatidylcholine (lysoPC) were applied singly or as mixed-lipid micelles to the apical surface of polarized Caco-2 cell monolayers. LysoPC increased L-IAP >10-fold over basal release. LysoPC released IAP into the apical medium more than other intestinal brush-border enzymes, 5'-nucleotidase, sucrase, aminopeptidase N, and lactase, without comparable lactate dehydrogenase release or cell injury. LysoPC increased human IAP mRNA levels by 1.5-fold in Caco-2 cells. Luminally applied lysoPC also increased release of IAP preferentially in mouse intestinal loops. These data show that lysoPC accelerates the formation of L-IAP from BBM-IAP, followed by enhanced IAP synthesis, suggesting the role that lysoPC might play in the turnover of brush-border proteins.

  7. Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation

    Directory of Open Access Journals (Sweden)

    Ciancaglini P.

    2006-01-01

    Full Text Available Endochondral calcification involves the participation of matrix vesicles (MVs, but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP during mineralization involves hydrolysis of inorganic pyrophosphate (PPi, it is unclear how the microenvironment of MV may affect the ability of TNAP to hydrolyze the variety of substrates present at sites of mineralization. We show that the implants contain high levels of TNAP capable of hydrolyzing p-nitrophenylphosphate (pNPP, ATP and PPi. The catalytic properties of glycosyl phosphatidylinositol-anchored, polidocanol-solubilized and phosphatidylinositol-specific phospholipase C-released TNAP were compared using pNPP, ATP and PPi as substrates. While the enzymatic efficiency (k cat/Km remained comparable between polidocanol-solubilized and membrane-bound TNAP for all three substrates, the k cat/Km for the phosphatidylinositol-specific phospholipase C-solubilized enzyme increased approximately 108-, 56-, and 556-fold for pNPP, ATP and PPi, respectively, compared to the membrane-bound enzyme. Our data are consistent with the involvement of MVs during ectopic calcification and also suggest that the location of TNAP on the membrane of MVs may play a role in determining substrate selectivity in this micro-compartment.

  8. Endotoxin- and ATP-neutralizing activity of alkaline phosphatase as a strategy to limit neuroinflammation

    Directory of Open Access Journals (Sweden)

    Huizinga Ruth

    2012-12-01

    Full Text Available Abstract Background Alkaline phosphatase (AP is a ubiquitously expressed enzyme which can neutralize endotoxin as well as adenosine triphosphate (ATP, an endogenous danger signal released during brain injury. In this study we assessed a potential therapeutic role for AP in inhibiting neuroinflammation using three complementary approaches. Methods Mice were immunized to induce experimental autoimmune encephalomyelitis (EAE and treated with AP for seven days during different phases of disease. In addition, serological assays to determine AP activity, endotoxin levels and endotoxin-reactive antibodies were performed in a cohort of multiple sclerosis (MS patients and controls. Finally, the expression of AP and related enzymes CD39 and CD73 was investigated in brain tissue from MS patients and control subjects. Results AP administration during the priming phase, but not during later stages, of EAE significantly reduced neurological signs. This was accompanied by reduced proliferation of splenocytes to the immunogen, myelin oligodendrocyte glycoprotein peptide. In MS patients, AP activity and isoenzyme distribution were similar to controls. Although endotoxin-reactive IgM was reduced in primary-progressive MS patients, plasma endotoxin levels were not different between groups. Finally, unlike AP and CD73, CD39 was highly upregulated on microglia in white matter lesions of patients with MS. Conclusions Our findings demonstrate that: 1 pre-symptomatic AP treatment reduces neurological signs of EAE; 2 MS patients do not have altered circulating levels of AP or endotoxin; and 3 the expression of the AP-like enzyme CD39 is increased on microglia in white matter lesions of MS patients.

  9. Enhanced cell adhesion on bioinert ceramics mediated by the osteogenic cell membrane enzyme alkaline phosphatase.

    Science.gov (United States)

    Aminian, Alieh; Shirzadi, Bahareh; Azizi, Zahra; Maedler, Kathrin; Volkmann, Eike; Hildebrand, Nils; Maas, Michael; Treccani, Laura; Rezwan, Kurosch

    2016-12-01

    Functional bone and dental implant materials are required to guide cell response, offering cues that provide specific instructions to cells at the implant/tissue interface while maintaining full biocompatibility as well as the desired structural requirements and functions. In this work we investigate the influence of covalently immobilized alkaline phosphatase (ALP), an enzyme involved in bone mineralization, on the first contact and initial cell adhesion. To this end, ALP is covalently immobilized by carbodiimide-mediated chemoligation on two highly bioinert ceramics, alpha-alumina (Al2O3) and yttria-stabilized zirconia (Y-TZP) that are well-established for load-bearing applications. The physicochemical surface properties are evaluated by profilometry, zeta potential and water contact angle measurements. The initial cell adhesion of human osteoblasts (HOBs), human osteoblast-like cells (MG-63) and mesenchymal stromal cells (hMSCs) was investigated. Cell adhesion was assessed at serum free condition via quantification of percentage of adherent cells, adhesion area and staining of the focal adhesion protein vinculin. Our findings show that after ALP immobilization, the Al2O3 and Y-TZP surfaces gained a negative charge and their hydrophilicity was increased. In the presence of surface-immobilized ALP, a higher cell adhesion, more pronounced cell spreading and a higher number of focal contact points were found. Thereby, this work gives evidence that surface functionalization with ALP can be utilized to modify inert materials for biological conversion and faster bone regeneration on inert and potentially load-bearing implant materials. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Interactive effects of temperature, ultraviolet radiation and food quality on zooplankton alkaline phosphatase activity.

    Science.gov (United States)

    Wolinski, Laura; Modenutti, Beatriz; Souza, Maria Sol; Balseiro, Esteban

    2016-06-01

    Ultraviolet Radiation (UVR) is a stressor for aquatic organisms affecting enzyme activities in planktonic populations because of the increase in reactive oxygen species. In addition, UVR exposure combined with other environmental factors (i.e. temperature and food quality) could have even higher detrimental effects. In this work, we aimed to determine the effect of UVR on somatic Alkaline Phosphatase Activity (APA) and Glutathione S-Transferase (GST) activity on the cladoceran Daphnia commutata under two different temperatures (10 °C and 20 °C) and under three food qualities (carbon:phosphorus ratios: 1150, 850 and 550). APA is a biomarker that is considered as a P deficiency indicator in zooplankton. Since recovery from UVR damage under dark conditions is an ATP depending reaction we also measured APA during recovery phases. We carried out a laboratory experiment combining different temperatures and food qualities with exposition to UVR followed by luminic and dark phases for recovery. In addition, we exposed organisms to H2O2, to establish if the response on APA to UVR was a consequence of the reactive oxygen species produced these short wavelengths. Our results showed that somatic APA was negatively affected by UVR exposure and this effect was enhanced under high temperature and low food quality. Consistently, GST activity was higher when exposed to UVR under both temperatures. The H2O2 experiments showed the same trend as UVR exposure, indicating that APA is affected mainly by oxidative stress than by direct effect of UVR on the enzyme. Finally, APA was affected in the dark phase of recovery confirming the P demands. These results enlighten the importance of food quality in the interacting effect of UVR and temperature, showing that C:P food ratio could determine the success or failure of zooplanktonic populations in a context of global change.

  11. Influence of bone and soft-tissue operations on serum concentrations of growth hormone, somatomedin C and alkaline phosphatase.

    Science.gov (United States)

    Casser, H R; Zilkens, K W; Forst, R; Brüggemann, A

    1990-01-01

    After animal experiments suggested there was an interaction between growth hormone and bone healing, our aim in this paper was to ascertain whether there were any changes or possible interaction between the serum level of growth hormone, somatomedin C and alkaline phosphatase while a fractured bone was healing. To this end, the serum concentrations of growth hormone, somatomedin C, alkaline phosphatase and calcium were ascertained both pre- and post-operatively in two groups of patients--one with bone operations, the other with soft-tissue operations--and the results were compared. Comparing the groups, we found that after bone operations there was no increase in the serum level of growth hormone, nor of somatomedin C. An increase would have implied that these two hormones are directly involved in bone regeneration. There was no change in the serum level of alkaline phosphatase or calcium after either bone or soft-tissue operations.

  12. Transferability studies for the AACC reference method and the IFCC method for measurement of alkaline phosphatase activity.

    Science.gov (United States)

    Tietz, N W; Rinker, A D; Burtis, C; Duncan, P; Ervin, K; Ewen, L; Hørder, M; Mathieu, M; Petitclerc, C; Grisley, D

    1984-05-01

    We present the results of measurements of alkaline phosphatase activity from interlaboratory transferability studies conducted by 12 laboratories in five countries. The variability, as demonstrated by within-day precision (CV less than or equal to 2.6%), between-day precision (CV less than or equal to 3.6%), and between-lab precision (CV less than or equal to 6.3%) establishes the transferability of this method for alkaline phosphatase. Some common errors encountered in enzyme measurements that affect the accuracy and precision of the measurements are listed.

  13. Purification and characterization of an alkaline phosphatase induced by phosphorus starvation in common bean (Phaseolus vulgaris L.) roots

    Energy Technology Data Exchange (ETDEWEB)

    Morales, L.; Gutierrez, N.; Maya, V.; Parra, C.; Martinez B, E.; Coello, P., E-mail: pcoello@servidor.unam.mx [UNAM, Facultad de Quimica, Departamento de Bioquimica, Ciudad Universitaria, 04510 Mexico D. F. (Mexico)

    2012-07-01

    Two phosphatase isoforms from roots of the common bean (Phaseolus vulgaris L.) showed an increase in activity in response to phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass of APIII was 35 kDa by both SDS-Page and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of ph 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phospho enol-pyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyro phosphatase activity of this enzyme, the hydrolysis of pyro phosphatase increased substantially in the presence of Mg{sup 2+}.

  14. Amino acid sequence of the cold-active alkaline phosphatase from Atlantic cod (Gadus morhua)

    DEFF Research Database (Denmark)

    Asgeirsson, Bjarni; Nielsen, Berit Noesgaard; Højrup, Peter

    2003-01-01

    Atlantic cod is a marine fish that lives at low temperatures of 0-10 degrees C and contains a cold-adapted alkaline phosphatase (AP). Preparations of AP from either the lower part of the intestines or the pyloric caeca area were subjected to proteolytic digestion, mass spectrometry and amino acid...... has the same variable residues as mammalian APs (His153 and His328 by E. coli AP numbering). General comparison of the amino acid composition with mammalian APs showed that cod AP contains fewer Cys, Leu, Met and Ser, but proportionally more Asn, Asp, Ile, Lys, Trp and Tyr residues. Three N......-linked glycosylation sites were found. The glycan structure was determined as complex biantennary in type with fucose and sialic acid attached, although a trace of complex tri-antennary structure was also observed. A three-dimensional model was obtained by homology modelling using the human placental AP scaffold. Cod...

  15. Chronic Cadmium Exposure Lead to Inhibition of Serum and Hepatic Alkaline Phosphatase Activity in Wistar Rats.

    Science.gov (United States)

    Treviño, Samuel; Andrade-García, Alejandra; Herrera Camacho, Irma; León-Chavez, Bertha Alicia; Aguilar-Alonso, Patricia; Flores, Gonzalo; Brambila, Eduardo

    2015-12-01

    Alkaline phosphatase (ALP) activity in the serum and liver from rats administered with cadmium (Cd) in drinking water was studied. After metal administration, Cd showed a time-dependent accumulation in the liver, meanwhile metallothionein had a maximum increase at 1 month, remaining in this level until the end of the study. On the other hand, serum and liver ALP activity was decreased after 3 months exposure. To determine if Cd produced an inhibition on enzyme, apo-ALP prepared from both nonexposed and exposed rats was reactivated with Zn, showing 60% more activity as compared with the enzyme isolated from nonexposed rats. In vitro assays showed that Cd-ALP was partially reactivated with Zn; however, in the presence of cadmium, Zn-ALP was completely inhibited. Kinetic studies indicate a noncompetitive inhibition by Cd; these results suggest that Cd can substitute Zn, and/or Cd can interact with nucleophilic ligands essential for the enzymatic activity.

  16. A correlational study of serum alkaline phosphatase level and developmental mandibular laterognathism.

    Science.gov (United States)

    Dai, Jiewen; Li, Hongliang; Yu, Hongbo; Si, Jiawen; Fang, Bing; Shen, Steve Guofang

    2015-03-01

    The abnormal cartilage/bone metabolism in unilateral condyle may be a direct factor that contributes to developmental mandibular laterognathism. However, although many molecules have been demonstrated to play crucial roles in the development of temporomandibular joints, the exact molecular mechanisms that lead to the disrupted condylar cartilage/bone development were greatly unknown. In this retrospective study, our findings revealed that serum alkaline phosphatase (ALP) level in adult patients with developmental mandibular laterognathism was lower than that in control subjects, and the serum ALP levels continue to reduce in adult patients (>20 years old). Although the exact relationship between the lower serum ALP level and developmental mandibular laterognathism is unclear, the findings further support the opinion that the condylar growth may sustain for a long time in the affected condyle in patients with developmental mandibular laterognathism and offer an alternative choice to use total serum ALP activity as a possible biomarker to assess condylar growth activity in patients with developmental mandibular laterognathism.

  17. Fluoride stimulates ( sup 3 H)thymidine incorporation and alkaline phosphatase production by human osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Khokher, M.A.; Dandona, P. (Royal Free Hospital, London (England))

    1990-11-01

    The effect of sodium fluoride on alkaline phosphatase (ALP) release and ({sup 3}H)thymidine uptake by human osteoblasts in culture was investigated. Sodium fluoride stimulated both ALP release and ({sup 3}H)thymidine uptake at concentrations of sodium fluoride greater than 250 mumol/L. This stimulation was similar in magnitude to that induced by 1,25-dihydroxycholecalciferol. The fluoride-induced increase in ALP was inhibited by verapamil, a calcium channel blocker. We conclude that sodium fluoride stimulates osteoblasts to proliferate and to release ALP. This stimulation by fluoride is dependent on calcium influx. Fluoride-induced stimulation of human osteoblasts may be relevant to its effect in enhancing bone formation in patients with osteoporosis.

  18. The Detection of Alkaline Phosphatase Using an Electrochemical Biosensor in a Single-Step Approach

    Directory of Open Access Journals (Sweden)

    Chung-Chiun Liu

    2009-10-01

    Full Text Available A one-step, single use, disposable Alkaline Phosphatase (ALP biosensor has been developed. It is based on the detection of phenol produced by an ALP enzymatic reaction. It can operate at 25 °C in a pH 10 medium. It measures ALP of 0–300 IU/L. The permissible concentrations of glucose, ascorbic acid and urea without interference are 10 mM/L, 5 mg/L and 400 mg/L, respectively. Experimental results are compared to those obtained by spectrophotometric measurements in bovine serum. Excellent linearity between the biosensor outputs and the ALP concentrations exists. The agreement between the measurements of this biosensor and the spectrophotometer is also outstanding.

  19. RELATION BETWEEN ALKALINE PHOSPHATASE IN GINGIGVAL CREVICULAR FLUID OF IMPLANT TEETH AND THE CURING RESULT

    Institute of Scientific and Technical Information of China (English)

    周正; 周坚; 邹石莹; 吴效民

    2001-01-01

    Objective. To discover the relation between alkaline phosphatase (ALP) in gingival crevicular fluid (GCF)of implant teeth aad the curing results.Methods. We measured the ALP level in GCF among 56 cases of implant teeth which included 2 failed cas-es, 5 cases with bad oral hygiene and gingivitis, and compared it with that in the normal group composed of 10persons.Results. The ALP levels in normal group and success implant group showed no difference. The ALP levelsin normal group and success with gingivitis group showed obvious difference. The ALP levels of the 2 failed cas-es are the highest of all.Conclusions. The ALP level in GCF is an important index in evaluating the curing result of the implantteeth.``

  20. Intestinal alkaline phosphatase: selective endocytosis from the enterocyte brush border during fat absorption

    DEFF Research Database (Denmark)

    Hansen, Gert Helge; Niels-Christiansen, Lise-Lotte; Immerdal, Lissi;

    2007-01-01

    Absorption of dietary fat in the small intestine is accompanied by a rise of intestinal alkaline phosphatase (IAP) in the serum and of secretion of IAP-containing surfactant-like particles from the enterocytes. In the present work, fat absorption was studied in organ cultured mouse intestinal...... clathrin-coated pits. By 60 min, IAP was seen in subapical endosomes and along membranes surrounding fat droplets. IAP is a well-known lipid raft-associated protein, and fat absorption was accompanied by a marked change in the density and morphology of the detergent-resistant membranes harboring IAP...... explants. By immunofluorescence microscopy, fat absorption caused a translocation of IAP from the enterocyte brush border to the interior of the cell, whereas other brush-border enzymes were unaffected. By electron microscopy, the translocation occurred by a rapid (5 min) induction of endocytosis via...

  1. Osteocalcin and bone-specific alkaline phosphatase in Asian elephants (Elephas maximus) at different ages.

    Science.gov (United States)

    Arya, Nlin; Moonarmart, Walasinee; Cheewamongkolnimit, Nareerat; Keratikul, Nutcha; Poon-Iam, Sawinee; Routh, Andrew; Bumpenpol, Pitikarn; Angkawanish, Taweepoke

    2015-11-01

    Bone turnover markers could offer a potential alternative means for the early diagnosis of metabolic bone disease in young growing elephants although the baseline of bone turnover markers in elephant is not well established. The aim of this study was to determine any relationship between the age of captive Asian elephants (Elephas maximus) and markers of bone formation. Serum samples from 24 female Asian elephants were collected to evaluate levels of two bone formation markers, namely, osteocalcin (OC) and bone-specific alkaline phosphatase (BAP). Both intact and N-terminal midfragment OC and BAP were negatively correlated with age. The findings demonstrate that younger elephants have a higher rate of bone turnover than older elephants. Use of these and additional bone markers could lead to the establishment of validated protocols for the monitoring of bone disease in elephants.

  2. A Disposable Alkaline Phosphatase-Based Biosensor for Vanadium Chronoamperometric Determination

    Science.gov (United States)

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia

    2014-01-01

    A chronoamperometric method for vanadium ion determination, based on the inhibition of the enzyme alkaline phosphatase, is reported. Screen-printed carbon electrodes modified with gold nanoparticles were used as transducers for the immobilization of the enzyme. The enzymatic activity over 4-nitrophenyl phosphate sodium salt is affected by vanadium ions, which results in a decrease in the chronoamperometric current registered. The developed method has a detection limit of 0.39 ± 0.06 μM, a repeatability of 7.7% (n = 4) and a reproducibility of 8% (n = 3). A study of the possible interferences shows that the presence of Mo(VI), Cr(III), Ca(II) and W(VI), may affect vanadium determination at concentration higher than 1.0 mM. The method was successfully applied to the determination of vanadium in spiked tap water. PMID:24569772

  3. A Disposable Alkaline Phosphatase-Based Biosensor for Vanadium Chronoamperometric Determination

    Directory of Open Access Journals (Sweden)

    Ana Lorena Alvarado-Gámez

    2014-02-01

    Full Text Available A chronoamperometric method for vanadium ion determination, based on the inhibition of the enzyme alkaline phosphatase, is reported. Screen-printed carbon electrodes modified with gold nanoparticles were used as transducers for the immobilization of the enzyme. The enzymatic activity over 4-nitrophenyl phosphate sodium salt is affected by vanadium ions, which results in a decrease in the chronoamperometric current registered. The developed method has a detection limit of 0.39 ± 0.06 µM, a repeatability of 7.7% (n = 4 and a reproducibility of 8% (n = 3. A study of the possible interferences shows that the presence of Mo(VI, Cr(III, Ca(II and W(VI, may affect vanadium determination at concentration higher than 1.0 mM. The method was successfully applied to the determination of vanadium in spiked tap water.

  4. Benomyl inhibits phosphorus transport but not fungal alkaline phosphatase activity in a Glomus–cucumber symbiosis

    DEFF Research Database (Denmark)

    Larsen, John; Thingstrup, Ida; Jakobsen, Iver

    1996-01-01

    Short-term effects of benomyl on the arbuscular mycorrhizal fungus Glomus caledonium (Nicol. & Gerd.) Trappe and Gerdeman associated with Cucumis sativus L. were studied by measuring effects on fungal P transport and on fungal alkaline phosphatase activity. Mycorrhizal plants were grown in three...... compartment systems where nylon mesh was used to separate n root-free hyphal compartment (HC) and a root + hyphal compartment(RHC) from The main root compartment (RC). Non-mycorrhizal control plants were grown in similar growth units. After 6 wk benomyl was applied to the plants in three ways: as soil...... drenches to RHC or HC, or as u spray to the leaves. Benomyl was added in three concentrations. Equal amounts of 32P and 33P were added to the HC and to the RHC respectively, immediately after the application of benomyl. Plants were harvested 4–6 d later. Hyphal transport of 32P from the HC was inhibited...

  5. A reference method for measurement of alkaline phosphatase activity in human serum.

    Science.gov (United States)

    Tietz, N W; Burtis, C A; Duncan, P; Ervin, K; Petitclerc, C J; Rinker, A D; Shuey, D; Zygowicz, E R

    1983-05-01

    We present an official AACC reference method for the measurement of alkaline phosphatase, the culmination of optimization experiments conducted by a group of independent laboratories. The details of this method and evaluation of factors affecting the measurement are described. A metal ion buffer has been incorporated that maintains optimal and constant concentrations of zinc(II) and magnesium(II) ions. Final reaction conditions are: pH (30 degrees C), 10.40 +/- 0.05; 2-amino-2-methyl-1-propanol buffer, 0.35 mol/L; 4-nitrophenyl phosphate, 16.0 mmol/L; magnesium acetate, 2.0 mmol/L; zinc sulfate, 1.0 mmol/L; and N-(2-hydroxyethyl)ethylenediaminetriacetic acid, 2.0 mmol/L.

  6. Multiple unfolding intermediates of human placental alkaline phosphatase in equilibrium urea denaturation.

    Science.gov (United States)

    Hung, H C; Chang, G G

    2001-12-01

    Alkaline phosphatase is an enzyme with a typical alpha/beta hydrolase fold. The conformational stability of the human placental alkaline phosphatase was examined with the chemical denaturant urea. The red shifts of fluorescence spectra show a complex unfolding process involving multiple equilibrium intermediates indicating differential stability of the subdomains of the enzyme. None of these unfolding intermediates were observed in the presence of 83 mM NaCl, indicating the importance of ionic interactions in the stabilization of the unfolding intermediates. Guanidinium chloride, on the other hand, could stabilize one of the unfolding intermediates, which is not a salt effect. Some of the unfolding intermediates were also observed in circular dichroism spectroscopy, which clearly indicates steady loss of helical structure during unfolding, but very little change was observed for the beta strand content until the late stage of the unfolding process. The enzyme does not lose its phosphate-binding ability after substantial tertiary structure changes, suggesting that the substrate-binding region is more resistant to chemical denaturant than the other structural domains. Global analysis of the fluorescence spectral change demonstrated the following folding-unfolding process of the enzyme: N I(1) I(2) I(3) I(4) I(5) D. These discrete intermediates are stable at urea concentrations of 2.6, 4.1, 4.7, 5.5, 6.6, and 7.7 M, respectively. These intermediates are further characterized by acrylamide and/or potassium iodide quenching of the intrinsic fluorescence of the enzyme and by the hydrophobic probes, 1-anilinonaphthalene-8-sulfonic acid and 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid. The stepwise unfolding process was interpreted by the folding energy landscape in terms of the unique structure of the enzyme. The rigid central beta-strand domain is surrounded by the peripheral alpha-helical and coil structures, which are marginally stable toward a chemical

  7. Correlation between magnesium and alkaline phosphatase from gingival crevicular fluid periodontal diseases

    Directory of Open Access Journals (Sweden)

    Nila Kasuma

    2015-09-01

    Full Text Available Background: Magnesium is one of the alkaline phosphatase (ALP cofactor. The amount of magnesium contained infoods affect ALP activity. Increased ALP activity will indicate the level of inflammation in periodontal disease. Elevated inflammation in periodontal disease will change gingivitis to periodontitis, where there has been damage to the bone ssupporting the teeth, and an increasing number of gingival crevicular fluid (GCF. The content of GCF consists of enzymatic and non-enzymatic. Changes in the composition of GCF occurs when the inflammation gets worse. Purpose: This study was aimed to prove the correlation between magnesium and ALP from GCF on periodontal disease. Method: This research involved 60 Minangkabaunese people with 20 healthy samples, 20 mild gingivitis samples, and 20 mild periodontitis samples. GCF was collected by absorbing method. Then ALP level in GCF was measured by using ELISA technique. Magnesium level in Minangkabaunese food was tested by Food Frequency Questionnaire (FFQ.Univariate analysis was performed to describe each variable. To see a normal distribution, Kolmogorov Smirnof Test was used (p>0.05. Unpaired T-test and Pearson correlation test was used to see correlation between ALP and magnesium level in Minangkabaunese food. Result: There is a significant correlation between the levels of ALP and magnesium level in Minangkabaunese food with periodontal disease (p=0.005. ALP is higest on mild periodontitis (137.74±23.01 ng/dl. Magnesium level normal control group is highest (250.14±32.34 mg and in mild periodontitis is the lowest (110.83±21.04 mg. Corelation between ALP and magnesium level indicates strong correlation with negative direction (r=- 0.907. Conclusion: There is correlation between the levels of alkaline phosphatase and magnesium level on periodontal disease. Increasing inflamation rate will elevate the ALP level.

  8. Fabrication of hydrogels with elasticity changed by alkaline phosphatase for stem cell culture.

    Science.gov (United States)

    Toda, Hiroyuki; Yamamoto, Masaya; Uyama, Hiroshi; Tabata, Yasuhiko

    2016-01-01

    The objective of this study is to design hydrogels whose elasticity can be changed by alkaline phosphatase (ALP) in cell culture and evaluate the effect of hydrogel elasticity on an osteogenic gene expression of cells. Hydrogels were prepared by the radical polymerization of acrylamide (AAm), N,N'-methylenebisacrylamide (BIS), and Phosmer™M containing phosphate groups (PE-PAAm hydrogels). The storage modulus of PE-PAAm hydrogels prepared was changed by the preparation conditions. When human mesenchymal stem cells (hMSC) were cultured on the ALP-responsive PE-PAAm hydrogels in the presence or absence of ALP, the morphology of hMSC was observed and one of the osteogenic differentiation markers, Runx2, was evaluated. By ALP addition into the culture medium, the morphology of hMSC was changed into an elongated shape without cell damage. ALP addition modified the level of Runx2 gene expression, which was influenced by the modulus of PE-PAAm hydrogels. It is concluded that the elasticity change of hydrogel substrates in cell culture had an influence on the Runx2 gene expression of hMSC. Stem cells sense the surface elasticity of culture substrates, and their differentiation fate is biologically modified by substrate properties. Most of experiments have been performed in static conditions during cell culture, while the in vivo microenvironment is dynamically changed. In this study, we established to design an enzyme-responsive hydrogel whose elasticity can be changed by alkaline phosphatase (ALP) in cell culture to mimic in vivo conditions. As a result, the cells were deformed and the gene expression level of an osteogenic maker, Runx2, was modified by ALP treatment. This is the novel report describing to demonstrate that the dynamic alteration of hydrogel substrate elasticity could modulate the osteoblastic gene expression of human MSC in vitro. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  9. Comparative Analysis of Salivary Alkaline Phosphatase in Post menopausal Women with and without Periodontitis

    Science.gov (United States)

    Sophia, Khumukcham; Sudhakar, Uma; Jayakumar, Parvathee; Mathew, Danny

    2017-01-01

    Introduction Alkaline phosphatase is an intracellular destruction enzyme in the periodontium, and it takes part in the normal turnover of the periodontal ligament, alveolar bone, and root cementum formation and maintenance. Aim The aim of this case control study was to evaluate the enzyme Alkaline Phosphatase (ALP) level in saliva of post menopausal women with and without chronic periodontitis. Materials and Methods In this study, 40 individuals, satisfying the study inclusion and exclusion criteria, were recruited. They were categorically divided, on the basis of gingival index, probing pocket depth and clinical attachment level, into two groups: Group I (post menopausal women with a clinically healthy periodontium, n=20); and Group II (post menopausal women with generalized chronic periodontitis, n=20). Clinical parameters assessed were Plaque Index (PI), Gingival Index (GI), Clinical Attachment Level (CAL) and Probing Pocket Depth (PPD). Unstimulated salivary samples were obtained in which the ALP concentration was measured using p-Nitrophenylphosphate, and 2-amino-2-methyl-1-propanol reagents in Beckman and Coulter, AU 480 auto analyser. Mann-Whitney U test was used to find statistical difference with respect to all clinical parameters such as PI, GI, CAL, PPD and salivary ALP levels. Results The mean ALP in saliva was found to be higher in Group II compared to Group I and the difference was statistically significant with the p-value of 0.008. Conclusion A noteworthy increase in the ALP concentration was seen in saliva in our study (Group II) may be due to increased periodontal inflammation in post menopausal women. Thus salivary ALP can be taken as an additional biomarker to early diagnosis, development and progression of periodontitis especially among post menopausal women. PMID:28274061

  10. Improvement of Student Understanding of How Kinetic Data Facilitates the Determination of Amino Acid Catalytic Function through an Alkaline Phosphatase Structure/Mechanism Bioinformatics Exercise

    Science.gov (United States)

    Grunwald, Sandra K.; Krueger, Katherine J.

    2008-01-01

    Laboratory exercises, which utilize alkaline phosphatase as a model enzyme, have been developed and used extensively in undergraduate biochemistry courses to illustrate enzyme steady-state kinetics. A bioinformatics laboratory exercise for the biochemistry laboratory, which complements the traditional alkaline phosphatase kinetics exercise, was…

  11. Improvement of Student Understanding of How Kinetic Data Facilitates the Determination of Amino Acid Catalytic Function through an Alkaline Phosphatase Structure/Mechanism Bioinformatics Exercise

    Science.gov (United States)

    Grunwald, Sandra K.; Krueger, Katherine J.

    2008-01-01

    Laboratory exercises, which utilize alkaline phosphatase as a model enzyme, have been developed and used extensively in undergraduate biochemistry courses to illustrate enzyme steady-state kinetics. A bioinformatics laboratory exercise for the biochemistry laboratory, which complements the traditional alkaline phosphatase kinetics exercise, was…

  12. Differentiation-dependent activation of the human intestinal alkaline phosphatase promoter by HNF-4 in intestinal cells

    DEFF Research Database (Denmark)

    Olsen, Line; Bressendorff, Simon; Troelsen, Jesper T

    2005-01-01

    of the enterocytes, we have conducted a computer-assisted cis-element search of the proximal human ALPI promoter sequence. A putative recognition site for the transcription factor hepatocyte nuclear factor (HNF)-4 was predicted at the positions from -94 to -82 in relation to the translational start site. The ability......The intestinal alkaline phosphatase gene (ALPI) encodes a digestive brush-border enzyme, which is highly upregulated during small intestinal epithelial cell differentiation. To identify new putative promoter motifs responsible for the regulation of ALPI expression during differentiation...... of HNF-4alpha to stimulate the expression from the ALPI promoter was investigated in the nonintestinal Hela cell line. Cotransfection with an HNF-4alpha expression vector demonstrated a direct activation of the ALPI promoter through this -94 to -82 element. EMSA showed that HNF-4alpha from nuclear...

  13. A fluorescence turn on assay for alkaline phosphatase based on the Cu(2+) catalyzed Fenton-like reaction.

    Science.gov (United States)

    Zhang, Qingfeng; Zhang, Cuiyun; Shahzad, Sohail Anjum; Yu, Cong

    2016-09-01

    A fluorescence turn-on assay was established for ALP (alkaline phosphatase) based on Cu(2+) catalyzed Fenton-like reaction and Graphene Oxide (GO). GO was utilized to quench the fluorescence of fluorescein (FAM) labeled single strand DNA (F-DNA). ALP can remove the phosphate group in sodium ascorbyl phosphate (SAP), and convert it into reducing ascorbate. Highly reactive hydroxyl radicals (·OH) were generated in the presence of ascorbate and Cu(2+) through the Fenton-like reaction. The reactive radicals generated in situ caused the cleavage of F-DNA into small fragments. When GO was added, the fluorescence emission of the sample without ALP was quenched and fluorescence emission recovered in the presence of ALP. The intensity of the recovered fluorescence was directly related to the concentration of ALP in the assay solution, and a sensitive and selective facile ALP assay is therefore established.

  14. Zein as biodegradable material for effective delivery of alkaline phosphatase and substrates in biokits and biosensors.

    Science.gov (United States)

    Jornet-Martínez, N; Campíns-Falcó, P; Hall, E A H

    2016-12-15

    A biodegradable material, zein, is proposed as a reagent delivery platform for biokits and biosensors based on alkaline phosphatase (ALP) activity/inhibition in the presence of phosphatase substrates. The immobilization and release of both the substrate and/or the active ALP, in a biodegradable and low-cost material such as zein, a prolamin from maize, and in combination with glycerol as plasticizer have been investigated. Three zein-based devices are proposed for several applications: (1) inorganic phosphorus estimation in water of different sources (river, lake, coastal water and tap water) with a detection limit of 0.2mg/L - compared to at least 1mg/L required by legislation, (2) estimation of ALP in saliva and (3) chlorpyrifos control in commercial preparations. The single-use kits developed are low cost, easy and fast to manufacture and are stable for at least 20 days at -20°C, so the zein film can preserve and deliver both the enzyme and substrates.

  15. Association between Serum Alkaline Phosphatase Level and Cerebral Small Vessel Disease.

    Directory of Open Access Journals (Sweden)

    Han-Bin Lee

    Full Text Available Serum alkaline phosphatase (ALP is a marker of vascular calcification. A high serum ALP level is associated with an increase in cardiovascular events, and predicts poor functional outcome in patients with stroke. We investigated whether serum ALP was associated with cerebral small vessel disease (cSVD and large cerebral artery stenosis (LCAS.We evaluated vascular risk factors, brain magnetic resonance images (MRIs, and MR angiograms from 1,011 neurologically healthy participants. The presence of silent lacunar infarction (SLI and moderate-to-severe cerebral white matter hyperintensities (MS-cWMH were evaluated as indices of cSVD on brain MRIs. Findings of extracranial arterial stenosis (ECAS or intracranial arterial stenosis (ICAS were considered to be indices of LCAS on MR angiograms.Subjects with SLI (odds ratio [OR]: 2.09; 95% confidence interval [CI]: 1.27-3.42; p = 0.004 and MS-cWMH (OR: 1.48; 95% CI; 1.03-2.13, p = 0.036 were significantly more likely to have ALP levels in the third tertile (ALP ≥ 195 IU/L than the first tertile (ALP ≤ 155 IU/L, after adjusting for cardiovascular risk factors. The mean serum ALP level was significantly higher in patients with SLI or MS-cWMH compared to patients without those findings. After adjustment for confounding factors, the multivariate model found that the statistical significance of serum ALP remained when the presence of SLI (OR: 1.05 per 10 IU/L increase in ALP; 95% CI: 1.02-1.08; p = 0.003 or MS-cWMH (OR: 1.03 per 10 IU/L increase in ALP; 95% CI: 1.00-1.06; p = 0.025 were added to the model. There were no differences in the proportions of patients with LCAS, ICAS, and ECAS across the serum ALP tertiles.Our study of neurologically healthy participants found a positive association between serum ALP level and indicators of cSVD, but no association between serum ALP level and the indicators of LCAS.

  16. Gamma-glutamyltransferase, aspartate aminotransferase and alkaline phosphatase as markers of alcohol consumption in out-patient alcoholics

    DEFF Research Database (Denmark)

    Gluud, C; Andersen, I; Dietrichson, O;

    1981-01-01

    Serum activity of gamma-glutamyltransferase, aspartate aminotransferase and alkaline phosphatase were determined in 316 patients attending an out-patients clinic for treatment of alcoholism. The activity of gamma-glutamyltransferase was raised in 34% and that of aspartate aminotransferase and alk...

  17. Synthesis of 3,3'-carbonyl-bis(chromones) and their activity as mammalian alkaline phosphatase inhibitors.

    Science.gov (United States)

    Miliutina, Mariia; Ejaz, Syeda Abida; Iaroshenko, Viktor O; Villinger, Alexander; Iqbal, Jamshed; Langer, Peter

    2016-01-14

    Hitherto unknown 3,3'-carbonyl-bis(chromones) 8, dimeric chromones bridged by a carbonyl group, were prepared by reaction of chromone-3-carboxylic acid chloride with 3-(dimethylamino)-1- (2-hydroxyphenyl)-2-propen-1-ones 9. The method is generally applicable for the synthesis of novel symmetrical or non-symmetrical products which were found to inhibit mammalian alkaline phosphatases.

  18. Effects of a human recombinant alkaline phosphatase on renal hemodynamics, oxygenation and inflammation in two models of acute kidney injury

    NARCIS (Netherlands)

    Peters, Esther; Ergin, Bülent; Kandil, Asli; Gurel-Gurevin, Ebru; van Elsas, Andrea; Masereeuw, Rosalinde; Pickkers, Peter; Ince, Can

    2016-01-01

    Two small clinical trials indicated that administration of bovine intestinal alkaline phosphatase (AP) improves renal function in critically ill patients with sepsis-associated acute kidney injury (AKI), for which the mechanism of action is not completely understood. Here, we investigated the

  19. Changes in Bone Alkaline Phosphatase and Procollagen Type-1 C-Peptide after Static and Dynamic Exercises

    Science.gov (United States)

    Kubo, Keitaro; Yuki, Kazuhito; Ikebukuro, Toshihiro

    2012-01-01

    We investigated the effects of two types of nonweight-bearing exercise on changes in bone-specific alkaline phosphatase (BAP) and pro-collagen type 1 C-peptide (P1P). BAP is a specific marker of bone synthesis, whereas P1P reflects synthesis of type 1 collagen in other organs as well as bone. Eight participants performed static and dynamic…

  20. In vitro response to alkaline phosphatase coatings immobilized onto titanium implants using electrospray deposition or polydopamine-assisted deposition

    NARCIS (Netherlands)

    Nijhuis, A.W.G.; Beucken, J.J.J.P van den; Jansen, J.A.; Leeuwenburgh, S.C.G.

    2014-01-01

    Immobilization of biomolecules onto implant surfaces is one of the most straightforward strategies to control the interaction between an implant and its biological environment. Recently, it was shown that the enzyme alkaline phosphatase (ALP) could be efficiently immobilized onto titanium implants i

  1. Electrochemical Detection of Alkaline Phosphatase in BALB/c Mouse Fetal Liver Stromal Cells with Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Xue Mei SUN; Dong LI; Zeng Liang BAI; Wen Rui JIN

    2004-01-01

    A method for determination of alkaline phosphatase (ALP) in BALB/c mouse fetal liver stromal cells has been described based on the catalytic reaction. After the cell extract is incubated with the substrate disodium phenyl phosphate, the reaction product phenol generated by ALP is determined by capillary electrophoresis with electrochemical detection.

  2. Effects of overlying water aeration on phosphorus fractions and alkaline phosphatase activity in surface sediment

    Institute of Scientific and Technical Information of China (English)

    Jianjun Chen; Shaoyong Lu; Yikun Zhao; Wei Wang; Minsheng Huang

    2011-01-01

    Microbial activity may influence phosphorus (P) deposit and release at the water sediment interface.The properties of DO (dissolved oxygen), pH, P fractions (TP, Ca-P, Fe-P, OP, IP), and APA (alkaline phosphatase activity) at the water sediment interface were measured to investigate microbial activity variations in surface sediment under conditions of two-month intermittent aeration in overlying water.Results showed that DO and TP of overlying water increased rapidly in the first week and then decreased gradually after 15 day of intermittent aeration.Microorganism metabolism in surface sediment increased pH and decreased DO and TP in the overlying water.After two-month intermittent aeration, APA and OP from surface sediment (0-2 crm) were both significantly higher than those from bottom sediment (6-8 cm) (p < 0.05), and surface sediment Fe-P was transferred to OP during the course of microorganism reproduction on the surface sediment.These results suggest that microbial activity and microorganism biomass from the surface sediment were higher than those from bottom sediment afar two-month intermittent aeration in the overlying water.

  3. Dynamic evolution of the LPS-detoxifying enzyme intestinal alkaline phosphatase in zebrafish and other vertebrates

    Directory of Open Access Journals (Sweden)

    Ye eYang

    2012-10-01

    Full Text Available Alkaline phosphatases (Alps are well-studied enzymes that remove phosphates from a variety of substrates. Alps function in diverse biological processes, including modulating host-bacterial interactions by dephosphorylating the Gram-negative bacterial cell wall component lipopolysaccharide (LPS. In animals, Alps are encoded by multiple genes characterized by either ubiquitous expression (named Alpls, for their liver expression, or their tissue-specific expression, for example in the intestine (Alpi. We previously characterized a zebrafish alpi gene (renamed here alpi.1 that is regulated by Myd88-dependent innate immune signaling and that is required to prevent a host’s excessive inflammatory reactions to its resident microbiota. Here we report the characterization of two new alp genes in zebrafish, alpi.2 and alp3. To understand their origins, we investigated the phylogenetic history of Alp genes in animals. We find that vertebrate Alp genes are organized in three clades with one of these clades missing from the mammals. We present evidence that these three clades originated during the two vertebrate genome duplications. We show that in zebrafish alpl is ubiquitously expressed, as it is in mammals, whereas the other three alps are specific to the intestine. Our phylogenetic analysis reveals that in contrast to Alpl, which has been stably maintained as a single gene throughout the vertebrates, the Alpis have been lost and duplicated multiple times independently in vertebrate lineages, likely reflecting the rapid and dynamic evolution of vertebrate gut morphologies, driven by changes in bacterial associations and diet.

  4. Differential expression of alkaline phosphatase gene in proliferating primary lymphocytes and malignant lymphoid cell lines.

    Science.gov (United States)

    Latheef, S A A; Devanabanda, Mallaiah; Sankati, Swetha; Madduri, Ramanadham

    2016-02-01

    Alkaline Phosphatase (APase) activity has been shown to be enhanced specifically in mitogen stimulated B lymphocytes committed to proliferation, but not in T lymphocytes. APase gene expression was analyzed in proliferating murine and human primary lymphocytes and human malignant cell lines using reverse transcriptase and real time PCR. In mitogen stimulated murine splenic lymphocytes, enhancement of APase activity correlated well with an increase in APase gene expression. However, in mitogen stimulated murine T lymphocytes and human PBL despite a vigorous proliferative response, no increase in APase enzyme activity or gene expression was observed. A constitutive expression of APase activity concomitant with APase gene expression was observed inhuman myeloma cell line, U266 B1. However, neither enzyme activity nor gene expression of APase were observed in human T cell lymphoma, SUPT-1. The results suggest a differential expression of APase activity and its gene in proliferating primary lymphocytes of mice and humans. The specific expression of APase activity and its gene only in human myeloma cells, but not in proliferating primary B cells can be exploited as a sensitive disease marker.

  5. Alkaline Phosphatase: The Next Independent Predictor of the Poor 90-Day Outcome in Alcoholic Hepatitis

    Directory of Open Access Journals (Sweden)

    Beata Kasztelan-Szczerbinska

    2013-01-01

    Full Text Available Aim. Determination of risk factors relevant to 90-day prognosis in AH. Comparison of the conventional prognostic models such as Maddrey’s modified discriminant function (mDF and Child-Pugh-Turcotte (CPT score with newer ones: the Glasgow Alcoholic Hepatitis Score (GAHS; Age, Bilirubin, INR, Creatinine (ABIC score, Model for End-Stage Liver Disease (MELD, and MELD-Na in the death prediction. Patients and Methods. The clinical and laboratory variables obtained at admission were assessed. The mDF, CPT, GAHS, ABIC, MELD, and MELD-Na scores’ different areas under the curve (AUCs and the best threshold values were compared. Logistic regression was used to assess predictors of the 90-day outcome. Results. One hundred sixteen pts fulfilled the inclusion criteria. Twenty (17.4% pts died and one underwent orthotopic liver transplantation (OLT within 90 days of follow-up. No statistically significant differences in the models‘ performances were found. Multivariate logistic regression identified CPT score, alkaline phosphatase (AP level higher than 1.5 times the upper limit of normal (ULN, and corticosteroids (CS nonresponse as independent predictors of mortality. Conclusions. The CPT score, AP > 1.5 ULN, and the CS nonresponse had an independent impact on the 90-day survival in AH. Accuracy of all studied scoring systems was comparable.

  6. EXPRESSION OF ALKALINE PHOSPHATASE DURING OSTEOGENIC DIFFERENTIATION OF RAT BONE MARROW STROMAL CELLS

    Directory of Open Access Journals (Sweden)

    AKBARI M

    2001-01-01

    Full Text Available Introduction: Bone marrow contains a population of stem cells capable of differentiating to osteoblast and forming the bone nodule by dexamethasone. Material and Methods: The stromal cells of bone marrow obtained from 4 to 6 weeks old Spruge-Dawely male rats were grown in primary culture for 7 days and subcultured for 18 days. The cells were cultured in either DMEM medium containing 15% fetal calf serum and antibiotics as the controls or the above medium supplemented with osteogenic supplements (OS: include 10 mM Na-beta glycerophosphate (Na-betaGp, 10 nM dexamethasone (Dex and 50 g/ml ascordic acid (AsA as the examined cultures. After 6, 12 and 18 days of grow up in subculture, the cultures were examined for mineralization and alkaline phosphatase (Apase expression. Results: Mesenchymal stem cells (MSCs in examined cultures underwent a dramatic change in cellular morphology and a significat increase in Apase activity by day 12. The deposition of a calcified matrix on the surface of the culture flasks became evident between days 12 and 18. Conclusion: The addition of osteogenic supplements (OS to MSCs cultures induced Apase expression that contributes to cellular differentiation and mineralization of extracellular matrix.

  7. Alkaline phosphatase levels in diagnostic peritoneal lavage fluid as a predictor of hollow visceral injury.

    Science.gov (United States)

    Jaffin, J H; Ochsner, M G; Cole, F J; Rozycki, G S; Kass, M; Champion, H R

    1993-06-01

    Isolated injuries to hollow viscera may result in equivocal diagnostic peritoneal lavage (DPL) findings. Small bowel injuries cause alkaline phosphatase (AP) levels to increase in DPL effluent. The goal of this study was to better define the role of AP levels in the evaluation of the injured abdomen. We prospectively measured AP levels in 672 patients undergoing DPL. These were retrospectively compared with the clinical findings. All 12 patients with small bowel injuries and three of four with large bowel injuries had an AP level > 10 IU/L. There was one patient with an AP level > 10 IU/L without clinically significant intra-abdominal injury. An AP level > 10 IU/L in the DPL effluent predicted injury requiring laparotomy with a specificity of 99.8% and a sensitivity of 94.7%. We recommend using AP levels only in the management of patients with equivocal findings on DPL who would otherwise not undergo laparotomy. This selective use of AP levels will improve the probability of early diagnosis of bowel injury without increasing the cost of care.

  8. Evaluation of serum sialic acid, heat stable alkaline phosphatase and fucose as markers of breast carcinoma.

    Science.gov (United States)

    Patel, P S; Baxi, B R; Adhvaryu, S G; Balar, D B

    1990-01-01

    Serum levels of total sialic acid (TSA), lipid bound sialic acid (LSA), heat stable alkaline phosphatase (HSAP) and fucose were measured in 39 patients with breast carcinoma, 14 patients with benign breast diseases and 35 healthy female individuals. Elevated levels of the four biomarkers in breast carcinoma were significant when compared with controls (p less than 0.001). Fucose levels were most sensitive (71.8%), while TSA levels were most specific (64.3%) for breast carcinoma. Sensitivity and specificity were 100% when combinations of LSA with fucose and TSA with HSAP were studied respectively. LSA was significantly elevated in infiltrating duct carcinoma patients compared with lobular carcinoma (p less than 0.001). TSA, HSAP and fucose also had lower mean values in lobular carcinoma as compared to infiltrating duct carcinoma. Increase in the levels of LSA and HSAP after surgical removal of the tumor in breast carcinoma occurred prior to the clinical evidence of the recurrence. The results indicate that the combination of the markers studied might be useful in breast cancer diagnosis and treatment monitoring.

  9. Titanium dioxide nanotube films: Preparation, characterization and electrochemical biosensitivity towards alkaline phosphatase.

    Science.gov (United States)

    Roman, Ioan; Trusca, Roxana Doina; Soare, Maria-Laura; Fratila, Corneliu; Krasicka-Cydzik, Elzbieta; Stan, Miruna-Silvia; Dinischiotu, Anca

    2014-04-01

    Titania nanotubes (TNTs) were prepared by anodization on different substrates (titanium, Ti6Al4V and Ti6Al7Nb alloys) in ethylene glycol and glycerol. The influence of the applied potential and processing time on the nanotube diameter and length is analyzed. The as-formed nanotube layers are amorphous but they become crystalline when subjected to subsequent thermal treatment in air at 550°C; TNT layers grown on titanium and Ti6Al4V alloy substrates consist of anatase and rutile, while those grown on Ti6Al7Nb alloy consist only of anatase. The nanotube layers grown on Ti6Al7Nb alloy are less homogeneous, with supplementary islands of smaller diameter nanotubes, spread across the surface. Better adhesion and proliferation of osteoblasts was found for the nanotubes grown on all three substrates by comparison to an unprocessed titanium plate. The sensitivity towards bovine alkaline phosphatase was investigated mainly by electrochemical impedance spectroscopy in relation to the crystallinity, the diameter and the nature of the anodization electrolyte of the TNT/Ti samples. The measuring capacity of the annealed nanotubes of 50nm diameter grown in glycerol was demonstrated and the corresponding calibration curve was built for the concentration range of 0.005-0.1mg/mL. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Rat enterocytes secrete SLPs containing alkaline phosphatase and cubilin in response to corn oil feeding.

    Science.gov (United States)

    Mahmood, Akhtar; Shao, Jian-su; Alpers, David H

    2003-08-01

    Surfactant-like particles (SLP) are unilamellar secreted membranes associated with the process of lipid absorption and isolated previously only from the apical surface of enterocytes. In this paper, the intracellular membrane has been isolated from corn oil-fed animals, identified by its content of the marker protein intestinal alkaline phosphatase (IAP). Another brush-border protein, cubilin, and its anchoring protein megalin have been identified as components of extracellular SLP, but only cubilin is present to any extent in intracellular SLP. During fat absorption, IAP is modestly enriched in intracellular SLP, but full-length cubilin (migrating at 210 kDa in fat-fed mucosal fractions) falls by one-half, although fragments of cubilin are abundant in the intracellular SLP. Both IAP and cubilin colocalize to the same cells during corn oil absorption and colocalize around lipid droplets. This localization is more intense during feeding of corn oil with Pluronic L-81, a detergent that allows uptake of fatty acids and monoglycerides from the lumen, but blocks chylomicron secretion. Confocal microscopy confirms the colocalization of IAP and the ligand for cubilin, intrinsic factor. Possible roles for cubilin in intracellular SLP include facilitating movement of the lipid droplet through the cell and binding to the basolateral membrane before reverse endocytosis.

  11. Curcumin and Chronic Kidney Disease (CKD: Major Mode of Action through Stimulating Endogenous Intestinal Alkaline Phosphatase

    Directory of Open Access Journals (Sweden)

    Siddhartha S. Ghosh

    2014-12-01

    Full Text Available Curcumin, an active ingredient in the traditional herbal remedy and dietary spice turmeric (Curcuma longa, has significant anti-inflammatory properties. Chronic kidney disease (CKD, an inflammatory disease, can lead to end stage renal disease resulting in dialysis and transplant. Furthermore, it is frequently associated with other inflammatory disease such as diabetes and cardiovascular disorders. This review will focus on the clinically relevant inflammatory molecules that play a role in CKD and associated diseases. Various enzymes, transcription factors, growth factors modulate production and action of inflammatory molecules; curcumin can blunt the generation and action of these inflammatory molecules and ameliorate CKD as well as associated inflammatory disorders. Recent studies have shown that increased intestinal permeability results in the leakage of pro-inflammatory molecules (cytokines and lipopolysaccharides from gut into the circulation in diseases such as CKD, diabetes and atherosclerosis. This change in intestinal permeability is due to decreased expression of tight junction proteins and intestinal alkaline phosphatase (IAP. Curcumin increases the expression of IAP and tight junction proteins and corrects gut permeability. This action reduces the levels of circulatory inflammatory biomolecules. This effect of curcumin on intestine can explain why, despite poor bioavailability, curcumin has potential anti-inflammatory effects in vivo and beneficial effects on CKD.

  12. Effect of Diazinon on Acid and Alkaline Phosphatase Activities in Plasma and Organs of Clarias gariepinus

    Directory of Open Access Journals (Sweden)

    I.R. Inyang

    2011-05-01

    Full Text Available The aim of this study was to determine the effect of the pesticide, diazinon, on phosphatases in the plasma and organs on Clarias gariepinus. Adult Clarias gariepinus were exposed in four replicates to varying sublethal concentrations diazinon (ranging from 1.00 to 10.0 mg/L in 30-day semi-static bioassays. Alkaline phoshatase (ALP and acid phosphate (ACP were determined in plasma and other organs (gastrointestinal tract - GIT, kidney, muscle, gill and liver of the fish after the experimental exposures. Dizinon did not cause any statistically significant difference on plasma ALP over the concentrations tested (p>0.05, but ACP showed significantly higher mean value at 10 mg/L compared to the control. ALP and ACP values in all the organs (GIT, intestinal tract, kidney, muscle, gill, liver decreased with increasing concentration of diazion. This indicates an evidence of inhibition of these enzymes in the organs by the toxicant, and therefore alteration of biochemical processes in C. gariepinus which can be used as bio-indicators of the effects of diazinon in the Niger Delta environment.

  13. Enrichment of thermosensitive chitosan hydrogels with glycerol and alkaline phosphatase for bone tissue engineering applications.

    Science.gov (United States)

    Douglas, Timothy E L; Krok-Borkowicz, Małgorzata; Macuda, Aleksandra; Pietryga, Krzysztof; Pamuła, Elżbieta

    2016-01-01

    Thermosensitive injectable chitosan hydrogels can be formed by neutralization of acidic chitosan solutions with sodium betaglycerophosphate (Na-β-GP) coupled with increasing temperature to body temperature. Such hydrogels have been considered for applications in bone regeneration. In this study, chitosan hydrogels were enriched with glycerol and the enzyme alkaline phosphatase (ALP) with a view to improving their suitability as materials for bone tissue engineering. Mineral formation was confirmed by infrared spectroscopy (FTIR) and increases in the mass fraction of the hydrogel not consisting of water. Incorporation of ALP in hydrogels followed by incubation in a solution containing calcium ions and glycerophosphate, a substrate for ALP, led to formation of calcium phosphate within the hydrogel. MG-63 osteoblast-like cells were cultivated in eluates from hydrogels containing ALP and without ALP at different dilutions and directly on the hydrogel samples. Hydrogels containing ALP exhibited superior cytocompatibility to ALP-free hydrogels. These results pave the way for the use of glycerol- and ALP-enriched hydrogels in bone regeneration.

  14. Activity of alkaline and acidic phosphatase in glandular cells of uterine endometrium of puerperal ewes after exposure to polychlorinated biphenyls

    OpenAIRE

    Valocky I.; Krajničakova Maria; Legath J.; Lenhardt L.; Ostro A.; Danko J.; Tkačikova L`udmila; Mojžišova Jana; Fialkovičova Maria; Mardzinova Silvia

    2005-01-01

    The study is focused on the observation of alkaline and acidic phosphatase activity in the glandular cells of uterine endometrium in puerperal ewes after exposure to polychlorinated biphenyls. Ewes of Slovak merino breed (n=25) divided into 2 groups were included in the experiment. The animals in the experimental group (n=14) and control group (n=11) were euthanised on day 17, 25 and 34 postpartum. The ewes in the experimental group were given per os capsules of the chemical preparation Delor...

  15. A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.

    Science.gov (United States)

    Chen, Wenbo; Liu, Chenxi; Xiao, Yutao; Zhang, Dandan; Zhang, Yongdong; Li, Xianchun; Tabashnik, Bruce E; Wu, Kongming

    2015-01-01

    Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry) proteins from the bacterium Bacillus thuringiensis (Bt) in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50) of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f) was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects.

  16. A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.

    Directory of Open Access Journals (Sweden)

    Wenbo Chen

    Full Text Available Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry proteins from the bacterium Bacillus thuringiensis (Bt in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50 of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects.

  17. PrognosticValue of PINP,BoneAlkaline Phosphatase, CTX-I, andYKL-40 in Patients With Metastatic Prostate Carcinoma

    DEFF Research Database (Denmark)

    Brasso, Klaus; Christensen, Ib Jarle; Johansen, Julia S

    2006-01-01

    Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma. Prostate. 2006 Apr 1;66(5):503-13. PMID: 16372331 [PubMed - indexed for MEDLINE]......Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma. Prostate. 2006 Apr 1;66(5):503-13. PMID: 16372331 [PubMed - indexed for MEDLINE]...

  18. Alkaline phosphatase inhibition based conductometric biosensor for phosphate estimation in biological fluids.

    Science.gov (United States)

    Upadhyay, Lata Sheo Bachan; Verma, Nishant

    2015-06-15

    Determination of phosphate ions concentration is very important from both, environmental and clinical point of view. In this study, a simple and novel conductometric biosensor for indirect determination of the phosphate ions in aqueous solution has been developed. The developed biosensor is based on the inhibition of immobilized alkaline phosphatase activity, in the presence of the phosphate ions. This is the first time we developed a mono-enzymatic biosensor for indirect estimation of phosphate ions. The developed biosensor showed a broad linear response (as compared to other reported biosensors) for phosphate ions in the range of 0.5-5.0 mM (correlation coefficient=0.995), with a detection limit of 50 µM. Different optimized parameters were obtained as the buffer concentration of 30 mM, substrate concentration of 1.0mM, and a pH of 9.0. All the optimized parameters were analyzed by analysis of variance, and were found to be statistically significant at a level of α=0.05. The developed biosensor is also suitable to determine the serum phosphate concentration, with a recovery of 86-104%, while a recovery of 102% was obtained from the water samples that were spiked with 500 µM phosphate. A relative standard deviation in the conductance response for five successive measurements (n=5) did not exceed 7%, with a shelf life of 30 days. With a lower detection limit and a higher recovery, the biosensor provides a facile approach for phosphate estimation in biological fluids.

  19. Wnt5a attenuates Wnt3a-induced alkaline phosphatase expression in dental follicle cells

    Energy Technology Data Exchange (ETDEWEB)

    Sakisaka, Yukihiko [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tsuchiya, Masahiro [Department of Oral Diagnosis, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tohoku Fukushi University, Sendai 989-3201 (Japan); Nakamura, Takashi [Department of Pediatric Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Liason Center for Innovative Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tamura, Masato [Department of Biochemistry and Molecular Biology, Hokkaido University Graduate School of Dentistry, Sapporo 060-8586 (Japan); Shimauchi, Hidetoshi [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Nemoto, Eiji, E-mail: e-nemoto@dent.tohoku.ac.jp [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)

    2015-08-01

    Wnt signaling regulates multiple cellular events such as cell proliferation, differentiation, and apoptosis through β-catenin-dependent canonical and β-catenin-independent noncanonical pathways. Canonical Wnt/β-catenin signaling can promote the differentiation of dental follicle cells, putative progenitor cells for cementoblasts, osteoblasts, and periodontal ligament cells, toward a cementoblast/osteoblast phenotype during root formation, but little is known about the biological significance of noncanonical Wnt signaling in this process. We identified the expression of Wnt5a, a representative noncanonical Wnt ligand, in tooth root lining cells (i.e. precementoblasts/cementoblasts) and dental follicle cells during mouse tooth root development, as assessed by immunohistochemistry. Silencing expression of the Wnt5a gene in a dental follicle cell line resulted in enhancement of the Wnt3a (a representative canonical Wnt ligand)-mediated increase in alkaline phosphatase (ALP) expression. Conversely, treatment with recombinant Wnt5a inhibited the increase in ALP expression, suggesting that Wnt5a signaling functions as a negative regulator of canonical Wnt-mediated ALP expression of dental follicle cells. Wnt5a did not affect the nuclear translocation of β-catenin as well as β-catenin-mediated transcriptional activation of T-cell factor (Tcf) triggered by Wnt3a, suggesting that Wnt5a inhibits the downstream part of the β-catenin-Tcf pathway. These findings suggest the existence of a feedback mechanism between canonical and noncanonical Wnt signaling during the differentiation of dental follicle cells. - Highlights: • Dental follicle cells express Wnt5a during tooth root development. • Silencing of Wnt5a enhances Wnt3a-mediated ALP expression of dental follicle cells. • Conversely, treatment with rWnt5a inhibited the increase in ALP expression. • Wnt5a functions as a negative regulator of Wnt3a-mediated ALP expression.

  20. Preoperative serum alkaline phosphatase: a predictive factor for early hypocalcaemia following parathyroidectomy of primary hyperparathyroidism

    Institute of Scientific and Technical Information of China (English)

    Sun Longhao; He Xianghui; Liu Tong

    2014-01-01

    Background Postoperative hypocalcemia is one of the most common complications following parathyroidectomy for primary hyperparathyroidism (PHPT).The aim of this study was to analyze the predictive value of biochemical parameters as indicators for episodes of hypocalcemia in patients undergoing parathyroidectomy for PHPT.Methods The patients with PHPT who underwent parathyroidectomy between February 2004 and February 2014 were studied retrospectively at a single medical center.The patients were divided into biochemical,clinical,and no postoperative hypocalcemia groups,based on different clinical manifestations.Potential risk factors for postoperative hypocalcemia were identified and investigated by univariate and multivariate Logistic regression analysis.Results Of the 139 cases,25 patients (18.0%) were diagnosed with postoperative hypocalcemia according to the traditional criterion.Univariate analysis revealed only alkaline phosphatase (ALP) and the small area under the curve (AUC) of receiver operating characteristics (ROC) curve for ALP demonstrates low accuracy in predicting the occurrence of postoperative hypocalcemia.Based on new criteria,22 patients were added to the postoperative hypocalcemia group and similar biochemical parameters were compared.The serum ALP was a significant independent risk factor for postoperative hypocalcemia (P=0.000) and its AUC of ROC curve was 0.783.The optimal cutoff point was 269 U/L and the sensitivity and specificity for prediction were 89.2% and 64.3%,respectively.Conclusions The risk of postoperative hypocalcemia after parathyroidectomy should be emphasized for patients with typical symptoms of hypocalcemia despite their serum calcium level is in normal or a little higher range.Serum ALP is a predictive factor for the occurrence of postoperative hypocalcemia.

  1. Comparative characterization of pulmonary surfactant aggregates and alkaline phosphatase isozymes in human lung carcinoma tissue.

    Science.gov (United States)

    Iino, Nozomi; Matsunaga, Toshiyuki; Harada, Tsuyoshi; Igarashi, Seiji; Koyama, Iwao; Komoda, Tsugikazu

    2007-05-01

    Alkaline phosphatase (AP) isozymes are surfactant-associated proteins (SPs). Since several different AP isozymes have been detected in the pneumocytes of lung cancer patients, we attempted to identify the relationship between pulmonary surfactant aggregate subtypes and AP isozymes. Pulmonary surfactant aggregates were isolated from carcinoma and non-carcinoma tissues of patients with non-small cell carcinoma of the lung. Upon analysis, ultraheavy, heavy, and light surfactant aggregates were detected in the non-carcinoma tissues, but no ultraheavy surfactant aggregates were found in the carcinoma tissues. Surfactant-associated protein A (SP-A) was detected as two bands (a 27-kDa band and a 54-kDa band) in the ultraheavy, heavy, and light surfactant aggregates found in the non-carcinoma tissues. Although both SP-A bands were detected in the heavy and light surfactant aggregates from adenocarcinoma tissues, the 54-kDa band was not detected in squamous cell carcinoma tissues. Liver AP (LAP) was detected in the heavy and light surfactant aggregates from both non-carcinoma and squamous carcinoma tissues, but not in heavy surfactant aggregates from adenocarcinoma tissues. A larger amount of bone type AP (BAP) was found in light surfactant aggregate fractions from squamous cell carcinomas than those from adenocarcinoma tissues or non-carcinoma tissues from patients with either type of cancer. LAP, BAP, and SP-A were identified immunohistochemically in type II pneumocytes from non-carcinoma tissues and adenocarcinoma cells, but no distinct SP-A staining was observed in squamous cell carcinoma tissues. The present study has thus revealed several differences in pulmonary surfactant aggregates and AP isozymes between adenocarcinoma tissue and squamous cell carcinoma tissue.

  2. The mechanism of mineralization and the role of alkaline phosphatase in health and disease.

    Science.gov (United States)

    Orimo, Hideo

    2010-02-01

    Biomineralization is the process by which hydroxyapatite is deposited in the extracellular matrix. Physiological mineralization occurs in hard tissues, whereas pathological calcification occurs in soft tissues. The first step of mineralization is the formation of hydroxyapatite crystals within matrix vesicles that bud from the surface membrane of hypertrophic chondrocytes, osteoblasts, and odontoblasts. This is followed by propagation of hydroxyapatite into the extracellular matrix and its deposition between collagen fibrils. Extracellular inorganic pyrophosphate, provided by NPP1 and ANKH, inhibits hydroxyapatite formation. Tissue-nonspecific alkaline phosphatase (TNAP) hydrolyzes pyrophosphate and provides inorganic phosphate to promote mineralization. Inorganic pyrophosphate, pyridoxal phosphate, and phosphoethanolamine are thought to be the physiologic substrates of TNAP. These accumulate in the event of TNAP deficiency, e.g., in cases of hypophosphatasia. The gene encoding TNAP is mapped to chromosome 1, consists of 12 exons, and possesses regulatory motifs in the 5'-untranslated region. Inhibition of TNAP enzymatic activity suppresses TNAP mRNA expression and mineralization in vitro. Hypophosphatasia is an inherited systemic bone disease characterized by hypomineralization of hard tissues. The phenotype of hypophosphatasia is varied. To date, more than 200 mutations in the TNAP gene have been reported. Knockout mice mimic the phenotypes of severe hypophosphatasia. Among the mutations in the TNAP gene, c.1559delT is frequent in the Japanese population. This frameshift mutation results in the expression of an abnormally long protein that is degraded in cells. DNA-based prenatal diagnosis using chorionic villus sampling has been developed, but requires thorough genetic counseling. Although hypophosphatasia is untreatable at present, the recent success of enzyme replacement therapy offers promise. The problems presented by impaired mineralization in age

  3. Intestinal alkaline phosphatase contributes to the reduction of severe intestinal epithelial damage.

    Science.gov (United States)

    Bol-Schoenmakers, Marianne; Fiechter, Daniëlle; Raaben, Willem; Hassing, Ine; Bleumink, Rob; Kruijswijk, Daniëlle; Maijoor, Kelly; Tersteeg-Zijderveld, Monique; Brands, Ruud; Pieters, Raymond

    2010-05-10

    Inflammatory bowel disease is characterized by chronic inflammation of the intestine and is accompanied by damage of the epithelial lining and by undesired immune responses towards enteric bacteria. It has been demonstrated that intestinal alkaline phosphatase (iAP) protects against the induction of inflammation, possibly due to dephosphorylation of lipopolysaccharide (LPS). The present study investigated the therapeutic potential of iAP in intestinal inflammation and epithelial damage. Intestinal epithelial damage was induced in C57BL/6 mice using detran sulfate sodium (DSS) and iAP was administered 4days after initial DSS exposure. Loss in body weight was significantly less in iAP-treated mice and accompanied with reduced colon damage (determined by combination of crypt loss, loss of goblet cells, oedema and infiltrations of neutrophils). Treatment with iAP was more effective in case of severe inflammation compared to situations of mild to moderate inflammation. Rectal administration of LPS into a moderate inflamed colon did not aggravate inflammation. Furthermore, soluble iAP did not lower LPS-induced nuclear factor-kappaB activation in epithelial cells in vitro but induction of cellular AP expression by butyrate resulted in decreased LPS response. In conclusion, the present study shows that oral iAP administration has beneficial effects in situations of severe intestinal epithelial damage, whereas in moderate inflammation endogenous iAP may be sufficient to counteract disease-aggravating effects of LPS. An approach including iAP treatment holds a therapeutic promise in case of severe inflammatory bowel disease. Copyright 2010 Elsevier B.V. All rights reserved.

  4. Alkaline phosphatase activity related to phosphorus stress of microphytoplankton in different trophic conditions

    Science.gov (United States)

    Ivančić, Ingrid; Pfannkuchen, Martin; Godrijan, Jelena; Djakovac, Tamara; Marić Pfannkuchen, Daniela; Korlević, Marino; Gašparović, Blaženka; Najdek, Mirjana

    2016-08-01

    The northern Adriatic (NA) is a favorable basin for studying the adaptive strategies of plankton to a variety of conditions along the steep gradients of environmental parameters over the year. Earlier studies identified phosphorus (P)-limitation as one of the key stresses within the NA that shape the biological response in terms of biodiversity and metabolic adjustments. A wide range of reports supports the notion that P-limitation is a globally important phenomenon in aquatic ecosystems. In this study P stress of marine microphytoplankton was determined at species level along a trophic gradient in the NA. In P-limitation all species with considerable contributions to the diatom community expressed alkaline phosphatase activity (APA), compared to only a few marginal dinoflagellate species. Nevertheless, APA expressing species did not always dominate the phytoplankton community, suggesting that APA is also an important strategy for species to survive and maintain active metabolism outside of their mass abundances. A symbiotic relationship could be supposed for diatoms that did not express APA themselves and probably benefited from APA expressed by attached bacteria. APA was not expressed by any microphytoplankton species during the autumn when P was not limiting, while most of the species did express APA during the P-limitation. This suggests that APA expression is regulated by orthophosphate availability. The methods employed in this study allowed the microscopic detection of APA for each microphytoplankton cell with simultaneous morphologic/taxonomic analysis. This approach uncovered a set of strategies to compete in P-limited conditions within the marine microphytoplankton community. This study confirms the role of P-limitation as a shaping factor in marine ecosystems.

  5. Mechanistic and Evolutionary Insights from Comparative Enzymology of Phosphomonoesterases and Phosphodiesterases across the Alkaline Phosphatase Superfamily.

    Science.gov (United States)

    Sunden, Fanny; AlSadhan, Ishraq; Lyubimov, Artem Y; Ressl, Susanne; Wiersma-Koch, Helen; Borland, Jamar; Brown, Clayton L; Johnson, Tory A; Singh, Zorawar; Herschlag, Daniel

    2016-11-02

    Naively one might have expected an early division between phosphate monoesterases and diesterases of the alkaline phosphatase (AP) superfamily. On the contrary, prior results and our structural and biochemical analyses of phosphate monoesterase PafA, from Chryseobacterium meningosepticum, indicate similarities to a superfamily phosphate diesterase [Xanthomonas citri nucleotide pyrophosphatase/phosphodiesterase (NPP)] and distinct differences from the three metal ion AP superfamily monoesterase, from Escherichia coli AP (EcAP). We carried out a series of experiments to map out and learn from the differences and similarities between these enzymes. First, we asked why there would be independent instances of monoesterases in the AP superfamily? PafA has a much weaker product inhibition and slightly higher activity relative to EcAP, suggesting that different metabolic evolutionary pressures favored distinct active-site architectures. Next, we addressed the preferential phosphate monoester and diester catalysis of PafA and NPP, respectively. We asked whether the >80% sequence differences throughout these scaffolds provide functional specialization for each enzyme's cognate reaction. In contrast to expectations from this model, PafA and NPP mutants with the common subset of active-site groups embedded in each native scaffold had the same monoesterase:diesterase specificities; thus, the >10(7)-fold difference in native specificities appears to arise from distinct interactions at a single phosphoryl substituent. We also uncovered striking mechanistic similarities between the PafA and EcAP monoesterases, including evidence for ground-state destabilization and functional active-site networks that involve different active-site groups but may play analogous catalytic roles. Discovering common network functions may reveal active-site architectural connections that are critical for function, and identifying regions of functional modularity may facilitate the design of new enzymes

  6. Serum alkaline phosphatase isoenzymes in laboratory beagle dogs detected by polyacrylamide-gel disk electrophoresis.

    Science.gov (United States)

    Hatayama, Kazuhisa; Nishihara, Yoshito; Kimura, Sayaka; Goto, Ken; Nakamura, Daichi; Wakita, Atsushi; Urasoko, Yoshinaka

    2011-10-01

    Serum alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. Itoh et al. (2002) reported that a commercially available polyacrylamide-gel (PAG) disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co., Ltd., Tokyo, Japan) for identifying serum ALP isoenzymes was useful for veterinary clinicopathological diagnosis in mongrel dogs. In the present study, based on the report of Itoh et al. (2002), we tried to expand the application range of this kit to laboratory beagle dogs which are commonly used in toxicity studies. In order to identify the origin of each ALP isoenzyme, tissue ALP extracts from the liver, bone and small intestine and serum samples were treated with neuraminidase, anti-small intestinal ALP antibody, ALP inhibitor levamisole and/or wheat germ agglutinin (WGA). The main serum ALP isoenzymes in 5-month-old intact beagle dogs were bone-derived (bone and atypical ALP: corresponding to human variant bone ALP) and they tended to decrease with age. However, liver-derived ALP isoenzyme greatly increased in the serum of cholestasis model dogs. The cholestasis model dogs also had a large molecular ALP detected in the resolving gel. This ALP could be originated from intestinal ALP or corticosteroid-induced ALP (CALP), because the activity remained even after levamisole inhibition. CALP was observed in intact laboratory beagle dogs with individual differences. These results suggest that the present method is a useful tool for detecting serum ALP isoenzymes in laboratory beagle dogs and concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity.

  7. Measurement of alkaline phosphatase in canine seminal plasma--an update.

    Science.gov (United States)

    Schäfer-Somi, S; Fröhlich, T; Schwendenwein, I

    2013-02-01

    In dogs, diagnosis of incomplete ejaculation and azoospermia can be made by measuring the activity of the enzyme alkaline phosphatase (AP) in seminal plasma. However, even though upper cut-off value of 5000 IU/l is given in the literature, results by different assays may vary considerably. Furthermore, no data exist concerning the stability of the enzyme during storage of frozen seminal plasma, and no recommendations for pre-analytic dilutions can be found. During the present study, we compared results from a conventional large scale wet chemistry analyzer to a widely used dry chemistry point of care system (POC) and established a best practice for pre-analytical dilutions. Furthermore, stability of enzyme activities in seminal plasma during storage at -18 °C for 24 h was evaluated. The average activity of AP in the 2nd fraction of normal ejaculates measured by Reflotron® was 107,328 IU/l. After 24 h of frozen storage, activities did not differ significantly (96,844 IU / l, p > 0.05). Fresh and frozen samples were analysed in parallel by the POC and conventional chemistry analyser, and the results compared that did not reveal a significant difference (p > 0.05). A dilution of seminal plasma with physiologic saline 1:100 prior to analysis was sufficient for the qualitative information whether AP activity is below or above 5000 IU/l. Present data show that AP measurement by a POC dry chemistry system is sufficiently accurate in diluted seminal plasma for the diagnosis of azoospermia and that seminal plasma can be stored frozen for 24h before analysis.

  8. Changes in Stoichiometry, Cellular RNA, and Alkaline Phosphatase Activity of Chlamydomonas in Response to Temperature and Nutrients.

    Science.gov (United States)

    Hessen, Dag O; Hafslund, Ola T; Andersen, Tom; Broch, Catharina; Shala, Nita K; Wojewodzic, Marcin W

    2017-01-01

    Phytoplankton may respond both to elevated temperatures and reduced nutrients by changing their cellular stoichiometry and cell sizes. Since increased temperatures often cause increased thermal stratification and reduced vertical flux of nutrients into the mixed zone, it is difficult to disentangle these drivers in nature. In this study, we used a factorial design with high and low levels of phosphorus (P) and high and low temperature to assess responses in cellular stoichiometry, levels of RNA, and alkaline phosphatase activity (APA) in the chlorophyte Chlamydomonas reinhardtii. Growth rate, C:P, C:N, N:P, RNA, and APA all responded primarily to P treatment, but except for N:P and APA, also temperature contributed significantly. For RNA, the contribution from temperature was particularly strong with higher cellular levels of RNA at low temperatures, suggesting a compensatory allocation to ribosomes to maintain protein synthesis and growth. These experiments suggest that although P-limitation is the major determinant of growth rate and cellular stoichiometry, there are pronounced effects of temperature also via interaction with P. At the ecosystem level, nutrients and temperature will thus interact, but temperatures would likely exert a stronger impact on these phytoplankton traits indirectly via its force on stratification regimes and vertical nutrient fluxes.

  9. Changes in Stoichiometry, Cellular RNA, and Alkaline Phosphatase Activity of Chlamydomonas in Response to Temperature and Nutrients

    Science.gov (United States)

    Hessen, Dag O.; Hafslund, Ola T.; Andersen, Tom; Broch, Catharina; Shala, Nita K.; Wojewodzic, Marcin W.

    2017-01-01

    Phytoplankton may respond both to elevated temperatures and reduced nutrients by changing their cellular stoichiometry and cell sizes. Since increased temperatures often cause increased thermal stratification and reduced vertical flux of nutrients into the mixed zone, it is difficult to disentangle these drivers in nature. In this study, we used a factorial design with high and low levels of phosphorus (P) and high and low temperature to assess responses in cellular stoichiometry, levels of RNA, and alkaline phosphatase activity (APA) in the chlorophyte Chlamydomonas reinhardtii. Growth rate, C:P, C:N, N:P, RNA, and APA all responded primarily to P treatment, but except for N:P and APA, also temperature contributed significantly. For RNA, the contribution from temperature was particularly strong with higher cellular levels of RNA at low temperatures, suggesting a compensatory allocation to ribosomes to maintain protein synthesis and growth. These experiments suggest that although P-limitation is the major determinant of growth rate and cellular stoichiometry, there are pronounced effects of temperature also via interaction with P. At the ecosystem level, nutrients and temperature will thus interact, but temperatures would likely exert a stronger impact on these phytoplankton traits indirectly via its force on stratification regimes and vertical nutrient fluxes. PMID:28167934

  10. Joint influence of temperature and ions of metals on level of activity alkaline phosphatase the mucous membrane of intestines beluga, the starlet and their hybrid

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    D. A. Bednyakov

    2010-01-01

    Full Text Available In work joint influence of ions of bivalent metals (Mn, Fe, Co, Ni, Cu and Zn and temperatures on level of activity alkaline phosphatase mucous membrane beluga, starlet and their hybrid is shown. Dependence of response of enzyme on action of ions of metals according to their position in a periodic table of chemical elements is shown. The given dependence remains and at temperature change incubation, only at low temperatures the activating effect of metals being in the period beginning is maximum, and at high, is maximum inhibiting effect of metals being in the period end.

  11. Disposable lateral flow-through strip for smartphone-camera to quantitatively detect alkaline phosphatase activity in milk.

    Science.gov (United States)

    Yu, Ling; Shi, ZhuanZhuan; Fang, Can; Zhang, YuanYuan; Liu, YingShuai; Li, ChangMing

    2015-07-15

    A disposable lateral flow-through strip was developed for smartphone to fast one-step quantitatively detect alkaline phosphatase (ALP) activity in raw milk. The strip comprises two functional components, a conjugation pad loaded with phosphotyrosine-coated gold nanoparticles (AuNPs@Cys-Try-p) and a testing line coated with anti-phosphotryosine antibody (anti-Tyr-p mAb). The dephosphorylation activity of ALP at the testing zone can be quantitatively assayed by monitoring the accumulated AuNPs-induced color changes by smartphone camera, thus providing a highly convenient portable detection method. A trace amount of ALP as low as 0.1UL(-1) with a linear dynamic range of 0.1-150UL(-1) (R(2)=0.999) in pasteurized milk and raw milk can be one-step detected by the developed flow-through strip within 10min, demonstrating the potential of smartphone-based portable sensing device for pathogen detection. This bio-hazards free lateral flow-through testing strip can be also used to fabricate rapid, sensitive and inexpensive enzyme or immunosensors for broad portable clinic diagnosis and food contamination analysis, particularly in point-of-care and daily food quality inspection. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Effects of garlic and diallyl trisulfide on the growth, photosynthesis, and alkaline phosphatase activity of the toxic cyanobacterium Microcystis aeruginosa.

    Science.gov (United States)

    Wang, Shoubing; Wang, Yuanan; Ma, Xiaoxue; Xu, Ziran

    2016-03-01

    To identify a botanical algicide and elucidate the response of cyanobacteria to the extract from higher plants, the effects of garlic and garlic-derived diallyl trisulfide on Microcystis aeruginosa were studied. Effects were evaluated by changes in cell density, chlorophyll a, maximum effective quantum yield (Fv/Fm), effective quantum yield (YII), non-photochemical quenching (NPQ), and rapid light curves of M. aeruginosa. In addition, alkaline phosphatase activity (APA) was measured when M. aeruginosa was incubated with diallyl trisulfide. Results indicated that the inhibition by garlic and diallyl trisulfide was significant. The 120-h 50 % effective concentrations of garlic and diallyl trisulfide (EC50) were 0.75 g L(-1) and 2.84 mg L(-1), respectively. Moreover, the inhibitory rate increased with increasing concentration and the growth of M. aeruginosa was inhibited by 90.0 % at the highest concentrations. We also show that the response of M. aeruginosa to stress could involve both impairment of the photosynthetic center PSII and alteration of APA. For example, at high garlic concentration (2.0 g L(-1)), Fv/Fm significantly decreased from 0.501 to 0.084 (p garlic as an environmentally friendly algicide.

  13. Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    Kriszta Molnár; (A)dám Vannay; Beáta Szebeni; Nóra Fanni Bánki; Erna Sziksz; (A)ron Cseh; Hajnalka Gy(o)rffy

    2012-01-01

    AIM:To investigate intestinal alkaline phosphatase (iAP) in the intestinal mucosa of children with inflammatory bowel disease (IBD).METHODS:Colonic biopsy samples were taken from 15 newly diagnosed IBD patients and from 10 healthy controls.In IBD patients,specimens were obtained both from inflamed and non-inflamed areas.The iAP mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blotting analysis,respectively.Tissue localization of iAP and Toll-like receptor (TLR) 4 was investigated by immunofluorescent staining.RESULTS:The iAP protein level in the inflamed mucosa of children with Crohn's disease (CD) and ulcerative colitis (UC) was significantly decreased when compared with controls (both P < 0.05).Similarly,we found a significantly decreased level of iAP protein in the inflamed mucosa in CD compared with non-inflamed mucosa in CD (P < 0.05).In addition,the iAP protein level in inflamed colonic mucosa in patients with UC was decreased compared with non-inflamed mucosa in patients with CD (P < 0.05).iAP protein levels in the non-inflamed mucosa of patients with CD were similar to controls.iAP mRNA expression in inflamed colonic mucosa of children with CD and UC was not significantly different from that in non-inflamed colonic mucosa with CD.Expression of iAP mRNA in patients with non-inflamed mucosa and in controls were similar.Co-localization of iAP with TLR4 showed intense staining with a dotted-like pattern.iAP was present in the inflamed and non-inflamed mucosa of patients with CD,UC,and in control biopsy specimens,irrespective of whether it was present in the terminal ileum or in the colon.However,the fluorescent signal of TLR4 was more pronounced in the colon compared with the terminal ileum in all groups studied.CONCLUSION:Lower than normal iAP protein levels in inflamed mucosa of IBD patients may indicate a role for iAP in inflammatory lesions in IBD.Based on our results,administration of exogenous

  14. Cord blood calcium, phosphate, magnesium, and alkaline phosphatase gestational age-specific reference intervals for preterm infants

    Directory of Open Access Journals (Sweden)

    Lyon Andrew W

    2011-08-01

    Full Text Available Abstract Background The objective was to determine the influence of gestational age, maternal, and neonatal variables on reference intervals for cord blood bone minerals (calcium, phosphate, magnesium and related laboratory tests (alkaline phosphatase, and albumin-adjusted calcium, and to develop gestational age specific reference intervals based on infants without influential pathological conditions. Methods Cross-sectional study. 702 babies were identified as candidates for this study in a regional referral neonatal unit. After exclusions (for anomalies, asphyxia, maternal magnesium sulfate administration, and death, relationships were examined between cord blood serum laboratory analytes (calcium, phosphate, magnesium, alkaline phosphatase, and albumin-adjusted calcium with gestation age and also with maternal and neonatal variables using multiple linear regression. Infants with influential pathological conditions were omitted from the development of gestational age specific reference intervals for the following categories: 23-27, 28-31, 32-34, 35-36 and > 36 weeks. Results Among the 506 preterm and 54 terms infants included in the sample. Phosphate, magnesium, and alkaline phosphatase in cord blood serum decreased with gestational age, calcium increased with gestational age. Those who were triplets, small for gestational age, and those whose mother had pregnancy-induced hypertension were influential for most of the analytes. The reference ranges for the preterm infants ≥ 36 weeks were: phosphate 1.5 to 2.6 mmol/L (4.5 to 8.0 mg/dL, calcium: 2.1 to 3.1 mmol/L (8.3 to 12.4 mg/dL; albumin-adjusted calcium: 2.3 to 3.2 mmol/L (9.1 to 12.9 mg/dL; magnesium 0.6 to 1.0 mmol/L (1.4 to 2.3 mg/dL, and alkaline phosphatase 60 to 301 units/L. Conclusions These data suggest that gestational age, as well as potentially pathogenic maternal and neonatal variables should be considered in the development of reference intervals for preterm infants.

  15. Treatment of PCR products with exonuclease I and heat-labile alkaline phosphatase improves the visibility of combined bisulfite restriction analysis

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Kousuke; Emoto, Noriko; Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Nagase, Takahide; Ohishi, Nobuya [Department of Respiratory Medicine, The University of Tokyo Hospital, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan); Takai, Daiya, E-mail: dtakai-ind@umin.ac.jp [Department of Clinical Laboratory, The University of Tokyo Hospital, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan)

    2010-08-27

    Research highlights: {yields} Incubating PCR products at a high temperature causes smears in gel electrophoresis. {yields} Smears interfere with the interpretation of methylation analysis using COBRA. {yields} Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. {yields} The elimination of smears improves the visibility of COBRA. -- Abstract: DNA methylation plays a vital role in the regulation of gene expression. Abnormal promoter hypermethylation is an important mechanism of inactivating tumor suppressor genes in human cancers. Combined bisulfite restriction analysis (COBRA) is a widely used method for identifying the DNA methylation of specific CpG sites. Here, we report that exonuclease I and heat-labile alkaline phosphatase can be used for PCR purification for COBRA, improving the visibility of gel electrophoresis after restriction digestion. This improvement is observed when restriction digestion is performed at a high temperature, such as 60 {sup o}C or 65 {sup o}C, with BstUI and TaqI, respectively. This simple method can be applied instead of DNA purification using spin columns or phenol/chloroform extraction. It can also be applied to other situations when PCR products are digested by thermophile-derived restriction enzymes, such as PCR restriction fragment length polymorphism (RFLP) analysis.

  16. Occurrence of matrix-bound phosphine in polar ornithogenic tundra ecosystems: effects of alkaline phosphatase activity and environmental variables.

    Science.gov (United States)

    Zhu, Renbin; Ma, Dawei; Ding, Wei; Bai, Bo; Liu, Yashu; Sun, Jianjun

    2011-09-01

    Phosphine (PH(3)), a reduced phosphorus compound, is a highly toxic and reactive atmospheric trace gas. In this study, a total of ten ornithogenic soil/sediment profiles were collected from tundra ecosystems of east Antarctica and Arctic, and matrix-bound phosphine (MBP), the phosphorus fractions and alkaline phosphatase activity (APA) were analyzed. High MBP concentrations were found in these profiles with the range from 39.59 ng kg(-1) dw to 11.77 μg kg(-1) dw. MBP showed a consistent vertical distribution pattern in almost all the soil profiles, and its concentrations increased at soil surface layers and then decreased with depths. MBP levels in the ornithogenic soils were two to three orders of magnitude lower than those in ornithogenic sediments. The yield of PH(3) as a fraction of total P in all the profiles ranged from 10(-5) to 10(-9) mgPH(3) mg(-1)P with higher mean PH(3) yield in the ornithogenic sediments. The ornithogenic soils showed high concentrations of total phosphorus (TP), organic phosphorus (OP), inorganic phosphorus (IP) and metal elements (Cu, Zn, Mn, Fe, Al and Ca) but low MBP levels, vice versa for the ornithogenic sediments. No correlation had been obtained between MBP concentrations and IP, OP and TP. There existed an exponential correlation (r=0.67, psoil/sediment moisture. MBP concentrations showed a significant positive correlation with APA (r=0.668, psoils/sediments. Our results indicated that MBP is an important gaseous link in the phosphorus biogeochemical cycles of ornithogenic tundra ecosystems in Antarctica and Arctic.

  17. Age- and diapause-related acid and alkaline phosphatase activities in the intestine and malpighian tubules of the Colorado potato beetle, Leptinotarsa decemlineata (Say).

    Science.gov (United States)

    Yi, S X; Adams, T S

    2001-03-01

    Specific activities for soluble (s) and membrane (m)-bound acid (ACP) and alkaline phosphatases (ALP) were determined in the midgut, hindgut, and Malpighian tubules for developing, prediapausing, and diapausing adult Colorado potato beetles, Leptinotarsa decemlineata (Say). High ACP activities were found in the hindgut and Malpighian tubules while high ALP activities were found in the Malpighian tubules. Variation in both ACP and ALP activities in each tissue reflects fluctuation in protein synthesis and secretion involved with digestion, excretion, and other unknown functions. Phosphatase activities in the tissues examined show the dynamic nature of diapause in this insect. Diapausing beetles showed increases in phosphatase activity after hormone treatments. JHA treatments increased s-ACP and m-ACP activities in all tissues but 20-HE did not increase activity in any tissue. Allatotropin tended to mimic the effects of JHA treatment. The s-ALP activity was also increased in all tissues whereas m-ALP was increased in the midgut and hindgut by JHA treatment. Malpighian tubule m-ALP activity was only increased by 20-HE treatments. Allatotropin was not as effective in increasing ALP activities as it was with ACP activities.

  18. The 1.4 A crystal structure of the large and cold-active Vibrio sp. alkaline phosphatase.

    Science.gov (United States)

    Helland, Ronny; Larsen, Renate Lie; Asgeirsson, Bjarni

    2009-02-01

    Alkaline phosphatase (AP) from the cold-adapted Vibrio strain G15-21 is among the AP variants with the highest known k(cat) value. Here the structure of the enzyme at 1.4 A resolution is reported and compared to APs from E. coli, human placenta, shrimp and the Antarctic bacterium strain TAB5. The Vibrio AP is a dimer although its monomers are without the long N-terminal helix that embraces the other subunit in many other APs. The long insertion loop, previously noted as a special feature of the Vibrio AP, serves a similar function. The surface does not have the high negative charge density as observed in shrimp AP, but a positively charged patch is observed around the active site that may be favourable for substrate binding. The dimer interface has a similar number of non-covalent interactions as other APs and the "crown"-domain is the largest observed in known APs. Part of it slopes over the catalytic site suggesting that the substrates may be small molecules. The catalytic serines are refined with multiple conformations in both monomers. One of the ligands to the catalytic zinc ion in binding site M1 is directly connected to the crown-domain and is closest to the dimer interface. Subtle movements in metal ligands may help in the release of the product and/or facilitate prior dephosphorylation of the covalent intermediate. Intersubunit interactions may be a major factor for promoting active site geometries that lead to the high catalytic activity of Vibrio AP at low temperatures.

  19. Acute effect of tea, wine, beer, and polyphenols on ecto-alkaline phosphatase activity in human vascular smooth muscle cells.

    Science.gov (United States)

    Negrão, Maria R; Keating, Elisa; Faria, Ana; Azevedo, Isabel; Martins, Maria J

    2006-07-12

    Alkaline phosphatase (ALP) is an ecto-enzyme widely distributed across species. It modulates a series of transmembranar transport systems, has an important role in bone mineralization, and can also be involved in vascular calcification. Polyphenol-rich diets seem to have protective effects on human health, namely, in the prevention of cardiovascular diseases. We aimed to investigate the effects of polyphenols and polyphenol-rich beverages upon membranar alkaline phosphatase (ecto-ALP) activity in intact human vascular smooth muscle cells (AALTR). The ecto-ALP activity was determined at pH 7.8, with p-nitrophenyl phosphate as the substrate, by absorbance spectrophotometry at 410 nm. Cell viability was assessed by the lactate dehydrogenase (LDH) method, and the polyphenol content of beverages was assessed using the Folin-Ciocalteu reagent. All polyphenols tested inhibited ecto-ALP activity, in a concentration-dependent way. Teas, wines, and beers also inhibited ecto-ALP activity, largely according to their polyphenol content. All tested compounds and beverages improved or did not change AALTR cell viability. Stout beer was an exception to the described behavior. Although more studies must be done, the inhibition of AALTR ecto-ALP activity by polyphenolic compounds and polyphenol-containing beverages may contribute to their cardiovascular protective effects.

  20. Which one of the two common reporter systems is more suitable for chemiluminescent enzyme immunoassay: alkaline phosphatase or horseradish peroxidase?

    Science.gov (United States)

    Yu, Songcheng; Yu, Fei; Liu, Lie; Zhang, Hongquan; Zhang, Zhenzhong; Qu, Lingbo; Wu, Yongjun

    2016-05-01

    Alkaline phosphatase and horseradish peroxidase are the most commonly used reporter systems in chemiluminescent enzyme immunoassay (CLEIA). Which one, therefore, would be better when establishing a CLEIA method for a new target substance? There was no standard answer. In this study, both reporters were compared systematically including luminescence kinetics, conjugation methods, optimal condition and detection performance, using two common drugs, SD-methoxy-pyrimidine and enrofloxacin, as determination objects. The results revealed that there was much difference between the luminescence kinetics of the two systems. However, there was little difference between these systems when detecting the same substance, including in optimal conditions and determination of performance. Both reporters were suitable for establishing chemiluminescent enzyme immunoassays. Therefore, the choice of alkaline phosphatase or horseradish peroxidase as the reporter system in chemiluminescent enzyme immunoassays depends on availability. Conversely, these two report systems could be applied in simultaneous analysis of multicomponents due to their different optical behaviors and similar performances. But attention should be paid to conjugation method and coating buffer, which affected the luminescent intensity of different determination targets.

  1. Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.

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    Juan Luis Jurat-Fuentes

    Full Text Available Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP were detected by two dimensional differential in-gel electrophoresis (2D-DIGE analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests.

  2. A study of the association between serum bone-specific alkaline phosphatase and serum phosphorus concentration or dietary phosphorus intake.

    Science.gov (United States)

    Haraikawa, Mayu; Tanabe, Rieko; Sogabe, Natsuko; Sugimoto, Aoi; Kawamura, Yuka; Michigami, Toshimi; Hosoi, Takayuki; Goseki-Sone, Masae

    2012-01-01

    Alkaline phosphatase (ALP) hydrolyzes a variety of monophosphate esters into phosphoric acid and alcohol at a high optimum pH (pH 8-10). Human ALPs are classified into four types: tissue-non specific (TNSALP, liver/bone/kidney), intestinal, placental, and germ cell types. Based on studies of hypophosphatasia (HPP), which is a systemic bone disease caused by the presence of either one or two pathologic mutations in ALPL that encodes TNSALP, TNSALP was suggested to be indispensable for skeletal mineralization. In this study, we explored the possibility that dietary nutrients contribute to regulate serum bone-specific ALP (BAP) activity. Serum biochemical parameters, such as serum ALP, BAP, osteocalcin, and fibroblast growth factor 23 (FGF23), were measured in healthy young subjects (n=193). Dietary nutrient intakes were measured based on 3-d food records before the day of blood examinations. The presence of a carrier of the deletion of T at nucleotide 1559 (c.1559delT), which has been reported to be the most frequent in Japanese HPP, was not detected in any subject. By the analysis of BAP activity and other biochemical parameters or dietary nutrient intakes, we obtained significant correlations between BAP activity and serum phosphorus (r=-0.165, p=0.022), calcium intake (mg/1,000 kcal/d) (r=-0.186, p=0.010), or phosphorus intake (mg/1,000 kcal/d) (r=-0.226, p=0.002). Further study on the regulation of BAP activity and calcium and/or phosphorus homeostasis will provide useful data for improving skeletal health.

  3. 1-step versus 2-step immobilization of alkaline phosphatase and bone morphogenetic protein-2 onto implant surfaces using polydopamine.

    Science.gov (United States)

    Nijhuis, Arnold W G; van den Beucken, Jeroen J J P; Boerman, Otto C; Jansen, John A; Leeuwenburgh, Sander C G

    2013-08-01

    Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the current study, a 1-step polydopamine-based approach was applied for alkaline phosphatase (ALP) and bone morphogenetic protein-2 (BMP-2) immobilization, and compared to the conventional 2-step polydopamine-based immobilization and plain adsorption. To this end, ALP and BMP-2 were immobilized onto titanium and polytetrafluoroethylene (PTFE) substrates. The absolute quantity and biological activity of immobilized ALP were assessed quantitatively to compare the three types of immobilization. Plain adsorption of both ALP and BMP-2 was inferior to both polydopamine-based immobilization approaches. ALP was successfully immobilized onto titanium and PTFE surfaces via the 1-step approach, and the immobilized ALP retained its enzymatic activity. Using the 1-step approach, the amount of immobilized ALP was increased twofold to threefold compared to the conventional 2-step immobilization process. In contrast, more BMP-2 was immobilized using the conventional 2-step immobilization approach. Retention of ALP and BMP-2 was measured over a period of 4 weeks and was found to be similar for the 1-step and 2-step methods and far superior to the retention of adsorbed biomolecules due to the formation of covalent linkages between catechol moieties and immobilized proteins. The biological behavior of ALP and BMP-2 coatings immobilized using polydopamine (1- and 2-step) as well as adsorption was assessed by culturing rat bone marrow cells, which revealed that the cell responses to the various experimental groups were not statistically different. In conclusion, the 1-step polydopamine-based immobilization method was shown to be more efficient for immobilization of ALP, whereas the conventional 2

  4. Alkaline Phosphatase Activity : an overlooked player on the phosphate behavior in macrotidal estuaries

    Science.gov (United States)

    Delmas, Daniel; Labry, Claire; Youenou, Agnes; Quere, Julien; Auguet, Jean Christophe; Montanie, Helene

    2014-05-01

    The non-conservative behavior of phosphate within the estuarine salinity gradient is essentially assigned to physico-chemical processes, such as desorption at low salinity and to benthic exchanges. Microbial phosphatase activity (APA), generally related to phosphate deficiency, is seldom studied in phosphate rich estuarine waters. In order to address the impact of microbial activity (bacterial abundance, production BSP, APA) on phosphate behavior, we studied these activities on a seasonal basis within the salinity gradient of two macrotidal estuaries presenting different levels of suspended solids. Whatever the season the Charente estuary is characterized by high levels of Suspended Particulate Matter (SPM > 1g.L-1), particularly in the Maximum Turbidity Zone (MTZ) located at the 5-10 psu. In this area characterized by high BSP and APA there is a significant increase of PO4 levels especially during summer. In the Aulne estuary the particle load is significantly lower (1/10) but high BSP and APA are equally recorded. In the highly turbid waters of the Charente estuary, active phytoplankton is virtually absent as pheopigments constitute up to 80% of the total pigments, particularly in the MTZ, therefore APA may essentially have a bacterial origin. In the Aulne estuary attached bacteria are dominant, both in numbers and production, and their distribution along the haline gradient perfectly follows those of APA and phosphate levels. These observations, associated with the very close relationships observed between APA, SPM and BSP, suggest that APA derive mainly from bacterial (attached) origin and operate at the expense of particulate phosphorus and hence contribute to PO4 regeneration, especially in spring and summer. Finally, as APA increased as PO4, whereas the reverse is observed in both fresh and marine waters, an original scheme for APA regulation, related to the large dominance of attached bacteria can be described for the estuarine waters.

  5. Differentiating Intracellular from Extracellular Alkaline Phosphatase Activity in Soil by Sonication

    NARCIS (Netherlands)

    Qin, S.P.; Hu, C.S.; Oenema, O.

    2013-01-01

    Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio

  6. A phage-displayed chicken single-chain antibody fused to alkaline phosphatase detects Fusarium pathogens and their presence in cereal grains

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Zu-Quan [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Li, He-Ping [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Zhang, Jing-Bo [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Huang, Tao [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Liu, Jin-Long; Xue, Sheng [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Wu, Ai-Bo [Institute for Agri-food Standards and Testing Technology, Laboratory of Quality and Safety Risk Assessment for Agro-products, Ministry of Agriculture, Shanghai Academy of Agricultural Sciences, 1000 Jinqi Road, Shanghai 201403 (China); Liao, Yu-Cai, E-mail: ycliao06@yahoo.com.cn [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); National Center of Plant Gene Research, Wuhan 430070 (China)

    2013-02-18

    Graphical abstract: A phage-displayed chicken scFv antibody, FvSG7, binds on the surface antigen of conidiospores and the mycelia of F. verticillioides. Its fusion with alkaline phosphatase (AP) through a 218 linker displayed a 4-fold higher affinity compared with the parent scFv antibody and efficiently detected toxigenic Fusarium pathogens in cereal grains. Highlights: ► Generation of a highly reactive scFv antibody against F. verticillioides. ► Localization of the antibody binding to the surface target of F. verticillioides. ► Expression of the antibody–alkaline phosphatase (AP) fusion linked by a 218 linker. ► The antibody–AP fusion has a higher affinity than the parental antibody. ► The antibody–AP fusion detects toxigenic Fusarium pathogens in cereal grains. -- Abstract: Fusarium and its poisonous mycotoxins are distributed worldwide and are of particular interest in agriculture and food safety. A simple analytical method to detect pathogens is essential for forecasting diseases and controlling mycotoxins. This article describes a proposed method for convenient and sensitive detection of Fusarium pathogens that uses the fusion of single-chain variable fragment (scFv) and alkaline phosphatase (AP). A highly reactive scFv antibody specific to soluble cell wall-bound proteins (SCWPs) of F. verticillioides was selected from an immunized chicken phagemid library by phage display. The antibody was verified to bind on the surface of ungerminated conidiospores and mycelia of F. verticillioides. The scFv–AP fusion was constructed, and soluble expression in bacteria was confirmed. Both the antibody properties and enzymatic activity were retained, and the antigen-binding capacity of the fusion was enhanced by the addition of a linker. Surface plasmon resonance measurements confirmed that the fusion displayed 4-fold higher affinity compared with the fusion's parental scFv antibody. Immunoblot analyses showed that the fusion had good binding

  7. High temperature and pressure alkaline electrolysis

    DEFF Research Database (Denmark)

    Allebrod, Frank; Chatzichristodoulou, Christodoulos; Mogensen, Mogens Bjerg

    2013-01-01

    the operational temperature and pressure to produce pressurized hydrogen at high rate (m3 H2·h-1·m-2 cell area) and high electrical efficiency. This work describes an exploratory technical study of the possibility to produce hydrogen and oxygen with a new type of alkaline electrolysis cell at high temperatures...... for immobilization of aqueous KOH solutions. Electrolysis cells with this electrolyte and metal foam based gas diffusion electrodes were successfully demonstrated at temperatures up to 250 °C at 40 bar. Different electro-catalysts were tested in order to reduce the oxygen and hydrogen overpotentials. Current...

  8. Tissue-nonspecific Alkaline Phosphatase Regulates Purinergic Transmission in the Central Nervous System During Development and Disease

    Directory of Open Access Journals (Sweden)

    Álvaro Sebastián-Serrano

    2015-01-01

    Full Text Available Tissue-nonspecific alkaline phosphatase (TNAP is one of the four isozymes in humans and mice that have the capacity to hydrolyze phosphate groups from a wide spectrum of physiological substrates. Among these, TNAP degrades substrates implicated in neurotransmission. Transgenic mice lacking TNAP activity display the characteristic skeletal and dental phenotype of infantile hypophosphatasia, as well as spontaneous epileptic seizures and die around 10 days after birth. This physiopathology, linked to the expression pattern of TNAP in the central nervous system (CNS during embryonic stages, suggests an important role for TNAP in neuronal development and synaptic function, situating it as a good target to be explored for the treatment of neurological diseases. In this review, we will focus mainly on the role that TNAP plays as an ectonucleotidase in CNS regulating the levels of extracellular ATP and consequently purinergic signaling.

  9. Investigating the kinetics of paramagnetic-beads linked alkaline phosphatase enzyme through microchannel resistance measurement in dielectric microchip.

    Science.gov (United States)

    Faure, Mathilde; Sotta, Bruno; Gamby, Jean

    2014-08-15

    Real time monitoring of electrolyte resistance changes during hydrolysis of 4-nitrophenylphosphate (pNPP) by alkaline phosphatase (ALP) bound on paramagnetic-beads was performed into a small dielectric channel. The reaction kinetic fit with a non-competitive substrate-inhibition equation. Michaelis-Menten apparent constant, KM(app), was determined as 0.33±0.06mM and the maximum apparent rate, Vmax(app) as 98±5pMs(-1). The detection limits were 15fM for ALP and 0.75mM for pNPP. This miniaturized device constitutes a powerful tool for analysis of interaction between ligands. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Copper sulfide nanoparticle-decorated graphene as a catalytic amplification platform for electrochemical detection of alkaline phosphatase activity.

    Science.gov (United States)

    Peng, Juan; Han, Xiao-Xia; Zhang, Qing-Chun; Yao, Hui-Qin; Gao, Zuo-Ning

    2015-06-01

    Copper sulfide nanoparticle-decorated graphene sheet (CuS/GR) was successfully synthesized and used as a signal amplification platform for electrochemical detection of alkaline phosphatase activity. First, CuS/GR was prepared through a microwave-assisted hydrothermal approach. The CuS/GR nanocomposites exhibited excellent electrocatalytic activity toward the oxidation of ALP hydrolyzed products such as 1-naphthol, which produced a current response. Thus, a catalytic amplification platform based on CuS/GR nanocomposite for electrochemical detection of ALP activity was designed using 1-naphthyl phosphate as a model substrate. The current response increased linearly with ALP concentration from 0.1 to 100 U L(-1) with a detection limit of 0.02 U L(-1). The assay was applied to estimate ALP activity in human serum samples with satisfactory results. This strategy may find widespread and promising applications in other sensing systems that involves ALP.

  11. Presence and patterns of alkaline phosphatase activity and phosphorus cycling in natural riparian zones under changing nutrient conditions

    Directory of Open Access Journals (Sweden)

    Peifang Wang

    2014-08-01

    Full Text Available Phosphorus (P is an important limiting nutrient in aquatic ecosystems and knowledge of P cycling is fundamental for reducing harmful algae blooms and other negative effects in water. Despite their importance, the characteristics of P cycling under changing nutrient conditions in shallow lakes were poorly investigated. In this study, in situ incubation experiments were conducted in a natural riparian zone in the main diversion channel used for water transfer into Lake Taihu (Wangyu River. Variations in microbial biomass, dissolved P fractions (organic and inorganic, and alkaline phosphatase activity (bulk APA and specific APA were determined after incubation with and without the addition of P and nitrogen (N (4 total water treatments: +P, +N, +NP, and control. Experiments were conducted during two seasons (late spring and early fall to account for natural differences in nutrient levels that may occur in situ. Our results demonstrated that low levels of DRP may not necessarily indicate P limitation. Phytoplankton exhibited “serial N limitation with P stress” in May, such that chlorophyll a (Chl a increased significantly with N addition, while the limiting nutrient shifted to P in October and phytoplankton biomass increased with P addition. Phytoplankton contributed greatly to APA production and was significantly influenced by P bioavailability, yet high levels of bulk APA were also not necessarily indicative of P limitation. In contrast to phytoplankton, bacteria were less P stressed. As a consequence of enhanced utilization of dissolved reactive P (DRP and dissolved organic P (DOP, +N treatment elevated APA significantly. By contrast, APA could be repressed to low values and phytoplankton converted a large portion of DRP to DOP with P addition. But this was not consistent with bacteria APA (bact-APA in the absence or presence of abundant phytoplankton biomass. The correlation between bulk APA and DRP was good at separate sites and discrepant

  12. Active site detection by spatial conformity and electrostatic analysis--unravelling a proteolytic function in shrimp alkaline phosphatase.

    Directory of Open Access Journals (Sweden)

    Sandeep Chakraborty

    Full Text Available Computational methods are increasingly gaining importance as an aid in identifying active sites. Mostly these methods tend to have structural information that supplement sequence conservation based analyses. Development of tools that compute electrostatic potentials has further improved our ability to better characterize the active site residues in proteins. We have described a computational methodology for detecting active sites based on structural and electrostatic conformity - CataLytic Active Site Prediction (CLASP. In our pipelined model, physical 3D signature of any particular enzymatic function as defined by its active sites is used to obtain spatially congruent matches. While previous work has revealed that catalytic residues have large pKa deviations from standard values, we show that for a given enzymatic activity, electrostatic potential difference (PD between analogous residue pairs in an active site taken from different proteins of the same family are similar. False positives in spatially congruent matches are further pruned by PD analysis where cognate pairs with large deviations are rejected. We first present the results of active site prediction by CLASP for two enzymatic activities - β-lactamases and serine proteases, two of the most extensively investigated enzymes. The results of CLASP analysis on motifs extracted from Catalytic Site Atlas (CSA are also presented in order to demonstrate its ability to accurately classify any protein, putative or otherwise, with known structure. The source code and database is made available at www.sanchak.com/clasp/. Subsequently, we probed alkaline phosphatases (AP, one of the well known promiscuous enzymes, for additional activities. Such a search has led us to predict a hitherto unknown function of shrimp alkaline phosphatase (SAP, where the protein acts as a protease. Finally, we present experimental evidence of the prediction by CLASP by showing that SAP indeed has protease activity in

  13. Active site detection by spatial conformity and electrostatic analysis--unravelling a proteolytic function in shrimp alkaline phosphatase.

    Science.gov (United States)

    Chakraborty, Sandeep; Minda, Renu; Salaye, Lipika; Bhattacharjee, Swapan K; Rao, Basuthkar J

    2011-01-01

    Computational methods are increasingly gaining importance as an aid in identifying active sites. Mostly these methods tend to have structural information that supplement sequence conservation based analyses. Development of tools that compute electrostatic potentials has further improved our ability to better characterize the active site residues in proteins. We have described a computational methodology for detecting active sites based on structural and electrostatic conformity - CataLytic Active Site Prediction (CLASP). In our pipelined model, physical 3D signature of any particular enzymatic function as defined by its active sites is used to obtain spatially congruent matches. While previous work has revealed that catalytic residues have large pKa deviations from standard values, we show that for a given enzymatic activity, electrostatic potential difference (PD) between analogous residue pairs in an active site taken from different proteins of the same family are similar. False positives in spatially congruent matches are further pruned by PD analysis where cognate pairs with large deviations are rejected. We first present the results of active site prediction by CLASP for two enzymatic activities - β-lactamases and serine proteases, two of the most extensively investigated enzymes. The results of CLASP analysis on motifs extracted from Catalytic Site Atlas (CSA) are also presented in order to demonstrate its ability to accurately classify any protein, putative or otherwise, with known structure. The source code and database is made available at www.sanchak.com/clasp/. Subsequently, we probed alkaline phosphatases (AP), one of the well known promiscuous enzymes, for additional activities. Such a search has led us to predict a hitherto unknown function of shrimp alkaline phosphatase (SAP), where the protein acts as a protease. Finally, we present experimental evidence of the prediction by CLASP by showing that SAP indeed has protease activity in vitro.

  14. Characterization of alkaline phosphatase activity in seminal plasma and in fresh and frozen-thawed stallion spermatozoa.

    Science.gov (United States)

    Bucci, Diego; Giaretta, Elisa; Spinaci, Marcella; Rizzato, Giovanni; Isani, Gloria; Mislei, Beatrice; Mari, Gaetano; Tamanini, Carlo; Galeati, Giovanna

    2016-01-15

    Alkaline phosphatase (AP) has been studied in several situations to elucidate its role in reproductive biology of the male from different mammalian species; at present, its role in horse sperm physiology is not clear. The aim of the present work was to measure AP activity in seminal plasma and sperm extracts from freshly ejaculated as well as in frozen-thawed stallion spermatozoa and to verify whether relationship exists between AP activity and sperm quality parameters. Our data on 40 freshly ejaculated samples from 10 different stallions demonstrate that the main source of AP activity is seminal plasma, whereas sperm extracts contribution is very low. In addition, we found that AP activity at physiological pH (7.0) is significantly lower than that observed at pH 8.0, including the optimal AP pH (pH 10.0). Alkaline phosphatase did not exert any effect on sperm-oocyte interaction assessed by heterologous oocyte binding assay. Additionally, we observed a thermal stability of seminal plasma AP, concluding that it is similar to that of bone isoforms. Positive correlations were found between seminal plasma AP activity and sperm concentration, whereas a negative correlation was present between both spermatozoa extracts and seminal plasma AP activity and seminal plasma protein content. A significant decrease in sperm extract AP activity was found in frozen-thawed samples compared with freshly ejaculated ones (n = 21), concomitantly with the decrease in sperm quality parameters. The positive correlation between seminal plasma AP activity measured at pH 10 and viability of frozen-thawed spermatozoa suggests that seminal plasma AP activity could be used as an additional predictive parameter for stallion sperm freezability. In conclusion, we provide some insights into AP activity in both seminal plasma and sperm extracts and describe a decrease in AP after freezing and thawing.

  15. Alkaline phosphatase activity-guided isolation of active compounds and new dammarane-type triterpenes from Cissus quadrangularis hexane extract.

    Science.gov (United States)

    Pathomwichaiwat, Thanika; Ochareon, Pannee; Soonthornchareonnon, Noppamas; Ali, Zulfiqar; Khan, Ikhlas A; Prathanturarug, Sompop

    2015-02-03

    The stem of Cissus quadrangularis L. (CQ) is used in traditional medicine to treat bone fractures and swelling. Anti-osteoporotic activity of CQ hexane extract has been reported, but the active compounds in this extract remain unknown. Thus, we aimed to identify the active compounds in CQ hexane extract using bioassay-guided isolation. The CQ hexane extract was fractionated sequentially with benzene, dichloromethane, ethyl acetate, and methanol. The examination of CQ extract and its fractions was guided by bioassays for alkaline phosphatase (ALP) activity during the differentiation of MC3T3-E1 osteoblastic cells. The cells were treated with or without the CQ extract and its fractions for a period of time, and then the stimulatory effect of the alkaline phosphatase enzyme, a bone differentiation marker, was investigated. The compounds obtained were structurally elucidated using spectroscopic techniques and re-evaluated for activity during bone differentiation. A total of 29 compounds were isolated, viz., triterpenes, fatty acid methyl esters, glycerolipids, steroids, phytols, and cerebrosides. Four new dammarane-type triterpenes were isolated for the first time from nature, and this report is the first to identify this group of compounds from the Vitaceae family. Seven compounds, viz., glycerolipids and squalene, stimulated ALP activity at a dose of 10μg/mL. Moreover, the synergistic effect of these compounds on bone formation was demonstrated. This report describes, for the first time, the isolation of active compounds from CQ hexane extract; these active compounds will be useful for the quality control of extracts from this plant used to treat osteoporosis. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  16. Lactoferrin up-regulates intestinal gene expression of brain-derived neurotrophic factors BDNF, UCHL1 and alkaline phosphatase activity to alleviate early weaning diarrhea in postnatal piglets.

    Science.gov (United States)

    Yang, Changwei; Zhu, Xi; Liu, Ni; Chen, Yue; Gan, Hexia; Troy, Frederic A; Wang, Bing

    2014-08-01

    The molecular mechanisms underlying how dietary lactoferrin (Lf) impacts gut development and maturation and protects against early weaning diarrhea are not well understood. In this study, we supplemented postnatal piglets with an Lf at a dose level of 155 and 285 mg/kg/day from 3 to 38 days following birth. Our findings show that the high dose of Lf up-regulated messenger RNA expression levels of genes encoding brain-derived neurotrophic factor (BDNF) and ubiquitin carboxy-terminal hydrolase L1 (ubiquitin thiolesterase (UCHL1) and, to a lesser extent, glial cell line-derived neurotrophic factor, in the duodenum (Pintestinal alkaline phosphatase activity (Pbrain-microbe axis that has not been previously reported.

  17. Evaluating the levels of salivary alkaline and acid phosphatase activities as biochemical markers for periodontal disease: A case series

    Directory of Open Access Journals (Sweden)

    Sarita Dabra

    2012-01-01

    Full Text Available Background: The purpose of this study was to determine the salivary levels of alkaline phosphatase (ALP and acid phosphatase (ACP activities in patients with periodontal disease and to evaluate the use of these enzymes as biochemical markers for periodontal tissue damage. Materials and Methods: In this prospective analytical study, we examined the activities of salivary ALP and ACP in patients with periodontal disease, before and after periodontal treatment. The experimental groups consisted of 20 gingivitis patients and 20 periodontitis patients and the control group had healthy subjects (20 samples. The stimulated saliva of the patient was collected in a sterile test tube and analyzed using Hitachi′s Diagnostic Automatic Analyser. Periodontal disease was determined based on clinical parameters such as gingival index, probing depth and clinical attachment loss. Patients with periodontal disease were under conventional periodontal treatment. The statistical analysis applied was Student′s t-test. Probabilities less than 0.05 (P < 0.05 were considered significant. Results: The obtained results showed statistically significant increased activities of ALP and ACP in saliva from patients with periodontal disease in relation to control group. A significant reduction in the enzyme levels was seen after conventional periodontal therapy. Conclusions: Based on these results, salivary ALP and ACP can be considered to be the biomarkers for evaluating periodontal tissue damage.

  18. Inhibition of the gut enzyme intestinal alkaline phosphatase may explain how aspartame promotes glucose intolerance and obesity in mice.

    Science.gov (United States)

    Gul, Sarah S; Hamilton, A Rebecca L; Munoz, Alexander R; Phupitakphol, Tanit; Liu, Wei; Hyoju, Sanjiv K; Economopoulos, Konstantinos P; Morrison, Sara; Hu, Dong; Zhang, Weifeng; Gharedaghi, Mohammad Hadi; Huo, Haizhong; Hamarneh, Sulaiman R; Hodin, Richard A

    2017-01-01

    Diet soda consumption has not been associated with tangible weight loss. Aspartame (ASP) commonly substitutes sugar and one of its breakdown products is phenylalanine (PHE), a known inhibitor of intestinal alkaline phosphatase (IAP), a gut enzyme shown to prevent metabolic syndrome in mice. We hypothesized that ASP consumption might contribute to the development of metabolic syndrome based on PHE's inhibition of endogenous IAP. The design of the study was such that for the in vitro model, IAP was added to diet and regular soda, and IAP activity was measured. For the acute model, a closed bowel loop was created in mice. ASP or water was instilled into it and IAP activity was measured. For the chronic model, mice were fed chow or high-fat diet (HFD) with/without ASP in the drinking water for 18 weeks. The results were that for the in vitro study, IAP activity was lower (p < 0.05) in solutions containing ASP compared with controls. For the acute model, endogenous IAP activity was reduced by 50% in the ASP group compared with controls (0.2 ± 0.03 vs 0.4 ± 0.24) (p = 0.02). For the chronic model, mice in the HFD + ASP group gained more weight compared with the HFD + water group (48.1 ± 1.6 vs 42.4 ± 3.1, p = 0.0001). Significant difference in glucose intolerance between the HFD ± ASP groups (53 913 ± 4000.58 (mg·min)/dL vs 42 003.75 ± 5331.61 (mg·min)/dL, respectively, p = 0.02). Fasting glucose and serum tumor necrosis factor-alpha levels were significantly higher in the HFD + ASP group (1.23- and 0.87-fold increases, respectively, p = 0.006 and p = 0.01). In conclusion, endogenous IAP's protective effects in regard to the metabolic syndrome may be inhibited by PHE, a metabolite of ASP, perhaps explaining the lack of expected weight loss and metabolic improvements associated with diet drinks.

  19. Changes in morphology of actin filaments and expression of alkaline phosphatase at 3D cultivation of MG-63 osteoblast-like cells on mineralized fibroin scaffolds.

    Science.gov (United States)

    Goncharenko, A V; Malyuchenko, N V; Moisenovich, A M; Kotlyarova, M S; Arkhipova, A Yu; Kon'kov, A S; Agapov, I I; Molochkov, A V; Moisenovich, M M; Kirpichnikov, M P

    2016-09-01

    3D cultivation of MG-63 osteoblast-like cells on mineralized fibroin scaffolds leads to an increase in the expression of alkaline phosphatase, an early marker of bone formation. Increased expression is associated with the actin cytoskeleton reorganization under the influence of 3D cultivation and osteogenic calcium phosphate component of the microcarrier.

  20. Investigations into the stabilisation of drugs by sugar glasses : II: Delivery of an inulin-stabilised alkaline phosphatase in the intestinal lumen via the oral route

    NARCIS (Netherlands)

    Eriksson, H.J.C.; Verweij, W.R.; Poelstra, K.; Hinrichs, W.L.J.; de Jong, G.J.; Somsen, G.W.; Frijlink, H.W.

    2003-01-01

    In this study the possibility to deliver the acid-sensitive enzyme alkaline phosphatase (AP) from calf intestine (CIAP) to the intestinal system by oral administration was investigated. Tablets were prepared and in vitro evaluated. Final proof of concept studies were performed in rats. This acid lab

  1. ANALYSIS OF A CONJUGATE BETWEEN ANTICARCINOEMBRYONIC ANTIGEN MONOCLONAL-ANTIBODY AND ALKALINE-PHOSPHATASE FOR SPECIFIC ACTIVATION OF THE PRODRUG ETOPOSIDE PHOSPHATE

    NARCIS (Netherlands)

    Haisma, Hidde; BOVEN, E; VANMUIJEN, M; DEVRIES, R; PINEDO, HM

    1992-01-01

    The selective targeting of tumours by enzymes conjugated to monoclonal antibodies (mAb) may be an ideal approach to convert relatively nontoxic prodrugs into active agents at the tumour site. We used the anti-carcinoembryonic antigen mAb BW431/26 conjugated to alkaline phosphatase (AP) and phosphory

  2. Characterization of human placental alkaline phosphatase by activity and protein assays, capillary electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    NARCIS (Netherlands)

    Eriksson, H J; Somsen, G W; Hinrichs, W L; Frijlink, H W; de Jong, G J

    2001-01-01

    Placental alkaline phosphatase (PLAP) that had been isolated from human placenta was further purified using subsequent ion-exchange chromatography (IEC), affinity chromatography (AC) and centrifugal membrane concentration (CMC). During the process, the PLAP samples from the different stages of purif

  3. Effect of noise exposure (85 dB ) on testicular adrenocortical steroidogenic key enzymes, acid and alkaline phosphatase activities of sex organs in mature albino rats

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Changes in the activities of △5-3β-hydroysteroid dehydrogenase (HSD) in testis and adrenal gland, 17β-hydroxysteroid dehydrogenase in testis, acid and alkaline phosphatase in testis, prostate and seminal vesicle were observed in noise exposed mature rats at the intensity of 85 dB for 8 h/day for 45 days. The results indicated that noise exposed group showed a significant diminution in the activities of androgenic key enzymes △5-3β and 17β-HSD, acid phosphatase in testis, prostate and seminal vesicle. There was a significant elevation in the activities of adrenal △5-3β-HSD, alkaline phosphatase in testis and other accessory sex organ in noise exposed group. Gonadosomatic, prostatosomatic and seminal vesiculo-somatic indexes were decreased significantly in noise exposed group. Therefore, it is evident that noise exposure at 85dB exerts a deleterious effect on testicular and adrenocortical activities.

  4. High Temperature and Pressure Alkaline Electrolysis

    DEFF Research Database (Denmark)

    Allebrod, Frank

    and oxygen with a new type of alkaline electrolysis cell at high temperatures and pressures. To perform measurements under high pressure and at elevated temperatures it was necessary to build a measurement system around an autoclave which could stand high temperatures up to 250 °C and pressures up to 200 bar...... as well as extremely caustic environments. Based on a literature study to identify resistant materials for these conditions, Inconel 600 was selected among the metals which are available for autoclave construction. An initial single atmosphere high temperature and pressure measurement setup was build...... comprising this autoclave. A second high temperature and pressure measurement setup was build based on experiences from the first setup in order to perform automatized measurements. The conductivity of aqueous KOH at elevated temperatures and high concentrations was investigated using the van der Pauw method...

  5. Clinical significance of γ-glutamyl transferase,alkaline phosphatase,high-sensitivity C-reactive protein in elderly patients with subtrochanteric fractures%老年股骨粗隆骨折患者γ-谷氨酰转肽酶、碱性磷酸酶、超敏C反应蛋白联合检测的临床意义

    Institute of Scientific and Technical Information of China (English)

    王涵; 叶玉芬; 周建球; 姚荣忠; 谭放花

    2012-01-01

    目的 探讨老年股骨粗隆骨折患者γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)、超敏C反应蛋白(hs-CRP)联合检测的临床意义.方法 回顾性分析67例老年股骨粗隆骨折患者的GGT、ALP、hs-CRP水平,并研究术后不同时间点各指标相对于术前的变化情况及不同降低水平的预后情况.结果 老年股骨粗隆骨折患者的GGT、ALP及hs-CRP水平高于健康对照者和其他部位骨折患者,且与病理参数有关;术后呈降低的趋势;老年股骨粗隆骨折患者中三项指标术后水平降幅较大者的Harris髋关节功能评分累积优良率高于降幅者低者,差异有统计学意义(P<0.01).结论 监测老年股骨粗隆骨折患者的血清GGT、ALP及hs-CRP水平及其术后的变化情况具有重要的临床意义.%Objective To explore the clinical significance of y glutamyl transferase(GGT) ,alkaline phosphatase(ALP) and high sensitivity C reactive protein(hs CRP) detection in elderly patients with subtrochanteric fractures. Methods GGT, ALP and hs CRP levels in 67 elderly patients with subtrochanteric fractures were retrospectively analyzed,and the changes of them before oper ation and at different postoperative time points as well as the prognosis of reducing level were also analyzed. Results GGT,ALP and hs CRP levels in patients with subtrochanteric fractures were higher than in health people and patients with other fractures,and were related to the pathological parameters. The three indicators showed a decreasing trend after operation. The Harris hip score ac cumulated good rates in high level decreasing groups were higher than those in low level decreasing groups(P<0. 01). Conclusion GGT,ALP and hs CRP levels and changes before and after surgery in elderly patients with subtrochanteric fractures might be with important clinical significance.

  6. Enzymatic methods for the determination of pollution in seawater using salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius.

    Science.gov (United States)

    Menzorova, Natalie I; Seitkalieva, Alexandra V; Rasskazov, Valerу A

    2014-02-15

    A new salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius (StAP) has been shown to have a unique property to hydrolyze substrate in seawater without loss of enzymatic activity. The enzyme has pH optimum at 8.0-8.5. Model experiments showed various concentrations of copper, zinc, cadmium and lead added to seawater or a standard buffer mixture to inhibit completely the enzyme activity at the concentrations of 15-150 μg/l. StAP sensitivity to the presence in seawater of metals, pesticides, detergents and oil products appears to be considerably less. Samples of seawater taken from aquatic areas of the Troitsy Bay of the Peter the Great Bay, Japan Sea have been shown to inhibit the enzyme activity; the same was shown for the samples of fresh waters. The phosphatase inhibition assay developed proved to be highly sensitive, technically easy-to use allowing to test a great number of samples.

  7. Assessment of the colorimetric and fluorometric assays for alkaline phosphatase activity in cow's, goat's, and sheep's milk.

    Science.gov (United States)

    Klotz, V; Hill, Art; Warriner, K; Griffiths, M; Odumeru, J

    2008-09-01

    Raw milk is a well-established vehicle for the carriage of human pathogens, and many regulatory bodies have consequently mandated compulsory pasteurization as a food safety intervention. The residual activity of alkaline phosphatase (ALP) has historically been used to verify the adequacy of pasteurization of cow's milk. However, there is uncertainty on how the current ALP standards and methods of analysis can be applied to sheep's and goat's milk, which naturally contain different levels of the enzyme than that found in cow's milk. The official ALP methods applied in Canada (colorimetric assay; MFO-3) and in the United States (Fluorophos) were assessed for their ability to detect enzyme activity in raw and pasteurized milk derived from cows, sheep, and goats. The detection limit and the limit of quantitation were 0.8 and 2.02 microg/ml phenol, respectively, for the MFO-3 method and 43 and 85 mU/liter, respectively, for the Fluorophos method. The average ALP levels in raw goat's, cow's, and sheep's milk were 165, 1,562, and 3,512 microg/ml phenol, respectively. Raw milk detection limits, which correspond to raw milk phosphatase levels, were 0.051, 0.485, and 0.023% in cow's, goat's, and sheep's milk, respectively, for the MFO-3 method and 0.007, 0.070, and 0.004%, respectively, for the Fluorophos method. Although both methods can be used for ALP determination in cow's, goat's, and sheep's milk, the Fluorophos assay was superior to the colorimetric MFO-3 method based on sensitivity and time required to complete the analysis.

  8. High temperature and pressure alkaline electrolysis

    DEFF Research Database (Denmark)

    Allebrod, Frank; Chatzichristodoulou, Christodoulos; Mogensen, Mogens Bjerg

    2013-01-01

    and pressures. Two measurement systems were built to perform measurements under high pressures and at elevated temperatures of up to 95 bar and 250 °C, respectively. The conductivity of aqueous KOH and aqueous KOH immobilized in a porous SrTiO3 structure were investigated at elevated temperatures and high...... the operational temperature and pressure to produce pressurized hydrogen at high rate (m3 H2·h-1·m-2 cell area) and high electrical efficiency. This work describes an exploratory technical study of the possibility to produce hydrogen and oxygen with a new type of alkaline electrolysis cell at high temperatures...... concentrations of the electrolyte using the van der Pauw method in combination with electrochemical impedance spectroscopy (EIS). Conductivity values as high as 2.9 S cm-1 for 45 wt% KOH aqueous KOH and 0.84 S cm-1 for the immobilized KOH of the same concentration were measured at 200 °C. Porous SrTiO3 was used...

  9. Ultrasensitive detection of cancer cells and glycan expression profiling based on a multivalent recognition and alkaline phosphatase-responsive electrogenerated chemiluminescence biosensor

    Science.gov (United States)

    Chen, Xiaojiao; He, Yao; Zhang, Youyu; Liu, Meiling; Liu, Yang; Li, Jinghong

    2014-09-01

    A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode interface, affording fast and highly sensitive ECL cytosensing and cell surface glycan evaluation. Combining the multivalent aptamer interface and ALP nanoprobes, the ECL cytosensor showed a detection limit of 38 CCRF-CEM cells per mL in human serum samples, broad dynamic range and excellent selectivity. In addition, the proposed biosensor provided a valuable insight into dynamic profiling of the expression of different glycans on cell surfaces, based on the carbohydrates recognized by lectins applied to the nanoprobes. This biosensor exhibits great promise in clinical diagnosis and drug screening.A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode

  10. Intestinal Alkaline Phosphatase: Potential Roles in Promoting Gut Health in Weanling Piglets and Its Modulation by Feed Additives - A Review.

    Science.gov (United States)

    Melo, A D B; Silveira, H; Luciano, F B; Andrade, C; Costa, L B; Rostagno, M H

    2016-01-01

    The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP) is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS), unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs) activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria) detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP's role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets.

  11. The relationship between the alkaline phosphatase network and the haematopoiesis in mice subjected to whole-body irradiation

    Directory of Open Access Journals (Sweden)

    Almohamad Khaled M.

    2014-08-01

    Full Text Available Purpose: To investigate the relationship between the alkaline phosphatase (ALP network of the marrow stroma and the haematopoietic regeneration after mice whole-body irradiation. Materials and methods: Three groups of mice were irradiated with a non-lethal ionising radiation dose: the fi rst one received an intraperitoneal injection of Levamisole, ALP inhibitor, 24 h before irradiation; the second one received an intraperitoneal injection of Lisinopril, haematopoiesis inhibitor, 24 h before irradiation; the third was left untreated, but irradiated. The fourth group, untreated and not irradiated, was the control. The total surface occupied by ALP positive processes, revealed by means of ALP cytochemistry in the marrow area, was evaluated semi-quantitively. Nucleated bone marrow cells were also counted. Results: ALP network began to increase 24 h after irradiation to reach a maximum after 72 h, when the bone marrow was almost become completely empty of the haematopoietic cells. This increase advances the haematopoietic recovery. This process was substantially delayed when the mice were injected with Levamisole 24 h before irradiation. On the contrary, ALP network increased strongly since the fi rst day after irradiation when the mice were injected with Lisinopril 24 h before irradiation. Conclusions: These data have indicated that the haematopoietic recovery and repopulation of the bone marrow were advanced by the ALP network recovery.

  12. Ultra-sensitive conductometric detection of heavy metals based on inhibition of alkaline phosphatase activity from Arthrospira platensis.

    Science.gov (United States)

    Tekaya, Nadèje; Saiapina, Olga; Ben Ouada, Hatem; Lagarde, Florence; Ben Ouada, Hafedh; Jaffrezic-Renault, Nicole

    2013-04-01

    This study is based on the conductometric measurement of alkaline phosphatase activity (APA) from the cyanobacterium, Arthrospira platensis, called Spirulina. Cyanobacterium cells were directly immobilized, by physical adsorption, on the ceramic part of gold interdigitated transducers. This activity was inhibited in the presence of heavy metals and a variation of the local conductivity was measured after addition of the substrate. The Michaelis-Menten constant (Km) was evaluated to be 0.75 mM through a calibration curve of the substrate, disodium 4-nitrophenylphosphate p-nitrophenyl phosphate (pNPP). Inhibition of APA was observed with cadmium and mercury with a detection limit of 10(-20) M. The half maximal inhibitory concentration (IC50) was determined at 10(-19) M for Cd(2+) and 10(-17) M for Hg(2+), and the binding affinity of heavy metal (Ki) was equal to the IC50. On the sensor surface, scanning electron microscopy (SEM) images revealed a remarkable evolution of the cyanobacterium's external surface that was attributable to the first defense mechanism against toxic heavy metals in trace. This effect was also confirmed through the important increase of response time τ(90%) recorded for APA response towards the substrate pNPP after cell exposure to metallic cations. Lifetime of the Spirulina-based biosensor was estimated to be more than 25 days.

  13. Assessment of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase activities in cow's milk as an indicator of subclinical mastitis.

    Science.gov (United States)

    Babaei, H; Mansouri-Najand, L; Molaei, M M; Kheradmand, A; Sharifan, M

    2007-05-01

    This study examined the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in the milk of lactating Holstein cows in association with subclinical mastitis (SCM). A total of 94 milk samples were collected from 58 lactating dairy cows representing stages of lactation from the second to the tenth week after calving. Those which were classified as positive by California mastitis test (CMT) were deemed to have subclinical mastitis. All the milk samples were skimmed by centrifugation at 10 000g at 0 degrees C and were used for enzyme activities estimations. The mean activities of LDH and ALP were higher in the milk from udders with SCM than in the milk from healthy udders (p CMT results and LDH and ALP values were seen at thresholds of > 180 IU/L and > 40 IU/L respectively (kappa values 0.65 and 0.79, respectively). However, the sensitivity of the tests for identifying SCM at these thresholds was higher for ALP (96.4%) than for LDH (68.5%). In this study, LDH and ALP tests were standardized for cow's milk and results showed that only the ALP test was reliable in the early diagnosis of subclinical mastitis.

  14. Yam (Dioscorea batatas) Root and Bark Extracts Stimulate Osteoblast Mineralization by Increasing Ca and P Accumulation and Alkaline Phosphatase Activity.

    Science.gov (United States)

    Kim, Suji; Shin, Mee-Young; Son, Kun-Ho; Sohn, Ho-Yong; Lim, Jae-Hwan; Lee, Jong-Hwa; Kwun, In-Sook

    2014-09-01

    Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within 0~10 μg/mL during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineralization (16~20 day) as appropriate. In study I, both yam root water and methanol extracts increased cell proliferation as concentration-dependent manner. Cellular collagen synthesis and alkaline phosphatase (ALP) activity, both the indicators of bone matrix protein and inorganic phosphate production for calcification respectively, were also increased by yam root water and methanol extract. Osteoblast calcification as cell matrix Ca and P accumulation was also increased by the addition of yam root extracts. In study II, yam bark extracts (water and hexane) increased osteoblast proliferation and differentiation, as collagen synthesis and ALP activity and osteoblast matrix Ca and P deposition. The study results suggested that both yam root and bark extracts stimulate osteogenic function in osteoblasts by stimulating bone matrix maturation by increasing collagen synthesis, ALP activity, and matrix mineralization.

  15. Effects of synthetic retinoids and retinoic acid isomers on the expression of alkaline phosphatase in F9 teratocarcinoma cells.

    Science.gov (United States)

    Gianni, M; Zanotta, S; Terao, M; Garattini, S; Garattini, E

    1993-10-15

    Expression of ALP in F9 teratocarcinoma cells is induced by all-trans retinoic acid (ATRA) (Gianni' et al., Biochem. J. 274: 673-678, 1991). The specific ligand for retinoic acid related receptors (RXRs), 9-cis retinoic acid (9-cis RA), and three synthetic analogs binding to the alpha, beta and gamma forms of the retinoic acid receptors (RARs), AM580, CD2019, and CD437, were used to study their effects on alkaline phosphatase (ALP) enzymatic activity and mRNA levels. At concentrations close to the Kd for their respective receptors, 9-cis RA, AM580 (the RAR alpha agonist) and CD437 (the RAR gamma agonist) clearly upregulate the expression of the ALP gene, whereas the effect of CD2019 (the RAR beta agonist) is very modest. A specific inhibitor of the RAR alpha, Ro 41-5253, completely blocks the induction of ALP triggered by AM580, while it has minor effects on the upregulation caused by ATRA, 9-cis RA, CD437 and CD2019. The induction of ALP observed with the various retinoids is inhibited by the contemporaneous treatment with dibutyryl cAMP. The levels of the RAR alpha and gamma transcripts are unaltered, while RAR beta mRNAs are induced by ATRA, AM580, CD437 and to a lower extent by 9-cis RA and CD2019.

  16. In vitro induction of alkaline phosphatase levels predicts in vivo bone forming capacity of human bone marrow stromal cells

    Directory of Open Access Journals (Sweden)

    Henk-Jan Prins

    2014-03-01

    Full Text Available One of the applications of bone marrow stromal cells (BMSCs that are produced by ex vivo expansion is for use in in vivo bone tissue engineering. Cultured stromal cells are a mixture of cells at different stages of commitment and expansion capability, leading to a heterogeneous cell population that each time can differ in the potential to form in vivo bone. A parameter that predicts for in vivo bone forming capacity is thus far lacking. We employed single colony-derived BMSC cultures to identify such predictive parameters. Using limiting dilution, we have produced sixteen single CFU-F derived BMSC cultures from human bone marrow and found that only five of these formed bone in vivo. The single colony-derived BMSC strains were tested for proliferation, osteogenic-, adipogenic- and chondrogenic differentiation capacity and the expression of a variety of associated markers. The only robust predictors of in vivo bone forming capacity were the induction of alkaline phosphatase, (ALP mRNA levels and ALP activity during in vitro osteogenic differentiation. The predictive value of in vitro ALP induction was confirmed by analyzing “bulk-cultured” BMSCs from various bone marrow biopsies. Our findings show that in BMSCs, the additional increase in ALP levels over basal levels during in vitro osteogenic differentiation is predictive of in vivo performance.

  17. Cellular response to zinc-containing organoapatite: an in vitro study of proliferation, alkaline phosphatase activity and biomineralization.

    Science.gov (United States)

    Storrie, Hannah; Stupp, Samuel I

    2005-09-01

    We present a series of experiments investigating the in vitro biological activity of zinc-containing organoapatite (ZnOA)-coated titanium meshes. ZnOA is a hydroxyapatite-based material that contains poly(l-lysine) and zinc ions and can be coated onto titanium by treating the metal surface with poly(amino acids) that allow for electrostatic bonding of the mineral to the titanium surface. Preosteoblastic mouse calyaria cells were cultured on ZnOA-coated titanium meshes in a three-dimensional (3D) bioreactor, which provides an in vitro culture environment that better simulates what cells experience in vivo, compared to traditional 2D cultures. Results of these studies show a time-dependent cascade of events leading to an earlier onset of alkaline phosphatase (ALP) expression and biomineralization of ZnOA-coated samples as compared to controls. After the observation of peak ALP levels in ZnOA-coated titanium samples, mineralized bone nodules were observed by scanning electron microscopy. Tetracycline staining confirmed that the observed mineral nodules were newly synthesized biomineral, and not due to the inorganic coating. ZnOA-coated titanium substrates represent a new class of materials for human repair that provide, mechanical stability, as well as chemical and biochemical signals to promote new bone growth.

  18. Serum total and bone alkaline phosphatase levels and their correlation with serum minerals over the lifespan of sheep.

    Science.gov (United States)

    Sousa, Cristina P; Azevedo, Jorge T; Silva, Amélia M; Viegas, Carlos A; Reis, Rui L; Gomes, Manuela E; Dias, Isabel R

    2014-06-01

    This study aimed to assess serum total alkaline phosphatase (ALP) and its bone isoform (BALP) levels during the ageing and in different physiologic states of sheep, in order to expand the knowledge about the variation of these biomarkers over the sheep lifespan. Ninety female sheep were divided into nine groups of various ages and physiological states (dry, lactation and pregnancy). Serum ALP, BALP and mineral levels were determined by commercial immunoassay, molecular absorbance spectrophotometry and chemical luminescence for BALP determination. Serum ALP and BALP decreased as sheep aged, and no statistically significant differences were obtained between ewes in different physiologic states. The continuous decline of serum BALP concentration along the sheep lifespan, namely in mature and old sheep, is a sign of decreasing bone turnover associated with ageing. Serum calcium concentrations increased slightly until 2 years of age and then showed a tenuous but statistically significant decrease in mature sheep, while serum phosphorus maintained an uninterrupted decrease as sheep matured. The knowledge of serum values of bone biomarkers throughout the sheep lifespan may be useful in preclinical orthopaedic research studies and for animal science studies using sheep.

  19. Valproic Acid Induces Hair Regeneration in Murine Model and Activates Alkaline Phosphatase Activity in Human Dermal Papilla Cells

    Science.gov (United States)

    Lee, Soung-Hoon; Yoon, Juyong; Shin, Seung Ho; Zahoor, Muhamad; Kim, Hyoung Jun; Park, Phil June; Park, Won-Seok; Min, Do Sik; Kim, Hyun-Yi; Choi, Kang-Yell

    2012-01-01

    Background Alopecia is the common hair loss problem that can affect many people. However, current therapies for treatment of alopecia are limited by low efficacy and potentially undesirable side effects. We have identified a new function for valproic acid (VPA), a GSK3β inhibitor that activates the Wnt/β-catenin pathway, to promote hair re-growth in vitro and in vivo. Methodology/ Principal Findings Topical application of VPA to male C3H mice critically stimulated hair re-growth and induced terminally differentiated epidermal markers such as filaggrin and loricrin, and the dermal papilla marker alkaline phosphatase (ALP). VPA induced ALP in human dermal papilla cells by up-regulating the Wnt/β-catenin pathway, whereas minoxidil (MNX), a drug commonly used to treat alopecia, did not significantly affect the Wnt/β-catenin pathway. VPA analogs and other GSK3β inhibitors that activate the Wnt/β-catenin pathway such as 4-phenyl butyric acid, LiCl, and BeCl2 also exhibited hair growth-promoting activities in vivo. Importantly, VPA, but not MNX, successfully stimulate hair growth in the wounds of C3H mice. Conclusions/ Significance Our findings indicate that small molecules that activate the Wnt/β-catenin pathway, such as VPA, can potentially be developed as drugs to stimulate hair re-growth. PMID:22506014

  20. In vitro response to alkaline phosphatase coatings immobilized onto titanium implants using electrospray deposition or polydopamine-assisted deposition.

    Science.gov (United States)

    Nijhuis, Arnold W G; van den Beucken, Jeroen J J P; Jansen, John A; Leeuwenburgh, Sander C G

    2014-04-01

    Immobilization of biomolecules onto implant surfaces is one of the most straightforward strategies to control the interaction between an implant and its biological environment. Recently, it was shown that the enzyme alkaline phosphatase (ALP) could be efficiently immobilized onto titanium implants in a single step using polydopamine. We hypothesized that such polydopamine-ALP coatings can enhance the early attachment of cells and increase mineralization. Therefore, the current study aimed at immobilization of ALP onto titanium by means of either one- or two-step polydopamine-assisted immobilization or electrospray deposition, the comparative characterization of these experimental substrates and subsequent cell behavioral analysis using primary osteoblast-like cells. Uncoated titanium and ALP-free polydopamine coatings served as controls. Despite significant ALP surface activity and lower water contact for angles ALP-containing surface modifications, only marginal effects on early cell behavior (i.e., cell spreading) and osteogenic differentiation (i.e., proliferation, differentiation and mineralization) were observed in comparison to uncoated titanium. Copyright © 2013 Wiley Periodicals, Inc.

  1. Low dietary copper increases fecal free radical production, fecal water alkaline phosphatase activity and cytotoxicity in healthy men.

    Science.gov (United States)

    Davis, Cindy D

    2003-02-01

    One possible dietary factor that may increase susceptibility to colon cancer is inadequate copper intake. The objective of this study was to investigate the effects of low and adequate copper intakes on copper nutriture and putative risk factors for colon cancer susceptibility in healthy men. Seventeen healthy free-living nonsmoking men aged 21-52 y completed a 13-wk controlled feeding study in a randomized crossover design. The basal diet contained 0.59 mg Cu/13.65 MJ. After a 1-wk equilibration period in which the men consumed the basal diet supplemented with 1.0 mg Cu/d, they were randomly assigned to receive either the basal diet or the basal diet supplemented with 2 mg Cu/d for 6 wk. After the first dietary period, the men immediately began to consume the other level of Cu for the last 6 wk. They collected their feces during the equilibration period and during the last 2 wk of the two dietary periods for free radical and fecal water analysis. Low dietary copper significantly (P copper significantly (P copper concentrations but did not affect fecal water volume, pH, iron or zinc concentrations. In contrast to the fecal analysis, hematological indicators of copper status were not significantly affected by the dietary treatments. These results suggest that low dietary copper adversely affects fecal free radical production and fecal water alkaline phosphatase activity, which are putative risk factors for colon cancer.

  2. Valproic acid induces hair regeneration in murine model and activates alkaline phosphatase activity in human dermal papilla cells.

    Directory of Open Access Journals (Sweden)

    Soung-Hoon Lee

    Full Text Available BACKGROUND: Alopecia is the common hair loss problem that can affect many people. However, current therapies for treatment of alopecia are limited by low efficacy and potentially undesirable side effects. We have identified a new function for valproic acid (VPA, a GSK3β inhibitor that activates the Wnt/β-catenin pathway, to promote hair re-growth in vitro and in vivo. METHODOLOGY/ PRINCIPAL FINDINGS: Topical application of VPA to male C3H mice critically stimulated hair re-growth and induced terminally differentiated epidermal markers such as filaggrin and loricrin, and the dermal papilla marker alkaline phosphatase (ALP. VPA induced ALP in human dermal papilla cells by up-regulating the Wnt/β-catenin pathway, whereas minoxidil (MNX, a drug commonly used to treat alopecia, did not significantly affect the Wnt/β-catenin pathway. VPA analogs and other GSK3β inhibitors that activate the Wnt/β-catenin pathway such as 4-phenyl butyric acid, LiCl, and BeCl(2 also exhibited hair growth-promoting activities in vivo. Importantly, VPA, but not MNX, successfully stimulate hair growth in the wounds of C3H mice. CONCLUSIONS/ SIGNIFICANCE: Our findings indicate that small molecules that activate the Wnt/β-catenin pathway, such as VPA, can potentially be developed as drugs to stimulate hair re-growth.

  3. Kinetic Isotope Effects for Alkaline Phosphatase Reactions: Implications for the Role of Active Site Metal Ions in Catalysis

    Science.gov (United States)

    Zalatan, Jesse G.; Catrina, Irina; Mitchell, Rebecca; Grzyska, Piotr K.; O’Brien, Patrick J.; Herschlag, Daniel; Hengge, Alvan C.

    2011-01-01

    Enzyme-catalyzed phosphoryl transfer reactions have frequently been suggested to proceed through transition states that are altered from their solution counterparts, with the alterations presumably arising from interactions with active site functional groups. In particular, the phosphate monoester hydrolysis reaction catalyzed by Escherichia coli alkaline phosphatase (AP) has been the subject of intensive scrutiny. Recent linear free energy relationship (LFER) studies suggest that AP catalyzes phosphate monoester hydrolysis through a loose transition state, similar to that in solution. To gain further insight into the nature of the transition state and active site interactions, we have determined kinetic isotope effects (KIEs) for AP-catalyzed hydrolysis reactions with several phosphate monoester substrates. The LFER and KIE data together provide a consistent picture for the nature of the transition state for AP-catalyzed phosphate monoester hydrolysis and support previous models suggesting that the enzymatic transition state is similar to that in solution. Moreover, the KIE data provides unique information regarding specific interactions between the transition state and the active site Zn2+ ions. These results provide strong support for a model in which electrostatic interactions between the bimetallo Zn2+ site and a nonbridging phosphate ester oxygen atom make a significant contribution to the large rate enhancement observed for AP-catalyzed phosphate monoester hydrolysis. PMID:17630738

  4. Kinetic isotope effects for alkaline phosphatase reactions: implications for the role of active-site metal ions in catalysis.

    Science.gov (United States)

    Zalatan, Jesse G; Catrina, Irina; Mitchell, Rebecca; Grzyska, Piotr K; O'brien, Patrick J; Herschlag, Daniel; Hengge, Alvan C

    2007-08-08

    Enzyme-catalyzed phosphoryl transfer reactions have frequently been suggested to proceed through transition states that are altered from their solution counterparts, with the alterations presumably arising from interactions with active-site functional groups. In particular, the phosphate monoester hydrolysis reaction catalyzed by Escherichia coli alkaline phosphatase (AP) has been the subject of intensive scrutiny. Recent linear free energy relationship (LFER) studies suggest that AP catalyzes phosphate monoester hydrolysis through a loose transition state, similar to that in solution. To gain further insight into the nature of the transition state and active-site interactions, we have determined kinetic isotope effects (KIEs) for AP-catalyzed hydrolysis reactions with several phosphate monoester substrates. The LFER and KIE data together provide a consistent picture for the nature of the transition state for AP-catalyzed phosphate monoester hydrolysis and support previous models suggesting that the enzymatic transition state is similar to that in solution. Moreover, the KIE data provides unique information regarding specific interactions between the transition state and the active-site Zn2+ ions. These results provide strong support for a model in which electrostatic interactions between the bimetallo Zn2+ site and a nonbridging phosphate ester oxygen atom make a significant contribution to the large rate enhancement observed for AP-catalyzed phosphate monoester hydrolysis.

  5. The role of alkaline phosphatase in intracellular lipid accumulation in the human hepatocarcinoma cell line, HepG2.

    Science.gov (United States)

    Chirambo, George M; van Niekerk, Chantal; Crowther, Nigel J

    2017-04-01

    Inhibition of tissue non-specific alkaline phosphatase (TNALP) decreases intracellular lipid accumulation in human preadipocytes and the murine preadipocyte cell line, 3T3-L1. Therefore, the current study was performed to determine if TNALP is required for intracellular lipid deposition in the human hepatocyte cell line, HepG2. Intracellular lipid accumulation, TNALP activity and peroxisome proliferator activated receptor (PPAR) γ gene expression were measured in HepG2 and 3T3-L1 cells in the presence and absence of the TNALP inhibitors levamisole and histidine. Sub-cellular TNALP activity was localized using cytochemical analysis. Both PPARγ gene expression and TNALP activity increased during intracellular lipid accumulation in HepG2 and 3T3-L1 cells. Inhibition of TNALP blocked intracellular lipid accumulation but did not alter expression of the PPARγ gene. In HepG2 cells, TNALP co-localized with adipophilin on the lipid droplet membrane. These data suggest a role for TNALP in lipid droplet formation, possibly downstream from PPARγ, within HepG2 and 3T3-L1 cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Synthesis of 2'(3')-O-DL-alanyl hexainosinic acid using T4 RNA ligase: suppression of the enzymic reverse transfer reaction by alkaline phosphatase.

    Science.gov (United States)

    Profy, A T; Lo, K M; Usher, D A

    1983-03-11

    2'(3')-O-DL-Alanyl (Ip)5I was synthesized by a new method. An alanine ortho ester of inosine 5'-phosphate was added to (Ip)4I using the ATP-independent reaction of T4 RNA ligase, and the product was converted smoothly to the desired ester. The enzymic reverse transfer reaction was conveniently suppressed by the dephosphorylation of the adenosine 5'-phosphate coproduct, catalyzed in situ by alkaline phosphatase.

  7. Resistance imparted by traditional Chinese medicines to the acute change of glutamic pyruvic transaminase, alkaline phosphatase and creatine kinase activities in rat blood caused by noise.

    Science.gov (United States)

    Zhu, Bei-Wei; Sun, Yu-Mei; Yun, Xia; Han, Song; Piao, Mei-Lan; Murata, Yoshiyuki; Tada, Mikiro

    2004-05-01

    The activities of serum glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP) and creatine kinase (CK) in rats injected or not with the Chinese medicines, Astragali, Rhodiolae and Ligusticum, were determined after noise exposure. Noise at 95 and 105 dB significantly increased the activities of GPT, ALP and CK, and showed a dependence on the exposure time. The injection of each medicine significantly suppressed the increased enzyme activities by 95 and 105 dB noise.

  8. Evaluation of alkaline phosphatase activity and availability of various P fractions for bean (Phaseolus vulgaris in some calcareous soils amended with municipal sewage sludge

    Directory of Open Access Journals (Sweden)

    T. Raeisi

    2014-07-01

    Full Text Available To evaluate the relationship of various P fractions and alkaline phosphatase activity with bean indices growing in 10 calcareous soils, amended with municipal sewage sludge from Chaharmahal-Va-Bakhtiari province, a greenhouse research was carried out. Soil samples were incubated for one month with sludge at a rate equivalent to 1% (w/w. Then, the P fractions, including P adsorbed by Fe and Al oxides (]NaOH+CB]-P, occluded P (CBD-P and P absorbed by Ca (HCl-P, were determined by Olsen and Summers' sequential fractionation procedure. Furthermore, total P, organic P and residual P were determined. Also, alkaline phosphatase activity was measured. A pot experiment in a completely randomized design with three replications in the ten soils was done to evaluate the bean plant indices. The results showed that the amount of P fractions decreased in the following order: HCl-P>residual-P>]NaOH+CB]-P > OP>CBD-P. The results also indicated that alkaline phosphatase activity was significantly correlated with CBD-P fraction, organic P and total P. In addition, significant correlations were found between ([NaOH+CB]-P and HCl-P and plant shoots. In general, the results of this research showed that P fractionation method appears to be a powerful tool to identify the P status and availability in the soils amended with sewage sludge.

  9. Microbial thiocyanate utilization under highly alkaline conditions.

    Science.gov (United States)

    Sorokin, D Y; Tourova, T P; Lysenko, A M; Kuenen, J G

    2001-02-01

    Three kinds of alkaliphilic bacteria able to utilize thiocyanate (CNS-) at pH 10 were found in highly alkaline soda lake sediments and soda soils. The first group included obligate heterotrophs that utilized thiocyanate as a nitrogen source while growing at pH 10 with acetate as carbon and energy sources. Most of the heterotrophic strains were able to oxidize sulfide and thiosulfate to tetrathionate. The second group included obligately autotrophic sulfur-oxidizing alkaliphiles which utilized thiocyanate nitrogen during growth with thiosulfate as the energy source. Genetic analysis demonstrated that both the heterotrophic and autotrophic alkaliphiles that utilized thiocyanate as a nitrogen source were related to the previously described sulfur-oxidizing alkaliphiles belonging to the gamma subdivision of the division Proteobacteria (the Halomonas group for the heterotrophs and the genus Thioalkalivibrio for autotrophs). The third group included obligately autotrophic sulfur-oxidizing alkaliphilic bacteria able to utilize thiocyanate as a sole source of energy. These bacteria could be enriched on mineral medium with thiocyanate at pH 10. Growth with thiocyanate was usually much slower than growth with thiosulfate, although the biomass yield on thiocyanate was higher. Of the four strains isolated, the three vibrio-shaped strains were genetically closely related to the previously described sulfur-oxidizing alkaliphiles belonging to the genus Thioalkalivibrio. The rod-shaped isolate differed from the other isolates by its ability to accumulate large amounts of elemental sulfur inside its cells and by its ability to oxidize carbon disulfide. Despite its low DNA homology with and substantial phenotypic differences from the vibrio-shaped strains, this isolate also belonged to the genus Thioalkalivibrio according to a phylogenetic analysis. The heterotrophic and autotrophic alkaliphiles that grew with thiocyanate as an N source possessed a relatively high level of cyanase

  10. A fluorometric assay for alkaline phosphatase activity based on β-cyclodextrin-modified carbon quantum dots through host-guest recognition.

    Science.gov (United States)

    Tang, Cong; Qian, Zhaosheng; Huang, Yuanyuan; Xu, Jiamin; Ao, Hang; Zhao, Meizhi; Zhou, Jin; Chen, Jianrong; Feng, Hui

    2016-09-15

    A convenient, reliable and highly sensitive assay for alkaline phosphatase (ALP) activity in the real-time manner is developed based on β-cyclodextrin-modified carbon quantum dots (β-CD-CQDs) nanoprobe through specific host-guest recognition. Carbon quantum dots were first functionalized with 3-aminophenyl boronic acid to produce boronic acid-functionalized CQDs, and then further modified with hydropropyl β-cyclodextrins (β-CD) through B-O bonds to form β-CD-CQDs nanoprobe. p-Nitrophenol phosphate disodium salt is used as the substrate of ALP, and can hydrolyze to p-nitrophenol under the catalysis of ALP. The resulting p-nitrophenol can enter the cavity of β-CD moiety in the nanoprobe due to their specific host-guest recognition, where photoinduced electron transfer process between p-nitrophenol and CQDs takes place to efficiently quench the fluorescence of the probe. The correlation between quenched fluorescence and ALP level can be used to establish quantitative evaluation of ALP activity in a broad range from 3.4 to 100.0U/L with the detection limit of 0.9U/L. This assay shows a high sensitivity to ALP even in the presence of a very high concentration of glucose. This study demonstrates a good electron donor/acceptor pair, which can be used to design general detection strategy through PET process, and also broadens the application of host-guest recognition for enzymes detection in clinical practice.

  11. Variaciones de la enzima fosfatasa alcalina en la pulpa dental Variations of alkaline phosphatase enzyme in the dental pulp

    Directory of Open Access Journals (Sweden)

    Zoraida Pons Pinillos

    2005-08-01

    Full Text Available En las últimas décadas, numerosas investigaciones se han dedicado al estudio de los mecanismos potenciales implicados en el desarrollo de la caries dental y su prevención, sin embargo, a pesar de haber disminuido gradualmente el índice de caries en la población, son muchos los pacientes que necesitan tratarse la caries dental, tal es así que continuamente se están utilizando diferentes materiales en la búsqueda de aquel que ante una agresión a la pulpa, ayude a una respuesta biológica de la misma, conservando de esta forma su integridad. De ahí la importancia de la actividad de la fosfatasa alcalina de la pulpa en el proceso carioso, como una reacción ante el hidróxido de calcio que continuamente se está usando en toda la red docente-asistencial del país. Se seleccionaron 50 dientes monorradiculares, con pulpa viva y con caries de segundo, tercer y cuarto grado y 50 dientes sanos de pacientes de diferentes edades. Se extrajo la pulpa de cada diente y se realizaron improntas (3 por cada muestra, una de las cuales se procesó para obtener orientación morfológica, y las otras 2 para valorar la actividad de la fosfatasa alcalina. Para esto se utilizaron 2 métodos: el de calcio cobalto y el de alpha naftol fosfato de Gomori. Como resultado, se obtuvo que la pulpa tiene más actividad enzimática en caries profunda y que la edad del paciente no determina el aumento o disminución de dicha actividad.In the last decades, numerous investigations have been made on the study of potential mechanisms involved in the development of dental caries and their prevention. However, in spite of the gradual reduction of dental caries in the population, a lot of patients need to have their dental caries treated and different materials are continuously used searching for one that before the aggression to the pulp helps it to give a biological response, conserving this way its integrity. That's why the activity of the alkaline phosphatase of the pulp

  12. Targeting the active site of the placental isozyme of alkaline phosphatase by phage-displayed scFv antibodies selected by a specific uncompetitive inhibitor

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    Kala Mrinalini

    2005-12-01

    Full Text Available Abstract Background The isozymes of alkaline phosphatase, the tissue non-specific, intestinal and placental, have similar properties and a high degree of identity. The placental isozyme (PLAP is an oncofetal antigen expressed in several malignancies including choriocarcinoma, seminoma and ovarian carcinoma. We had earlier attempted to isolate PLAP-specific scFv from a synthetic human immunoglobulin library but were unable to do so, presumably because of the similarity between the isozymes. In this work, we have employed a PLAP-specific uncompetitive inhibitor, L-Phe-Gly-Gly, to select isozyme specific scFvs. An uncompetitive inhibitor binds to the enzyme in the presence of substrate and stabilizes the enzyme-substrate complex. Several uncompetitive inhibitors have varying degrees of isozyme specificity for human alkaline phosphatase isozymes. A specific uncompetitive inhibitor would be able to unmask conformational differences between the otherwise very similar molecules. Also, such inhibitors would be directed to regions at/close to the active site of the enzyme. In this work, the library was first incubated with PLAP and the bound clones then eluted by incubation with L-Phe-Gly-Gly along with the substrate, para-nitro phenyl phosphate (pNPP. The scFvs were then studied with regard to the biochemical modulation of their binding, isozyme specificity and effect on enzyme activity. Results Of 13 clones studied initially, the binding of 9 was inhibited by L-Phe-Gly-Gly (with pNPP and 2 clones were inhibited by pNPP alone. Two clones had absolute and 2 clones had partial specificity to PLAP. Two clones were cross-reactive with only one other isozyme. Three scFv clones, having an accessible His6-tag, were purified and studied for their modulation of enzyme activity. All the three scFvs inhibited PLAP activity with the kinetics of competitive inhibition. Cell ELISA could demonstrate binding of the specific scFvs to the cell surface expressed PLAP

  13. Unraveling the Alkaline Phosphatase Inhibition, Anticancer, and Antileishmanial Potential of Coumarin-Triazolothiadiazine Hybrids: Design, Synthesis, and Molecular Docking Analysis.

    Science.gov (United States)

    Ibrar, Aliya; Zaib, Sumera; Jabeen, Farukh; Iqbal, Jamshed; Saeed, Aamer

    2016-07-01

    A series of new coumarin-triazolothiadiazine hybrid compounds (5a-j) was designed and synthesized by using the molecular hybridization concept. The cyclocondensation reaction involves the coumarinyl 4-amino-1,2,4-triazole and a range of bromo-acetophenones, delivering the desired products in good yields. The structures of the synthesized compounds were established on the basis of spectro-analytical data. The prepared compounds were evaluated against alkaline phosphatase (ALP) where compound 5j incorporating bis-coumarinyl motifs at the 3- and 6-positions of the heteroaromatic core turned out to be a potent inhibitor with an IC50 value of 1.15 ± 1.0 µM. The synthesized compounds were also tested against Leishmania major and 5h was the lead member with an IC50 value of 0.89 ± 0.08 μM. Anticancer activity was also determined using kidney fibroblast (BHK-21) and lung carcinoma (H-157) cancer cell lines. Compound 5i showed highest cytotoxic potential against H-157 cells with an IC50 value of 1.01 ± 0.12 μM, which is an improved inhibition compared to the standards (vincristine and cisplatin) used in this assay. Molecular docking studies were carried out on the synthesized library of coumarin-triazolothiadiazine hybrids against ALP. Almost all of the compounds showed strong interactions with the key residues of the active site of the receptor. In case of compounds 5a-c, 5h, and 5j, docking results positively complemented the experimental screening. These results provided substantial evidence for the further development of these compounds as potent inhibitors of ALP.

  14. Effects of Dihydroartemisinin and Artemether on the Growth, Chlorophyll Fluorescence, and Extracellular Alkaline Phosphatase Activity of the Cyanobacterium Microcystis aeruginosa

    Science.gov (United States)

    Wang, Shoubing; Xu, Ziran

    2016-01-01

    Increased eutrophication in the recent years has resulted in considerable research focus on identification of methods for preventing cyanobacterial blooms that are rapid and efficient. The objectives of this study were to investigate the effects of dihydroartemisinin and artemether on the growth of Microcystis aeruginosa and to elucidate its mode of action. Variations in cell density, chlorophyll a, soluble protein, malondialdehyde, extracellular alkaline phosphatase activity (APA), and chlorophyll fluorescence parameters (Fv/Fm, ΦPSII, ETR, rapid light curves, fast chlorophyll fluorescence curves on fluorescence intensity, and relative variable fluorescence) were evaluated by lab-cultured experiments. Our results demonstrated that both dihydroartemisinin and artemether inhibited the growth of M.aeruginosa by impairing the photosynthetic center in photosystem II and reducing extracellular APA, with a higher sensitivity exhibited toward artemether. The inhibitory effects of dihydroartemisinin on M.aeruginosa increased with concentration, and the maximum growth inhibitory rate was 42.17% at 24 mg·L-1 after 120h exposure, whereas it was 55.72% at 6 mg·L-1 artemetherafter 120h exposure. Moreover, the chlorophyll fluorescence was significantly inhibited (p<0.05) after 120h exposure to 12 and 24 mg·L-1 dihydroartemisinin. Furthermore, after 120h exposure to 6 mg·L-1 artemether, Fv/Fm, ΦPSII, ETR and rETRmax showed a significant decrease (p<0.01) from initial values of 0.490, 0.516, 17.333, and 104.800, respectively, to 0. One-way analysis of variance showed that 6 mg·L-1 artemether and 24 mg·L-1 dihydroartemisinin had significant inhibitory effects on extracellular APA (p<0.01). The results of this study would be useful to further studies to validate the feasibility of dihydroartemisinin and artemether treatment to inhibit overall cyanobacterial growth in water bodies, before this can be put into practice. PMID:27755566

  15. Probing the origin of the compromised catalysis of E. coli alkaline phosphatase in its promiscuous sulfatase reaction.

    Science.gov (United States)

    Catrina, Irina; O'Brien, Patrick J; Purcell, Jamie; Nikolic-Hughes, Ivana; Zalatan, Jesse G; Hengge, Alvan C; Herschlag, Daniel

    2007-05-02

    The catalytic promiscuity of E. coli alkaline phosphatase (AP) and many other enzymes provides a unique opportunity to dissect the origin of enzymatic rate enhancements via a comparative approach. Here, we use kinetic isotope effects (KIEs) to explore the origin of the 109-fold greater catalytic proficiency by AP for phosphate monoester hydrolysis relative to sulfate monoester hydrolysis. The primary 18O KIEs for the leaving group oxygen atoms in the AP-catalyzed hydrolysis of p-nitrophenyl phosphate (pNPP) and p-nitrophenylsulfate (pNPS) decrease relative to the values observed for nonenzymatic hydrolysis reactions. Prior linear free energy relationship results suggest that the transition states for AP-catalyzed reactions of phosphate and sulfate esters are "loose" and indistinguishable from that in solution, suggesting that the decreased primary KIEs do not reflect a change in the nature of the transition state but rather a strong interaction of the leaving group oxygen atom with an active site Zn2+ ion. Furthermore, the primary KIEs for the two reactions are identical within error, suggesting that the differential catalysis of these reactions cannot be attributed to differential stabilization of the leaving group. In contrast, AP perturbs the KIE for the nonbridging oxygen atoms in the reaction of pNPP but not pNPS, suggesting a differential interaction with the transferred group in the transition state. These and prior results are consistent with a strong electrostatic interaction between the active site bimetallo Zn2+ cluster and one of the nonbridging oxygen atoms on the transferred group. We suggest that the lower charge density of this oxygen atom on a transferred sulfuryl group accounts for a large fraction of the decreased stabilization of the transition state for its reaction relative to phosphoryl transfer.

  16. Ejaculation training, seminal alkaline phosphatase and semen preservation through cooling in a milk-based extender in domestic cats.

    Science.gov (United States)

    Valiente, Carla; de la Sota, Pablo E; Arauz, Sandra; Gobello, Cristina

    2014-04-01

    The purpose of this report is to describe (1) the training of domestic cats in ejaculation into an artificial vagina (AV), (2) alkaline phosphatase (AP) concentrations in whole ejaculates, and (3) the in vitro effect of a skimmed-milk plus egg yolk (SM-Y) extender on feline spermatozoa incubated at 4ºC. Five post-pubertal cats were trained to ejaculate into an AV three times a week for 20 mins in the presence of a teaser queen. Fifty AV-obtained ejaculates were macro- and microscopically assessed, and the AP therein measured by optimized colorimetry. Eighty AV-obtained ejaculates were pooled, diluted in SM-Y extender [80% (v/v) skimmed milk, 20% (v/v) egg yolk, and antibiotics], stored at 4°C and evaluated daily for 6 days. All the animals could be trained to ejaculate, although the interval up to the first AV ejaculation varied from 1.5 to 5.5 months (mean 3.9 months). The final performance at collection ranged from excellent to poor and was inversely related to the training period required in all cases. The mean AP concentration in whole ejaculates was 20,645.6 ± 4405U/l, which was not correlated with the concentration of spermatozoa. Most seminal parameters [(%); total (77 ± 2.3) and progressive (62.7 ± 3.4) motility, live sperm (91.8 ± 1.2), intact plasmalemma (83.5 ± 2.6), normal acrosomes (83.5 ± 2.6), pH (6.6 ± 0.0) and osmolarity (mOsm/l; 321 ± 5.2)], though decreasing during storage in the cold, remained within values compatible with in vivo fertilization for 2 days.

  17. A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae.

    Science.gov (United States)

    Fernandez, Luisa E; Aimanova, Karlygash G; Gill, Sarjeet S; Bravo, Alejandra; Soberón, Mario

    2006-02-15

    A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide-displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI-ALP (GPI-anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI-ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI-ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI-ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae.

  18. Tissue-nonspecific alkaline phosphatase is required for the calcification of collagen in serum: a possible mechanism for biomineralization.

    Science.gov (United States)

    Price, Paul A; Toroian, Damon; Chan, Wai Si

    2009-02-13

    Previous studies have shown that the type I collagen of tendon and demineralized bone both calcify rapidly in serum. The speed, collagen matrix-type specificity, and extent of the re-calcification of demineralized bone in serum suggest that the serum calcification activity identified in these studies may participate in normal biomineralization. Because of its presence in serum and its long history of association with the normal mineralization of the collagen matrix of bone, tissue-nonspecific alkaline phosphatase (TNAP) is an obvious candidate for a protein that could be a component of serum calcification activity, and experiments were therefore carried out to test this possibility. These experiments show that the inactivation of TNAP in serum prevents collagen calcification, and that the addition of physiological levels of purified TNAP restores the ability of TNAP-deficient serum to calcify collagen. Additional experiments show that the role of TNAP in collagen calcification is to activate a serum nucleator of apatite crystal formation. Based on these and earlier studies, the mechanism of collagen calcification in serum requires at least four elements as follows. 1) A matrix (collagen fibrils) that is accessible to small apatite crystals but not large molecules ( Toroian, D., Lim, J. E., and Price, P. A. (2007) J. Biol. Chem. 282, 22437-22447 ). 2) A large serum nucleator that generates small crystals, some of which diffuse into the fibrils. 3) A source of TNAP to activate the serum nucleator. 4) A large protein (fetuin) that selectively inhibits growth of crystals remaining in solution, thereby ensuring that only crystals within fibrils grow ( Toroian, D. T., and Price, P. A. (2008) Calcif. Tissue Int. 82, 116-126 ).

  19. The antioxidant effects of vitamin C on liver enzymes: aspartate aminotransferase, alanine aminotranferease, alkaline phosphatase and gamma-glutamyltransferase activities in rats under Paraquat insult

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    Benjamin Nnamdi Okolonkwo

    2013-06-01

    Full Text Available Paraquat (PQ is a bipyridylium herbicide; applied around trees in orchards and between crop rows to control broad-leaved and grassy weeds. Its oxidation results in the formation of superoxides which causes damage to cellular components. In this study, we determined the antioxidant effect vitamin C has on the liver enzymes [aspartate aminotransferase (SGOT, alanine aminotranferease (SGPT, alkaline phosphatase (ALP, and gamma-glutamyltransferase (GGT] of rats under this toxic insult. Male rats in groups (A, B, C and D were intraperitoneally injected with different sublethal increasing doses (0, 0.02, 0.04 and 0.06 g/kg body weigh of PQ respectively on monthly basis. Subsequently, the subgroups (A2, B2, C2 and D2 were given orally, 200 mg/L vitamin C, while the subgroups A1, B1, C1, and D1, received only water. Four animals per subgroup were decapitated on monthly basis and blood samples taken for enzyme assay. The parameters studied were - SGOT, SGPT, ALP and GGT - liver enzymes. The dose and time dependent PQ toxicity effect resulted in highly elevated Liver enzymes activities. The subgroups on vitamin C had significantly lower enzyme activities when compared to the same subgroups on only PQ insult. But the values were high when compared to the control subgroups (A1 and A2. These results were indication that vitamin C when given at moderate doses and maintained for a longer period could be a life saving adjunct to toxic insult.

  20. Alkaline phosphatases and aminopeptidases are altered in a Cry11Aa resistant strain of Aedes aegypti.

    Science.gov (United States)

    Lee, Su-Bum; Aimanova, Karlygash G; Gill, Sarjeet S

    2014-11-01

    Bacillus thuringiensis subsp. israelensis (Bti) is widely used for the biological control of mosquito populations. However, the mechanism of Bti toxins is still not fully understood. To further elucidate the mechanism of Bti toxins, we developed an Aedes aegypti resistant strain that shows high-level resistance to Cry11Aa toxin. After 27 selections with Cry11Aa toxin, the larvae showed a 124-fold resistance ratio for Cry11Aa (strain G30). G30 larvae showed cross-resistance to Cry4Aa (66-fold resistance), less to Cry4Ba (13-fold), but not to Cry11Ba (2-fold). Midguts from these resistant larvae did not show detectable difference in the processing of the Cry11Aa toxin compared to that in susceptible larvae (WT). Brush border membrane vesicles (BBMV) from resistant larvae bound slightly less Cry11Aa compared to WT BBMV. To identify potential proteins associated with Cry11A resistance, not only transcript changes in the larval midgut were analyzed using Illumina sequencing and qPCR, but alterations of previously identified receptor proteins were investigated using immunoblots. The transcripts of 375 genes were significantly increased and those of 208 genes were down regulated in the resistant larvae midgut compared to the WT. None of the transcripts for previously identified receptors of Cry11Aa (Aedes cadherin, ALP1, APN1, and APN2) were altered in these analyses. The genes for the identified functional receptors in resistant larvae midgut did not contain any mutation in their sequences nor was there any change in their transcript expression levels compared to WT. However, ALP proteins were expressed at reduced levels (∼ 40%) in the resistant strain BBMV. APN proteins and their activity were also slightly reduced in resistance strain. The transcript levels of ALPs (AAEL013330 and AAEL015070) and APNs (AAEL008158, AAEL008162) were significantly reduced. These results strongly suggest that ALPs and APNs could be associated with Cry11Aa resistance in Ae. aegypti

  1. Effect Modifying Role of Serum Calcium on Mortality-Predictability of PTH and Alkaline Phosphatase in Hemodialysis Patients: An Investigation Using Data from the Taiwan Renal Registry Data System from 2005 to 2012.

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    Yen-Chung Lin

    Full Text Available Predicting mortality in dialysis patients based on low intact parathyroid hormone levels is difficult, because aluminum intoxication, malnutrition, older age, race, diabetes, or peritoneal dialysis may influence these levels. We investigated the clinical implications of low parathyroid hormone levels in relation to the mortality of dialysis patients using sensitive, stratified, and adjusted models and a nationwide dialysis database. We analyzed data from 2005 to 2012 that were held on the Taiwan Renal Registry Data System, and 94,983 hemodialysis patients with valid data regarding their intact parathyroid levels were included in this study. The patient cohort was subdivided based on the intact parathyroid hormone and alkaline phosphatase levels. The mean hemodialysis duration within this cohort was 3.5 years. The mean (standard deviation age was 62 (14 years. After adjusting for age, sex, diabetes, the hemodialysis duration, serum albumin levels, hematocrit levels, calcium levels, phosphate levels, and the hemodialysis treatment adequacy score, the single-pool Kt/V, the crude and adjusted all-cause mortality rates increased when alkaline phosphatase levels were higher or intact parathyroid hormone levels were lower. In general, at any given level of serum calcium or phosphate, patients with low intact parathyroid hormone levels had higher mortality rates than those with normal or high iPTH levels. At a given alkaline phosphatase level, the hazard ratio for all-cause mortality was 1.33 (p 9.5 mg/dL, but in the group with intact parathyroid hormone levels > 300 pg/mL and serum calcium levels > 9.5 mg/dL, the hazard ratio was 0.92 (95% confidence interval 0.85-1.01. Hence, maintaining albumin-corrected high serum calcium levels at > 9.5 mg/dL may correlate with poor prognoses for patients with low intact parathyroid hormone levels.

  2. Evidence of associations between feto-maternal vitamin D status, cord parathyroid hormone and bone-specific alkaline phosphatase, and newborn whole body bone mineral content.

    Science.gov (United States)

    Dror, Daphna K; King, Janet C; Fung, Ellen B; Van Loan, Marta D; Gertz, Erik R; Allen, Lindsay H

    2012-02-01

    In spite of a high prevalence of vitamin D inadequacy in pregnant women and neonates, relationships among vitamin D status (25(OH)D), parathyroid hormone (PTH), bone specific alkaline phosphatase (BALP), and whole body bone mineral content (WBBMC) in the newborn are poorly characterized. The purpose of the present study was to investigate the relationships between maternal and cord 25(OH)D, PTH, BALP, and WBBMC in newborns in a multiethnic population in Oakland, California and to evaluate the predictive value of the biochemical indices as indicators of WBBMC. Maternal and cord blood were collected from 80 mother-infant pairs and infant WBBMC was measured by dual energy X-ray absorptiometry 8-21 days post-birth. Cord PTH and BALP were each inversely correlated with infant WBBMC (r = -0.28, p = 0.01 and r = -0.26, p = 0.02) and with cord 25(OH)D (r = -0.24, p = 0.03 and r = -0.34, p = 0.002), while cord 25(OH)D and unadjusted or weight-adjusted WBBMC were not significantly correlated with one other. In multivariate regression modeling, infant WBBMC was most strongly predicted by infant weight (p feto-maternal 25(OH)D, cord PTH and BALP, and early infant WBBMC, though neither feto-maternal 25(OH)D nor the measured biochemical indices were suitable indicators of WBBMC.

  3. ALP (Alkaline Phosphatase) Test

    Science.gov (United States)

    ... Paget's disease or other bone conditions, such as vitamin D deficiency. If ALP results are increased but ... be seen temporarily after blood transfusions or heart bypass surgery. A deficiency in zinc may cause decreased ...

  4. Alkaline phosphatase activity: new assay for the Reflotron system. Results of the evaluation in eight clinical laboratories.

    Science.gov (United States)

    Schumann, G; Dominick, H C; Hellmann, D; Klauke, R; Möckesch, M; Stekel, H; von Schenck, H; Kraft, M; Nagel, R; Hänseler, E

    2001-01-01

    A new reagent carrier, Reflotron ALP, has been developed for the Reflotron system, allowing easy and rapid measurement (in less than 3 minutes) of alkaline phosphatase (ALP) activity in capillary blood, venous blood, heparinized plasma or serum. The evaluation of the analytical performance of the assay was carried out at eight clinical laboratories. The study of the imprecision using the measurements in human samples resulted in coefficients of variation ranging from 1.3% to 4.6% (within-run) and from 3.2% to 4.0% (day-to-day). The analytical specificity of the Reflotron ALP assay agrees well with ALP methods using a N-methyl-D-glucamine buffer solution. The calibration of the Reflotron ALP assay, however, is related to the reference intervals for ALP methods using a diethanolamine buffer solution. Method comparisons were performed with the ALP method on Hitachi instruments using diethanolamine buffer. Reflotron ALP measurements in blood and plasma in 157 randomly selected split samples showed excellent agreement (slope: 0.99; intercept: 0.7 U/l; median bias: 2.3%; median difference from the comparison method: -0.3%). Specimens from pregnant women and adolescents were excluded from this study. Differing values were obtained in a method comparison using 48 samples containing predominantly the ALP bone isoform (slope: 0.81; intercept: 31.5 U/l; median bias: 5.7%; median difference from the comparison method: -12.2%). Regression analysis of the results from 21 sera with prevailing placental ALP gave a slope of 1.51, and an intercept of -41.1 U/l (median bias: 8.6%; median difference from the comparison method: 35.6%). Reflotron ALP was compared with three different wet chemistry procedures using different buffer compounds: N-methyl-D-glucamine or diethanolamine or 2-amino-2-methyl-1-propanol. In samples containing predominantly ALP isoforms not of liver origin, the measurements with N-methyl-D-glucamine buffer gave the best fit with respect to Reflotron. In an

  5. Enzymatically mediated bioprecipitation of heavy metals from industrial wastes and single ion solutions by mammalian alkaline phosphatase.

    Science.gov (United States)

    Chaudhuri, Gouri; Shah, Gaurav A; Dey, Pritam; S, Ganesh; Venu-Babu, P; Thilagaraj, W Richard

    2013-01-01

    The study was aimed at investigating the potential use of calf intestinal alkaline phosphatase (CIAP) enzyme in the removal of heavy metals (Cd(2+), Ni(2+), Co(2+) and Cr(3+/6+)) from single ion solutions as well as tannery and electroplating effluents. CIAP mediated bioremediation (white biotechnology) is a novel technique that is eco-friendly and cost effective unlike the conventional chemical technologies. Typical reactions containing the enzyme (CIAP) and p-nitrophenyl phosphate (pNPP) as substrate in Tris-HCl buffer (pH 8 and 11) and either single ion metal solutions (250 ppm and 1000 ppm) or effluents from tannery or electroplating industry were incubated at 37°C for 30 min, 60 min and 120 min. The inorganic phosphate (P(i)) generated due to catalytic breakdown of pNPP complexes free metal ions as metal-phosphate and the amount of metal precipitated was derived by estimating the reduction in the free metal ion present in the supernatant of reactions employing atomic absorption spectrophotometer (AAS). Better precipitation of metal was obtained at pH 11 than at pH 8 and between the two concentrations of different metals tested, an initial metal concentration of 250 ppm in the reaction gave more precipitation than with 1000 ppm. Experimental data showed that at pH 11, the percentage of removal of metal ions (for an initial concentration of 250 ppm) was in the following order: Cd(2+) (80.99%) > Ni(2+) (64.78%) > Cr(3+) > (46.15%) > Co(2+) (36.47%) > Cr(6+) (32.33%). The overall removal of Cr(3+) and Cr(6+) from tannery effluent was 32.77% and 37.39% respectively in 120 min at pH 11. Likewise, the overall removal of Cd(2+), Co(2+) and Ni(2+) from electroplating effluent was 50.42%, 13.93% and 38.64% respectively in 120 min at pH 11. The study demonstrates that bioprecipitation by CIAP may be a viable and environmental friendly method for clean-up of heavy metals from tannery and electroplating effluents.

  6. The cellular prion protein interacts with the tissue non-specific alkaline phosphatase in membrane microdomains of bioaminergic neuronal cells.

    Directory of Open Access Journals (Sweden)

    Myriam Ermonval

    Full Text Available BACKGROUND: The cellular prion protein, PrP(C, is GPI anchored and abundant in lipid rafts. The absolute requirement of PrP(C in neurodegeneration associated to prion diseases is well established. However, the function of this ubiquitous protein is still puzzling. Our previous work using the 1C11 neuronal model, provided evidence that PrP(C acts as a cell surface receptor. Besides a ubiquitous signaling function of PrP(C, we have described a neuronal specificity pointing to a role of PrP(C in neuronal homeostasis. 1C11 cells, upon appropriate induction, engage into neuronal differentiation programs, giving rise either to serotonergic (1C11(5-HT or noradrenergic (1C11(NE derivatives. METHODOLOGY/PRINCIPAL FINDINGS: The neuronal specificity of PrP(C signaling prompted us to search for PrP(C partners in 1C11-derived bioaminergic neuronal cells. We show here by immunoprecipitation an association of PrP(C with an 80 kDa protein identified by mass spectrometry as the tissue non-specific alkaline phosphatase (TNAP. This interaction occurs in lipid rafts and is restricted to 1C11-derived neuronal progenies. Our data indicate that TNAP is implemented during the differentiation programs of 1C11(5-HT and 1C11(NE cells and is active at their cell surface. Noteworthy, TNAP may contribute to the regulation of serotonin or catecholamine synthesis in 1C11(5-HT and 1C11(NE bioaminergic cells by controlling pyridoxal phosphate levels. Finally, TNAP activity is shown to modulate the phosphorylation status of laminin and thereby its interaction with PrP. CONCLUSION/SIGNIFICANCE: The identification of a novel PrP(C partner in lipid rafts of neuronal cells favors the idea of a role of PrP in multiple functions. Because PrP(C and laminin functionally interact to support neuronal differentiation and memory consolidation, our findings introduce TNAP as a functional protagonist in the PrP(C-laminin interplay. The partnership between TNAP and PrP(C in neuronal cells may

  7. Osteoblast response (initial adhesion and alkaline phosphatase activity following exposure to a barrier membrane/enamel matrix derivative combination

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    Thangakumaran S

    2009-01-01

    Full Text Available Background and Objective: The enamel matrix derivative (EMD has been used in combination with barrier membranes to optimize regeneration in vertical osseous defects. However, the osteoblast response when exposed to the EMD/barrier membrane combination has not yet been evaluated. The osteoblast behavior when exposed to a combination of regenerative materials must be evaluated to fully understand their effect on bone regeneration. Therefore, the present study was undertaken to estimate the initial adhesion and alkaline phosphatase (ALP activity of an osteoblast cell line (SaOS-2 when exposed to four commercially available resorbable membranes and determine if the addition of EMD had any modulatory effect on osteoblast behavior. Materials and Methods: 5 x 104 SaOS-2 cells between passages 7-10 were cultured in two 24-well culture plates. Plate A was used for the adhesion assay and Plate B was used for the ALP assay. A MTT (3-[4, 5-dimethylthiazolyl-2]-2, 5-diphenyltetrazolium bromide assay was done after 24 hours to determine the adhesion of the osteoblastic cells to four barrier membranes: 1 a non cross-linked porcine Type I and III collagen membrane (BG, 2 a weakly cross-linked Type I collagen membrane (HG, 3 a glutaraldehyde cross-linked bovine Type I collagen (BM, and 4 a resorbable polymer membrane (CP. Osteoblast differentiation was studied using an ALP assay with p-nitro phenyl phosphate as the substrate at 24 hours, 72 hours, and 1 week. A total of 50 µg/ml of EMD dissolved in 10 mM acetic acid was added into each well and the entire experimental protocol outlined above was repeated. Results: The osteoblast adhesion to collagen barriers showed a statistically insignificant reduction following the addition of EMD. Adhesion to the polymer barrier, although significantly lower when compared with collagen barriers, was unaffected by the addition of EMD. ALP activity after 1 week among the various groups was as follows: EMD alone (75.59±2

  8. Effects of Intercropping with Potato Onion on the Growth of Tomato and Rhizosphere Alkaline Phosphatase Genes Diversity

    Science.gov (United States)

    Wu, Xia; Wu, Fengzhi; Zhou, Xingang; Fu, Xuepeng; Tao, Yue; Xu, Weihui; Pan, Kai; Liu, Shouwei

    2016-01-01

    Background and Aims: In China, excessive fertilization has resulted in phosphorus (P) accumulation in most greenhouse soils. Intercropping can improve the efficiency of nutrient utilization in crop production. In this study, pot experiments were performed to investigate the effects of intercropping with potato onion (Allium cepa L. var. aggregatum G. Don) on tomato (Solanum lycopersicum L.) seedlings growth and P uptake, the diversity of rhizosphere phosphobacteria and alkaline phosphatase (ALP) genes in phosphorus-rich soil. Methods: The experiment included three treatments, namely tomato monoculture (TM), potato onion monoculture (OM), and tomato/potato onion intercropping (TI-tomato intercropping and OI-potato onion intercropping). The growth and P uptake of tomato and potato onion seedlings were evaluated. The dilution plating method was used to determine the population of phosphate-solubilizing bacteria (PSB) and phosphate-mineralizing bacteria (PMB). The genomic DNAs of PSB and PMB in the rhizosphere of tomato and potato onions were extracted and purified, and then, with the primer set of 338f /518r, the PCR amplification of partial bacterial 16S rDNA sequence was performed and sequenced to determine the diversities of PSB and PMB. After extracting the total genomic DNAs from the rhizosphere, the copy numbers and diversities of ALP genes were investigated using real-time PCR and PCR-DGGE, respectively. Results: Intercropping with potato onion promoted the growth and P uptake of tomato seedlings, but inhibited those of potato onion. After 37 days of transplanting, compared to the rhizosphere of TM, the soil pH increased, while the electrolytic conductivity and Olsen P content decreased (p < 0.05) in the rhizosphere of TI. The populations and diversities of PSB, PMB, and ALP genes increased significantly in the rhizosphere of TI, compared to the rhizosphere of TM. Conclusion: The results indicated that intercropping with potato onion promoted the growth and P

  9. Synthesis of benzofuran derivatives as selective inhibitors of tissue-nonspecific alkaline phosphatase: effects on cell toxicity and osteoblast-induced mineralization.

    Science.gov (United States)

    Marquès, Stéphanie; Buchet, René; Popowycz, Florence; Lemaire, Marc; Mebarek, Saïda

    2016-03-01

    Tissue-nonspecific alkaline phosphatase (TNAP) by hydrolyzing pyrophosphate, an inhibitor of apatite formation, promotes extracellular matrix calcification during bone formation and growth, as well as during ectopic calcification under pathological conditions. TNAP is a target for the treatment of soft tissue pathological ossification. We synthesized a series of benzofuran derivatives. Among these, SMA14, displayed TNAP activity better than levamisole. SMA14 was found to be not toxic at doses of up to 40μM in osteoblast-like Saos-2 cells and primary osteoblasts. As probed by Alizarin Red staining, this compound inhibited mineral formation in murine primary osteoblast and in osteoblast-like Saos-2 cells.

  10. Patient pools and the use of "patient means" are valuable tools in quality control illustrated by a bone-specific alkaline phosphatase assay

    DEFF Research Database (Denmark)

    Hinge, Maja; Lund, Erik D.; Brandslund, Ivan;

    2016-01-01

    BACKGROUND: Quality control (QC) is an essential part of clinical biochemistry to ensure that laboratory test results are reliable and correct. Those tests without a defined reference method constitute a special challenge, as is the case with bone-specific alkaline phosphatase (BAP). METHODS...... AND RESULTS: The present study reports an example where a shift in a BAP assay was detected by use of a patient pool and supported by a retrospective calculation of "patient mean", while the external QC and specific assay control material were unaffected by the shift. CONCLUSIONS: Patient pools and the use...

  11. 骨碱性磷酸酶与碱性磷酸酶在佝偻病诊断中的比较%Bone alkaline phosphatase versus serum alkaline phosphatase in the diagnosis of rickets

    Institute of Scientific and Technical Information of China (English)

    林岚; 陈力; 张春玲; 杨延萍; 肖玉联

    2011-01-01

    Objective To explore the significance of quantitative and qualitative measurements of bone alkaline phosphatase ( BAP-E and BAP-CIT ) and detection of serum AP in the diagnosis of rickets. Methods Serum AP level was measured, quantitative BAP was detected by ELISA, and qualitative BAP was determined by CIT in 55 children with rickets and 26 control subjects. Results AP level and BAP-E were ( 164.38 ± 23.87 ) U/L and ( 57.99 ± 7.67 ) U/L in controls, and were ( 209.50 ± 44.28 ) U/L, ( 80.38 ± 15.75 ) U/L, ( 206.06 ± 30.80 ) U/L, ( 78.05± 11.69) U/L, ( 1000.47 ± 464.65) U/L, and (283.22 ± 96.42) U/L in mild or severe rickets ,respectively. AP and BAP-E were elevated significantly in children with rickets ( P< 0.05 ), and there were significant differences between the severe groups and the mild groups ( P < 0.05 ). AP positively related with BAP-E ( r= 0.921, P< 0.01 ) and BAP-CIT ( r= 0.49, P< 0.01 ). In the control group,BAP-CIT within the normal range was only in one patient and that beyond the normal range was in the remaining patients. Conclusions AP and BAP-E are positively correlated with the severity of rickets.AP may be used in severe patients. BAP-CIT should be used carefully in the diagnosis of rickets.%目的 了解定量骨碱性磷酸酶( BAP-E)、定性骨碱性磷酸酶(BAP-CIT)、血清碱性磷酸酶(AP)三个指标在佝偻病诊断中的意义。方法 检测55例佝偻病患儿、26例对照婴幼儿血清AP、BAP-E、BAP-CIT,BAP-E采用ELISA法,BAP-CIT用全血干化学和免疫浓缩技术。结果 佝偻病患儿的AP和BAP-E较对照组(164.38 U/L±23.87 U/L,57.99 U/L±7.67 U/L)明显增高(P<0.05),中、重度组佝偻病患儿的AP和BAP-E( 1000.47 U/L±464.65U/L,283.22 U/L±96.42 U/L)较轻度组(209.50 U/L±44.28 U/L,80.38 U/L±15.75 U/L;206.06 U/L±30.80 U/L,78.05 U/L±11.69 U/L)明显增高(P<0.05),对照组BAP-CIT仅1例在试剂盒的正常范围≤200 U/L,余皆在异常范围(200-300) U/L,AP

  12. Serum proteins, trace metals and phosphatases in psoriasis

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    Bhatnagar M

    1994-01-01

    Full Text Available Serum proteins, zinc, copper, acid phosphatase (AcPase and alkaline phosphatase (AlPase were studied in both active and remission phases of psoriasis. Data were compared with healthy controls, ?1, ? and ? globulins were high in active phase while ?1 and ? globulins were at par in remission phase. Serum copper was low but zinc and alkaline phosphatase were significantly high in both active and remission phases of the disease. Acid phosphatase level was at par in all the experimental groups. Study suggests a positive correlation of globulin, zinc and Alpase in active and remission phase of psoriasis.

  13. Splenomegaly, elevated alkaline phosphatase and mutations in the SRSF2/ASXL1/RUNX1 gene panel are strong adverse prognostic markers in patients with systemic mastocytosis.

    Science.gov (United States)

    Jawhar, M; Schwaab, J; Hausmann, D; Clemens, J; Naumann, N; Henzler, T; Horny, H-P; Sotlar, K; Schoenberg, S O; Cross, N C P; Fabarius, A; Hofmann, W-K; Valent, P; Metzgeroth, G; Reiter, A

    2016-12-01

    We evaluated the impact of clinical and molecular characteristics on overall survival (OS) in 108 patients with indolent (n=41) and advanced systemic mastocytosis (SM) (advSM, n=67). Organomegaly was measured by magnetic resonance imaging-based volumetry of the liver and spleen. In multivariate analysis of all patients, an increased spleen volume ⩾450 ml (hazard ratio (HR), 5.2; 95% confidence interval (CI), (2.1-13.0); P=0.003) and an elevated alkaline phosphatase (AP; HR 5.0 (1.1-22.2); P=0.02) were associated with adverse OS. The 3-year OS was 100, 77, and 39%, respectively (P<0.0001), for patients with 0 (low risk, n=37), 1 (intermediate risk, n=32) or 2 (high risk, n=39) parameters. For advSM patients with fully available clinical and molecular data (n=60), univariate analysis identified splenomegaly ⩾1200 ml, elevated AP and mutations in the SRSF2/ASXL1/RUNX1 (S/A/R) gene panel as significant prognostic markers. In multivariate analysis, mutations in S/A/R (HR 3.2 (1.1-9.6); P=0.01) and elevated AP (HR 2.6 (1.0-7.1); P=0.03) remained predictive adverse prognostic markers for OS. The 3-year OS was 76 and 38%, respectively (P=0.0003), for patients with 0-1 (intermediate risk, n=28) or 2 (high risk, n=32) parameters. We conclude that splenomegaly, elevated AP and mutations in the S/A/R gene panel are independent of the World Health Organization classification and provide the most relevant prognostic information in SM patients.

  14. The synthesis of Phosphate-repressible alkaline phosphatase do not appear to be regulated by ambient pH in the filamentous mould Neurospora crassa

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    Nozawa Sérgio R.

    2002-01-01

    Full Text Available In order to investigate further the adaptive response of moulds to ambient pH, we have measured by ELISA the pho-2-encoded Pi-repressible alkaline phosphatase synthesised by Neurospora crassa. We showed that the 74A and pho-2A strains of this mould secrete similar amounts of the pho-2-encoded enzyme irrespective of ambient pH, when both the preg and pgov genes are not functional, i.e., in strains nuc-2+ growing under Pi-starvation. This suggests that pho-2, which is responsive to Pi starvation via the action of genes nuc-2, preg, pgov and nuc-1, is not a gene responsive to ambient pH and that the differential glycosylation observed for the Pi-repressible alkaline phosphatase retained by the mycelium at pH 5.6 or secreted into the growth medium at pH 8.0 is the genetic response to ambient pH sensing in N. crassa.

  15. Inhibition of Alkaline Phosphatase from Pearl Oyster Pinctada fucata by o-Phthalaldehyde: Involvement of Lysine and Histidine Residues at the Active Site

    Institute of Scientific and Technical Information of China (English)

    CHEN Hongtao; XIE Liping; YU Zhenyan; ZHANG Rongqing

    2005-01-01

    Alkaline phosphatase from Pinctada fucata was inactivated by o-phthalaldehyde (OPA). The inactivation followed pseudo first-order kinetics with a second rate constant of 0.167 (mmol/L)-1·min-1 at pH 7.5 and 25°C. A Tsou's plot analysis showed that inactivation occurred upon formation of one isoindole group. The OPA-modified enzyme lost the ability to bind with the specific affinity column and the presence of substrates or competitive inhibitors protected the enzyme from inactivation. The results revealed that the OPA-reaction site was at the enzyme substrate binding site. Prior modification of the enzyme by lysine or histidine specific reagent abolished formation of the isoindole derivatives, suggesting that lysine and histidine residues were involved in the OPA-induced inactivation. Taken together, OPA inactivated the alkaline phosphatase from Pinctada fucata by cross-linking lysine and histidine residues at the active site and formed an isoindole group at the substrate binding site of the enzyme.

  16. Profile of serum alkaline phosphatase after inoculation of mononuclear cells and bone morphogenetic protein in the repair of osteochondral defects in rabbits

    Directory of Open Access Journals (Sweden)

    Luiz Augusto de Souza

    2011-12-01

    Full Text Available In this study, serum alkaline phosphatase activity was measured in response to the repair of osteochondral defects in twenty-four New Zealand rabbits. The animals were divided into three groups: a control (GC, those treated with bone marrow mononuclear cells (GCM and those that received mononuclear cells with autologous bone morphogenetic protein (BMP + GCM. After exposing the trochlear groove of the left stifle joint, a wedge-shaped segment was removed. Later, the defect was filled with an osteochondral autograft preserved in 98% glycerin. For the GC group, only the bone graft was performed. For the GCM, in addition to the graft, 2x106 seed mononuclear cells were implanted. For the GCM + BMP, the same number of cells, associated with 1μg of bone morphogenetic protein, were intraarticularly administered. The osteoblastic response was measured by analyzing the serum alkaline phosphatase on day 0 (preoperative 3, 15, 30, and 45 after surgery, and by radiographic examinations. Analysis of variance in randomized blocks, factorial and Tukey’s test (p = 0.05 were made. The overall mean GCM was superior to the other groups and the highest rates were among the 15th and 45th days postoperatively. The discrepancy in values between individuals of the same group casts doubts on the veracity of the test.

  17. Boron Induces Early Matrix Mineralization via Calcium Deposition and Elevation of Alkaline Phosphatase Activity in Differentiated Rat Bone Marrow Mesenchymal Stem Cells

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    Bent-al-hoda Movahedi Najafabadi

    2016-04-01

    Full Text Available Objective: Boron (B is essential for plant development and might be an essential micronutrient for animals and humans. This study was conducted to characterize the impact of boric acid (BA on the cellular and molecular nature of differentiated rat bone marrow mesenchymal stem cells (BMSCs. Materials and Methods: In this experimental study, BMSCs were extracted and expanded to the 3rd passage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM complemented with osteogenic media as well as 6 ng/ml and 6 μg/ml of BA. After 5, 10, 15 and 21 days the viability and the level of mineralization was determined using MTT assay and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic area of the cells were studied with the help of fluorescent dye. The concentration of calcium, activity of alanine transaminase (ALT, aspartate transaminase (AST, lactate dehydrogenase (LDH and alkaline phosphatase (ALP as well as sodium and potassium levels were also evaluated using commercial kits and a flame photometer respectively. Results: Although 6 μg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased the osteogenic ability of the cell significantly throughout the treatment. In addition it was observed that B treatment caused the early induction of matrix mineralization compared to controls. Conclusion: Although more investigation is required, we suggest the prescription of a very low concentration of B in the form of BA or foods containing BA, in groups at high risk of osteoporosis or in the case of bone fracture.

  18. Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells: roles of tumor necrosis factor-alpha and oncostatin M derived from macrophages.

    Science.gov (United States)

    Shioi, Atsushi; Katagi, Miwako; Okuno, Yasuhisa; Mori, Katsuhito; Jono, Shuichi; Koyama, Hidenori; Nishizawa, Yoshiki

    2002-07-12

    Inflammatory cells such as macrophages and T lymphocytes play an important role in vascular calcification associated with atherosclerosis and cardiac valvular disease. In particular, macrophages activated with cytokines derived from T lymphocytes such as interferon-gamma (IFN-gamma) may contribute to the development of vascular calcification. Moreover, we have shown the stimulatory effect of 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) on in vitro calcification through increasing the expression of alkaline phosphatase (ALP), an ectoenzyme indispensable for bone mineralization, in vascular smooth muscle cells. Therefore, we hypothesized that macrophages may induce calcifying phenotype, especially the expression of ALP in human vascular smooth muscle cells (HVSMCs) in the presence of IFN-gamma and 1,25(OH)2D3. To test this hypothesis, we used cocultures of HVSMCs with human monocytic cell line (THP-1) or peripheral blood monocytes (PBMCs) in the presence of IFN-gamma and 1,25(OH)2D3. THP-1 cells or PBMCs induced ALP activity and its gene expression in HVSMCs and the cells with high expression of ALP calcified their extracellular matrix by the addition of beta-glycerophosphate. Thermostability and immunoassay showed that ALP induced in HVSMCs was bone-specific enzyme. We further identified tumor necrosis factor-alpha (TNF-alpha) and oncostatin M (OSM) as major factors inducing ALP in HVSMCs in the culture supernatants of THP-1 cells. TNF-alpha and OSM, only when applied together, increased ALP activities and in vitro calcification in HVSMCs in the presence of IFN-gamma and 1,25(OH)2D3. These results suggest that macrophages may contribute to the development of vascular calcification through producing various inflammatory mediators, especially TNF-alpha and OSM.

  19. Extracellular Matrix Proteins, Alkaline Phosphatase and Pyrophosphate as Molecular Determinants of Bone, Tooth, Kidney and Vascular Calcification

    Science.gov (United States)

    McKee, Marc D.

    2008-09-01

    Progress in biomineralization research in recent years has identified, characterized and described functions for key noncollagenous extracellular matrix proteins regulating crystal growth in the skeleton and dentition. Some of these same proteins expressed in soft tissues undergoing pathologic calcification also inhibit ectopic crystal growth. In addition to extracellular matrix proteins regulating matrix mineralization, the enzyme tissue-nonspecific alkaline phosphatase—which is highly expressed by cells in mineralized tissues—cleaves pyrophosphate, an anionic small-molecule inhibitor of mineralization. Together with the required mineral ion availability necessary for crystal growth, these molecular determinants appear to function in limiting the spread of pathologic calcification seen in soft tissues such as blood vessels and kidneys. Osteopontin, in particular, is a potent calcification inhibitor that accumulates in mineralized tissues and in calcified deposits during vascular calcification and nephrolithiasis/urolithiasis. Additional research is required to establish the exact temporal sequence in which the molecular determinants of pathologic calcification appear relative to mineral crystal growth in different tissues, and to establish their relationship (if any) to the activation of osteogenic differentiation programs.

  20. Electron micrographic study of precipitates formed by interaction of silicic acid and alkaline phosphatase: contribution to a study of silica urolithiasis in cattle.

    Science.gov (United States)

    Bailey, C B; Cheng, K J; Costerton, J W

    1982-12-01

    Association of alkaline phosphatase with silicic acid in precipitates formed in dilute solution was studied as a model for the nonspecific reaction between silicic acid and protein. Precipitates contained 68-83% of the silicic acid and 52-83% of the enzyme in the original mixture and were in the form of aggregates of roundish particles 150-800 nm in diameter. Enzyme protein formed a tightly bound layer on the surface of particles formed in solutions of freshly prepared silicic acid. The similarity between the ultrastructural features of precipitates from solutions of silicic acid and of internal portions of siliceous urinary calculi from cattle suggests that deposition of silica during development of such calculi is due, at least in part, to the interaction of protein with silicic acid in urine.

  1. Microchannel conductivity measurements in microchip for on line monitoring of dephosphorylation rates of organic phosphates using paramagnetic-beads linked alkaline phosphatase.

    Science.gov (United States)

    Kechadi, Mohammed; Sotta, Bruno; Gamby, Jean

    2015-01-01

    This paper presents the use of polymer coated microelectrodes for the realtime conductivity monitoring in a microchannel photoablated through the polymer without contact. Based on this strategy, a small conductometry sensor has been developed to record in time conductivity variation when an enzymatic reaction occurs through the channel. The rate constant determination, k2, for the dephosphorylation of organic phosphate-alkaline phosphatase-superparamagnetic beads complex using chemically different substrates such as adenosine monoesterphosphate, adenosine diphosphate and adenosine triphosphate was taken as an example to demonstrate selectivity and sensivity of the detection scheme. The k2 value measured for each adenosine phosphate decreases from 39 to 30 s(-1) in proportion with the number (3, 2 and 1) of attached phosphate moiety, thus emphasizing the steric hindrance effect on kinetics.

  2. Partial Enteral Nutrition Preserves Elements of Gut Barrier Function, Including Innate Immunity, Intestinal Alkaline Phosphatase (IAP) Level, and Intestinal Microbiota in Mice.

    Science.gov (United States)

    Wan, Xiao; Bi, Jingcheng; Gao, Xuejin; Tian, Feng; Wang, Xinying; Li, Ning; Li, Jieshou

    2015-08-03

    Lack of enteral nutrition (EN) during parenteral nutrition (PN) leads to higher incidence of infection because of gut barrier dysfunction. However, the effects of partial EN on intestina linnate immunity, intestinal alkaline phosphatase (IAP) and microbiota remain unclear. The mice were randomized into six groups to receive either standard chow or isocaloric and isonitrogenous nutritional support with variable partial EN to PN ratios. Five days later, the mice were sacrificed and tissue samples were collected. Bacterial translocation, the levels of lysozyme, mucin 2 (MUC2), and IAP were analyzed. The composition of intestinal microbiota was analyzed by 16S rRNA pyrosequencing. Compared with chow, total parenteral nutrition (TPN) resulted in a dysfunctional mucosal barrier, as evidenced by increased bacterial translocation (p microbiota (p intestinal microbiota.

  3. Using a Personal Glucose Meter and Alkaline Phosphatase for Point-of-Care Quantification of Galactose-1-Phosphate Uridyltransferase in Clinical Galactosemia Diagnosis.

    Science.gov (United States)

    Zhang, Jingjing; Xiang, Yu; Novak, Donna E; Hoganson, George E; Zhu, Junjie; Lu, Yi

    2015-10-01

    The personal glucose meter (PGM) was recently shown to be a general meter to detect many targets. Most studies, however, focus on transforming either target binding or enzymatic activity that cleaves an artificial substrate into the production of glucose. More importantly, almost all reports exhibit their methods by using artificial samples, such as buffers or serum samples spiked with the targets. To expand the technology to even broader targets and to validate its potential in authentic, more complex clinical samples, we herein report expansion of the PGM method by using alkaline phosphatase (ALP) that links the enzymatic activity of galactose-1-phosphate uridyltransferase to the production of glucose, which allows point-of-care galactosemia diagnosis in authentic human clinical samples. Given the presence of ALP in numerous enzymatic assays for clinical diagnostics, the methods demonstrated herein advance the field closer to point-of-care detection of a wide range of targets in real clinical samples.

  4. Variation of Alkaline Phosphatase Activity in Sediments of Shrimp Culture Ponds and Its Relationship with the Contents of C, N and P

    Institute of Scientific and Technical Information of China (English)

    SU Yuepeng; MA Shen; DONG Shuanglin

    2005-01-01

    Nine enclosures (5 m × 5 m) were built in a Fenneropenaeus chinensis culture pond of Rushan Gulf in April, 2001.The probiotics and BIO ENERGIZER solution were applied for disparate treatments. Variations of alkaline phosphatase activity (APA) and its relationship with the contents of C, N and P in sediments were studied. Results show that APA of sediments increases from 3.096 nmol g-1min-1 to 5.407 nmol g-1nin-1 in culture period; the bacteria biomass is not the only factor to determine APA; the contents of total P and total organic carbon have a significant positive correlation with APA, while that of total nitrogen has a negative correlation. In addition, the contents of inorganic P and organic P are not regular with APA. By comparison, TOC shows a more significant coherence with APA, meaning that organic pollution in sediments affects APA remarkably.

  5. Labelling of T cell subsets under field conditions in tropical countries. Adaptation of the immuno-alkaline phosphatase staining method for blood smears.

    Science.gov (United States)

    Lisse, I M; Whittle, H; Aaby, P; Normark, M; Gyhrs, A; Ryder, L P

    1990-05-08

    Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could be transferred abroad without antigenic damage. Identical total CD4 and CD8 counts were obtained on venous and capillary blood, when compared using a FACS analyser. Although the AP method gave somewhat higher total CD4 and CD8 counts, the ratio remained the same. The major advantages of the method are: (i) no expensive equipment is required, (ii) only minute amounts of blood are needed, and (iii) slides can be stored for long periods before labelling and can be preserved for later reading. The method is suitable for community studies where there is a need for assessing the immune status of the population.

  6. Aspartic acid-484 of nascent placental alkaline phosphatase condenses with a phosphatidylinositol glycan to become the carboxyl terminus of the mature enzyme.

    Science.gov (United States)

    Micanovic, R; Bailey, C A; Brink, L; Gerber, L; Pan, Y C; Hulmes, J D; Udenfriend, S

    1988-01-01

    A carboxyl-terminal chymotryptic peptide from mature human placental alkaline phosphatase was purified by HPLC and monitored by a specific RIA. Sequencing and amino acid assay showed that the carboxyl terminus of the peptide was aspartic acid, representing residue 484 of the proenzyme as deduced from the corresponding cDNA. Further analysis of the peptide showed it to be a peptidoglycan containing one residue of ethanolamine, one residue of glucosamine, and two residues of neutral hexose. The inositol glycan is apparently linked to the alpha carboxyl group of the aspartic acid through the ethanolamine. Location of the inositol glycan on Asp-484 of the proenzyme indicates that a 29-residue peptide is cleaved from the nascent protein during the post-translational condensation with the phosphatidylinositol-glycan. PMID:3422741

  7. A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

    Science.gov (United States)

    Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario

    2013-03-01

    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders.

  8. Acid and alkaline phosphatase activities in a novel phosphorothionate (RPR-11) treated male and female rats. Evidence of dose and time-dependent response.

    Science.gov (United States)

    Rahman, M F; Siddiqui, M K; Jamil, K

    2000-08-01

    The effect of a novel phosphorothionate, the methyl ester of 2-butenoic acid-3-diethoxy phosphinothioyl (RPR-II) was studied on membrane bound target enzymes Acid (AcP) and Alkaline (AkP) Phosphatases in different tissues of male and female albino Wistar rats. Three sub-chronic doses 0.014 (low), 0.028 (medium) and 0.042 (high)mg/kg-1 were administered to the rats daily for a period of 90 days. The long term and repeated administration of RPR-II caused significant increase of AcP and AkP in serum and kidney (AcP), whereas these enzymes simultaneously decreased significantly in liver, kidney (female rat AkP) and lung tissues in both male and female rats after 45 and 90 days of treatment. However, the kidney AcP increased significantly in both the sexes which is suggestive of an increase in synthesis of this enzyme which may be an adaptive mechanism to the toxicant stress. The changes in serum, liver, kidney and lung of both male and female rats by this compound were statistically significant when compared with two way Anova showing that they are dose and time dependent. The alterations in male rats were statistically insignificant when compared with female rats showing no sexual dimorphism by this compound. Recovery was observed after 28 days of post treatment (withdrawal study) indicating reversal of the toxic symptoms once the toxicant is removed. High degree negative correlation was observed for serum versus liver and lung and in other cases substantial correlation was observed. The changes observed in these enzymes showed that liver was most susceptible followed by lung and kidney. There are marker enzymes and their increase in different tissues might be due to the increased permeability of plasma membrane or cellular necrosis, showing the stress condition of the treated rats. This investigation elucidates the effect of these biomarker enzymes which increased in blood, might be due to the necrosis of liver, kidney and lung tissues by this compound.

  9. Chitosan nanofiber scaffold improves bone healing via stimulating trabecular bone production due to upregulation of the Runx2/osteocalcin/alkaline phosphatase signaling pathway

    Science.gov (United States)

    Ho, Ming-Hua; Yao, Chih-Jung; Liao, Mei-Hsiu; Lin, Pei-I; Liu, Shing-Hwa; Chen, Ruei-Ming

    2015-01-01

    Osteoblasts play critical roles in bone formation. Our previous study showed that chitosan nanofibers can stimulate osteoblast proliferation and maturation. This translational study used an animal model of bone defects to evaluate the effects of chitosan nanofiber scaffolds on bone healing and the possible mechanisms. In this study, we produced uniform chitosan nanofibers with fiber diameters of approximately 200 nm. A bone defect was surgically created in the proximal femurs of male C57LB/6 mice, and then the left femur was implanted with chitosan nanofiber scaffolds for 21 days and compared with the right femur, which served as a control. Histological analyses revealed that implantation of chitosan nanofiber scaffolds did not lead to hepatotoxicity or nephrotoxicity. Instead, imaging analyses by X-ray transmission and microcomputed tomography showed that implantation of chitosan nanofiber scaffolds improved bone healing compared with the control group. In parallel, microcomputed tomography and bone histomorphometric assays further demonstrated augmentation of the production of new trabecular bone in the chitosan nanofiber-treated group. Furthermore, implantation of chitosan nanofiber scaffolds led to a significant increase in the trabecular bone thickness but a reduction in the trabecular parameter factor. As to the mechanisms, analysis by confocal microscopy showed that implantation of chitosan nanofiber scaffolds increased levels of Runt-related transcription factor 2 (Runx2), a key transcription factor that regulates osteogenesis, in the bone defect sites. Successively, amounts of alkaline phosphatase and osteocalcin, two typical biomarkers that can simulate bone maturation, were augmented following implantation of chitosan nanofiber scaffolds. Taken together, this translational study showed a beneficial effect of chitosan nanofiber scaffolds on bone healing through stimulating trabecular bone production due to upregulation of Runx2-mediated alkaline

  10. Bifunctional coating based on carboxymethyl chitosan with stable conjugated alkaline phosphatase for inhibiting bacterial adhesion and promoting osteogenic differentiation on titanium

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Dong; Neoh, Koon Gee, E-mail: chenkg@nus.edu.sg; Kang, En-Tang

    2016-01-01

    Graphical abstract: - Highlights: • Alkaline phosphatase was immobilized on carboxymethyl chitosan coating on Ti. • The coating is bifunctional; resists bacterial adhesion and enhances cell functions. • Osteogenic differentiation of osteoblasts and stem cells is enhanced on the coating. • The coating remains stable and functional after ethanol treatment and autoclaving. - Abstract: In this work, alkaline phosphatase (ALP) was covalently immobilized on carboxymethyl chitosan (CMCS)-coated polydopamine (PDA)-functionalized Ti to achieve a bifunctional surface. Our results showed ∼89% reduction in Staphylococcus epidermidis adhesion on this surface compared to that on pristine Ti. The ALP-modified Ti supported cell proliferation, and significantly enhanced cellular ALP activity and calcium deposition of osteoblasts, human mesenchymal stem cells (hMSCs) and human adipose-derived stem cells (hADSCs). The extent of enhancement in the functions of these cells is dependent on the surface density of immobilized ALP. The substrate prepared using an ALP solution of 50 μg/cm{sup 2} resulted in 44%, 54% and 129% increase in calcium deposited by osteoblasts, hMSCs and hADSCs, respectively, compared to those cultured on pristine Ti. The ALP-modified substrates also promoted the osteogenic differentiation of hMSCs and hADSCs by up-regulating gene expressions of runt-related transcription factor 2 (RUNX2), osterix (OSX), and osteocalcin (OC) in the two types of stem cells. The surface-immobilized ALP was stable after being subjected to 1 h immersion in 70% ethanol and autoclaving at 121 °C for 20 min. However, the enzymatic bioactivity of the surface-immobilized ALP was reduced by about 50% after these substrates were immersed in phosphate buffered saline (PBS) or PBS containing lysozyme for 14 days.

  11. Comparison of diazo-coupling, formazan, and silver staining techniques for visualizing alkaline phosphatase isoenzymes after electrophoresis in homogeneous-pore and gradient-pore polyacrylamide gels.

    Science.gov (United States)

    Hodson, A W; Skillen, A W

    1988-03-01

    Three techniques for visualization of alkaline phosphatase after polyacrylamide-gel electrophoresis are compared. These are diazo-dye simultaneous coupling with the substrate sodium naphthyl phosphate and 5-chloro-2-toluene diazonium chloride; formazan precipitation with the substrate 5-bromo-4-chloro-3-indolyl phosphate and 3-[4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; and silver staining with the substrate sodium glycerophosphate. Each staining technique was tested with gradient-pore and homogeneous-pore acrylamide-gel electrophoresis. The main factors assessed are sensitivity; separation of the human serum alkaline phosphatase isoenzymes of the liver, bone, and intestinal types; and differences in substrate affinity, as well as the complexity of each technique. Using the three techniques only minor differences in substrate affinity are evident. There is some nonspecific staining with the diazo-coupling technique but not with the formazan and silver staining techniques. The differences, in the mobility of the liver, bone, and intestinal isoenzymes achieved by homogeneous-pore gel electrophoresis are sufficient to allow them to be clearly distinguished. However, only very small differences in mobility are found with gradient-pore gel electrophoresis, but the sharper bands in this medium allow much smaller amounts of activity to be detected. As little as 160 microU of enzyme can be visualized by the diazo technique. Silver staining gives an approximately fourfold increase in sensitivity over the formazan technique, which in turn gives a fourfold increase over the diazo technique. An important aspect of the silver staining technique is the potential of increasing sensitivity much further by improvements in the photographic physical development stage.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Calcium-phosphate biomineralization induced by alkaline phosphatase activity in Escherichia coli: localization, kinetics and potential signatures in the fossil record

    Directory of Open Access Journals (Sweden)

    Julie eCosmidis

    2015-12-01

    Full Text Available Bacteria are thought to play an important role in the formation of calcium-phosphate minerals composing marine phosphorites, as supported by the common occurrence of fossil microbes in these rocks. Phosphatase enzymes may play a key role in this process. Indeed, they may increase the supersaturation with respect to Ca-phosphates by releasing orthophosphate ions following hydrolysis of organic phosphorus. However, several questions remain unanswered about the cellular-level mechanisms involved in this model, and its potential signatures in the mineral products. We studied Ca-phosphate precipitation by different strains of Escherichia coli which were genetically modified to differ in the abundance and cellular localization of the alkaline phosphatase (PHO A produced. The mineral precipitated by either E. coli or purified PHO A was invariably identified as a carbonate-free non-stoichiometric hydroxyapatite. However, the bacterial precipitates could be discriminated from the ones formed by purified PHO A at the nano-scale. PHO A localization was shown to influence the pattern of Ca-phosphate nucleation and growth. Finally, the rate of calcification was proved to be consistent with the PHO A enzyme kinetics. Overall, this study provides mechanistic keys to better understand phosphogenesis in the environment, and experimental references to better interpret the microbial fossil record in phosphorites.

  13. Calcium-phosphate biomineralization induced by alkaline phosphatase activity in Escherichia coli: localization, kinetics and potential signatures in the fossil record

    Science.gov (United States)

    Cosmidis, Julie; Benzerara, Karim; Guyot, François; Skouri-Panet, Fériel; Duprat, Elodie; Férard, Céline; Guigner, Jean-Michel; Babonneau, Florence; Coelho, Cristina

    2015-12-01

    Bacteria are thought to play an important role in the formation of calcium-phosphate minerals composing marine phosphorites, as supported by the common occurrence of fossil microbes in these rocks. Phosphatase enzymes may play a key role in this process. Indeed, they may increase the supersaturation with respect to Ca-phosphates by releasing orthophosphate ions following hydrolysis of organic phosphorus. However, several questions remain unanswered about the cellular-level mechanisms involved in this model, and its potential signatures in the mineral products. We studied Ca-phosphate precipitation by different strains of Escherichia coli which were genetically modified to differ in the abundance and cellular localization of the alkaline phosphatase (PHO A) produced. The mineral precipitated by either E. coli or purified PHO A was invariably identified as a carbonate-free non-stoichiometric hydroxyapatite. However, the bacterial precipitates could be discriminated from the ones formed by purified PHO A at the nano-scale. PHO A localization was shown to influence the pattern of Ca-phosphate nucleation and growth. Finally, the rate of calcification was proved to be consistent with the PHO A enzyme kinetics. Overall, this study provides mechanistic keys to better understand phosphogenesis in the environment, and experimental references to better interpret the microbial fossil record in phosphorites.

  14. Loss of skeletal mineralization by the simultaneous ablation of PHOSPHO1 and alkaline phosphatase function: a unified model of the mechanisms of initiation of skeletal calcification.

    Science.gov (United States)

    Yadav, Manisha C; Simão, Ana Maria Sper; Narisawa, Sonoko; Huesa, Carmen; McKee, Marc D; Farquharson, Colin; Millán, José Luis

    2011-02-01

    Endochondral ossification is a carefully orchestrated process mediated by promoters and inhibitors of mineralization. Phosphatases are implicated, but their identities and functions remain unclear. Alkaline phosphatase (TNAP) plays a crucial role promoting mineralization of the extracellular matrix by restricting the concentration of the calcification inhibitor inorganic pyrophosphate (PP(i)). Mutations in the TNAP gene cause hypophosphatasia, a heritable form of rickets and osteomalacia. Here we show that PHOSPHO1, a phosphatase with specificity for phosphoethanolamine and phosphocholine, plays a functional role in the initiation of calcification and that ablation of PHOSPHO1 and TNAP function prevents skeletal mineralization. Phospho1(-/-) mice display growth plate abnormalities, spontaneous fractures, bowed long bones, osteomalacia, and scoliosis in early life. Primary cultures of Phospho1(-/-) tibial growth plate chondrocytes and chondrocyte-derived matrix vesicles (MVs) show reduced mineralizing ability, and plasma samples from Phospho1(-/-) mice show reduced levels of TNAP and elevated plasma PP(i) concentrations. However, transgenic overexpression of TNAP does not correct the bone phenotype in Phospho1(-/-) mice despite normalization of their plasma PP(i) levels. In contrast, double ablation of PHOSPHO1 and TNAP function leads to the complete absence of skeletal mineralization and perinatal lethality. We conclude that PHOSPHO1 has a nonredundant functional role during endochondral ossification, and based on these data and a review of the current literature, we propose an inclusive model of skeletal calcification that involves intravesicular PHOSPHO1 function and P(i) influx into MVs in the initiation of mineralization and the functions of TNAP, nucleotide pyrophosphatase phosphodiesterase-1, and collagen in the extravesicular progression of mineralization.

  15. Development of a single-chain variable fragment-alkaline phosphatase fusion protein and a sensitive direct competitive chemiluminescent enzyme immunoassay for detection of ractopamine in pork

    Energy Technology Data Exchange (ETDEWEB)

    Dong Jiexian; Li Zhenfeng; Lei Hongtao; Sun Yuanming [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Ducancel, Frederic [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Xu Zhenlin [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Boulain, Jean-Claude [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Yang Jinyi; Shen Yudong [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Wang Hong, E-mail: gzwhongd@63.com [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China)

    2012-07-29

    Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: Black-Right-Pointing-Pointer The scFv-AP fusion protein against ractopamine (RAC) was produced. Black-Right-Pointing-Pointer A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. Black-Right-Pointing-Pointer The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. Black-Right-Pointing-Pointer Recovery tests from pork samples were studied. Black-Right-Pointing-Pointer Good accuracy was obtained. - Abstract: A rapid, sensitive chemiluminescent enzyme immunoassay (CLEIA) for ractopamine (RAC) based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein was developed. The scFv gene was prepared by cloning the heavy- and light-chain variable region genes (V{sub H} and V{sub L}) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling V{sub H} and V{sub L} genes with a linker by means of splicing overlap extension polymerase chain reaction. The resulting scFv gene was inserted into the expression vector pLIP6/GN containing AP to produce the fusion protein in Escherichia coli strain BL21. The purified scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) protocol for detection of RAC. The average concentration required for 50% inhibition of binding and the limit of detection of the assay were 0.25 {+-} 0.03 and 0.02 {+-} 0.004 ng mL{sup -1}, respectively, and the linear response range extended from 0.05 to 1.45 ng mL{sup -1}. The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The results showed a good correlation between

  16. Cationized dextran nanoparticle-encapsulated CXCR4-siRNA enhanced correlation between CXCR4 expression and serum alkaline phosphatase in a mouse model of colorectal cancer

    Directory of Open Access Journals (Sweden)

    Abedini F

    2012-07-01

    Full Text Available Fatemeh Abedini,1 Hossein Hosseinkhani,2 Maznah Ismail,1,3 Abraham J Domb,4 Abdul Rahman Omar,1,5 Pei Pei Chong,1,2 Po-Da Hong,3 Dah-Shyong Yu,6 Ira-Yudovin Farber41Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Selangor, 2Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan, 3Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Selangor, Malaysia, 4Institute of Drug Research, The Center for Nanoscience and Nanotechnology, School of Pharmacy-Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel, 5Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia, 6Nanomedicine Research Center, National Defense Medical Center, Taipei, TaiwanPurpose: The failure of colorectal cancer treatments is partly due to overexpression of CXCR4 by tumor cells, which plays a critical role in cell metastasis. Moreover, serum alkaline phosphatase (ALP levels are frequently elevated in patients with metastatic colorectal cancer. A polysaccharide, dextran, was chosen as the vector of siRNA. Spermine was conjugated to oxidized dextran by reductive amination process to obtain cationized dextran, so-called dextran-spermine, in order to prepare CXCR4-siRNAs/dextran-spermine nanoparticles. The fabricated nanoparticles were used in order to investigate whether downregulation of CXCR4 expression could affect serum ALP in mouse models of colorectal cancer.Methods: Colorectal cancer was established in BALB/C mice following injection of mouse colon carcinoma cells CT.26WT through the tail vein. CXCR4 siRNA for two sites of the target gene was administered following injection of naked siRNA or siRNA encapsulated into nanoparticles.Results: In vivo animal data revealed that CXCR4 silencing by dextran-spermine nanoparticles significantly downregulated CXCR4 expression compared with naked CXCR4 siRNA. Furthermore, there was

  17. High-performance alkaline polymer electrolyte for fuel cell applications

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Jing; Lu, Shanfu; Li, Yan; Huang, Aibin; Zhuang, Lin; Lu, Juntao [College of Chemistry and Molecular Sciences, Hubei Key Lab. of Electrochemical Power Sources, Wuhan University (China)

    2010-01-22

    Although the proton exchange membrane fuel cell (PEMFC) has made great progress in recent decades, its commercialization has been hindered by a number of factors, among which is the total dependence on Pt-based catalysts. Alkaline polymer electrolyte fuel cells (APEFCs) have been increasingly recognized as a solution to overcome the dependence on noble metal catalysts. In principle, APEFCs combine the advantages of and alkaline fuel cell (AFC) and a PEMFC: there is no need for noble metal catalysts and they are free of carbonate precipitates that would break the waterproofing in the AFC cathode. However, the performance of most alkaline polyelectrolytes can still not fulfill the requirement of fuel cell operations. In the present work, detailed information about the synthesis and physicochemical properties of the quaternary ammonia polysulfone (QAPS), a high-performance alkaline polymer electrolyte that has been successfully applied in the authors' previous work to demonstrate an APEFC completely free from noble metal catalysts (S. Lu, J. Pan, A. Huang, L. Zhuang, J. Lu, Proc. Natl. Acad. Sci. USA 2008, 105, 20611), is reported. Monitored by NMR analysis, the synthetic process of QAPS is seen to be simple and efficient. The chemical and thermal stability, as well as the mechanical strength of the synthetic QAPS membrane, are outstanding in comparison to commercial anion-exchange membranes. The ionic conductivity of QAPS at room temperature is measured to be on the order of 10{sup -2} S cm{sup -1}. Such good mechanical and conducting performances can be attributed to the superior microstructure of the polyelectrolyte, which features interconnected ionic channels in tens of nanometers diameter, as revealed by HRTEM observations. The electrochemical behavior at the Pt/QAPS interface reveals the strong alkaline nature of this polyelectrolyte, and the preliminary fuel cell test verifies the feasibility of QAPS for fuel cell applications. (Abstract Copyright [2010

  18. Arbuscular mycorrhizal fungal diversity, root colonization, and soil alkaline phosphatase activity in response to maize-wheat rotation and no-tillage in North China.

    Science.gov (United States)

    Hu, Junli; Yang, Anna; Zhu, Anning; Wang, Junhua; Dai, Jue; Wong, Ming Hung; Lin, Xiangui

    2015-07-01

    Monitoring the effects of no-tillage (NT) in comparison with conventional tillage (CT) on soil microbes could improve our understanding of soil biochemical processes and thus help us to develop sound management strategies. The objective of this study was to compare the species composition and ecological function of soil arbuscular mycorrhizal (AM) fungi during the growth and rotation of crops under NT and CT. From late June 2009 to early June 2010, 32 topsoil (0-15 cm) samples from four individual plots per treatment (CT and NT) were collected at both the jointing and maturation stages of maize (Zea mays L.) and wheat (Triticum aestivum L.) from a long-term experimental field that was established in an Aquic Inceptisol in North China in June 2006. The AM fungal spores were isolated and identified and then used to calculate species diversity indices, including the Shannon- Wiener index (H'), Evenness (E), and Simpson's index (D). The root mycorrhizal colonization and soil alkaline phosphatase activity were also determined. A total of 34 species of AM fungi within nine genera were recorded. Compared with NT, CT negatively affected the soil AM fungal community at the maize sowing stage, leading to decreases in the average diversity indices (from 2.12, 0.79, and 0.82 to 1.79, 0.72, and 0.74 for H', E, and D, respectively), root mycorrhizal colonization (from 28% to 20%), soil alkaline phosphatase activity (from 0.24 to 0.19 mg/g/24 h) and available phosphorus concentration (from 17.4 to 10.5 mg/kg) at the maize jointing stage. However, reductions in diversity indices of H', E, and D were restored to 2.20, 0.81, and 0.84, respectively, at the maize maturation stage. CT should affect the community again at the wheat sowing stage; however, a similar restoration in the species diversity of AM fungi was completed before the wheat jointing stage, and the highest Jaccard index (0.800) for similarity in the species composition of soil AM fungi between CT and NT was recorded at

  19. Effects of Myriophyllum spicatum on Kinetic Characteristics of Alkaline Phosphatase in Water and Sediments%狐尾藻对水体和沉积物中碱性磷酸酶动力学特征的影响

    Institute of Scientific and Technical Information of China (English)

    赵海超; 王圣瑞; 金相灿; 焦立新

    2011-01-01

    The enzyme mechanism of an inactivating eutrophic lake by submerged plants was determined by analyzing the kinetic parameter variations of alkaline phosphataae in the overlying water, interstitial water and sediments under indoor simulating conditions with the plant Myriophyllum spicatum. The results indicated that under the experimental condition Vmax of the alkaline phosphatase, the overlying water, the interstitial water and the sediments were all reduced by planting Myriophyllum spicatum. Myriophyllum spicatum inactivated the alkaline phosphatase in the overlying water and sediments obviously, and mainly inactivated the dissolved alkaline phosphatase Vmax in the interstitial water. The effect of Myriophyllum spicatum on alkaline phosphatase in the soil was higher than that in the sediment of same nutrition. The Vmax and Km of the alkaline phosphatase in the overlying water of the soil substrate were higher than those of the sediment substrate, and the Vmax and Km of the alkaline phosphatase in the soil were lower than those in the sediment. The Vmax of the alkaline phosphatase in the sediments ascended opposite to that in the overlying water. The alkaline phosphatase in the interstitial water had obvious seasonal variation and characteristics, and Vmax attained the highest value from July to August.%在室内模拟条件下栽培狐尾藻,通过对上覆水、间隙水和沉积物中碱性磷酸酶动力学参数变化的分析,揭示了沉水植物对湖泊富营养化影响的酶学机制.结果表明:在试验条件下,栽培狐尾藻使上覆水、间隙水和沉积物中的碱性磷酸酶的最大反应速率(Vmax)均有所降低;狐尾藻对上覆水和底质中碱性磷酸酶反应速率及亲和力的抑制作用比较明显,对间隙水主要是抑制溶解性碱性磷酸酶的Vmax;狐尾藻对土壤中碱性磷酸酶的影响比同一营养水平的沉积物大,与沉积物相比,土壤作底质时上覆水中碱性磷酸酶的Vmax和K(ms)(米氏

  20. Intestinal Alkaline Phosphatase: Potential Roles in Promoting Gut Health in Weanling Piglets and Its Modulation by Feed Additives — A Review

    Science.gov (United States)

    Melo, A. D. B.; Silveira, H.; Luciano, F. B.; Andrade, C.; Costa, L. B.; Rostagno, M. H.

    2016-01-01

    The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP) is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS), unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs) activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria) detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP’s role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets. PMID:26732323

  1. Intestinal Alkaline Phosphatase: Potential Roles in Promoting Gut Health in Weanling Piglets and Its Modulation by Feed Additives — A Review

    Directory of Open Access Journals (Sweden)

    A. D. B. Melo

    2016-01-01

    Full Text Available The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS, unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP’s role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets.

  2. Prokaryotic responses to ammonium and organic carbon reveal alternative CO2 fixation pathways and importance of alkaline phosphatase in the mesopelagic North Atlantic

    Directory of Open Access Journals (Sweden)

    Federico Baltar

    2016-10-01

    Full Text Available To decipher the response of mesopelagic prokaryotic communities to input of nutrients, we tracked changes in prokaryotic abundance, extracellular enzymatic activities, heterotrophic production, dark dissolved inorganic carbon (DIC fixation, community composition (16S rRNA sequencing and community gene expression (metatranscriptomics in 3 microcosm experiments with water from the mesopelagic North Atlantic. Responses in 3 different treatments amended with thiosulfate, ammonium or organic matter (i.e. pyruvate plus acetate were compared to unamended controls. The strongest stimulation was found in the organic matter enrichments, where all measured rates increased >10-fold. Strikingly, in the organic matter treatment, the dark DIC fixation rates —assumed to be related to autotrophic metabolisms— were equally stimulated as all the other heterotrophic-related parameters. This increase in DIC fixation rates was paralleled by an up-regulation of genes involved in DIC assimilation via anaplerotic pathways. Alkaline phosphatase was the metabolic rate most strongly stimulated and its activity seemed to be related to cross-activation by nonpartner histidine kinases, and/or the activation of genes involved in the regulation of elemental balance during catabolic processes. These findings suggest that episodic events such as strong sedimentation of organic matter into the mesopelagic might trigger rapid increases of originally rare members of the prokaryotic community, enhancing heterotrophic and autotrophic carbon uptake rates, ultimately affecting carbon cycling. Our experiments highlight a number of fairly unstudied microbial processes of potential importance in mesopelagic waters that require future attention.

  3. Bifunctional coating based on carboxymethyl chitosan with stable conjugated alkaline phosphatase for inhibiting bacterial adhesion and promoting osteogenic differentiation on titanium

    Science.gov (United States)

    Zheng, Dong; Neoh, Koon Gee; Kang, En-Tang

    2016-01-01

    In this work, alkaline phosphatase (ALP) was covalently immobilized on carboxymethyl chitosan (CMCS)-coated polydopamine (PDA)-functionalized Ti to achieve a bifunctional surface. Our results showed ∼89% reduction in Staphylococcus epidermidis adhesion on this surface compared to that on pristine Ti. The ALP-modified Ti supported cell proliferation, and significantly enhanced cellular ALP activity and calcium deposition of osteoblasts, human mesenchymal stem cells (hMSCs) and human adipose-derived stem cells (hADSCs). The extent of enhancement in the functions of these cells is dependent on the surface density of immobilized ALP. The substrate prepared using an ALP solution of 50 μg/cm2 resulted in 44%, 54% and 129% increase in calcium deposited by osteoblasts, hMSCs and hADSCs, respectively, compared to those cultured on pristine Ti. The ALP-modified substrates also promoted the osteogenic differentiation of hMSCs and hADSCs by up-regulating gene expressions of runt-related transcription factor 2 (RUNX2), osterix (OSX), and osteocalcin (OC) in the two types of stem cells. The surface-immobilized ALP was stable after being subjected to 1 h immersion in 70% ethanol and autoclaving at 121 °C for 20 min. However, the enzymatic bioactivity of the surface-immobilized ALP was reduced by about 50% after these substrates were immersed in phosphate buffered saline (PBS) or PBS containing lysozyme for 14 days.

  4. Uptake of nickel from 316L stainless steel into contacting osteoblastic cells and metal ion interference with BMP-2-induced alkaline phosphatase.

    Science.gov (United States)

    Mölders, Martina; Felix, Joachim; Bingmann, Dieter; Hirner, Alfred; Wiemann, Martin

    2007-11-01

    Bone cells contacting nickel (Ni)-containing implant materials may be affected by Ni species via disturbed signaling pathways involved in bone cell development. Here we analyze effects of the Ni-containing steel 316L and major metal constituents thereof on bone morphogenetic protein-2 (BMP-2)-induced alkaline phosphatase (ALP) of MC3T3-E1 cells. While cells grew normally on 316L, cellular Ni content increased 10-fold vs. control within 4 days. With respect to the major components of 316L, Ni2+ (3-50 microM) was most inhibitory to BMP-2-induced ALP, whereas even 50 microM Fe3+, Cr3+, Mo5+, or Mn2+ had no such effect. In line with this, BMP-2-induced ALP was significantly reduced in cells on 316L. This effect was not prevented by the metal ion chelator diethylenetriaminepentaacetic acid (DTPA). Instead, DTPA abolished the stimulatory effect of BMP-2 on ALP, pointing to chelatable metal ions involved. Zn2+, as one possible candidate, antagonized the Ni2+ inhibition of BMP-2-induced ALP in both MC3T3-E1 and human bone marrow stromal cells. Results show that cells contacting 316L steel are exposed to increased concentrations of Ni which suffice to impair BMP-2-induced ALP activity. Zn2+, as a competitor of this inhibition, may help to restore normal osteoblastic function and bone development under these conditions.

  5. The utility of alkaline phosphatase measurement as a screening test for rickets in breast-fed infants and toddlers: a study from the puget sound pediatric research network.

    Science.gov (United States)

    Taylor, James A; Richter, Monica; Done, Stephen; Feldman, Kenneth W

    2010-12-01

    To determine if alkaline phosphatase (AP) levels are a useful screening test for rickets, the authors measured serum AP levels in children 6 to 15 months old who were predominantly breast-fed for > 6 months without vitamin D supplementation. Radiographs were obtained on children with elevated AP levels to determine the presence of rickets. AP levels were obtained on 246 children; levels were elevated in 33 (13.4%). Rickets was present in 4 of 18 children with elevated levels on whom radiographs were obtained. The sensitivity and specificity of AP levels as a test for rickets was maximal at a cutoff value of 552 U/L. Using this cutoff value, the specificity of AP levels as a test for rickets was 97.4%, and the positive predictive value (PPV) was 40.0%. These results suggest that AP levels may be a useful screening test for rickets in children who are breast-fed for prolonged periods without vitamin D supplementation.

  6. A single electrochemical biosensor for detecting the activity and inhibition of both protein kinase and alkaline phosphatase based on phosphate ions induced deposition of redox precipitates.

    Science.gov (United States)

    Shen, Congcong; Li, Xiangzhi; Rasooly, Avraham; Guo, Linyan; Zhang, Kaina; Yang, Minghui

    2016-11-15

    Protein kinase (PKA) and alkaline phosphatase (ALP) are clinically relevant enzymes for a number of diseases. In this work, we developed a new simple electrochemical biosensor for the detection of the activity and inhibition of both PKA and ALP. One common feature of the PKA and ALP catalyzing process is that PKA can hydrolysis adenosine-5'-triphosphate (ATP) and ALP can hydrolysis pyrophosphate, both reactions produce phosphate ions, and the amount of phosphate ion produced is proportional to enzyme activity. Our assay is based on the principle that phosphate ions react with molybdate to form redox molybdophosphate precipitates on the electrode surface, thus generating electrochemical current. The detection limit for PKA and ALP were much lower than existing assays. The biosensor has good specificity and was used to measure drug-stimulated PKA from lysates of HeLa cells. We also evaluated the use of the biosensor as a screening tool for enzyme inhibitors. To the best of our knowledge, this is the first report of a biosensor capable of detecting the activity of both PKA and ALP. This tool has the potential to simplify PKA and ALP clinical measurement, thereby improving diagnostics of relevant diseases. It also may serve as the basis for a simple screening method for new enzyme inhibitors for disease treatment.

  7. Evaluation of Anti-aging Compounds Using the Promoters of Elastin and Fibrillin-1 Genes Combined with a Secreted Alkaline Phosphatase Reporter in Normal Human Fibroblasts.

    Science.gov (United States)

    Lin, Chih-Chien; Yang, Chao-Hsun; Kuo, Wan-Ting; Chen, Cheng-Yu

    2015-01-01

    Elastic fibers are major constituents of the extracellular matrix (ECM) in dynamic tissues in the human body, and regulation of elastin and fibrillin-1 expression mediates the formation of these fibers. Traditional assays for the measurement of elastin and fibrillin-1, such as western blotting, Luna staining and immunostaining, are relatively complex and time-consuming. Thus, a relatively simple assay system that also provides rational results is urgently needed. In the study, we aimed to develop a human cell-based assay system that can be used to analyze functional compounds using the promoters of elastin (ELN) and fibrillin-1 (FBN1) genes integrated with a secreted alkaline phosphatase (SEAP) reporter in normal human fibroblast cells. We used this system to assess anti-aging compounds. We used several regulators of elastinogenesis, including retinol, coenzyme Q10, deoxyArbutin and Elestan(TM) (Manilkara multinervis leaf extract), to verify the efficacy of this assay system. Our results demonstrate that this assay system can be used as a fast and realistic method for identifying anti-aging components for future use in foods, cosmetics and drugs.

  8. Naked-eye sensitive detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) based on a horseradish peroxidase catalytic colorimetric system with Cu(ii).

    Science.gov (United States)

    Shi, Dongmin; Sun, Yue; Lin, Lin; Shi, Chunjun; Wang, Guangfeng; Zhang, Xiaojun

    2016-10-07

    In this paper, a novel colorimetric method for the detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) was designed based on a Cu(2+)-horseradish peroxidase (HRP)-3,3',5,5'-tetra-methylbenzidine (TMB)-H2O2 system. In the presence of ALP, l-ascorbic acid-2-phosphate (AAP) could be hydrolyzed to ascorbic acid which could reduce Cu(2+) to Cu(+) to inhibit the enzymatic activity of HRP in the colorimetric system. The change in absorbance was found to be proportional to the ALP concentration with a linear detection range and a limit of detection of 5.4 mU mL(-1). In the presence of PPi, because Cu(2+) was chelated by PPi, the conversion of Cu(ii) by AA was effectively inhibited. The color of the HRP-TMB-H2O2 system with Cu(2+) showed blue. The HRP-TMB-H2O2 system with the Cu(2+) colorimetric system could also detect PPi with a satisfying result. In summary, this method possesses sensitivity, reproducibility, and cost-effectiveness without labelling and separation and the use of a colorimetric method is more in line with the requirements of on-site detection and green chemistry.

  9. Alkaline phosphatases are involved in the response of Aedes aegypti larvae to intoxication with Bacillus thuringiensis subsp. israelensis Cry toxins.

    Science.gov (United States)

    Stalinski, Renaud; Laporte, Frédéric; Després, Laurence; Tetreau, Guillaume

    2016-03-01

    Bacillus thuringiensis subsp. israelensis (Bti) is a natural pathogen of dipterans widely used as a biological insecticide for mosquito control. To characterize the response of mosquitoes to intoxication with Bti, the transcriptome profile of Bti-exposed susceptible Aedes aegypti larvae was analysed using Illumina RNA-seq. Gene expression of 11 alkaline phosphatases (ALPs) was further investigated by real time quantitative polymerase chain reaction and ALP activity was measured in the susceptible strain and in four strains resistant to a single Bti Cry toxin or to Bti. These strains were unexposed or exposed to their toxin of selection. Although all resistant strains constitutively exhibited a higher level of transcription of ALP genes than the susceptible strain, they showed a lower total ALP activity. The intoxication with different individual Cry toxins triggered a global pattern of ALP gene under-transcription in all the one-toxin-resistant strains but involving different specific sets of ALPs in each resistant phenotype. Most of the ALPs involved are not known Cry-binding proteins. RNA interference experiment demonstrated that reducing ALP expression conferred increased the survival of larvae exposed to Cry4Aa, confirming the involvement of ALP in Cry4Aa toxicity.

  10. Copper(II) complexes with cyanoguanidine and o-phenanthroline: Theoretical studies, in vitro antimicrobial activity and alkaline phosphatase inhibitory effect

    Science.gov (United States)

    Martínez Medina, Juan J.; Islas, María S.; López Tévez, Libertad L.; Ferrer, Evelina G.; Okulik, Nora B.; Williams, Patricia A. M.

    2014-01-01

    Calculations based on density functional methods are carried out for two Cu(II) complexes with cyanoguanidine (cnge) and o-phenanthroline (o-phen): [Cu(o-phen)2(cnge)](NO3)2ṡ2H2O (1) and [Cu(o-phen)(cnge)(H2O)(NO3)2] (2). The calculated geometrical parameters are in agreement with the experimental values. The results of Atoms in Molecules (AIM) topological analysis of the electron density indicate that the Cu-N(phen) bonds in complex (1) have lower electron density, suggesting that those bonds are stronger in complex (2). Moreover, the ionic character of the Cu-N bond in the complex (1) is slightly stronger than that in the complex (2) and this situation would explain the fact that only complex (2) was stable in water solution. For this reason, the antimicrobial and enzymatic assays were performed using complex (2). It is well known that the increased use of antibiotics has resulted in the development of resistant bacterial and fungal strains. In this context, the study of novel antimicrobial agents has an enormous importance and metal complexes represent an interesting alternative for the treatment of infectious diseases. The aim of this work is to prove the modification of some biological properties like antimicrobial activity or alkaline phosphatase inhibitory activity upon copper complexation.

  11. The evaluation of 25-hydroxy vitamin D, calcium, phosphate and alkaline phosphatase levels in epileptic children under antiepileptic medication

    Directory of Open Access Journals (Sweden)

    Keyhani doost Z

    2011-01-01

    Full Text Available "n 800x600 Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif";} Background: Epilepsy is a common disease in the pediatric neurology. There are frequent anti-epileptic drugs which are used in management of epilepsy. Anti-epileptic drugs may have some complications on bone and vitamin-D metabolism. In this study we aimed to evaluate vitamin-D metabolism in epileptic children."n"nMethods: The study was a prospective and cross sectional one. A total 89 epileptic children who were taking anti-epileptic drugs for longer than six months with no underlying disorder in Imam Khomeini and Bahrami Hospitals in Tehran, Iran were enrolled in our study"n"nResults: Forty nine boys and 40 girls were enrolled in this study; mean age of the patients was 7.8±2.1 years. Mean duration of anti-epileptic drug therapy was 2.3 years (SD=0.4, 70 of patients were under monotherapy and 19 were under polytherapy. None of the patients had signs of rickets. Serum calcium and phosphor levels were within normal ranges. Serum alkaline phosphates levels were increased more than two times in 43%. 42% had vitamin-D deficiency (25-OH Vit D<10 ng/ml and another 33% had vitamin-D insufficiency (10<25-oh Vit D<20 ng/ml. 29 patients (32% were taking prophylactic supplemental Vit D (200-400 IU/day. There was significant difference between patients taking supplemental vitamin-D as prophylaxis and patients who did not (p=0.04. There was no significant difference in vitamin-D levels between patients according to age, gender or different drugs."n"nConclusion: Periodic

  12. Assessment of the alkaline phosphatase level in gingival crevicular fluid, as a biomarker to evaluate the effect of scaling and root planing on chronic periodontitis: An in vivo study

    Science.gov (United States)

    Kunjappu, Jimly James; Mathew, Vinod Babu; Hegde, Shashikanth; Kashyap, Rajesh; Hosadurga, Rajesh

    2012-01-01

    Context: Clinical evaluation of gingivitis and/or periodontitis does not predict the progression or remission of the disease. Due to this diagnostic constraint, clinicians assume that the pathology has an increased risk of progression and plan treatments, despite the knowledge that all inflamed sites are not necessarily progressing. Extensive research has been carried out on gingival crevicular fluid (GCF) components that might serve as potential diagnostic markers for periodontitis. Among them alkaline phosphatase (ALP) levels in GCF has shown promise as a diagnostic marker. Aim: This study compares the levels of GCF alkaline phosphatase in patients with chronic periodontitis before and after scaling and root planing. Materials and Methods: This study is an in vivo longitudinal study conducted on twenty patients with localized periodontitis. The GCF was collected from the affected site prior to scaling and root planing and ALP level estimated. The probing depth and plaque index at the site were also measured for correlation. Patients were recalled after 7, 30, and 60 days for reassessment. Results: The GCF ALP values showed a sustained, statistically significant decrease after treatment. There was a positive correlation with probing depth but not with plaque index measured at each interval. Conclusion: The assessment of level of periodontal disease and effect of mechanical plaque control on the progression and regression of the disease can be evaluated precisely by the corresponding GCF ALP levels. Thus, alkaline phosphatase level is not only a biomarker for the pathology but also an indicator of prognosis of periodontitis. PMID:22438644

  13. 碱性磷酸酶试剂开瓶的稳定性研究%Study on stability of alkaline phosphatase reagent after the packaging was opened

    Institute of Scientific and Technical Information of China (English)

    段爱军; 段爱华

    2015-01-01

    Objective:To study the stability of alkaline phosphatase(ALP) after the packaging was opened and to provide the data for purchasing right amount reageng. Methods:The method of synchronous measurement was used and five kits(S0, S1, S2, S3, S4) were chosen. ALP routine test was performed using the kit S4 at eight a.m. in first day. The remaining reagent was covered tightly and preserved at 2-8 ℃. Then S3, S2, S1 and S0 kits were opened at the same time in second, third, fourth and fifth day using the same method, respectively. Five of the same parameters for ALP detection channel (ALP0, ALP1, ALP2, ALP3, ALP4, corresponding to S0, S1, S2, S3, S4) were also set in the instrument according to the reagent instruction. The detection channels were calibrated using the calibration materials(BioSino Bio-technology and Science Inc) in Selectra-E fully automatic biochemical analyzer. Results: Compared with the results obtained by S0, the results from S3 and S4 showed statistically significance(P0.05). Conclusion: The ALP reagent is stable in 48 hours after the packaging was opened and the laboratory should choose suitable packing size of reagent kit according to the dosage to use.%目的:研究碱性磷酸酶(alkaline phosphatase, ALP)试剂开瓶后的稳定性,为实验室合理购买试剂包规格提供依据。方法:采用同步测量设计方案,首先选择5个试剂盒,分别记为S0、S1、S2、S3、S4,第1天上午8点启用试剂S4进行ALP常规检测,未用完试剂盖紧盖,置2~8℃保存备用;第2天、第3天、第4天上午相同时间、相同方式启用S3、S2、S1;第5天启用试剂S0;在仪器上按试剂说明书设置5个相同参数的ALP检测通道,记为ALP0、ALP1、ALP2、ALP3、ALP4,对应试剂为S0、S1、S2、S3、S4;在荷兰威图Selectra-E全自动生化仪上用校准品(中生北控生物科技有限公司)校准各个通道;测量样品并进行结果比较分析。结果:试剂 S3、S4与试剂 S0

  14. 妊娠妇女不同孕期血清碱性磷酸酶变化%Variation of serum alkaline phosphatase in pregnant in different stage

    Institute of Scientific and Technical Information of China (English)

    周洁

    2016-01-01

    Objective To explore the variation of serum alkaline phosphatase in Pregnancy in dif‐ferent stage .Methods Unicel Dxc800 fully automatic biochemical anaylzer was used to detect the con‐centrations of serum ALP in 522 pregnant and 157 control .Results The concentrations of serum ALP in control group ,early pregnancy ,midtrimester and late pregnancy group were (66 .58 ± 44 .09) U /L , (61 .64 ± 17 .86)U /L ,(79 .98 ± 26 .38)U /L ,and (202 .89 ± 68 .94 )U /L respectively .Statistic analysis dem‐onstrated that there was no difference between early pregnancy group and control one( P > 0 .05) .The concentrations of serum ALP in midtrimester pregnancy and late pregnancy groups were hiher than those in early pregnancy and control groups significantly ,and the concentration of serum ALP in late pregnancy group was hiher than that in midtrimester pregnancy group significantly ( P < 0 .05) .Condu‐sion The serum level of alkaline phosphatase in pregnant increased with gestational prolongation .%①目的探讨不同孕周妊娠期妇女血清碱性磷酸酶的变化。②方法采用贝克曼(Unicel Dxc800)全自动生化分析仪检测522例妊娠早、中、晚期妇女血清碱性磷酸酶值,同时选取同期157例健康育龄非妊娠妇女作为对照,方差分析不同组间血清碱性磷酸酶值差异。③结果健康育龄非妊娠妇女及妊娠期妇女孕早期、孕中期和孕晚期血清碱性磷酸酶浓度分别为(66.58±44.09)U /L 、(61.64±17.86)U /L 、(79.98±26.38)U /L 、(202.89±68.94)U /L 。统计学分析结果表明:孕早期血清碱性磷酸酶水平与对照组比较差异无统计学意义( P >0.05);孕中期、孕晚期高于孕早期及对照组,孕晚期高于孕中期,差异均具有统计学意义( P 均<0.05)。④结论妇女孕期血清碱性磷酸酶水平随孕周数增加呈逐渐上升趋势。

  15. Evaluation of a chemiluminescence method for measuring alkaline phosphatase activity in whole milk of multiple species and bovine dairy drinks: interlaboratory study.

    Science.gov (United States)

    Salter, Robert S; Fitchen, John

    2006-01-01

    Alkaline phosphatase (ALP) is a ubiquitous enzyme in milk with time-temperature destruction similar to that of certain pathogens destroyed in pasteurization. Measurement of ALP to indicate proper pasteurization is a common practice. Recently the public health level for ALP was decreased to 350 mU/L, a level below the sensitivity of older colorimetric ALP methods. This study was conducted within the structure of the International Dairy Federation and the International Organization for Standardization to evaluate the reproducibility of the chemiluminescence method (Charm PasLite) for ALP at 50, 100, 350, and 500 mU/L in whole milk of multiple species to meet new regulations in the United States and proposed regulations in the European Union (EU). Fifteen laboratories from 8 countries evaluated bovine, goat, sheep, and buffalo milk, bovine skim milk, 20% fat cream, and 2% fat chocolate milk. At ALP levels of 350 and 500 mU/L, the average relative standard deviation for repeatability (RSDr) was 7.5%, and the average relative standard deviation of reproducibility was (RSDR) 15%. For ALP at 100 and 50 mU/L, the average RSDr values were 10.5 and 12.6%, respectively, and the average RSDR values were 18 and 25%, respectively. The limit of detection was 20 mU/L. Results are comparable to those obtained with other enzymatic photo-activated system methods such as the fluorometric method. Results indicate that the method is suitable for measuring ALP in the milk of multiple species and in dairy drinks at U.S. and proposed EU levels.

  16. Partial characterization and response under hyperregulating conditions of Na+-K+ ATPase and levamisole-sensitive alkaline phosphatase activities in chela muscle of the euryhaline crab Cyrtograpsus angulatus

    Directory of Open Access Journals (Sweden)

    Silvina Andrea Pinoni

    2008-03-01

    Full Text Available The occurrence, characteristics and response to changes in environmental salinity of Na+-K+ ATPase and levamisole-sensitive alkaline phosphatase (AP activities were studied in chela muscle of the euryhaline crab Cyrtograpsus angulatus. Chela muscle exhibited an Na+-K+ ATPase activity which was strongly dependent on ATP concentration, pH and temperature of the reaction mixture. Maximal activity was found at 1 mM ATP, 30-37°C and pH 7.4. Levamisole-sensitive AP activity was characterised at physiological pH 7.4 and at pH 8.0. I50 for levamisole-sensitive AP activity was 8.8 mM and 8.0 mM at pH 7.4 and 8.0, respectively. At both pH levels, levamisole-sensitive AP activity exhibited Michaelis-Menten kinetics (Km=3.451 mM and 6.906 mM at pH 7.4 and 8.0, respectively. Levamisole-sensitive AP activities were strongly affected by temperature, exhibiting a peak at 37ºC. In crabs acclimated to low salinity (10; hyperegulating conditions, Na+-K+ ATPase activity and levamisole-sensitive AP activity at the physiological pH were higher than in 35 psu (osmoconforming conditions. The response to low salinity suggests that both activities could be components of muscle regulatory mechanisms at the biochemical level secondary to hyperegulation of C. angulatus. The study of these activities under hyperegulating conditions contributes to a better understanding of the complexity of biochemical mechanisms underlying the adaptive process of euryhaline crabs.

  17. Display of E. coli Alkaline Phosphatase pIII or pVIII Fusions on Phagemid Surfaces Reveals Monovalent Decoration with Active Molecules

    Science.gov (United States)

    Weichel, Michael; Jaussi, Rolf; Rhyner, Claudio; Crameri, Reto

    2008-01-01

    Active alkaline phosphatase of Escherichia coli (PhoA, EC 3.1.3.1) was displayed via the leucine zipper element of the Jun-Fos heterodimer on the surface of filamentous phage and the kinetic parameters Km and kcat were determined. The phoA gene was cloned downstream of fos while jun was inserted upstream of pIII or pVIII, alternatively, in the pJuFo phagemid vector. Both fusion genes are regulated by independent lacZ promoters. PhoA displayed on the phagemid pIII surface exhibited a Km of 11.2 µM with 4-nitrophenyl phosphate as substrate, which is consistent with data published for soluble PhoA. Based on these data we calculated the decoration of pJuFo phagemid with PhoA using the minor and major coat proteins pIII and pVIII as fusion partners under variable inducing conditions. We found that, even if the promoters are fully induced at a concentration of 1000 µM IPTG, the phagemids display maximally one copy of PhoA-Fos-Jun-coat protein fusion, irrespective of whether the protein is presented via pIII or pVIII. However, since PhoA is displayed in a native-like fashion, as deduced from the kinetic parameters of the enzymatic reaction, the pJuFo technology provides a versatile tool for the functional screening of complex cDNA libraries displayed on the phagemids' surface. PMID:18949073

  18. Incorporation of alkaline phosphatase into layer-by-layer polyelectrolyte films on the surface of affi-gel heparin beads: physicochemical characterization and evaluation of the enzyme stability.

    Science.gov (United States)

    Derbal, Lylia; Lesot, Hervé; Voegel, Jean Claude; Ball, Vincent

    2003-01-01

    The preparation of functionalized beads in the micrometer size range that can be used to probe the action of immobilized biomolecules on cell cultures during controlled periods of time is of fundamental importance in cell biology. However, the preparation and characterization of such particles is tedious because of their fast sedimentation. It is hence difficult to prepare such beads in a reproducible manner. This highlights the need to prepare an important batch of functionnalized particles and to store them under conditions where the loss of biological activity is minimized. The aim of this paper was to immobilize alkaline phosphatase (AP) as a model enzyme on the surface of Affi-gel heparin beads functionnalized by means of a layer-by-layer (LBL) film made of poly-l-glutamic (PGA) acid and poly-l-lysine (PLL). The enzyme has been adsorbed either on the top of the LBL film or embedded under five polyelectrolyte layers. When embedded, the enzyme was not released in buffer and retained more than 30% of its initial activity after 3 months of storage at 4 degrees C. However, when the enzyme was adsorbed on top of the LBL film, about 80% of the adsorbed enzyme was released in the buffer after a few days of storage. Longer storage did not lead to any further desorption and the remaining enzyme displayed the same evolution of its activity with time as the embedded enzyme. The time evolution of the enzyme activity on the beads is compared with that in solution alone and in the presence of PGA and PLL separately.

  19. Tetramisole and Levamisole Suppress Neuronal Activity Independently from Their Inhibitory Action on Tissue Non-specific Alkaline Phosphatase in Mouse Cortex.

    Science.gov (United States)

    Nowak, Lionel G; Rosay, Benoît; Czégé, Dávid; Fonta, Caroline

    2015-01-01

    Tissue non-specific alkaline phosphatase (TNAP) may be involved in the synthesis of GABA and adenosine, which are the main inhibitory neurotransmitters in cortex. We explored this putative TNAP function through electrophysiological recording (local field potential ) in slices of mouse somatosensory cortex maintained in vitro. We used tetramisole, a well documented TNAP inhibitor, to block TNAP activity. We expected that inhibiting TNAP with tetramisole would lead to an increase of neuronal response amplitude, owing to a diminished availability of GABA and/or adenosine. Instead, we found that tetramisole reduced neuronal response amplitude in a dose-dependent manner. Tetramisole also decreased axonal conduction velocity. Levamisole had identical effects. Several control experiments demonstrated that these actions of tetramisole were independent from this compound acting on TNAP. In particular, tetramisole effects were not stereo-specific and they were not mimicked by another inhibitor of TNAP, MLS-0038949. The decrease of axonal conduction velocity and preliminary intracellular data suggest that tetramisole blocks voltage-dependent sodium channels. Our results imply that levamisole or tetramisole should not be used with the sole purpose of inhibiting TNAP in living excitable cells as it will also block all processes that are activity-dependent. Our data and a review of the literature indicate that tetramisole may have at least four different targets in the nervous system. We discuss these results with respect to the neurological side effects that were observed when levamisole and tetramisole were used for medical purposes, and that may recur nowadays due to the recent use of levamisole and tetramisole as cocaine adulterants.

  20. Down regulation of a gene for cadherin, but not alkaline phosphatase, associated with Cry1Ab resistance in the sugarcane borer Diatraea saccharalis.

    Directory of Open Access Journals (Sweden)

    Yunlong Yang

    Full Text Available The sugarcane borer, Diatraea saccharalis, is a major target pest of transgenic corn expressing Bacillus thuringiensis (Bt proteins (i.e., Cry1Ab in South America and the mid-southern region of the United States. Evolution of insecticide resistance in such target pests is a major threat to the durability of transgenic Bt crops. Understanding the pests' resistance mechanisms will facilitate development of effective strategies for delaying or countering resistance. Alterations in expression of cadherin- and alkaline phosphatase (ALP have been associated with Bt resistance in several species of pest insects. In this study, neither the activity nor gene regulation of ALP was associated with Cry1Ab resistance in D. saccharalis. Total ALP enzymatic activity was similar between Cry1Ab-susceptible (Cry1Ab-SS and -resistant (Cry1Ab-RR strains of D. saccharalis. In addition, expression levels of three ALP genes were also similar between Cry1Ab-SS and -RR, and cDNA sequences did not differ between susceptible and resistant larvae. In contrast, altered expression of a midgut cadherin (DsCAD1 was associated with the Cry1Ab resistance. Whereas cDNA sequences of DsCAD1 were identical between the two strains, the transcript abundance of DsCAD1 was significantly lower in Cry1Ab-RR. To verify the involvement of DsCAD1 in susceptibility to Cry1Ab, RNA interference (RNAi was employed to knock-down DsCAD1 expression in the susceptible larvae. Down-regulation of DsCAD1 expression by RNAi was functionally correlated with a decrease in Cry1Ab susceptibility. These results suggest that down-regulation of DsCAD1 is associated with resistance to Cry1Ab in D. saccharalis.

  1. Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B{sub 1} detection in cereal

    Energy Technology Data Exchange (ETDEWEB)

    Shu, Mei [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Xu, Yang, E-mail: xuyang@ncu.edu.cn [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Liu, Xing [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228 (China); Li, Yanping; He, Qinghua [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Tu, Zhui [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Fu, Jinheng [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Gee, Shirley J.; Hammock, Bruce D. [Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616 (United States)

    2016-06-14

    A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB{sub 1} was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB{sub 1} were 2.69 and 0.35 ng mL{sup −1}, respectively, with a linear range of 0.93–7.73 ng mL{sup −1}. The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL{sup −1}, and the IC{sub 50} was 0.89 ± 0.09 ng mL{sup −1} with a linear range of 0.29–2.68 ng mL{sup −1}. Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB{sub 1} contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems. - Highlights: • Ab2β−Nb−AP has the potential to replace chemically-coupled probes. • Ab2β−Nb−AP is homogeneous enzyme-labelled antigen can be prepared reproducibly. • We developed a green and rapid one-step competitive enzyme immunoassay. • The sensitivity of one-step CLIA was 9-folds higher than two-step ELISA.

  2. Mechanical loading induced expression of bone morphogenetic protein-2,alkaline phosphatase activity,and collagen synthesis in osteoblastic MC3T3-E1 cells

    Institute of Scientific and Technical Information of China (English)

    LU Hong-fei; MAI Zhi-hui; XU Ye; WANG Wei; AI Hong

    2012-01-01

    Background Bone morphogenetic protein(BMP)-2,alkaline phosphatase(ALP),and collagen typeⅠ?are known to play a critical role in the process of bone remodeling.However,the relationship between mechanical strain and the expression of BMP-2,ALP,and COL-Ⅰ?in osteoblasts was still unknown.The purpose of this study was to investigate the effects of different magnitudes of mechanical strain on osteoblast morphology and on the expression of BMP-2,ALP,and COL-Ⅰ.Methods Osteoblast-like cells were flexed at four deformation rates(0,6%,12%,and 18% elongation).The expression of BMP-2 mRNA,ALP,and COL-Ⅰ?in osteoblast-like cells were determined by real-time quantitative reverse transcription polymerase chain reaction,respectively.The results were subjected to analysis of variance(ANOVA)using SPSS 13.0 statistical software.Results The cells changed to fusiform and grew in the direction of the applied strain after the mechanical strain was loaded.Expression level of the BMP-2,ALP,and COL-Ⅰ?increased magnitude-dependently with mechanical loading in the experimental groups,and the 12% elongation group had the highest expression(P<0.05).Conclusion Mechanical strain can induce morphological change and a magnitude-dependent increase in the expression of BMP-2,ALP,and COL-Ⅰ?mRNA in osteoblast-like cells,which might influence bone remodeling in orthodontic treatment.

  3. Ultrasensitive electroanalysis of low-level free microRNAs in blood by maximum signal amplification of catalytic silver deposition using alkaline phosphatase-incorporated gold nanoclusters.

    Science.gov (United States)

    Si, Yanmei; Sun, Zongzhao; Zhang, Ning; Qi, Wei; Li, Shuying; Chen, Lijun; Wang, Hua

    2014-10-21

    An ultrasensitive sandwich-type analysis method has been initially developed for probing low-level free microRNAs (miRNAs) in blood by a maximal signal amplification protocol of catalytic silver deposition. Gold nanoclusters (AuNCs) were first synthesized and in-site incorporated into alkaline phosphatase (ALP) to form the ALP-AuNCs. Unexpectedly, the so incorporated AuNCs could dramatically enhance the catalysis activities of ALP-AuNCs versus native ALP. A sandwiched hybridization protocol was then proposed using ALP-AuNCs as the catalytic labels of the DNA detection probes for targeting miRNAs that were magnetically caught from blood samples by DNA capture probes, followed by the catalytic ligation of two DNA probes complementary to the targets. Herein, the ALP-AuNC labels could act as the bicatalysts separately in the ALP-catalyzed substrate dephosphorylation reaction and the AuNCs-accelerated silver deposition reaction. The signal amplification of ALP-AuNCs-catalyzed silver deposition was thereby maximized to be measured by the electrochemical outputs. The developed electroanalysis strategy could allow for the ultrasensitive detection of free miRNAs in blood with the detection limit as low as 21.5 aM, including the accurate identification of single-base mutant levels in miRNAs. Such a sandwich-type analysis method may circumvent the bottlenecks of the current detection techniques in probing short-chain miRNAs. It would be tailored as an ultrasensitive detection candidate for low-level free miRNAs in blood toward the diagnosis of cancer and the warning or monitoring of cancer metastasis in the clinical laboratory.

  4. A disulfide bond-containing alkaline phosphatase triggers a BdbC-dependent secretion stress response in Bacillus subtilis

    NARCIS (Netherlands)

    Darmon, Elise; Dorenbos, Ronald; Meens, Jochen; Freudl, Roland; Antelmann, Haike; Hecker, Michael; Kuipers, Oscar P.; Bron, Sierd; Quax, Wim J.; Dubois, Jean-Yves F.; van Dijl, Jan Maarten

    2006-01-01

    The gram-positive bacterium Bacillus subtilis secretes high levels of proteins into its environment. Most of these secretory proteins are exported from the cytoplasm in an unfolded state and have to fold efficiently after membrane translocation. As previously shown for alpha-amylases of Bacillus

  5. 1-Step Versus 2-Step Immobilization of Alkaline Phosphatase and Bone Morphogenetic Protein-2 onto Implant Surfaces Using Polydopamine

    NARCIS (Netherlands)

    Nijhuis, A.W.G.; Beucken, J.J.J.P van den; Boerman, O.C.; Jansen, Jan; Leeuwenburgh, S.C.G.

    2013-01-01

    Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the cu

  6. 1-Step Versus 2-Step Immobilization of Alkaline Phosphatase and Bone Morphogenetic Protein-2 onto Implant Surfaces Using Polydopamine

    NARCIS (Netherlands)

    Nijhuis, A.W.G.; Beucken, J.J.J.P van den; Boerman, O.C.; Jansen, Jan; Leeuwenburgh, S.C.G.

    2013-01-01

    Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the

  7. Manganese Dioxide with High Specific Surface Area for Alkaline Battery

    Institute of Scientific and Technical Information of China (English)

    HUANG You-ju; LIN Yu-li; LI Wei-shan

    2012-01-01

    The authors reported a facile method for the synthesis of manganese dioxide without any template and catalyst at a low-temperature.The prepared sample was characterized with X-ray diffraction(XRD),scanning electron microscopy(SEM),Brunauer-Emmett-Teller(BET) surface analysis,Fourier transform infrared(FTIR) spectrometry,cyclic voltammetry,altemative current(AC) impedance test and battery discharge test.It is found that the prepared sample belongs to α-MnO2 and has a microsphere morphology and a large BET surface area.The electrochemical characterization indicates that the prepared sample displays a larger electrochemical capacitance than the commercial electrolytic manganese dioxides(EMD) in Na2SO4 solution,and exhibits larger discharge capacity than EMD,especially at a high rate discharge condition when it is used as cathode of alkaline Zn/MnO2 battery.

  8. Short-term effect of vermicompost application on biological properties of an alkaline soil with high lime content from Mediterranean region of Turkey.

    Science.gov (United States)

    Uz, Ilker; Tavali, Ismail Emrah

    2014-01-01

    This study was conducted to investigate direct short-term impact of vermicompost on some soil biological properties by monitoring changes after addition of vermicompost as compared to farmyard manure in an alkaline soil with high lime content from semiarid Mediterranean region of Turkey. For this purpose, mixtures of soil and organic fertilizers in different doses were incubated under greenhouse condition. Soil samples collected in regular intervals were analyzed for biological parameters including dehydrogenase, β-glucosidase, urease, alkaline phosphatase activities, and total number of aerobic mesophilic bacteria. Even though soil dehydrogenase activity appeared to be dose-independent based on overall evaluation, organic amendments were found to elevate dehydrogenase activity when sampling periods are evaluated individually. β-glucosidase, urease, alkaline phosphatase activity, and aerobic mesophilic bacterial numbers in vermicompost treatments fluctuated but remained significantly above the control. A slight but statistically significant difference was detected between organic amendments in terms of urease activity. Vermicompost appeared to more significantly increase bacterial number in soil. Clearly, vermicompost has a potential to be used as an alternative to farmyard manure to improve and maintain soil biological activity in alkaline calcareous soils from the Mediterranean region of Turkey. Further studies are needed to assess its full potential for these soils.

  9. Calcium aluminate cement hydration in a high alkalinity environment

    Directory of Open Access Journals (Sweden)

    Palomo, Á.

    2009-03-01

    Full Text Available The present paper forms part of a broader research project that aims primarily to devise new cementitious products via the alkali activation of silico-aluminous materials. This work addresses the possibility of using small percentages of calcium aluminate cement (CAC as a source of reactive aluminium. For this reason, a preliminary review was needed of the behaviour of CACs in highly alkaline media (2, 8 and 12M NaOH solutions. Two, 28- and 180-day mechanical strength was determined and the reaction products were characterized with XRD and FTIR. The water-hydrated CAC was used as the control.The results obtained showed that CAC hardening took place much more slowly in highly alkaline media than in water. Nonetheless, the 28-day compressive strength obtained, ≥80MPa. As main reaction products, to ambient temperature and from the two days of cured, cubic aluminate C3AH6, and AH3 polymorphs are formed, instead of the usual hexagonal aluminatos (CAH10 and C2AH8 that are formed in the normal hydrate with water.El presente trabajo forma parte de una amplia investigación cuyo objetivo principal es el de elaborar nuevos materiales con propiedades cementantes mediante la activación alcalina de materiales de naturaleza silito-aluminosa. En estos estudios se contempla la posibilidad de utilizar pequeños porcentajes de cemento de aluminato de calcio (CAC como fuente de aluminio reactivo. Por ello inicialmente se ha estudiado el comportamiento de los CAC en medios fuertemente alcalinos (disoluciones de NaOH 2M, 8M y 12M. Se determinaron las resistencias mecánicas a 2, 28 y 180 días y se realizó una caracterización de los productos de reacción formados por DRX, FTIR. Como sistema de referencia se consideró la hidratación del CAC con agua.Los resultados obtenidos muestran que en medios fuertemente alcalinos se retrasan los procesos de rápido endurecimiento de CAC con agua. No obstante a 28 días se obtienen valores de resistencia a compresión

  10. 骨形态发生蛋白7诱导骨膜细胞碱性磷酸酶的表达*%Bone morphogenetic protein-7 induces the expression of alkaline phosphatase in periosteal cells

    Institute of Scientific and Technical Information of China (English)

    廖家成; 贝抗胜; 连银川

    2013-01-01

      背景:关于骨形态发生蛋白7作为刺激因子诱导细胞成骨的报道目前较少见。目的:观察骨膜细胞经骨形态发生蛋白7诱导后碱性磷酸酶的表达。  方法:取材于成人胫骨骨膜,常规细胞培养法行骨膜细胞体外培养,分为实验组和对照组,分别加入骨形态发生蛋白7加成骨细胞培养辅助剂和单纯成骨细胞培养辅助剂,相差显微镜观察骨膜细胞形态特征及超微结构。每组分别在第7,14,21天设3个时间点,每个时间点设3个样本,采用碱性磷酸酶试剂盒法检测成骨细胞特异性标志物碱性磷酸酶表达情况。  结果与结论:骨膜细胞经分组培养后,第7天时,实验组和对照组骨膜细胞均有明显增殖,碱性磷酸酶的可被检测出,但量不多,细胞外形为梭形,实验组比对照组检测的碱性磷酸酶数量稍多;第14天时,实验组及对照组骨膜细胞均显著增殖,细胞外形由梭形变为宽梭形,实验组比对照组检测的碱性磷酸酶数量明显增多。第21天时,实验组及对照组骨膜细胞均增殖,其中实验组细胞增殖明显,细胞外形为宽梭形,实验组比对照组检测的碱性磷酸酶数量显著增多。经过统计学分析由骨形态发生蛋白7诱导的骨膜细胞的成骨标志物碱性磷酸酶阳性率明显高于对照组(P OBJECTIVE:To study the expression of alkaline phosphatase of periosteal cel s after induced by bone morphogenetic protein-7 in vitro. METHODS:Periosteal cel s were obtained from adult tibial periosteum, and then the periosteal cel s were cultured by routine method in vitro. The cel s were divided into experimental group and control group, and then cultured with bone morphogenetic protein-7 plus osteoblast culture adjuvants and simple osteoblast culture adjuvants, respectively. The phase contrast microscope was used to observe the morphology and ultrastructure of periosteal

  11. Growth hormone (GH) treatment increases serum insulin-like growth factor binding protein-3, bone isoenzyme alkaline phosphatase and forearm bone mineral content in young adults with GH deficiency of childhood onset

    DEFF Research Database (Denmark)

    Juul, A; Pedersen, S A; Sørensen, S;

    1994-01-01

    Recent studies have demonstrated that growth hormone (GH)-deficient adults have a markedly decreased bone mineral content compared to healthy adults. However, there are conflicting results regarding the effects of GH treatment on bone mineral content in GH-deficient adults. Therefore, we evaluated...... the effect of GH treatment on a marker of bone formation (bone alkaline phosphatase), hepatic excretory function and distal forearm bone mineral content in GH-deficient adults. Growth hormone was administered subcutaneously in 21 adults (13 males and 8 females) with GH deficiency of childhood onset for 4...

  12. An in vitro study of alkaline phosphatase sensitivity to mixture of aflatoxin B1 and fumonisin B1 in the hepatopancreas of coastal lagoon wild and farmed shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Pérez-Acosta, Jesús A; Burgos-Hernandez, Armando; Velázquez-Contreras, Carlos A; Márquez-Ríos, Enrique; Torres-Arreola, Wilfrido; Arvizu-Flores, Aldo A; Ezquerra-Brauer, J Marina

    2016-08-01

    This study aimed to establish the combined effect of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on wild Litopenaeus vannamei hepatopancreas alkaline phosphatase (AP) activity compared with that of farmed shrimp. AP activity in hepatopancreas extract was confirmed by several specific inhibitor assays. AP activity of wild shrimp was higher than that of farmed shrimp (p vannamei was sensitive to the presence of both mycotoxins; however, AP is more sensitive to the combination of AFB1 + FB1 suggesting a possible synergistic or potentiating inhibitory effect.

  13. High osmolarity glycerol response PtcB phosphatase is important for Aspergillus fumigatus virulence.

    Science.gov (United States)

    Winkelströter, Lizziane K; Bom, Vinícius Leite Pedro; de Castro, Patrícia Alves; Ramalho, Leandra Naira Zambelli; Goldman, Maria Helena S; Brown, Neil Andrew; Rajendran, Ranjith; Ramage, Gordon; Bovier, Elodie; Dos Reis, Thaila Fernanda; Savoldi, Marcela; Hagiwara, Daisuke; Goldman, Gustavo H

    2015-04-01

    Aspergillus fumigatus is a fungal pathogen that is capable of adapting to different host niches and to avoid host defenses. An enhanced understanding of how, and which, A. fumigatus signal transduction pathways are engaged in the regulation of these processes is essential for the development of improved disease control strategies. Protein phosphatases are central to numerous signal transduction pathways. To comprehend the functions of protein phosphatases in A. fumigatus, 32 phosphatase catalytic subunit encoding genes were identified. We have recognized PtcB as one of the phosphatases involved in the high osmolarity glycerol response (HOG) pathway. The ΔptcB mutant has both increased phosphorylation of the p38 MAPK (SakA) and expression of osmo-dependent genes. The ΔptcB strain was more sensitive to cell wall damaging agents, had increased chitin and β-1,3-glucan, and impaired biofilm formation. The ΔptcB strain was avirulent in a murine model of invasive pulmonary aspergillosis. These results stress the importance of the HOG pathway in the regulation of pathogenicity determinants and virulence in A. fumigatus.

  14. Maternal antibiotic-induced early changes in microbial colonization selectively modulate colonic permeability and inducible heat shock proteins, and digesta concentrations of alkaline phosphatase and TLR-stimulants in swine offspring.

    Directory of Open Access Journals (Sweden)

    Marie-Edith Arnal

    Full Text Available Elevated intake of high energy diets is a risk factor for the development of metabolic diseases and obesity. High fat diets cause alterations in colonic microbiota composition and increase gut permeability to bacterial lipopolysaccharide, and subsequent low-grade chronic inflammation in mice. Chronic inflammatory bowel diseases are increasing worldwide and may involve alterations in microbiota-host dialog. Metabolic disorders appearing in later life are also suspected to reflect changes in early programming. However, how the latter affects the colon remains poorly studied. Here, we hypothesized that various components of colonic physiology, including permeability, ion exchange and protective inducible heat shock proteins (HSP are influenced in the short- and long-terms by early disturbances in microbial colonization. The hypothesis was tested in a swine model. Offspring were born to control mothers (n = 12 or mothers treated with the antibiotic (ATB amoxicillin around parturition (n = 11. Offspring were slaughtered between 14 and 42 days of age to study short-term effects. For long-term effects, young adult offspring from the same litters consumed a normal or a palm oil-enriched diet for 4 weeks between 140 and 169 days of age. ATB treatment transiently modified maternal fecal microbiota although the minor differences observed for offspring colonic microbiota were nonsignificant. In the short-term, consistently higher HSP27 and HSP70 levels and transiently increased horseradish peroxidase permeability in ATB offspring colon were observed. Importantly, long-term consequences included reduced colonic horseradish peroxidase permeability, and increased colonic digesta alkaline phosphatase (AP and TLR2- and TLR4-stimulant concentrations in rectal digesta in adult ATB offspring. Inducible HSP27 and HSP70 did not change. Interactions between early ATB treatment and later diet were noted for paracellular permeability and concentrations of colonic

  15. Activation of fly ashes by the high temperature and high alkalinity in ASR tests

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    High temperature and high alkalinity are typical testing conditions to accelerate the appraisal process of the suppressing effect of fly ashes on alkali silica reaction(ASR),but the reaction mechanism of fly ashes would be quite different under such conditions compared to the normal condition of temperature and alkalinity.To make a reasonable analysis of the suppressing effect of fly ashes,13 types of fly ashes were tested in this paper by both the accelerated mortar bar test method and the 60°C accelerated concrete prism test method.The results showed that the effect of fly ashes would be magnified under the condition of high temperature and high alkalinity.The XRD analysis showed that all the phases of fly ash could react with the hot alkaline solution except for mullite and a small amount of quartz.Fly ash could be significantly activated by the 80°C 1 mol/L NaOH solution,and form mainly C-S-H phase and P type zeolite,but its effect on inhibiting ASR was exaggerated then.According to the mortar strength test and the ASR suppressing test results,C-S-H phase contributed to mortar strength,but its amount did not decide the ASR suppressing effect of fly ash.

  16. Alkaline Earth Core Level Photoemission Spectroscopy of High-Temperature Superconductors

    Science.gov (United States)

    Vasquez, R.

    1993-01-01

    This paper examines photoemission measurements of the alkaline Earth core levels of high-temperature superconductors and related materials, models that seek to explain the large negative shifts observed relative to the corresponding alkaline Earth metals, and the effect of lattice site disorder on the core level spectra and the presence or absence of intrinsic surface peaks.

  17. Alkaline Earth Core Level Photoemission Spectroscopy of High-Temperature Superconductors

    Science.gov (United States)

    Vasquez, R.

    1993-01-01

    This paper examines photoemission measurements of the alkaline Earth core levels of high-temperature superconductors and related materials, models that seek to explain the large negative shifts observed relative to the corresponding alkaline Earth metals, and the effect of lattice site disorder on the core level spectra and the presence or absence of intrinsic surface peaks.

  18. Detection of phosphatase activity in aquatic and terrestrial cyanobacterial strains

    Directory of Open Access Journals (Sweden)

    Babić Olivera B.

    2013-01-01

    Full Text Available Cyanobacteria, as highly adaptable microorganisms, are characterized by an ability to survive in different environmental conditions, in which a significant role belongs to their enzymes. Phosphatases are enzymes produced by algae in relatively large quantities in response to a low orthophosphate concentration and their activity is significantly correlated with their primary production. The activity of these enzymes was investigated in 11 cyanobacterial strains in order to determine enzyme synthesis depending on taxonomic and ecological group of cyanobacteria. The study was conducted with 4 terrestrial cyanobacterial strains, which belong to Nostoc and Anabaena genera, and 7 filamentous water cyanobacteria of Nostoc, Oscillatoria, Phormidium and Microcystis genera. The obtained results showed that the activity of acid and alkaline phosphatases strongly depended on cyanobacterial strain and the environment from which the strain originated. Higher activity of alkaline phosphatases, ranging from 3.64 to 85.14 μmolpNP/s/dm3, was recorded in terrestrial strains compared to the studied water strains (1.11-5.96 μmolpNP/s/dm3. The activity of acid phosphatases was higher in most tested water strains (1.67-6.28 μmolpNP/s/dm3 compared to the activity of alkaline phosphatases (1.11-5.96 μmolpNP/s/dm3. Comparing enzyme activity of nitrogen fixing and non-nitrogen fixing cyanobacteria, it was found that most nitrogen fixing strains had a higher activity of alkaline phosphatases. The data obtained in this work indicate that activity of phosphatases is a strain specific property. The results further suggest that synthesis and activity of phosphatases depended on eco-physiological characteristics of the examined cyanobacterial strains. This can be of great importance for the further study of enzymes and mechanisms of their activity as a part of cyanobacterial survival strategy in environments with extreme conditions. [Projekat Ministarstva nauke Republike

  19. Altered alkaline phosphatase activity in obese Zucker rats liver respect to lean Zucker and Wistar rats discussed in terms of all putative roles ascribed to the enzyme

    Directory of Open Access Journals (Sweden)

    V. Bertone

    2011-02-01

    Full Text Available Biliary complications often lead to acute and chronic liver injury after orthotopic liver transplantation (OLT. Bile composition and secretion depend on the integrated action of all the components of the biliary tree, starting from hepatocytes. Fatty livers are often discarded as grafts for OLT, since they are extremely vulnerable to conventional cold storage (CS. However, the insufficiency of donors has stimulated research to improve the usage of such marginal organs as well as grafts. Our group has recently developed a machine perfusion system at subnormothermic temperature (20°C; MP20 that allows a marked improvement in preservation of fatty and even of normal rat livers as compared with CS. We sought to evaluate the response of the biliary tree of fatty liver to MP20, and a suitable marker was essential to this purpose. Alkaline phosphatase (AlkP, EC 3.1.3.1, frequently used as marker of membrane transport in hepatocytes and bile ducts, was our first choice. Since no histochemical data were available on AlkP distribution and activity in fatty liver, we have first settled to investigate AlkP activity in the steatotic liver of fatty Zucker rats (fa/fa, using as controls lean Zucker (fa/+ and normal Wistar rats. The AlkP reaction in Wistar rats was in accordance with the existing data and, in particular, was present in bile canaliculi of hepatocytes in the periportal region and midzone, in the canals of Hering and in small bile ducts but not in large bile ducts. In lean ZR liver the AlkP reaction in Hering canals and small bile ducts was similar to Wistar rat liver but hepatocytes had lower canalicular activity and besides presented moderate basolateral reaction. The difference between lean Zucker and Wistar rats, both phenotypically normal animals, could be related to the fact that lean Zucker rats are genotypically heterozygous for a recessive mutated allele. In fatty liver, the activity in ductules and small bile ducts was unchanged, but

  20. 19416例健康成人碱性磷酸酶活性分析%19416 cases of serum alkaline phosphatase activity analysis of healthy adults

    Institute of Scientific and Technical Information of China (English)

    吉利; 连丽丽; 孙淑艳; 孙小慧

    2011-01-01

    目的 分析健康成人血清ALP活性,探讨ALP水平在成人性别和年龄的分布规律.方法 随机选择2008年来吉大一院健康体检各项指标正常的健康人19416例,对其血清ALP活性进行回顾性统计分析.结果 21-50岁男性ALP值显著高于同年龄组女性(P<0.01);51岁以上男性ALP值显著低于同年龄段女性(P<0.01).男性20-30岁ALP水平显著高于31岁以上各年龄组,31岁以上各年龄组间差异无统计学意义;女性20-40岁年龄段ALP水平差异无统计学意义,40岁以后,ALP水平随年龄增长而增高.结论 ALP在性别和年龄的分布规律,反映了健康成人骨代谢情况,雌激素可降低ALP活性,为研究雌激素对成骨细胞标志物ALP的影响提供科学的依据.%bjective To Analyze the activity of Alkaline phosphatase (ALP) in serum of healthy adults ,in terms of discussing the distribution rule of ALP levels in gender and age .Methods Making retrospective analysis of the ALP activity data of healthy people who has normal medical examination results from the physical examination center our hospital in 2008 .Results The ALP levels of 21-30,31-40 and 41-50 age group male were significantly higher than female of the same age group ( P<0.01 ) ;the ALP levels of 51-60 and older than 60 age groups male were significantly lower than those of the same age female ( P<0.01 ) .The variance analysis results of ALP levels between different age groups of male and female respectively showed that the ALP levds of 21-30 yearold male were obviously hgher than those of all age groups above 31 year-old and the male ALP results above 31 age groups has no different significance in statistics ;the ALP levels of 20-40 year-old female has no different significance in statistics and the other age groups has statistical significance the female ALP levels increased from 40 year-old as age grew .Conclusion Ths research obtained the distribution rule of adults serum ALP activity in gender and

  1. A constructed alkaline consortium and its dynamics in treating alkaline black liquor with very high pollution load.

    Directory of Open Access Journals (Sweden)

    Chunyu Yang

    Full Text Available BACKGROUND: Paper pulp wastewater resulting from alkaline extraction of wheat straw, known as black liquor, is very difficult to be treated and causes serious environmental problems due to its high pH value and chemical oxygen demand (COD pollution load. Lignin, semicellulose and cellulose are the main contributors to the high COD values in black liquor. Very few microorganisms can survive in such harsh environments of the alkaline wheat straw black liquor. A naturally developed microbial community was found accidentally in a black liquor storing pool in a paper pulp mill of China. The community was effective in pH decreasing, color and COD removing from the high alkaline and high COD black liquor. FINDINGS: Thirty-eight strains of bacteria were isolated from the black liquor storing pool, and were grouped as eleven operational taxonomy units (OTUs using random amplified polymorphic DNA-PCR profiles (RAPD. Eleven representative strains of each OTU, which were identified as genera of Halomonas and Bacillus, were used to construct a consortium to treat black liquor with a high pH value of 11.0 and very high COD pollution load of 142,600 mg l(-1. After treatment by the constructed consortium, about 35.4% of color and 39,000 mg l(-1 (27.3% COD(cr were removed and the pH decreased to 7.8. 16S rRNA gene polymerase chain reaction denaturant gradient gel electrophoresis (PCR-DGGE and gas chromatography/mass spectrometry (GC/MS analysis suggested a two-stage treatment mechanism to elucidate the interspecies collaboration: Halomonas isolates were important in the first stage to produce organic acids that contributed to the pH decline, while Bacillus isolates were involved in the degradation of lignin derivatives in the second stage under lower pH conditions. CONCLUSIONS/SIGNIFICANCE: Tolerance to the high alkaline environment and good controllability of the simple consortium suggested that the constructed consortium has good potential for black liquor

  2. Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acid-etched (MSLA), laser-treated, and laser and acid-treated Ti surfaces

    Science.gov (United States)

    Li, Lin-Jie; Kim, So-Nam

    2016-01-01

    PURPOSE In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P.05). CONCLUSION This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies. PMID:27350860

  3. QM/MM analysis suggests that Alkaline Phosphatase (AP) and Nucleotide pyrophosphatase/phosphodiesterase slightly tighten the transition state for phosphate diester hydrolysis relative to solution: implication for catalytic promiscuity in the AP superfamily

    Science.gov (United States)

    Hou, Guanhua

    2011-01-01

    Several members of the Alkaline Phosphatase (AP) superfamily exhibit a high level of catalytic proficiency and promiscuity in structurally similar active sites. A thorough characterization of the nature of transition state for different substrates in these enzymes is crucial for understanding the molecular mechanisms that govern those remarkable catalytic properties. In this work, we study the hydrolysis of a phosphate diester, MpNPP−, in solution, two experimentally well-characterized variants of AP (R166S AP, R166S/E322Y AP) and wild type Nucleotide pyrophosphatase/phosphodiesterase (NPP) by QM/MM calculations in which the QM method is an approximate density functional theory previously parameterized for phosphate hydrolysis (SCC-DFTBPR). The general agreements found between these calculations and available experimental data for both solution and enzymes support the use of SCC-DFTBPR/MM for a semi-quantitative analysis of the catalytic mechanism and nature of transition state in AP and NPP. Although phosphate diesters are cognate substrates for NPP but promiscuous substrates for AP, the calculations suggest that their hydrolysis reactions catalyzed by AP and NPP feature similar synchronous transition states that are slightly tighter in nature compared to that in solution, due in part to the geometry of the bimetallic zinc motif. Therefore, this study provides the first direct computational support to the hypothesis that enzymes in the AP superfamily catalyze cognate and promiscuous substrates via similar transition states to those in solution. Our calculations do not support the finding of recent QM/MM studies by López-Canut and coworkers, who suggested that the same diester substrate goes through a much looser transition state in NPP/AP than in solution, a result likely biased by the large structural distortion of the bimetallic zinc site in their simulations. Finally, our calculations for different phosphate diester orientations and phosphorothioate diesters

  4. The parathyroid hormone-related protein is secreted during the osteogenic differentiation of human dental follicle cells and inhibits the alkaline phosphatase activity and the expression of DLX3.

    Science.gov (United States)

    Klingelhöffer, C; Reck, A; Ettl, T; Morsczeck, C

    2016-08-01

    The dental follicle is involved in tooth eruption and it expresses a great amount of the parathyroid hormone-related protein (PTHrP). PTHrP as an extracellular protein is required for a multitude of different regulations of enchondral bone development and differentiation of bone precursor cells and of the development of craniofacial tissues. The dental follicle contains also precursor cells (DFCs) of the periodontium. Isolated DFCs differentiate into periodontal ligament cells, alveolar osteoblast and cementoblasts. However, the role of PTHrP during the human periodontal development remains elusive. Our study evaluated the influence of PTHrP on the osteogenic differentiation of DFCs under in vitro conditions for the first time. The PTHrP protein was highly secreted after 4days of the induction of the osteogenic differentiation of DFCs with dexamethasone (2160.5pg/ml±345.7SD. in osteogenic differentiation medium vs. 315.7pg/ml±156.2SD. in standard cell culture medium; Student's t Test: pPTHrP inhibited the alkaline phosphatase activity and the expression of the transcription factor DLX3, but the depletion of PTHrP did not support the differentiation of DFCs. Previous studies have shown that Indian Hedgehog (IHH) induces PTHrP and that PTHrP, in turn, inhibits IHH via a negative feedback loop. We showed that SUFU (Suppressor Of Fused Homolog) was not regulated during the osteogenic differentiation in DFCs. So, neither the hedgehog signaling pathway induced PTHrP nor PTHrP suppressed the hedgehog signaling pathway during the osteogenic differentiation in DFCs. In conclusion, our results suggest that PTHrP regulates independently of the hedgehog signaling pathway the osteogenic differentiated in DFCs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Inhibitors of the Yersinia protein tyrosine phosphatase through high throughput and virtual screening approaches.

    Science.gov (United States)

    Hu, Xin; Vujanac, Milos; Southall, Noel; Stebbins, C Erec

    2013-02-15

    The bacterial protein tyrosine phosphatase YopH is an essential virulence determinant in Yersinia pestis and a potential antibacterial drug target. Here we report our studies of screening for small molecule inhibitors of YopH using both high throughput and in silico approaches. The identified inhibitors represent a diversity of chemotypes and novel pTyr mimetics, providing a starting point for further development and fragment-based design of multi-site binding inhibitors. We demonstrate that the applications of high throughput and virtual screening, when guided by structural binding mode analysis, is an effective approach for identifying potent and selective inhibitors of YopH and other protein phosphatases for rational drug design. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. [ATPase and phosphatase activity of drone brood].

    Science.gov (United States)

    Bodnarchuk, L I; Stakhman, O S

    2004-01-01

    Most researches on insect enzymes concern carbohydrate and nitrogenous exchange. Data on ATPase activity for larval material of drone brood are absent in the available literature. The drone brood is one of the least investigated apiproducts. Allowing for the important role of ATPase in the vital functions of the insect cells our work was aimed at the study of ATPase of the drone blood activity and that of alkaline and acid phosphatases. When studying liophylised preparations of the drone brood homogenate we have found out high activity of Mg2+, Na+, K+-, Ca2+- and Mg2+-ATPase and of alkaline and acid phosphatase, that is the possible explanation of the high-intensity power and plastic processes proceeding during growth and development of larvae.

  7. Effects of Different Fertilizer on Soil Urease and Alkaline Phosphatase Activity%不同肥料对土壤脲酶和碱性磷酸酶活性的影响

    Institute of Scientific and Technical Information of China (English)

    周俊国; 杨鹏鸣

    2012-01-01

    Ureas and alkaline phosphatase activity of soil in different fertilizer was studied in this experiment. The results shewed urea, chicken manure, straw increased soil urease activity by 15. 38 % , 55.94 % , 27.97 % and soil alkaline phosphatase activity by 2.31 % , 16.20 % , 10.42 % respectively compared with the control in the Ⅰ fertilization levrl. Three fertiliters byd soil urease activity by 24. 48 % ,67.83 % ,46.15 % and alkaline phosphatase activity by 9.72% , 28.01 % , 37.04 % reapectively in the Ⅱ fertilisation level. Urea, chicken manure, straw increased soil urease activity by 37.76 % , 86. 71 % , 62.24 % , and soil alkaline phosphatase activity by 28.47 % , 65.74 % , 58.10 % respectively in the Ⅲ fertilization level. The affect order for soil urease aclivity by 3 fertilizers in different fertilizer levels was chicken manure, straw,urea.The affecting order for soil alkaline phosphstase activity was chicken manure, stravt, urea in Ⅱ fertilization level. The affecting order for soil alkaline phoaphatase activity was straw, chicken manure, urea in Ⅰ and Ⅲ fertilization level.%本实验对不同施肥条件下土壤脲酶活性、碱性磷酸酶活性进行了系统研究.结果表明,和对照相比,尿素、鸡粪、秸秆在Ⅰ施肥水平上,使土壤脲(酶)活性分别增加15.38%、55.94%、27.97%,土壤碱性磷酸酶活性分别增加2.31%、16.20%、10.42%;在Ⅱ施肥水平上使土壤脲酶活性分别增加24.48%,67.83%,46.15%,使土壤碱性磷酸酶活性分别增加9.72%、28.01%和37.04%.在Ⅲ施肥水平上使土壤脲酶分别增加37.76%、86.71%、62.24%,土壤碱性磷酸酶活性分别增加28.47%、65.74%和58.10%.3种不同施肥水平对土壤腺酶活性影响顺序依次是鸡粪、秸秆、尿素.在Ⅱ施肥水平上,对土壤碱性磷酸酶活性的影响大小依次是鸡粪、秸秆、尿素.在Ⅰ、Ⅲ施肥水平上是秸秆、鸡粪、尿素.

  8. Influence of triethyl phosphate on phosphatase activity in shooting range soil: Isolation of a zinc-resistant bacterium with an acid phosphatase.

    Science.gov (United States)

    Story, Sandra; Brigmon, Robin L

    2017-03-01

    Phosphatase-mediated hydrolysis of organic phosphate may be a viable means of stabilizing heavy metals via precipitation as a metal phosphate in bioremediation applications. We investigated the effect of triethyl phosphate (TEP) on soil microbial-phosphatase activity in a heavy-metal contaminated soil. Gaseous TEP has been used at subsurface sites for bioremediation of organic contaminants but not applied in heavy-metal contaminated areas. Little is known about how TEP affects microbial activity in soils and it is postulated that TEP can serve as a phosphate source in nutrient-poor groundwater and soil/sediments. Over a 3-week period, TEP amendment to microcosms containing heavy-metal contaminated soil resulted in increased activity of soil acid-phosphatase and repression of alkaline phosphatase, indicating a stimulatory effect on the microbial population. A soil-free enrichment of microorganisms adapted to heavy-metal and acidic conditions was derived from the TEP-amended soil microcosms using TEP as the sole phosphate source and the selected microbial consortium maintained a high acid-phosphatase activity with repression of alkaline phosphatase. Addition of 5mM zinc to soil-free microcosms had little effect on acid phosphatase but inhibited alkaline phosphatase. One bacterial member from the consortium, identified as Burkholderia cepacia sp., expressed an acid-phosphatase activity uninhibited by high concentrations of zinc and produced a soluble, indigo pigment under phosphate limitation. The pigment was produced in a phosphate-free medium and was not produced in the presence of TEP or phosphate ion, indicative of purple acid-phosphatase types that are pressed by bioavailable phosphate. These results demonstrate that TEP amendment was bioavailable and increased overall phosphatase activity in both soil and soil-free microcosms supporting the possibility of positive outcomes in bioremediation applications.

  9. Hydrogen embrittlement on {alpha}-iron in high alkaline environment

    Energy Technology Data Exchange (ETDEWEB)

    Hu, R.; Habashi, M.; Galland, J. [Ecole Central Paris, Chatenay-Malabry (France)

    1994-12-31

    The partial pressure of hydrogen in concrete`s pore is very low. This hydrogen is due to the chemical reaction between the silica fumes and the alkaline solutions filling the concrete`s pore. Silica fumes are added in the concrete to increase its compression resistance. If the hydrogen pressure is low, the risk of hydrogen embrittlement is also low. However, for constructional works destined to endure more than 50 years, is this risk negligible? To answer this question, the authors have studied the hydrogen embrittlement on {alpha}-iron in alkaline solutions, in the pH range 9.5 to 13.3, presenting the liquids found in the concrete`s pores after different aging, periods. Cathodic charging has been performed for low current densities in the range 0.25 to 90 A/m{sup 2} simulating several partial pressures of hydrogen on the {alpha}-iron surface with and without EDTA inhibitor. The deformation rate was 2.5{times}10{sup {minus}5} s{sup {minus}1}. Finally {alpha}-iron samples and tensile specimens have been immersed in a mixture of silica fumes and an alkaline solution at pH 13.3 in an autoclave during 1,000 hours with the aim to measure the outgassed quantity of hydrogen under vacuum at 600C and to measure also the hydrogen embrittlement. The main conclusions of this study are as following: (1) Hydrogen embrittlement is promoted by oxide Fe{sub 3}O{sub 4} film rupture and/or hydroxide Fe(OH){sub 2}. This mechanism is efficient for current densities equivalent to a cathodic potential lower or equal to {minus}1V/NHE. (2) Silica fumes in contact with a solution of pH 13.3 provoke hydrogen release and its diffusion into the {alpha}-iron, but this quantity is not enough to embrittle it.

  10. Evaluation of High Solids Alkaline Pretreatment of Rice Straw

    OpenAIRE

    Cheng, Yu-Shen; Zheng, Yi; Yu, Chao Wei; Dooley, Todd M.; Jenkins, Bryan M.; VanderGheynst, Jean S.

    2010-01-01

    Fresh-harvested, air-dried rice straw was pretreated at a water content of 5 g H2O/g straw using sodium hydroxide (NaOH) and compared to pretreatment at 10 g H2O/g straw by hydrated lime (Ca(OH)2). Full factorial experiments including parallel wash-only treatments were completed with both sources of alkali. The experiments were designed to measure the effects of alkaline loading and pretreatment time on delignification and sugar yield upon enzymatic hydrolysis. Reaction temperature was held c...

  11. Water-free Alkaline Polymer-inorganic Acid Complexes with High Conductivity at Ambient Temperature

    Institute of Scientific and Technical Information of China (English)

    O.V.Chervakov; M.V.Andriianova; V.V.Riabenko; A.V.Markevich; E.M.Shembel; D.Meshri

    2007-01-01

    1 Results Recently increased interest is shown to proton conducting materials based on the alkaline polymer-inorganic acid complexes that is caused by a possibility of their application as the high-temperature electrolyte systems for various electrochemical devices (fuel cells,sensors,lithium power sources etc.).Complexes of inorganic acids with the alkaline polymers (polybenzimidazoles[1],polyvinylpyridines[2]) are characterized by high ionic conductivity at ambient temperatures (up to 10-2 Ω-1·cm-1) a...

  12. Evaluation of high solids alkaline pretreatment of rice straw.

    Science.gov (United States)

    Cheng, Yu-Shen; Zheng, Yi; Yu, Chao Wei; Dooley, Todd M; Jenkins, Bryan M; VanderGheynst, Jean S

    2010-11-01

    Fresh-harvested, air-dried rice straw was pretreated at a water content of 5 g H(2)O/g straw using sodium hydroxide (NaOH) and compared to pretreatment at 10 g H(2)O/g straw by hydrated lime (Ca(OH)(2)). Full factorial experiments including parallel wash-only treatments were completed with both sources of alkali. The experiments were designed to measure the effects of alkaline loading and pretreatment time on delignification and sugar yield upon enzymatic hydrolysis. Reaction temperature was held constant at 95 degrees C for lime pretreatment and 55 degrees C for NaOH pretreatment. The range of delignification was 13.1% to 27.0% for lime pretreatments and was 8.6% to 23.1% for NaOH pretreatments. Both alkaline loading and reaction time had significant positive effects (p pretreatment at 95 degrees C, but there was little effect observed at 55 degrees C. Post-pretreatment washing of biomass was not necessary for subsequent enzymatic hydrolysis. Maximum glucose yields were 176.3 mg/g dried biomass (48.5% conversion efficiency of total glucose) in lime-pretreated and unwashed biomass and were 142.3 mg/g dried biomass (39.2% conversion efficiency of total glucose) in NaOH-pretreated and unwashed biomass.

  13. Hydroxide Self-Feeding High-Temperature Alkaline Direct Formate Fuel Cells.

    Science.gov (United States)

    Li, Yinshi; Sun, Xianda; Feng, Ying

    2017-03-11

    Conventionally, both the thermal degradation of the anion-exchange membrane and the requirement of additional hydroxide for fuel oxidation reaction hinder the development of the high-temperature alkaline direct liquid fuel cells. The present work addresses these two issues by reporting a polybenzimidazole-membrane-based direct formate fuel cell (DFFC). Theoretically, the cell voltage of the high-temperature alkaline DFFC can be as high as 1.45 V at 90 °C. It has been demonstrated that a proof-of-concept alkaline DFFC without adding additional hydroxide yields a peak power density of 20.9 mW cm(-2) , an order of magnitude higher than both alkaline direct ethanol fuel cells and alkaline direct methanol fuel cells, mainly because the hydrolysis of formate provides enough OH(-) ions for formate oxidation reaction. It was also found that this hydroxide self-feeding high-temperature alkaline DFFC shows a stable 100 min constant-current discharge at 90 °C, proving the conceptual feasibility.

  14. Growth hormone (GH) treatment increases serum insulin-like growth factor binding protein-3, bone isoenzyme alkaline phosphatase and forearm bone mineral content in young adults with GH deficiency of childhood onset

    DEFF Research Database (Denmark)

    Juul, A; Pedersen, S A; Sørensen, S

    1994-01-01

    Recent studies have demonstrated that growth hormone (GH)-deficient adults have a markedly decreased bone mineral content compared to healthy adults. However, there are conflicting results regarding the effects of GH treatment on bone mineral content in GH-deficient adults. Therefore, we evaluated...... the effect of GH treatment on a marker of bone formation (bone alkaline phosphatase), hepatic excretory function and distal forearm bone mineral content in GH-deficient adults. Growth hormone was administered subcutaneously in 21 adults (13 males and 8 females) with GH deficiency of childhood onset for 4...... months in a double-blind, placebo-controlled GH trial, while 13 of the patients then received further GH for an additional 14 months. Serum insulin-like growth factor I (IGF-I) increased significantly from 100 to 279 micrograms/l and IGF binding protein-3 (IGFBP-3) from 1930 to 3355 micrograms/l after 4...

  15. Analysis of detection results of bone alkaline phosphatase in 2 184 children in Baoji area%宝鸡地区2184例儿童骨碱性磷酸酶结果的分析

    Institute of Scientific and Technical Information of China (English)

    乔正梅; 葛君琍; 赵秋剑; 张利强

    2014-01-01

    目的:评价骨碱性磷酸酶对儿童佝偻病早期诊断及疗效评价的临床价值,通过骨碱性磷酸酶的检测了解该地区儿童佝偻病的发病率。方法选择2013年1~12月来宝鸡市中心医院体检的2184例儿童,按年龄分成婴儿、幼儿、学龄前、学龄期4组,采集指血,采用北京中生金域诊断技术有限公司提供的骨源性碱性磷酸酶试剂盒检测末梢血中骨碱性磷酸酶水平。结果2184例受检儿童中,临床佝偻病(NBAP>250 U/L)的检出率为10.1%,且年龄越小检出率越高;亚临床佝偻病(NBAP 200~250 U/L)的检出率为65.5%,说明宝鸡地区大部分儿童处于亚健康状态;春冬季检出率高于夏秋季;NBAP 检出率无性别差异。结论儿童为生长发育较快时期,易引起维生素 D 缺乏,儿童骨碱性磷酸酶活性检测对维生素 D 缺乏引起的佝偻病的早期诊断有重要的临床价值。预防应从围产期开始,及时检测儿童骨碱性磷酸酶水平,以便早诊断、早治疗。%Objective To evaluate the clinical value of bone alkaline phosphatase in early diagnosis and treatment effect of chil-dren′rickets and to understand the incidence rate of rickets in children in the local region by the detection of neonatal bone alkaline phosphatase(NBAP).Methods 2 184 children with physical examination in our hospital from January to December 2013 were se-lected and divided into the infants,young children,preschool and school-age children groups according to age.The finger blood was collected for detecting the bone alkaline phosphatase in the peripheral blood by the bone alkaline phosphatase reagent kit provided by Beijing Jinyu diagnosis technology Co.,Ltd.Results Among detected 2 184 children,the detection rate of clinical rickets(NBAP>250 U/L)was 10.1 %,moreover the smaller the age,the higher the detection rate;the detection rate of subclinical rickets(NBAP 200~250 U/L)was 65

  16. Human PHOSPHO1 exhibits high specific phosphoethanolamine and phosphocholine phosphatase activities

    Science.gov (United States)

    2004-01-01

    Human PHOSPHO1 is a phosphatase enzyme for which expression is upregulated in mineralizing cells. This enzyme has been implicated in the generation of Pi for matrix mineralization, a process central to skeletal development. PHOSPHO1 is a member of the haloacid dehalogenase (HAD) superfamily of Mg2+-dependent hydrolases. However, substrates for PHOSPHO1 are, as yet, unidentified and little is known about its activity. We show here that PHOSPHO1 exhibits high specific activities toward phosphoethanolamine (PEA) and phosphocholine (PCho). Optimal enzymic activity was observed at approx. pH 6.7. The enzyme shows a high specific Mg2+-dependence, with apparent Km values of 3.0 μM for PEA and 11.4 μM for PCho. These results provide a novel mechanism for the generation of Pi in mineralizing cells from PEA and PCho. PMID:15175005

  17. Characterization of Laboratory Prepared Concrete Pastes Exposed to High Alkaline and High Sodium Salt Solutions

    Energy Technology Data Exchange (ETDEWEB)

    Langton, C. A. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2016-06-30

    The objective of this study was to identify potential chemical degradation mechanisms for the Saltstone Disposal Unit (SDU) concretes, which over the performance life of the structures may be exposed to highly alkaline sodium salt solutions containing sulfate, hydroxide, and other potentially corrosive chemicals in salt solution and saltstone flush water, drain water, leachate and / or pore solution. The samples analyzed in this study were cement pastes prepared in the SIMCO Technologies, Inc. concrete laboratory. They were based on the paste fractions of the concretes used to construct the Saltstone Disposal Units (SDUs). SDU 1 and 4 concrete pastes were represented by the PV1 test specimens. The paste in the SDU 2, 3, 5, and 6 concrete was represented by the PV2 test specimens. SIMCO Technologies, Inc. selected the chemicals and proportions in the aggressive solutions to approximate proportions in the saltstone pore solution [2, 3, 5, and 6]. These test specimens were cured for 56 days in curing chamber before being immersed in aggressive solutions. After exposure, the samples were frozen to prevent additional chemical transport and reaction. Selected archived (retrieved from the freezer) samples were sent to the Savannah River National Laboratory (SRNL) for additional characterization using x-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive x-ray (EDX) spectroscopy. Characterization results are summarized in this report. In addition, a correlation between the oxide composition of the pastes and their chemical durability in the alkaline salt solutions is provided.

  18. Shifts in leaf N:P stoichiometry during rehabilitation in highly alkaline bauxite processing residue sand.

    Science.gov (United States)

    Goloran, Johnvie B; Chen, Chengrong; Phillips, Ian R; Elser, James J

    2015-10-07

    Large quantities of sodic and alkaline bauxite residue are produced globally as a by-product from alumina refineries. Ecological stoichiometry of key elements [nitrogen (N) and phosphorus (P)] plays a critical role in establishing vegetation cover in bauxite residue sand (BRS). Here we examined how changes in soil chemical properties over time in rehabilitated sodic and alkaline BRS affected leaf N to P stoichiometry of native species used for rehabilitation. Both Ca and soil pH influenced the shifts in leaf N:P ratios of the study species as supported by consistently significant positive relationships (P stoichiometry can effectively provide a meaningful assessment on understanding nutrient limitation and productivity of native species used for vegetating highly sodic and alkaline BRS, and is a crucial indicator for assessing ecological rehabilitation performance.

  19. Direct determination of phosphatase activity from physiological substrates in cells.

    Directory of Open Access Journals (Sweden)

    Zhongyuan Ren

    Full Text Available A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min(-1 mg(-1 for PPi, to 56 ± 11 nmol min(-1 mg(-1 for AMP, to 79 ± 23 nmol min(-1 mg(-1 for beta-glycerophosphate and to 73 ± 15 nmol min(-1 mg(-1 for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole--a TNAP inhibitor--served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes.

  20. High Temperature Alkaline Electrolysis Cells with Metal Foam Based Gas Diffusion Electrodes

    DEFF Research Database (Denmark)

    Chatzichristodoulou, Christodoulos; Allebrod, Frank; Mogensen, Mogens Bjerg

    2016-01-01

    Alkaline electrolysis cells operating at 250°C and 40 bar are able to convert electrical energy into hydrogen at very high efficiencies and power densities. In the present work we demonstrate the application of a PTFE hydrophobic network and Ag nanowires as oxygen evolution electrocatalyst...

  1. Critical parameters in cost-effective alkaline extraction for high protein yield from leaves

    NARCIS (Netherlands)

    Zhang, C.; Sanders, J.P.M.; Bruins, M.E.

    2014-01-01

    Leaves are potential resources for feed or food, but their applications are limited due to a high proportion of insoluble protein and inefficient processing. To overcome these problems, parameters of alkaline extraction were evaluated using green tea residue (GTR). Protein extraction could be maximi

  2. Synthesis of High-Quality Graphene through Electrochemical Exfoliation of Graphite in Alkaline Electrolyte

    OpenAIRE

    Tripathi, Prashant; Patel, Ch. Ravi Prakash; Shaz, M. A.; Srivastava, O N

    2013-01-01

    Owing to wide variety of applications of graphene, high-quality and economical way of synthesizing graphene is highly desirable. In this study, we report a cost effective and simple approach to production of high-quality graphene. Here the synthesis route is based on electrochemical exfoliation of graphite. Instead of using strong acids (which oxidise and damage the geometrical topology of graphene), we have used alkaline solution (KOH dissolved in water) as electrolyte. TEM analysis shows th...

  3. Microbial communities associated with the anthropogenic, highly alkaline environment of a saline soda lime, Poland.

    Science.gov (United States)

    Kalwasińska, Agnieszka; Felföldi, Tamás; Szabó, Attila; Deja-Sikora, Edyta; Kosobucki, Przemysław; Walczak, Maciej

    2017-07-01

    Soda lime is a by-product of the Solvay soda process for the production of sodium carbonate from limestone and sodium chloride. Due to a high salt concentration and alkaline pH, the lime is considered as a potential habitat of haloalkaliphilic and haloalkalitolerant microbial communities. This artificial and unique environment is nutrient-poor and devoid of vegetation, due in part to semi-arid, saline and alkaline conditions. Samples taken from the surface layer of the lime and from the depth of 2 m (both having pH ~11 and ECe up to 423 dS m(-1)) were investigated using culture-based (culturing on alkaline medium) and culture-independent microbiological approaches (microscopic analyses and pyrosequencing). A surprisingly diverse bacterial community was discovered in this highly saline, alkaline and nutrient-poor environment, with the bacterial phyla Proteobacteria (representing 52.8% of the total bacterial community) and Firmicutes (16.6%) showing dominance. Compared to the surface layer, higher bacterial abundance and diversity values were detected in the deep zone, where more stable environmental conditions may occur. The surface layer was dominated by members of the genera Phenylobacterium, Chelativorans and Skermanella, while in the interior layer the genus Fictibacillus was dominant. The culturable aerobic, haloalkaliphilic bacteria strains isolated in this study belonged mostly to the genus Bacillus and were closely related to the species Bacillus pseudofirmus, B. cereus, B. plakortidis, B. thuringensis and B. pumilus.

  4. Stimulus Response of Au-NPs@GMP-Tb Core-Shell Nanoparticles: Toward Colorimetric and Fluorescent Dual-Mode Sensing of Alkaline Phosphatase Activity in Algal Blooms of a Freshwater Lake.

    Science.gov (United States)

    Zhang, Xiaolei; Deng, Jingjing; Xue, Yumeng; Shi, Guoyue; Zhou, Tianshu

    2016-01-19

    In this study, we demonstrate a colorimetric and fluorescent dual-mode method for alkaline phosphatase activity (APA) sensing in freshwater lake with stimuli-responsive gold nanoparticles@terbium-guanosine monophosphate (Au-NPs@GMP-Tb) core-shell nanoparticles. Initially, the core-shell nanoparticles were fabricated based on Au-NPs decorated with a fluorescent GMP-Tb shell. Upon being excited at 290 nm, the as-formed Au-NPs@GMP-Tb core-shell nanoparticles emit green fluorescence, and the decorated GMP-Tb shell causes the aggregation of Au-NPs. However, the addition of ALP destroys GMP-Tb shell, resulting in the release of Au-NPs from the shell into the solvent. As a consequence, the aggregated Au-NPs solubilizes with the changes in the UV-vis spectrum of the dispersion, and in the meantime, the fluorescence of GMP-Tb shell turns off, which constitutes a new mechanism for colorimetric and fluorescent dual-mode sensing of APA. With the method developed here, we could monitor the dynamic change of APA during an algal bloom of a freshwater lake, both by the naked eye and further confirmed by fluorometric determination. This study not only offers a new method for on-site visible detection of APA but also provides a strategy for dual-mode sensing mechanisms by the rational design of the excellent optical properties of Au-NPs and the adaptive inclusion properties of the luminescent infinite coordination polymers.

  5. A Chronoamperometric Screen Printed Carbon Biosensor Based on Alkaline Phosphatase Inhibition for W(VI) Determination in Water, Using 2-Phospho-l-Ascorbic Acid Trisodium Salt as a Substrate

    Science.gov (United States)

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia

    2015-01-01

    This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3), a repeatability of 9.4% (n = 3) and a detection limit of 0.29 ± 0.01 μM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case) and a dynamic range from 0.6 to 30 μM. This study was performed by means of a Lineweaver–Burk plot, showing a mixed kinetic inhibition. PMID:25621602

  6. A chronoamperometric screen printed carbon biosensor based on alkaline phosphatase inhibition for W(IV) determination in water, using 2-phospho-L-ascorbic acid trisodium salt as a substrate.

    Science.gov (United States)

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia

    2015-01-22

    This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3), a repeatability of 9.4% (n = 3) and a detection limit of 0.29 ± 0.01 µM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case) and a dynamic range from 0.6 to 30 µM. This study was performed by means of a Lineweaver-Burk plot, showing a mixed kinetic inhibition.

  7. A Chronoamperometric Screen Printed Carbon Biosensor Based on Alkaline Phosphatase Inhibition for W(VI Determination in Water, Using 2-Phospho-l-Ascorbic Acid Trisodium Salt as a Substrate

    Directory of Open Access Journals (Sweden)

    Ana Lorena Alvarado-Gámez

    2015-01-01

    Full Text Available This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3, a repeatability of 9.4% (n = 3 and a detection limit of 0.29 ± 0.01 µM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case and a dynamic range from 0.6 to 30 µM. This study was performed by means of a Lineweaver–Burk plot, showing a mixed kinetic inhibition.

  8. Alkaline phosphatase expression/activity and multilineage differentiation potential are the differences between fibroblasts and orbital fat-derived stem cells--a study in animal serum-free culture conditions.

    Science.gov (United States)

    Martins, Thaís Maria da Mata; de Paula, Ana Cláudia Chagas; Gomes, Dawidson Assis; Goes, Alfredo Miranda

    2014-10-01

    Human orbital fat tissues are a potential source to isolate stem cells for the development of regenerative medicine therapies. For future safe clinical application of these cells, it is critical to establish animal component-free culture conditions as well as to clearly define the stem cell population characteristics differentiating them from other cell types, such as fibroblasts. Therefore, the present study aimed to compare phenotypic and functional characteristics of orbital fat-derived stem cells (OFSCs) and fibroblasts resident in the eyelid skin in donor-matched samples grown in culture medium supplemented with pooled allogeneic human serum (HS) replacing fetal bovine serum (FBS). We first investigated the proliferative effects of OFSCs on HS, and then we compared the alkaline phosphatase (AP) expression and activity, immunophenotypic profile, and in vitro multilineage differentiation potential of OFSCs side-by-side with fibroblasts. The results showed that HS enhanced OFSCs proliferation without compromising their immunophenotype, AP activity, and osteogenic, adipogenic, and chondrogenic differentiation capacities. In contrast to OFSCs, the fibroblasts did not exhibit AP expression and activity and did not have multilineage differentiation potential. The results enabled us to successfully distinguish OFSCs from fibroblasts populations, suggesting that AP expression/activity and multilineage differentiation assays can be used reliably to discriminate mesenchymal stem cells from fibroblasts. Our findings also support the feasibility of pooled allogeneic HS as a safer and more effective alternative to FBS for clinical applications.

  9. Oxidative Alkaline leaching of Americium from simulated high-level nuclear waste sludges

    Energy Technology Data Exchange (ETDEWEB)

    Reed, Wendy A.; Garnov, Alexander Yu.; Rao, Linfeng; Nash, Kenneth L.; Bond, Andrew H.

    2004-01-23

    Oxidative alkaline leaching has been proposed to pre-treat the high-level nuclear waste sludges to remove some of the problematic (e.g., Cr) and/or non-radioactive (e.g., Na, Al) constituents before vitrification. It is critical to understand the behavior of actinides, americium and plutonium in particular, in oxidative alkaline leaching. We have studied the leaching behavior of americium from four different sludge simulants (BiPO{sub 4}, BiPO{sub 4 modified}, Redox, PUREX) using potassium permanganate and potassium persulfate in alkaline solutions. Up to 60% of americium sorbed onto the simulants is leached from the sludges by alkaline persulfate and permanganate. The percentage of americium leached increases with [NaOH] (between 1.0 and 5.0 M). The initial rate of americium leaching by potassium persulfate increases in the order BiPO{sub 4} sludge < Redox sludge < PUREX sludge. The data are most consistent with oxidation of Am{sup 3+} in the sludge to either AmO{sub 2}{sup +} or AmO{sub 2}{sup 2+} in solution. Though neither of these species is expected to exhibit long-term stability in solution, the potential for mobilization of americium from sludge samples would have to be accommodated in the design of any oxidative leaching process for real sludge samples.

  10. “补肾健脾”针刺法对绝经后骨质疏松症模型血清碱性磷酸酶水平的影响%Effect of acupuncture method of"invigorating the kidney and strengthening the spleen"on the level of serum alkaline phosphatase in postmenopausal osteoporosis model

    Institute of Scientific and Technical Information of China (English)

    李灿; 黎喜平; 陈久毅; 沈骏; 秧荣昆; 王建

    2016-01-01

    group(P<0.05).Compared with the model group,the serum ALP levels in the acupuncture group and the estrogen group were lower than those in the model group(P<0.01).Conclusion:The content of alkaline phosphatase in serum of ovariectomized rats was increased,format a high conversion osteoporosis model,after estrogen replacement and acupuncture treatment,the content of serum ALP decreased,both of the two treatments can resist the high conversion of bone metabolism,to provide part of the clinical effect of acupuncture method of"invigorating the kidney and strengthening the spleen"in the treatment of postmenopausal osteoporosis.

  11. A facile chemical route for recovery of high quality zinc oxide nanoparticles from spent alkaline batteries.

    Science.gov (United States)

    Deep, Akash; Sharma, Amit L; Mohanta, Girish C; Kumar, Parveen; Kim, Ki-Hyun

    2016-05-01

    Recycling of spent domestic batteries has gained a great environmental significance. In the present research, we propose a new and simple technique for the recovery of high-purity zinc oxide nanoparticles from the electrode waste of spent alkaline Zn-MnO2 batteries. The electrode material was collected by the manual dismantling and mixed with 5M HCl for reaction with a phosphine oxide reagent Cyanex 923® at 250°C for 30min. The desired ZnO nanoparticles were restored from the Zn-Cyanex 923 complex through an ethanolic precipitation step. The recovered particle product with about 5nm diameter exhibited fluorescent properties (emission peak at 400nm) when excited by UV radiation (excitation energy of 300nm). Thus, the proposed technique offered a simple and efficient route for recovering high purity ZnO nanoparticles from spent alkaline batteries.

  12. Effects of DEP and DOP exposure on the acid phosphatase and alkaline phosphatase in the%DEP、DOP暴露对鲤鱼的酸碱性磷酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    祁明亮; 朱敏; 赵晓祥

    2012-01-01

    . As a result, we found that DEP and DOP may cause distortion of the carp in bodies and make them upside-down and head-up, with obvious scoliosis. It shows that the % h - LC50 is 41.50 mg/L and 7.95 mg/L. And, then, we tried three different concentrations of DEP and DOP (1/10 LC50, 1/5 LC50, 1/2.5 LC50) with a control group (0 mg/L) con-fected to measure the activities of ACP and ALP in carp' s liver and muscles. The results of our experiments show that the effects of DEP and DOP on carp' s iiver and muscle tissue tend to be inhibited by the high concentrations (16.60 mg/L, 3.20 mg/L), whereas the low concentrations(4.15 mg/L, 0.80 mg/L) were induced. The effects of DEP on the ALP activity of carp liver and muscle tissue also indicate inhibition of high concentrations(16.60 mg/L, 3.20 mg/L) and induction of low concentrations(4.15 mg/L, 0.80 mg/L). At the same time, the effects of DOP on the carp' s liver reveal that: as the concentration increases, the ALP activity tends to decrease, showing a clear dose-response relationship. Thus, ACP and ALP may imply a significant role in resisting the toxic effects of phthalales, though the reason for it needs further studies.

  13. High Iridium concentration of alkaline rocks of Deccan and implications to K/T boundary

    Indian Academy of Sciences (India)

    P N Shukla; N Bhandari; Anirban Das; A D Shukla; J S Ray

    2001-06-01

    We report here an unusually high concentration of iridium in some alkali basalts and alkaline rocks of Deccan region having an age of about 65Ma, similar to the age of the Cretaceous-Tertiary boundary. The alkali basalts of Anjar, in the western periphery of Deccan province, have irid-ium concentration as high as 178pg/g whereas the alkaline rocks and basalts associated with the Amba Dongar carbonatite complex have concentrations ranging between 8 and 80 pg/g. Some of these values are more than an order of magnitude higher than the concentration in the tholeiiticbasalts of Deccan, indicating the significance of alkaline magmatism in the iridium inventory at the Cretaceous-Tertiary boundary. Despite higher concentration, their contribution to the global inventory of iridium in the Cretaceous-Tertiary boundary clays remains small. The concentration of iridium in uorites from Amba Dongar was found to be < 30 pg/g indicating that iridium is not incorporated during their formation in hydrothermal activity.

  14. Spatial-temporal Variations of Alkaline Phosphatase Activity in the Kuilei Lake and Their Influencing Factors%傀儡湖水体中碱性磷酸酶活性的时空变化及其影响因素

    Institute of Scientific and Technical Information of China (English)

    张成艳; 成小英; 王建军; 张路; 杜应旸

    2013-01-01

    Alkaline phosphatase activity (APA) plays an important role in the biogeochemical cycling of phosphorus.The spatial-temporal distribution of APA and related environmental variables were studied in the Kuilei Lake.The research found significant seasonal and regional variations of APA in the water body,with lower APAs in outlet waters and higher in the estuarine.APAs were relatively low in winter (January),ranging from 2.06 to 17.33 nmol mL-1 h-1,and high in summer (July),ranging from 5.78 to 13.52 nmol mL-1 h-1.Correlation analyses indicated that APAs were significantly related to particulate organic nitrogen,total nitrogen and NO2--N.Other nitrogen forms (NO3--N and NH4+-N) were also related,though not significantly.The results implied that the factors related to nitrogen are the primary factors affecting APAs.%碱性磷酸酶在磷的生物地球化学循环过程中发挥着重要作用.以傀儡湖为研究区域,对其水体中碱性磷酸酶活性(Alkaline phosphatase activity,APA)及其理化性质的时空分布进行分析.结果表明,傀儡湖水体APA呈时空异质性:河道较低,河口则易出现较高的APA;全湖的APA存在较强的季节性变化,冬季(1月)最低,变化范围为2.06-17.33nmol mL-1h-1;夏季(7月)最高,变化范围为5.78-13.52 nmol mL-1 h-1.APA与环境因子相关性分析显示,APA与颗粒态有机氮浓度关系最为密切,与总氮含量的相关性其次,随后显著相关的因子为NO2--N,此外是NO3--N和NH4+-N.这一结果表明与氮元素有关的环境因子可能是影响APA的主要因素.

  15. Field-Evolved Mode 1 Resistance of the Fall Armyworm to Transgenic Cry1Fa-Expressing Corn Associated with Reduced Cry1Fa Toxin Binding and Midgut Alkaline Phosphatase Expression.

    Science.gov (United States)

    Jakka, Siva R K; Gong, Liang; Hasler, James; Banerjee, Rahul; Sheets, Joel J; Narva, Kenneth; Blanco, Carlos A; Jurat-Fuentes, Juan L

    2015-12-04

    Insecticidal protein genes from the bacterium Bacillus thuringiensis (Bt) are expressed by transgenic Bt crops (Bt crops) for effective and environmentally safe pest control. The development of resistance to these insecticidal proteins is considered the most serious threat to the sustainability of Bt crops. Resistance in fall armyworm (Spodoptera frugiperda) populations from Puerto Rico to transgenic corn producing the Cry1Fa insecticidal protein resulted, for the first time in the United States, in practical resistance, and Bt corn was withdrawn from the local market. In this study, we used a field-collected Cry1Fa corn-resistant strain (456) of S. frugiperda to identify the mechanism responsible for field-evolved resistance. Binding assays detected reduced Cry1Fa, Cry1Ab, and Cry1Ac but not Cry1Ca toxin binding to midgut brush border membrane vesicles (BBMV) from the larvae of strain 456 compared to that from the larvae of a susceptible (Ben) strain. This binding phenotype is descriptive of the mode 1 type of resistance to Bt toxins. A comparison of the transcript levels for putative Cry1 toxin receptor genes identified a significant downregulation (>90%) of a membrane-bound alkaline phosphatase (ALP), which translated to reduced ALP protein levels and a 75% reduction in ALP activity in BBMV from 456 compared to that of Ben larvae. We cloned and heterologously expressed this ALP from susceptible S. frugiperda larvae and demonstrated that it specifically binds with Cry1Fa toxin. This study provides a thorough mechanistic description of field-evolved resistance to a transgenic Bt crop and supports an association between resistance and reduced Cry1Fa toxin binding and levels of a putative Cry1Fa toxin receptor, ALP, in the midguts of S. frugiperda larvae.

  16. Detection and analysis on bone alkaline phosphatase activity of 2540 infants%2540例婴幼儿骨源性碱性磷酸酶活性检测分析

    Institute of Scientific and Technical Information of China (English)

    盛巍

    2012-01-01

    目的:调查长兴地区0~5岁婴幼儿骨健康状况.方法:使用骨源性碱性磷酸酶(BAP)试剂盒对2009~2010年来长兴妇幼保健院门诊就诊的2 540例0~5岁婴幼儿进行检测.结果:在2 540例0~5岁婴幼儿中,BAP活性正常的1 215例,占总人数的47.83%;BAP活性处于预防水平的1 196例,占总人数的47.09%;BAP活性处于治疗水平的129例,占总人数的5.08%.结论:长兴地区约50.00%的婴幼儿骨骼处于亚健康状态,易患佝偻病,需及早预防.%Objective: To investigate the bone health status of 0 - 5 - year - old infants in Changxing county. Methods: Bone alkaline phosphatase ( BAP) activity of 2 540 infants aged 0-5 years who went to outpatient department of the hospital from 2009 to 2010 was detected by BAP kit. Results; Among2 540 infante aged 0 -5 years old, I 215 infants were found with normal BAP activity, accounting for 47. 83% ; BAP activity of 1 196 infants was at prevention level, accounting for 47.09% ; BAP activity of 129 infants was at treatment level, accounting for 5.08%. Conclusion: The bones of about 50.00% of the infants in Changxing county were subhealthy, the infants are susceptible to rickets, and prevention should be earned out early.

  17. High-rate capability of zinc anodes in alkaline primary cells

    Science.gov (United States)

    Huot, Jean-Yves; Malservisi, Martin

    This work is devoted to the electrochemical aspects of high-power testing of primary alkaline LR6 ("AA") cells and to the factors influencing cell performance, namely the corresponding zinc anode behaviour under such high-rate conditions. The influence of the high-rate testing regime, such as the discharge mode and the end-potential, on zinc utilisation in alkaline cells has been monitored and its behaviour has been isolated by means of a pseudo-reference electrode. As anticipated, anode formulation, including zinc alloy composition and size distribution, is found to affect the cell's discharge curve and the corresponding zinc electrode potential and utilisation. The effects of these parameters on the discharge curve are discussed in terms of three stages of discharge. Finally, the high-rate capability of commercial LR6 cells is analysed in terms of zinc anode formulation. It was concluded that zinc electrode polarisation is very small and is relatively independent of manufacturer, of zinc anode formulation and of zinc alloying. On the other hand, metallic zinc utilisation remains very low under high-rate conditions.

  18. Crystallographic studies on shrimp alkaline phosphatase

    NARCIS (Netherlands)

    Backer, M.M.E. (Maaike Maria Eva)

    2003-01-01

    The earth has large cold areas, such as mountains, oceans and (ant)arctic regions, in which organisms have evolved to survive. This adaptation happens at a molecular level. The question is, how do proteins adjust such that they function at low temperatures? "Cold-active" or "cold-adapted" enzymes ha

  19. Alkaline electrolysis cell at high temperature and pressure of 250 °C and 42 bar

    DEFF Research Database (Denmark)

    Allebrod, Frank; Chatzichristodoulou, Christodoulos; Mogensen, Mogens Bjerg

    2013-01-01

    A new type of alkaline electrolysis cells with nickel foam based gas diffusion electrodes and KOH (aq) immobilized in mesoporous SrTiO3 has been developed and tested at temperatures and pressures up to 250 °C and 42 bar, respectively. Current densities of 1.0 A cm−2 have been measured at a cell...... voltage of 1.5 V without the use of expensive noble metal catalysts. High electrical efficiency and current density combined with relatively small production costs may lead to both reduced investment and operating costs for hydrogen and oxygen production....

  20. Use of highly alkaline conditions to improve cost-effectiveness of algal biotechnology.

    Science.gov (United States)

    Canon-Rubio, Karen A; Sharp, Christine E; Bergerson, Joule; Strous, Marc; De la Hoz Siegler, Hector

    2016-02-01

    Phototrophic microorganisms have been proposed as an alternative to capture carbon dioxide (CO2) and to produce biofuels and other valuable products. Low CO2 absorption rates, low volumetric productivities, and inefficient downstream processing, however, currently make algal biotechnology highly energy intensive, expensive, and not economically competitive to produce biofuels. This mini-review summarizes advances made regarding the cultivation of phototrophic microorganisms at highly alkaline conditions, as well as other innovations oriented toward reducing the energy input into the cultivation and processing stages. An evaluation, in terms of energy requirements and energy return on energy invested, is performed for an integrated high-pH, high-alkalinity growth process that uses biofilms. Performance in terms of productivity and expected energy return on energy invested is presented for this process and is compared to previously reported life cycle assessments (LCAs) for systems at near-neutral pH. The cultivation of alkaliphilic phototrophic microorganisms in biofilms is shown to have a significant potential to reduce both energy requirements and capital costs.

  1. Phosphatase activity and organic phosphorus turnover on a high Arctic glacier

    Directory of Open Access Journals (Sweden)

    M. Stibal

    2009-05-01

    Full Text Available Arctic glacier surfaces harbour abundant microbial communities consisting mainly of heterotrophic and photoautotrophic bacteria. The microbes must cope with low concentrations of nutrients and with the fact that both the dissolved and debris-bound nutrient pools are dominated by organic phases. Here we provide evidence that phosphorus (P is deficient in the supraglacial environment on a Svalbard glacier, we quantify the enzymatic activity of phosphatases in the system and we estimate the contribution of the microbes to the cycling of the dominant organic P in the supraglacial environment. Incubation of cryoconite debris revealed significant phosphatase activity in the samples (19–67 nmol MUP g−1 h−1. It was inhibited by inorganic P during incubations and had its optimum at around 30°C. The phosphatase activity measured at near-in situ temperature and substrate concentration suggests that the available dissolved organic P can be turned over by microbes within ~3–11 h on the glacier surface. By contrast, the amount of potentially bioavailable debris-bound organic P is sufficient for a whole ablation season. However, it is apparent that some of this potentially bioavailable debris-bound P is not accessible to the microbes.

  2. High-Strength / High Alkaline Resistant Fe-Phosphate Glass Fibers as Concrete Reinforcement

    Energy Technology Data Exchange (ETDEWEB)

    Mariano Velez

    2008-03-31

    Calcium-iron-phosphate glasses were developed whose chemical durabilities in alkaline solutions (pH 13) were comparable or superior to those of commercial alkaline-resistant (AR) silica-based glasses. However, the tensile strength of Ca-Fe-phosphate fibers, after being exposed to alkaline environments, including wet Portland cement pastes, is lower than that of current AR silicate fibers. Another series of Ca-Fe-phosphate glasses were developed with excellent chemical durability in strong acidic solutions (H2SO4, HF), indicating potential applications where silica-based fibers degrade very quickly, including E-glass. The new Ca-Fe-phosphate glasses can be melted and processed 300 to 500°C lower than silica-based glasses. This offers the possibility of manufacturing glass fibers with lower energy costs by 40-60% and the potential to reduce manufacturing waste and lower gas emissions. It was found that Ca-Fe-phosphate melts can be continuously pulled into fibers depending on the slope of the viscosity-temperature curve and with viscosity ~100 poise, using multi-hole Pt/Rh bushings.

  3. A newly high alkaline lipase: an ideal choice for application in detergent formulations

    Directory of Open Access Journals (Sweden)

    Cherif Slim

    2011-11-01

    Full Text Available Abstract Background Bacterial lipases received much attention for their substrate specificity and their ability to function in extreme environments (pH, temperature.... Many staphylococci produced lipases which were released into the culture medium. Reports of thermostable lipases from Staphylococcus sp. and active in alkaline conditions are not previously described. Results A newly soil-isolated Staphylococcus sp. strain ESW secretes an induced lipase in the culture medium. The effects of temperature, pH and various components in a detergent on the activity and stability of Staphylococcus sp. lipase (SL1 were studied in a preliminary evaluation for use in detergent formulation solutions. The enzyme was highly active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 12.0. The relative activity at pH 13.0 was about 60% of that obtained at pH 12.0. It exhibited maximal activity at 60°C. This novel lipase, showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and relative stability towards oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various commercial solid and liquid detergents. Conclusions These properties added to the high activity in high alkaline pH make this novel lipase an ideal choice for application in detergent formulations.

  4. Regional scale hydrological and biogeochemical processes controlling high biodiversity of a groundwater fed alkaline fen

    Science.gov (United States)

    van der Zee, Sjoerd E. A. T. M.; (D. G.) Cirkel, Gijsbert; (J. P. M) witte, Flip

    2014-05-01

    The high floral biodiversity of groundwater fed fens and mesotrophic grasslands depends on the different chemical signatures of the shallow rainwater fed topsoil water and the slightly deeper geochemically affected groundwater. The relatively abrupt gradients between these two layers of groundwater enable the close proximity of plants that require quite different site factors and have different rooting depths. However, sulphur inflow into such botanically interesting areas is generally perceived as a major threat to biodiversity. Although in Europe atmospheric deposition of sulphur has decreased considerably over the last decades, groundwater pollution by sulphate may still continue due to pyrite oxidation in soil as a result of excessive fertilisation. Inflowing groundwater rich in sulphate can change biogeochemical cycling in nutrient-poor wetland ecosystems because of 'so called' internal eutrophication as well as the accumulation of dissolved sulphide, which is phytotoxic. Complementary to conventions, we propose that upwelling sulphate rich groundwater may, in fact, promote the conservation of rare and threatened alkaline fens: excessive fertilisation and pyrite oxidation also produces acidity, which invokes calcite dissolution, and increased alkalinity and hardness of the inflowing groundwater. For a very species-rich wetland nature reserve, we show that sulphate is reduced and effectively precipitated as iron sulphides, when this calcareous and sulphate rich groundwater flows upward through the organic soil of the investigated nature reserve. Also, we show that sulphate reduction occurs simultaneously with an increase in alkalinity production, which in our case results in active calcite precipitation in the soil. In spite of the occurring sulphate reduction, we found no evidence for internal eutrophication. Extremely low phosphorous concentration in the pore water could be attributed to a high C:P ratio of soil organic matter and co-precipitation with

  5. [Effect of VAM fungi on phosphatase activity in maize rhizosphere].

    Science.gov (United States)

    Song, Y; Li, X; Feng, G

    2001-08-01

    The effect of VAM fungi on phosphatase activity in maize rhizosphere was examined by pot culture experiment, in which, three-compartment-pots were used, the central compartment being separated from the outer two by a nylon net with 30 microns mesh. Plants were harvested 70 days after planting. Soil acid and alkaline phosphatase were measured at different distances from root surface. The results showed that VAM increased the activities of soil acid and alkaline phosphatase in the rhizosphere. It was found that different phosphorous sources had different effects on phosphatase activity.

  6. Phosphatase activity and organic phosphorus turnover on a high Arctic glacier

    Directory of Open Access Journals (Sweden)

    M. Stibal

    2009-02-01

    Full Text Available Arctic glacier surfaces harbor abundant microbial communities consisting mainly of heterotrophic and photoautotrophic bacteria. The microbes must cope with very low concentrations of nutrients and with the fact that both the dissolved and debris-bound nutrient pools are dominated by organic phases. Here we provide evidence that phosphorus (P is deficient and limiting in the supraglacial environment on a Svalbard glacier, we show how the microbial community responds to the P stress and we quantify the contribution of the microbes to the cycling of the dominant organic P in the supraglacial environment. Incubation of cryoconite debris revealed significant phosphatase activity in the samples (19–67 nmol MUP g−1 h−1, which was controlled by the concentration of inorganic P during incubations and had its optimum at around 30°C. The phosphatase activity rates measured at near-in situ temperature and substrate concentration imply that the available dissolved organic P can be turned over by microbes within ~3–11 h on the glacier surface. By contrast, the amount of potentially bioavailable debris-bound organic P is sufficient for a whole ablation season. However, it is apparent that some of this potentially bioavailable debris-bound P is not accessible to the microbes.

  7. High Temperature and Pressure Alkaline Electrochemical Reactor for Conversion of Power to Chemicals

    DEFF Research Database (Denmark)

    Chatzichristodoulou, Christodoulos

    2016-01-01

    Moving away from fossil fuels requires harvesting more and more intermittent renewable energy resources and establishing a sustainable system for the production of chemicals. This brings forward the need for efficient large scale energy storage technologies 1-3 and technologies for the conversion...... densities. This work will provide an overview of our efforts to develop components of such high temperature alkaline electrochemical reactors for different applications. Low-cost large-scale production methods have been successfully employed for the production of ceramic diaphragms and full cells...... of renewable electricity to chemicals. Electrochemical reactors can play a crucial role in this endeavor, since they can efficiently and reversibly transform electricity to high-value chemicals, and thus serve as energy storage and recovery devices for balancing the grid, while offering a means...

  8. Trichoderma harzianum transformant has high extracellular alkaline proteinase expression during specific mycoparasitic interactions

    Directory of Open Access Journals (Sweden)

    Goldman Maria Helena S.

    1998-01-01

    Full Text Available The mycoparasite Trichoderma harzianum produces an alkaline proteinase that may be specifically involved in mycoparasitism. We have constructed transformant strains of this fungus that overexpress this alkaline proteinase. Some of the transformants were assessed for alkaline proteinase activity, and those with higher activity than the wild type were selected for further studies. One of these transformant strains produced an elevated and constitutive pbr1 mRNA level during mycoparasitic interactions with Rhizoctonia solani.

  9. Thermal poling of alkaline earth boroaluminosilicate glasses with intrinsically high dielectric breakdown strength

    Science.gov (United States)

    Smith, Nicholas J.; Lanagan, Michael T.; Pantano, Carlo G.

    2012-04-01

    Per the rectification model of thermal poling, it has been proposed that intrinsic breakdown strength plays a strong limiting role in the internal DC fields supported by the glass from the poling process. One might therefore hypothesize proportionately larger second-order nonlinearity (SON) in glasses with intrinsically high dielectric breakdown strength. We test these ideas by thermal poling of two different commercial alkali-free alkaline-earth boroaluminosilicate display glasses—one with barium only (AF45 from Schott), and the other with a mixture of alkaline-earth ions (OA-10 G from NEG). Not only are such compositions relevant from a commercial standpoint, they are also interesting in that they have been recently shown to exhibit remarkably high intrinsic dielectric breakdown strengths of 11-14 MV/cm. Quantitative Maker fringe and stack Maker-fringe measurements provide an accurate evaluation of the poling-induced SON susceptibilities, and indicate maximum χ(2) values of 0.44 and 0.26 pm/V in these glasses. These values are comparable to those reported for silica and other multicomponent glasses. Thus, the hypothesis that higher χ(2) would be observed in high intrinsic breakdown strength glasses was not validated. Based on our application of the rectification model, internal fields of the order 2-4 MV/cm were calculated, which are well below the measured intrinsic breakdown strengths at room temperature. The most plausible explanation for these observations is nonlinear electronic conduction effects taking place within the depletion region at the poling temperature, limiting internal fields to a fraction of the breakdown field.

  10. High-Efficiency Artificial Photosynthesis Using a Novel Alkaline Membrane Cell

    Science.gov (United States)

    Narayan, Sri; Haines, Brennan; Blosiu, Julian; Marzwell, Neville

    2009-01-01

    A new cell designed to mimic the photosynthetic processes of plants to convert carbon dioxide into carbonaceous products and oxygen at high efficiency, has an improved configuration using a polymer membrane electrolyte and an alkaline medium. This increases efficiency of the artificial photosynthetic process, achieves high conversion rates, permits the use of inexpensive catalysts, and widens the range of products generated by this type of process. The alkaline membrane electrolyte allows for the continuous generation of sodium formate without the need for any additional separation system. The electrolyte type, pH, electrocatalyst type, and cell voltage were found to have a strong effect on the efficiency of conversion of carbon dioxide to formate. Indium electrodes were found to have higher conversion efficiency compared to lead. Bicarbonate electrolyte offers higher conversion efficiency and higher rates than water solutions saturated with carbon dioxide. pH values between 8 and 9 lead to the maximum values of efficiency. The operating cell voltage of 2.5 V, or higher, ensures conversion of the carbon dioxide to formate, although the hydrogen evolution reaction begins to compete strongly with the formate production reaction at higher cell voltages. Formate is produced at indium and lead electrodes at a conversion efficiency of 48 mg of CO2/kilojoule of energy input. This efficiency is about eight times that of natural photosynthesis in green plants. The electrochemical method of artificial photosynthesis is a promising approach for the conversion, separation and sequestration of carbon dioxide for confined environments as in space habitats, and also for carbon dioxide management in the terrestrial context. The heart of the reactor is a membrane cell fabricated from an alkaline polymer electrolyte membrane and catalyst- coated electrodes. This cell is assembled and held in compression in gold-plated hardware. The cathode side of the cell is supplied with carbon

  11. High-resolution crystal structure of the PDZ1 domain of human protein tyrosine phosphatase PTP-Bas.

    Science.gov (United States)

    Lee, Sang-Ok; Lee, Mi-Kyung; Ku, Bonsu; Bae, Kwang-Hee; Lee, Sang Chul; Lim, Heon M; Kim, Seung Jun; Chi, Seung-Wook

    2016-09-23

    Protein tyrosine phosphatase-Basophil (PTP-Bas) is a membrane-associated protein tyrosine phosphatase with five PDZ domains and is involved in apoptosis, tumorigenesis, and insulin signaling. The interaction between PTP-Bas and tandem-PH-domain-containing protein 1/2 (TAPP1/2) plays an essential role in the regulation of insulin signaling. Despite its high sequence homology with the other PDZ domains, only the PDZ1 domain of PTP-Bas showed distinct binding specificity for TAPP1/2. Although the interaction between PTP-Bas PDZ1 and TAPP1/2 is a therapeutic target for diabetes, the structural basis for the interaction has not been elucidated. In the present study, we determined the crystal structure of the PTP-Bas PDZ1 domain at 1.6 Å resolution. In addition, we calculated the structural models of complexes of PTP-Bas PDZ1 and the C-terminal peptides of TAPP1/2 (referred to as TAPP1p/2p). Structural comparison with the PTP-Bas PDZ2/RA-GEF2 peptide complex revealed a structural basis for distinct binding specificity of PTP-Bas PDZ1 for TAPP1p/2p peptides. Our high-resolution crystal structure of PTP-Bas PDZ1 will serve as a useful template for rational structure-based design of novel anti-diabetes therapeutics.

  12. Phosphate-solubility and phosphatase activity in Gangetic alluvial soil as influenced by organophosphate insecticide residues.

    Science.gov (United States)

    Majumder, Shyam Prasad; Das, Amal Chandra

    2016-04-01

    An experiment was conducted under laboratory conditions to investigate the effect of four organophosphate insecticides, viz. monocrotophos, profenophos, quinalphos and triazophos at their field application rates (0.75, 1.0, 0.5 and 0.6 kg a.i.ha(-1), respectively), on the growth and activities of phosphate solubilizing microorganisms in relation to availability of insoluble phosphates in the Gangetic alluvial soil of West Bengal, India. The proliferation of phosphate solubilizing microorganisms was highly induced with profenophos (38.3%), while monocrotophos exerted maximum stimulation (20.8%) towards the solubility of insoluble phosphates in soil. The phosphatase activities of the soil (both acid phosphatase and alkaline phosphatase) were significantly increased due to the incorporation of the insecticides in general, and the augmentation was more pronounced with quinalphos (43.1%) followed by profenophos (27.6%) for acid phosphatase, and with monocrotophos (25.2%) followed by profenophos (16.1%) for alkaline phosphatase activity in soil. The total phosphorus was highly retained by triazophos (19.9%) followed by monocrotophos (16.5%), while incorporation of triazophos and quinalphos manifested greater availability of water soluble phosphorus in soil.

  13. [Phosphatase activity in Amoeba proteus at pH 9.0].

    Science.gov (United States)

    Sopina, V A

    2007-01-01

    In the free-living amoeba Amoeba proteus (strain B), after PAAG disk-electrophoresis of the homogenate supernatant, at using 1-naphthyl phosphate as a substrate and pH 9.0, three forms of phosphatase activity were revealed; they were arbitrarily called "fast", "intermediate", and "slow" phosphatases. The fast phosphatase has been established to be a fraction of lysosomal acid phosphatase that preserves some low activity at alkaline pH. The question as to which particular class the intermediate phosphatase belongs to has remained unanswered: it can be both acid phosphatase and protein tyrosine phosphatase (PTP). Based on data of inhibitor analysis, large substrate specificity, results of experiments with reactivation by Zn ions after inactivation with EDTA, other than in the fast and intermediate phosphatases localization in the amoeba cell, it is concluded that only slow phosphatase can be classified as alkaline phosphatase (EC 3.1.3.1).

  14. 秦皇岛近海褐潮高发区浮游植物的碱性磷酸酶活性分析%Alkaline phosphatase activity of phytoplankton in Qin-huangdao coastal waters with frequent brown tide occurrences

    Institute of Scientific and Technical Information of China (English)

    刘晓红; 覃仙玲; 蔡阳扬; 欧林坚; 吕颂辉

    2016-01-01

    Alkaline phosphatase (AP) is a type of ectoenzyme that is expressed by phytoplankton under phosphorus (P) stress states and can hydrolyze dissolved organic P (DOP). AP has been widely used to indicate the P stress status of phytoplankton in the sea. In this study, the AP activity (APA) of phytoplankton during the brown tide of Aureococcus anophagefferens that occurred near the coastal waters of Qinghuangdao was investigated in July 2013. The P status of phytoplankton and the capability of phytoplankton in using DOP during the bloom were analyzed. The results show that the density of A. anophagefferens was as high as 108 cells/L when the bloom occurred in the coastal area of Qinhuangdao. DOP was the main P source for phytoplankton growth. The densities of A. ano-phagefferens showed positive correlations with NO3−, DOP, dissolved inorganic P, and so forth. Phytoplankton ex-pressed abundant AP, and the average APA in water reached as high as 217.72 nmol/(µg·h)chla ± 90.86 nmol/(µg·h)chla (350.44 nmol/(L·h) ± 130.57 nmol/(L·h)). APA increased significantly with phytoplankton biomass. Our results suggest that A. anophagefferens experienced severe P stress or even P limitation when the brown tide occurred. The bioavailability of P, particularly DOP, may play a key role in the occurrence and persistence of the brown tide of A. anophagefferens in the coastal waters of Qinhuangdao.%碱性磷酸酶(Alkaline phosphatase, AP)是浮游植物在磷胁迫状态下表达的一种水解有机磷源的胞外酶,可用于指示海区浮游植物的磷胁迫状态。本研究于2013年7月,对秦皇岛近海抑食金球藻(Aureococcus anophagefferens)褐潮发生期间浮游植物的碱性磷酸酶活性(AP activity, APA)进行研究,结合其他理化参数,分析藻华发生时浮游植物的磷营养状态及其对海水中磷源的水解与利用情况。结果表明,褐潮发生时,抑食金球藻细胞密度高达108个/L,溶解有机磷(Dissolved organic phosphorus

  15. Effect of phenylmercuric acetate injections on phosphatase activity in chickens resistant and susceptible to Leukosis

    Energy Technology Data Exchange (ETDEWEB)

    Miller, V.L.; Bearse, G.E.; Csonka, E.

    1972-01-01

    The weighted means of liver and kidney alkaline phosphatase activity was greater in three strains of chickens classified as susceptible to limphoid leukosis than in five strains classified as resistant. On the same basis, four strains classified as susceptible to Marek's disease had more liver alkaline phosphatase activity than four strains classified as resistant. The weighted means of liver and kidney acid phosphatase activity were not different among the same strains of chickens classified similarly. Kidney alkaline phosphatase activity was the most generally inhibited by phenylmercuric acetate injections, followed by liver acid and alkaline phosphatase. Kidney acid phosphatase activity was enhanced by phenylmercuric acetate injections in three strains of chickens classified as resistant to both lymphoid leukosis and Marek's disease. Liver acid phosphatase activity was depressed in three strains classed as resistant to lymphoid leukosis.

  16. 环境因子对湖泊沉积物碱性磷酸酶活性的影响%Impact of Environmental Factors on Alkaline Phosphatase Activity of Sediment in Lakes

    Institute of Scientific and Technical Information of China (English)

    张翠英; 徐德兰; 万蕾; 琚淑明; 高明侠

    2013-01-01

    Alkaline phosphatase activity(APA) of sediment in Yunlong Lake and Luoma Lake was determined, and effects of physical and chemical characteristics of water, seasonal dynamics and aquatic plants on APA of different lakes were analyzed. Results showed that APA was significantly positive correlation with pH value (r=0.220, P<0.05) and temperature (r=0.478, P<0.01) respectively, but DO had little effect on APA Seasonal change of APA in Yunlong Lake was obvious, with the highest APA of 330.27 mg/(kg·h) in summer and the lowest of 154.65 mg/(kg·h) in spring. pH value varied from 8.12 to 8.50, temperature varied from 3.3 t to 26.5 ℃, DO varied from 6.84 mg/L to 8.47 mg/L. Aquatic plants had great effect on APA of Luoma Lake, APA of grass area was greater than that of no grass area, and seasonal change of APA was obvious in grass area. In brief, physical and chemical characteristics of water, seasonal dynamics and aquatic plants have great effect on APA.%文章对云龙湖和骆马湖2种不同类型湖泊沉积物的碱性磷酸酶进行了测定,分析了水体理化特性、季节变化及水生植物对湖泊沉积物碱性磷酸酶活性的影响,结果表明:pH值、温度对沉积物碱性磷酸酶活性影响较大,呈显著正相关(r=0.220,P<0.05; r=0.478,P<0.01),而DO值的影响较小,相关不显著;随着季节的变化,云龙湖沉积物碱性磷酸酶活性变化较明显,其变化规律为夏季>秋季>冬季>春季,夏季最高,为330.27 mg/(kg·h),春季最低,为154.65 mg/(kg·h);随着季节的变化,水体pH、DO变化较小,分别为8.12~8.50、6.84~8.47 mg/L,温度变化较大,为3.3~26.5℃;水生植物对骆马湖沉积物碱性磷酸酶活性影响较大,有草区均大于无草区且有草区碱性磷酸酶活性随季节变化显著.因此,水体理化特性、季节变化及水生植物等环境因子对不同湖泊沉积物中碱性磷酸酶活性有重要影响.

  17. Value of detection of calcium in whole blood and bone alkaline phosphatase in the diagnosis of rickets%全血钙和骨碱性磷酸酶检测在佝偻病诊断中的应用

    Institute of Scientific and Technical Information of China (English)

    韦孟兰

    2013-01-01

    目的 探讨全血钙和骨碱性磷酸酶(BALP)在儿童维生素D缺乏性佝偻病中的诊断价值.方法 对755例体检儿童采末梢血,用全血干化学法和免疫浓缩技术检测BALP,全血钙检测采用原子吸收光谱分析法.结果 755例体检儿童中BALP≤200 U/L 138例,200~250 U/L 386例,≥250U/L231例.血钙<1.55 mmol/L 79例,≥1.55mmol/L676例.BALP异常检出率为81.7%(617/755),血钙异常检出率为10.5%(79/755).血钙在各年龄组和男女性别间差异均无统计学意义.BALP在男女性别间差异无统计学意义,在不同年龄组间差异有统计学意义,1岁以下组BALP异常检出率最高,3~5岁组最低.大部分儿童处于维生素D轻度缺乏或缺乏状态,且随着年龄增长发病率有逐渐降低的趋势.结论 BALP的检测在维生素D缺乏性佝偻病诊断中较血钙测定敏感,更能反映人体钙营养水平.为更好地预防、控制和治疗佝偻病,婴幼儿应定期做相关项目的健康体检.%Objective To investigate the diagnostic value of calcium in whole blood and bone alkaline phosphatase (BALP) in vitamin D deficiency rickets in children. Methods Peripheral blood was collected from 755 children. Levels of BALP were detected based on dry chemical and immune concentrated technique, and calcium was detected by atomic absorption spectroscopy. Results BALP levels were showed ≤200 U/L in 138 children, 200-250 U/L in 386 children, and ≥ 250 U/L in 231 children. Calcium was found <1.55 mmo1/L in 79 children and ≥ 1.55 mmol/L in 676 children. The detection rate of abnormal BALP was 81.7% (617/755), and that of abnormal calcium was 0.5% (79/755). Calcium showed no statistically significant difference between different age groups and gender groups. The BALP showed no statistically significant difference between males and females, but significant differences between different age groups. The detection rate of abnormal BALP was highest in age group that smaller than one

  18. Highly efficient anode catalyst with a Ni@PdPt core–shell nanostructure for methanol electrooxidation in alkaline media

    Institute of Scientific and Technical Information of China (English)

    Pei-shu Yu; Chun-tao Liu; Bo Feng; Jia-feng Wan; Li Li; Chun-yu Du

    2015-01-01

    To enhance the electrocatalytic activity of anode catalysts used in alkaline-media direct methanol fuel cells (DMFCs), a Ni@PdPt electrocatalyst was successfully prepared using a three-phase-transfer method. The Ni@PdPt electrocatalyst was characterized by X-ray dif-fraction (XRD), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), and high-resolution TEM (HRTEM) techniques. The experimental results indicate that the average particle size of the core–shell-structured Ni@PdPt electrocatalyst is approxi-mately 5.6 nm. The Ni@PdPt electrocatalyst exhibits a catalytic activity 3.36 times greater than that of PdPt alloys for methanol oxidation in alkaline media. The developed Ni@PdPt electrocatalyst offers a promising alternative as a highly electrocatalytically active anode catalyst for alkaline DMFCs.

  19. 泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化%Changes of digestive enzymes and alkaline phosphatase specific activities in Misgurnus anguillicaudatus during the development of early larval ontogeny

    Institute of Scientific and Technical Information of China (English)

    张云龙; 樊启学; 彭聪; 胡培培; 宗克金; 宋林; 沈凡

    2013-01-01

    The specific activities of several digestive enzymes (pensin, trypsin, lipase, amylase) and alkaline phosphatase of Misgurnus anguillicaudatus were determined from hatching to 30 DAH ( days after hatching). Pepsin activity could not be detected during the entire period of the experiment. In contrast, trypsin performed a high level activity after hatching , and its specific activity increased significantly after first feeding, peaking at 6 DAH, and decreased abruptly after that. The variation of lipase and amylase shared a similar pattern, their specific activities had two obvious peaks around the time of the transition of endogenous to exdogenous feeding and the beginning of the transportation of larval stage to juvenile stage. From 2 to 6 DAH, the specific activity of alkaline phosphatase significantly increased and then declined to a roughly stable level. This study demonstrated that M. anguillicaudatus has only a structural not functional stomach during the larval stage. The period of 2 to 6 DAH was the stage of the maturation of digestive gut in M. anguillicaudatus larvae and the crucial step forward the digestion mode of adult M. anguillicaudatus. The steadiness of these specific activities suggested that M. anguillicaudatus was efficient to utilize carbohydrate and lipid in food.%研究了泥鳅(Misgurnus anguillicaudatus)从孵化至30 DAH(日龄,Days after hatching)几种消化酶及碱性磷酸酶比活力的变化情况.胃蛋白酶直至30 DAH仍未检出活性.而胰蛋白酶表现出较高的比活力,其比活力在初次摄食之后显著上升,6 DAH达到最大值之后开始显著降低(P<0.05).脂肪酶与淀粉酶的变化模式相似,在内源性营养向外源性营养转变及仔鱼向稚鱼转变这两个时间段出现两个高峰值.碱性磷酸酶比活力在2-6 DAH显著上升(P<0.05),之后开始下降并趋于平稳.研究表明,泥鳅在仔稚鱼阶段只具有结构性的胃而缺乏分泌细胞的分化.2-6 DAH是泥鳅仔鱼肠

  20. Effect of Proteolysis with Alkaline Protease Following High Hydrostatic Pressure Treatment on IgE Binding of Buckwheat Protein.

    Science.gov (United States)

    Lee, Chaeyoon; Lee, Wonhui; Han, Youngshin; Oh, Sangsuk

    2017-03-01

    Buckwheat is a popular food material in many Asian countries and it contains major allergenic proteins. This study was performed to analyze the effects of hydrolysis with alkaline protease following high hydrostatic pressure (HHP) treatment on the IgE binding of buckwheat protein. Extracted buckwheat protein was treated with HHP at 600 MPa for 30 min and hydrolyzed with alkaline protease for 240 min. IgE binding was examined using an enzyme-linked immunosorbent assay (ELISA) with serum samples from 14 patients who were allergic to buckwheat. Depending on the serum samples, HHP treatment of buckwheat protein without enzymatic hydrolysis decreased the IgE binding by 8.9% to 73.2% or increased by 31% to 78%. The IgE binding of buckwheat protein hydrolyzed with alkaline protease decreased by 73.8% to 100%. The IgE binding of buckwheat protein hydrolyzed with alkaline protease following HHP treatment decreased by 83.8% to 100%. This suggested that hydrolysis with alkaline protease following HHP treatment could be applied to reduce the IgE binding of buckwheat protein. © 2017 Institute of Food Technologists®.

  1. Estudio de la fosfatasa ácida y alcalina en suelos de la Región Pampeana Norte del área sojera argentina Study of acid and alkaline phosphatase in soils of the Pampean North Region from argentine soybean area

    Directory of Open Access Journals (Sweden)

    Leticia Andrea Fernández

    2008-07-01

    ón de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po.Transformation of organic phosphorus (Po into soluble inorganic phosphorus (Pi is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB and cultivated fungi (CF was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5 CFU g-1 soil, while alkaline activity was 5.80 10(5 CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5 mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

  2. Alkaline earth silicate wools - A new generation of high temperature insulation.

    Science.gov (United States)

    Brown, Robert C; Harrison, Paul T C

    2012-11-01

    Intensive study of the natural asbestiform minerals that cause human diseases, and the consequent understanding of their hazardous characteristics, has enabled the development of manufactured fibres whose physical and/or chemical properties, in particular as they relate to biopersistence, have been adjusted to minimize possible harm to health. A strong driver for the developmentof new high temperature insulation materials wasthe perception of the toxicity of refractory ceramic fibres (RCF)and their classification in the EU as a category 2 carcinogen under Directive 67/548/EEC. Such classification carries with it the requirement for substitution by less hazardous materials. This paper focuses on the development of alkaline earth silicate (AES) wools as a new class of high temperature insulation with the capability of such substitution in a number of applications. These wools have only a low potential to cause harm because they do not persist in lung tissue once deposited, and have produced minimal effects in experimental test systems. AES wools are increasingly being used in a wide range of high temperature applications.

  3. Biohydrogen Fermentation from Sucrose and Piggery Waste with High Levels of Bicarbonate Alkalinity

    Directory of Open Access Journals (Sweden)

    Jeongdong Choi

    2015-03-01

    Full Text Available This study examined the influence of biohydrogen fermentation under the high bicarbonate alkalinity (BA and pH to optimize these critical parameters. When sucrose was used as a substrate, hydrogen was produced over a wide range of pH values (5–9 under no BA supplementation; however, BA affected hydrogen yield significantly under different initial pHs (5–10. The actual effect of high BA using raw piggery waste (pH 8.7 and BA 8.9 g CaCO3/L showed no biogas production or propionate/acetate accumulation. The maximum hydrogen production rate (0.32 L H2/g volatile suspended solids (VSS-d was observed at pH 8.95 and 3.18 g CaCO3/L. BA greater than 4 g CaCO3/L also triggered lactate-type fermentation, leading to propionate accumulation, butyrate reduction and homoacetogenesis, potentially halting the hydrogen production rate. These results highlight that the substrate with high BA need to amend adequately to maximize hydrogen production.

  4. Effect of puerarin combined with estradiol on bone tissue and serum calcium, phosphorus, and alkaline phosphatase in ovariectomized rats%葛根素联合雌二醇对去卵巢大鼠骨组织及血钙、磷和碱性磷酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    李海; 王金花; 黄海玲; 陈建海; 黎飚

    2013-01-01

    , 50 mg/kg, once a day) , estradiol group (the rats were treated with subcutaneous injection of estradiol, 200 μg/kg, twice a week) , low -dose puerarin combined with estradiol group (combined therapy group) (the rats were treated with subcutaneous injection of estradiol, 100 μg/kg, twice a week; subcutaneous injection of puerarin, 25 mg/kg, once a day) . At 4, 8, 12 , and 20 weeks after start of the experiment, 6 rats were randomly selected, femur smears were prepared to observe bone tissue; the blood samples were obtained to detect serum levels of calcium, phosphorus, and alkaline phosphatase; the data were analyzed statistically. Results : The pathological changes of osteoporosis were observed at 4, 8, 12, and 20 weeks in ovariectomized model group, the serum levels of calcium and phosphorus in ovariectomized model group were statistically significantly lower than those in sham - operation group (P 0. 05 ) . Bone tissue, serum levels of calcium, phosphorus, and alkaline phosphatase in combined therapy group basically returned to normal (P > 0.05), compared with puerarin group and estradiol group, there was no statistically significant difference (P > 0. 05) . Conclusion ; The curative effect of low - dose estradiol combined with puerarin for treatment of osteoporosis in ovariectomized rats is similar to curative effects of relatively high - dose puerarin or estradiol.

  5. Characterization of high temperature-tolerant rhizobia isolated from Prosopis juliflora grown in alkaline soil.

    Science.gov (United States)

    Kulkarni, Suneeta; Nautiyal, Chandra Shekhar

    1999-10-01

    A method was developed for the fast screening and selection of high-temperature tolerant rhizobial strains from root nodules of Prosopis juliflora growing in alkaline soils. The high-temperature tolerant rhizobia were selected from 2,500 Rhizobium isolates with similar growth patterns on yeast mannitol agar plates after 72 h incubation at 30 and 45 degrees C, followed by a second screening at 47.5 degrees C. Seventeen high-temperature tolerant rhizobial strains having distinguishable protein band patterns were finally selected for further screening by subjecting them to temperature stress up to 60 degrees C in yeast mannitol broth for 6 h. The high-temperature tolerant strains were NBRI12, NBRI329, NBRI330, NBRI332, and NBRI133. Using this procedure, a large number of rhizobia from root nodules of P. juliflora were screened for high-temperature tolerance. The assimilation of several carbon sources, tolerance to high pH and salt stress, and ability to nodulate P. juliflora growing in a glasshouse and nursery of the strains were studied. All five isolates had higher plant dry weight in the range of 29.9 to 88.6% in comparison with uninoculated nursery-grown plants. It was demonstrated that it is possible to screen in nature for superior rhizobia exemplified by the isolation of temperature-tolerant strains, which established effective symbiosis with nursery-grown P. juliflora. These findings indicate a correlation between strain performance under in vitro stress in pure culture and strain behavior under symbiotic conditions. Pure culture evaluation may be a useful tool in search for Rhizobium strains better suited for soil environments where high temperature, pH, and salt stress constitutes a limitation for symbiotic biological nitrogen fixation.

  6. Anti-obesity effect of alkaline reduced water in high fat-fed obese mice.

    Science.gov (United States)

    Ignacio, Rosa Mistica Coles; Kang, Tae-Young; Kim, Cheol-Su; Kim, Soo-Ki; Yang, Young-Chul; Sohn, Joon-Hyung; Lee, Kyu-Jae

    2013-01-01

    Whether or not alkaline reduced water (ARW) has a positive effect on obesity is unclear. This study aims to prove the positive effect of ARW in high-fat (HF) diet-induced obesity (DIO) in C57BL/6 mice model. Toward this, obesity was induced by feeding the C57BL/6 male mice with high-fat diet (w/w 45% fat) for 12 weeks. Thereafter, the animals were administered with either ARW or tap water. Next, the degree of adiposity and DIO-associated parameters were assessed: clinico-pathological parameters, biochemical measurements, histopathological analysis of liver, the expression of cholesterol metabolism-related genes in the liver, and serum levels of adipokine and cytokine. We found that ARW-fed mice significantly ameliorated adiposity: controlled body weight gain, reduced the accumulation of epididymal fats and decreased liver fats as compared to control mice. Accordingly, ARW coordinated the level of adiponectin and leptin. Further, mRNA expression of cytochrome P450 (CYP)7A1 was upregulated. In summary, our data shows that ARW intake inhibits the progression of HF-DIO in mice. This is the first note on anti-obesity effect of ARW, clinically implying the safer fluid remedy for obesity control.

  7. Enhanced coagulation for high alkalinity and micro-polluted water: the third way through coagulant optimization.

    Science.gov (United States)

    Yan, Mingquan; Wang, Dongsheng; Qu, Jiuhui; Ni, Jinren; Chow, Christopher W K

    2008-04-01

    Conventional coagulation is not an effective treatment option to remove natural organic matter (NOM) in water with high alkalinity/pH. For this type of water, enhanced coagulation is currently proposed as one of the available treatment options and is implemented by acidifying the raw water and applying increased doses of hydrolyzing coagulants. Both of these methods have some disadvantages such as increasing the corrosive tendency of water and increasing cost of treatment. In this paper, an improved version of enhanced coagulation through coagulant optimization to treat this kind of water is demonstrated. A novel coagulant, a composite polyaluminum chloride (HPAC), was developed with both the advantages of polyaluminum chloride (PACl) and the additive coagulant aids: PACl contains significant amounts of highly charged and stable polynuclear aluminum hydrolysis products, which is less affected by the pH of the raw water than traditional coagulants (alum and ferric salts); the additives can enhance both the charge neutralization and bridging abilities of PACl. HPAC exhibited 30% more efficiency than alum and ferric salts in dissolved organic carbon (DOC) removal and was very effective in turbidity removal. This result was confirmed by pilot-scale testing, where particles and organic matter were removed synergistically with HPAC as coagulant by sequential water treatment steps including pre-ozonation, coagulation, flotation and sand filtration.

  8. The Effects of High Alkaline Fly Ash on Strength Behaviour of a Cohesive Soil

    Directory of Open Access Journals (Sweden)

    A. Binal

    2016-01-01

    Full Text Available Contemporarily, there are 16 coal-burning thermal power plants currently operating in Turkey. This number is expected to rise to 46 in the future. Annually, about 15 million tons of fly ash are removed from the existing thermal power plants in Turkey, but a small proportion of it, 2%, is recyclable. Turkey’s plants are fired by lignite, producing Class C fly ash containing a high percentage of lime. Sulfate and alkali levels are also higher in Class C fly ashes. Therefore, fly ash is, commonly, unsuitable as an additive in cement or concrete in Turkey. In this study, highly alkaline fly ash obtained from the Yeniköy thermal power plants is combined with soil samples in different proportions (5%, 10%, 15%, 20%, and 25% and changes in the geomechanical properties of Ankara clay were investigated. The effect of curing time on the physicomechanical properties of the fly ash mixed soil samples was also analyzed. The soil classification of Ankara clay changed from CH to MH due to fly ash additives. Free swelling index values showed a decrease of 92.6%. Direct shear tests on the cohesion value of Ankara clay have shown increases by multiples of 15.85 and 3.01 in internal friction angle values. The California bearing ratio has seen a more drastic increase in value (68.7 times for 25% fly ash mix.

  9. Assessment of commercially available ion exchange materials for cesium removal from highly alkaline wastes

    Energy Technology Data Exchange (ETDEWEB)

    Brooks, K.P.; Kim, A.Y.; Kurath, D.E.

    1996-04-01

    Approximately 61 million gallons of nuclear waste generated in plutonium production, radionuclide removal campaigns, and research and development activities is stored on the Department of Energy`s Hanford Site, near Richland, Washington. Although the pretreatment process and disposal requirements are still being defined, most pretreatment scenarios include removal of cesium from the aqueous streams. In many cases, after cesium is removed, the dissolved salt cakes and supernates can be disposed of as LLW. Ion exchange has been a leading candidate for this separation. Ion exchange systems have the advantage of simplicity of equipment and operation and provide many theoretical stages in a small space. The organic ion exchange material Duolite{trademark} CS-100 has been selected as the baseline exchanger for conceptual design of the Initial Pretreatment Module (IPM). Use of CS-100 was chosen because it is considered a conservative, technologically feasible approach. During FY 96, final resin down-selection will occur for IPM Title 1 design. Alternate ion exchange materials for cesium exchange will be considered at that time. The purpose of this report is to conduct a search for commercially available ion exchange materials which could potentially replace CS-100. This report will provide where possible a comparison of these resin in their ability to remove low concentrations of cesium from highly alkaline solutions. Materials which show promise can be studied further, while less encouraging resins can be eliminated from consideration.

  10. Pediatric reference intervals of serum alkaline phosphatase for healthy Han population in Changchun%长春市汉族儿童血清碱性磷酸酶参考区间的建立

    Institute of Scientific and Technical Information of China (English)

    王迪; 杨春; 周琪; 许建成

    2015-01-01

    目的:建立0~14岁长春市汉族儿童血清碱性磷酸酶(ALP)的参考区间。方法采用日立7600-210全自动生化分析仪检测4211名健康体检儿童(男2090名,女2121名)血清 ALP。弃离群值后,判断数据是否正态分布。One-Way ANOVA 比较组间差异及确定是否需性别、年龄分组。非参数方法计算参考值的2.5百分位数和97.5百分位数,R 语言计算90%置信区间。结果长春市儿童 ALP 参考区间有年龄及性别差异。婴儿出生1个月内 ALP 水平较低,此后开始增高,1个月至11岁儿童 ALP 水平相对稳定且无性别差异。12岁后儿童 ALP 水平逐渐出现性别差异,12~14岁女孩 ALP 水平逐渐下降,而12~14岁男孩 ALP 水平高于同年龄段女孩。年龄、性别合并后的参考区间包括0~30 d、1~12月、1~10岁、11岁、12~14岁(男)、12岁(女)、13岁(女)及14岁(女)。结论建立儿童年龄、性别相关的血清 ALP 参考区间对儿童预防保健及疾病状况分析具有临床应用价值。%Objective To establish pediatric reference intervals of serum alkaline phosphatase (ALP) for healthy children with an age range of 0-14 years old in Changchun. Methods A total of 4 211 healthy children (2 090 males and 2 121 females) were enrolled in Changchun. ALP was performed on Hitachi 7600-210 automatic biochemical analyzer. After outlier data exclusion, data were estimated to or not to follow Gaussian distributions. One-way ANOVA was used to compare the differences of gender and age groups. The 2.5th and 97.5th percentiles of ALP were calculated by nonparametric method and 90%confidence intervals were computed by R language. Results The study showed there were apparent age and gender variations of the reference intervals for ALP. After a temporary low level in newborns, there was an increase from 1 month. The reference intervals of ALP were relatively stable and there was no significant gender

  11. Thermodynamics of a Ca(2+)-dependent highly thermostable alkaline protease from a haloalkliphilic actinomycete.

    Science.gov (United States)

    Gohel, S D; Singh, S P

    2015-01-01

    An alkaline protease from salt-tolerant alkaliphilic actinomycetes, Nocardiopsis alba OK-5 was purified by a single-step hydrophobic interaction chromatography and characterized. The purified protease with an estimated molecular mass of 20 kDa was optimally active at 70 °C in 0-3 M NaCl and 0-100 mM Ca(2+) displaying significant stability at 50-80 °C. The enzyme was stable at 80 °C in 100 mM Ca(2+) with Kd of 17 × 10(-3) and t1/2 of 32 min. The activation energy (Ea), enthalpy (ΔH*), and entropy (ΔS*) for the protease deactivation calculated in the presence of 200 mM Ca(2+) were 38.15 kJ/mol, 35.49 kJ/mol and 183.48 J/mol, respectively. The change in free energy (ΔG*) for protease deactivation at 60 °C in 200 mM Ca(2+) was 95.88 kJ/mol. Decrease in ΔH* reflected reduced cooperativity of deactivation and unfolding. The enzyme was intrinsically stable that counteracted heat denaturation by a weak cooperativity during the unfolding. Further, the enzyme was highly stable in the presence of various cations, surfactants, H2O2, β-mercaptoethanol, and commercial detergents. The compatibility of the enzyme with various cations, surfactants, and detergent matrices suggests its suitability as an additive in the detergents and peptide synthesis.

  12. Relação tireóide-gônadas e níveis plasmáticos de fósforo, cálcio e fosfatase alcalina em ratas Relationship between thyroid, gonads and plasmatic levels of phosphorus, calcium and alkaline phosphatase in rats

    Directory of Open Access Journals (Sweden)

    R. Serakides

    2000-12-01

    Full Text Available A relação tireóide-gônadas-metabolismo ósseo foi estudada em ratas Wistar adultas, castradas ou intactas e mantidas em estado hipertireóideo ou eutireóideo por períodos de 30, 60 e 90 dias. Foram utilizadas como características do metabolismo ósseo o cálcio, o fósforo e a atividade da fosfatase alcalina plasmáticos, correlacionando-os com os valores de estrógeno, de progesterona e de T4 livre. Verificou-se que o hipogonadismo e o hipertireoidismo alteram as características plasmáticas do metabolismo ósseo. O hipertireoidismo induz hiperfosfatemia e hipocalcemia, o hipogonadismo tem pouca influência sobre o fósforo, mas potencializa a hiperfosfatemia e a hipocalcemia desencadeadas pelo hipertireoidismo. Com relação à fosfatase alcalina, conclui-se que o hipertireoidismo reduz o efeito do hipogonadismo sobre a atividade dessa enzima.The interrelation between thyroid, gonads and osseous metabolism was studied in either intact or castrated adult female rats kept under hyperthyroidism or euthyroidism for 30, 60, or 90 days. Plasmatic levels of phosphorus, calcium, and alkaline phosphatase were measured to assess the osseous metabolism. These characteristics were correlated to the levels of estrogen, progesterone, and free T4. Either hypogonadism or hyperthyroidism interfered with the plasmatic characteristics of osseous metabolism. Hyperphosphatemia and hypocalcemia were induced by hyperthyroidism, whereas the hypogonadism had little effect on the levels of phosphorus, but it had a potencialization effect on the hyperphosphatemia and hypocalcemia induced by hyperthyroidism. The effect of hypogonadism on the alkaline phosphatase activity was reduced by the hyperthyroidism.

  13. 骨碱性磷酸酶检测技术诊治小儿佝偻病在浙江农村推广的适宜性评估%Evaluation of diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection in township-level health organizations of Zhejiang rural areas

    Institute of Scientific and Technical Information of China (English)

    朱雯; 朱炜; 杨再峰; 王红妹; 张扬; 沈毅; 李鲁; 姒健敏; 蒋健敏; 夏志俊; 蒋海瑛

    2009-01-01

    Objective: To evaluate the health technology of diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection in township-level health organizations of Zhejiang rural areas. Methods: The health technology was evaluated by the assessment model for selection and assessment of rural appropriate health technology in Zhejiang province. Results: The weighted score of first-level indicators were 8.05, 8.06, 6.32, 18.13, 18.88, 4.84 and 4.36 respectively. The total score was 68.64. Maturity: good; need: fair; convenience: good; safety: good; validity: very good; economic attributes: fair; sociality and ethicality: good. Conclusion: Diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection is suitable for dissemination and application in township-level health organizations in Zhejiang rural areas.%目的:对浙江省农村乡级卫生机构在小儿佝偻病诊治中应用骨碱性磷酸酶检测技术进行系统全面的评估.方法:使用"浙江省农村卫生适宜技术筛选评估指标体系"进行评估.结果:7个一级指标的加权分分别为8.05、8.06、6.32、18.13、18.88、4.84、4.36,总分为68.64;该技术成熟性较好;需要性一般;简便性较好;安全性较好;有效性很好;经济性一般;社会和伦理适应性较好.结论:该技术适宜在浙江省乡级农村卫生机构推广应用.

  14. Electromembrane recycling of highly mineralized alkaline blowdown water from evaporative water treatment plants at thermal power stations

    Science.gov (United States)

    Chichirova, N. D.; Chichirov, A. A.; Lyapin, A. I.; Minibaev, A. I.; Silov, I. Yu.; Tolmachev, L. I.

    2016-12-01

    Thermal power stations (TPS) are the main source of highly mineralized effluents affecting the environment. An analysis of their water systems demonstrates that alkaline effluents prevail at TPSs. Extraction of an alkali from highly mineralized effluents can make the recycling of effluents economically feasible. A method is proposed of electromembrane recycling of liquid alkaline highly mineralized wastes from TPSs. The process includes electromembrane apparatuses of two types, namely, a diffusion dialysis extractor (DDE) intended for extraction of the alkali from a highly mineralized solution having a complex composition and an electrodialysis concentrator for increasing the concentration of the extracted solution to a value suitable for use in water treatment plants at TPSs. For implementation of the first process (i.e. the extraction of alkali from alkaline-salt solution) various membranes from various manufacturers were studied: CM-PAD and AM-PAD (Ralex, Czechia), MK-40, MA-40, MA-41, MA-414, and MB-2 (OOO OKhK "Shchekinoazot", Russia), AR103-QDF and CR61-CMP (Ionies Inc., USA). The experiments demonstrate that the acceptable degree of separation of the alkali and the salt is achieved in a pair of cation-exchange membranes with the efficiency of separation being higher without an electric field. The highest efficiency was attained with Russian-made membranes (MK-40, OOO OKhK "Shchekinoazot"). A full scale experiment on recycling of highly-mineralized blowdown water from the evaporating water treatment system at the Kazan cogeneration power station No. 3 (TETs-3) was performed in a pilot unit consisting of two electromembrane apparatuses made by UAB "Membraninės Technologijos LT". In the experiments every ton of blowdown water yielded 0.1 t of concentrated alkaline solution with an alkali content of up to 4 wt % and 0.9 t of the softened salt solution suitable for the reuse in the TPS cycle. The power rate is 6 kWh / ton of blowdown water.

  15. Phage & phosphatase: a novel phage-based probe for rapid, multi-platform detection of bacteria.

    Science.gov (United States)

    Alcaine, S D; Pacitto, D; Sela, D A; Nugen, S R

    2015-11-21

    Genetic engineering of bacteriophages allows for the development of rapid, highly specific, and easily manufactured probes for the detection of bacterial pathogens. A challenge for novel probes is the ease of their adoption in real world laboratories. We have engineered the bacteriophage T7, which targets Escherichia coli, to carry the alkaline phosphatase gene, phoA. This inclusion results in phoA overexpression following phage infection of E. coli. Alkaline phosphatase is commonly used in a wide range of diagnostics, and thus a signal produced by our phage-based probe could be detected using common laboratory equipment. Our work demonstrates the successful: (i) modification of T7 phage to carry phoA; (ii) overexpression of alkaline phosphatase in E. coli; and (iii) detection of this T7-induced alkaline phosphatase activity using commercially available colorimetric and chemilumiscent methods. Furthermore, we demonstrate the application of our phage-based probe to rapidly detect low levels of bacteria and discern the antibiotic resistance of E. coli isolates. Using our bioengineered phage-based probe we were able to detect 10(3) CFU per mL of E. coli in 6 hours using a chemiluminescent substrate and 10(4) CFU per mL within 7.5 hours using a colorimetric substrate. We also show the application of this phage-based probe for antibiotic resistance testing. We were able to determine whether an E. coli isolate was resistant to ampicillin within 4.5 hours using chemiluminescent substrate and within 6 hours using a colorimetric substrate. This phage-based scheme could be readily adopted in labs without significant capital investments and can be translated to other phage-bacteria pairs for further detection.

  16. Marked Transient Alkaline Phosphatemia Following Pediatric Liver Transplantation

    Science.gov (United States)

    Koneru, Baburao; Carone, Eduardo; Malatack, J. Jeffrey; Esquivel, Carlos O.; Starzl, Thomas E.

    2010-01-01

    An isolated marked transient rise in serum alkaline phosphatase levels in otherwise healthy children is a well-documented occurrence. However, in children undergoing liver transplantation, elevated alkaline phosphatase values raise the possibility of biliary obstruction, rejection, or both. During a 6-year period, 6 of 278 children undergoing liver transplantation exhibited a similar phenomenon as an isolated abnormality. None had rejection, biliary obstruction, or other allograft dysfunction during a long follow-up. Eventually and without intervention, the alkaline phosphatase levels returned to normal. These instructive cases suggest that caution be used in advocating Invasive procedures if elevated alkaline phosphatase levels are an isolated abnormality, and close observation with noninvasive testing is recommended. PMID:2658549

  17. Structural Genomics of Protein Phosphatases

    Energy Technology Data Exchange (ETDEWEB)

    Almo,S.; Bonanno, J.; Sauder, J.; Emtage, S.; Dilorenzo, T.; Malashkevich, V.; Wasserman, S.; Swaminathan, S.; Eswaramoorthy, S.; et al

    2007-01-01

    The New York SGX Research Center for Structural Genomics (NYSGXRC) of the NIGMS Protein Structure Initiative (PSI) has applied its high-throughput X-ray crystallographic structure determination platform to systematic studies of all human protein phosphatases and protein phosphatases from biomedically-relevant pathogens. To date, the NYSGXRC has determined structures of 21 distinct protein phosphatases: 14 from human, 2 from mouse, 2 from the pathogen Toxoplasma gondii, 1 from Trypanosoma brucei, the parasite responsible for African sleeping sickness, and 2 from the principal mosquito vector of malaria in Africa, Anopheles gambiae. These structures provide insights into both normal and pathophysiologic processes, including transcriptional regulation, regulation of major signaling pathways, neural development, and type 1 diabetes. In conjunction with the contributions of other international structural genomics consortia, these efforts promise to provide an unprecedented database and materials repository for structure-guided experimental and computational discovery of inhibitors for all classes of protein phosphatases.

  18. Early containment of high-alkaline solution simulating low-level radioactive waste stream in clay-bearing blended cement

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A.A. [Westinghouse Hanford Co., Richland, WA (United States); Olson, R.A.; Tennis, P.D. [Northwestern Univ., Evanston, IL (United States). Center for Advanced Cement-Based Materials] [and others

    1995-04-01

    Portland cement blended with fly ash and attapulgite clay was mixed with high-alkaline solution simulating low-level radioactive waste stream at a one-to-one weight ratio. Mixtures were adiabatically and isothermally cured at various temperatures and analyzed for phase composition, total alkalinity, pore solution chemistry, and transport properties as measured by impedance spectroscopy. Total alkalinity is characterized by two main drops. The early one corresponds to a rapid removal of phosphorous, aluminum, sodium, and to a lesser extent potassium solution. The second drop from about 10 h to 3 days is mainly associated with the removal of aluminum, silicon, and sodium. Thereafter, the total alkalinity continues descending, but at a lower rate. All pastes display a rapid flow loss that is attributed to an early precipitation of hydrated products. Hemicarbonate appears as early as one hour after mixing and is probably followed by apatite precipitation. However, the former is unstable and decomposes at a rate that is inversely related to the curing temperature. At high temperatures, zeolite appears at about 10 h after mixing. At 30 days, the stabilized crystalline composition Includes zeolite, apatite and other minor amounts of CaCO{sub 3}, quartz, and monosulfate Impedance spectra conform with the chemical and mineralogical data. The normalized conductivity of the pastes shows an early drop, which is followed by a main decrease from about 12 h to three days. At three days, the permeability of the cement-based waste as calculated by Katz-Thompson equation is over three orders of magnitude lower than that of ordinary portland cement paste. However, a further decrease in the calculated permeability is questionable. Chemical stabilization is favorable through incorporation of waste species into apatite and zeolite.

  19. High-Throughput Spheroid Screens Using Volume, Resazurin Reduction, and Acid Phosphatase Activity.

    Science.gov (United States)

    Ivanov, Delyan P; Grabowska, Anna M; Garnett, Martin C

    2017-01-01

    Mainstream adoption of physiologically relevant three-dimensional models has been slow in the last 50 years due to long, manual protocols with poor reproducibility, high price, and closed commercial platforms. This chapter describes high-throughput, low-cost, open methods for spheroid viability assessment which use readily available reagents and open-source software to analyze spheroid volume, metabolism, and enzymatic activity. We provide two ImageJ macros for automated spheroid size determination-for both single images and images in stacks. We also share an Excel template spreadsheet allowing users to rapidly process spheroid size data, analyze plate uniformity (such as edge effects and systematic seeding errors), detect outliers, and calculate dose-response. The methods would be useful to researchers in preclinical and translational research planning to move away from simplistic monolayer studies and explore 3D spheroid screens for drug safety and efficacy without substantial investment in money or time.

  20. Combination of Successive Alkalinity Producing System (SAPS) and Aeration for Passive Treatment of Highly Acidic Mine Drainage

    Science.gov (United States)

    Oh, C.; Ji, S.

    2015-12-01

    Passive treatment system has been widely used for remediation of mine drainage since its advantage of low installation and maintenance cost. The system, however, has also a disadvantage in assuring remediation and management efficiency if the drainage is highly acidic mine drainage. To remediate acid mine drainage (AMD) especially showing high acidity, passive treatment system which consists of successive alkalinity producing system (SAPS) and subsequent aeration pond was proposed and its mechanisms and efficiency was evaluated in this research. Target AMD was obtained from Waryong coal mine and showed typical characteristics of AMD having high metal concentration and low pH (acidity > 300 mg/L as CaCO3). Four experimental cases were conducted; untreated, treated with SAPS, treated with aeration, treated with SAPS and aeration to compare role and mechanism of each unit. Between organic matter and limestone layer which constitute SAPS, the former eliminated most of Fe(III) and Al in the AMD so that the latter was kept from being clogged by precipitates. Net acidity of the AMD rapidly decreased by supplement of alkalinity at the limestone layer. A primary function of SAPS, producing alkalinity constantly without clogging, was attained due to addition a portion of limestone particle into the organic matter layer. The discharge from SAPS had low ORP and DO values because of an anaerobic environment formed at the organic matter layer although its alkalinity was increased. This water quality was unfavorable for Fe(II) to be oxidized. Installation of aeration pond after SAPS, therefore, could be effective way of enhancing oxidation rate of Fe(II). Among the experimental cases, the combination of SAPS and aeration pond was only able to remediate the AMD. This concluded that to remediate highly acidic mine drainage with passive treatment system, three critical conditions were required; pre-precipitation of Fe(III) and Al at organic matter layer in SAPS, constant alkalinity

  1. Synthesis of Highly Selective Submicromolar Microcystin‐Based Inhibitors of Protein Phosphatase (PP)2A over PP1

    Science.gov (United States)

    Fontanillo, Miriam; Zemskov, Ivan; Häfner, Maximilian; Uhrig, Ulrike; Salvi, Francesca; Simon, Bernd; Wittmann, Valentin

    2016-01-01

    Abstract Research and therapeutic targeting of the phosphoserine/threonine phosphatases PP1 and PP2A is hindered by the lack of selective inhibitors. The microcystin (MC) natural toxins target both phosphatases with equal potency, and their complex synthesis has complicated structure–activity relationship studies in the past. We report herein the synthesis and biochemical evaluation of 11 MC analogues, which was accomplished through an efficient strategy combining solid‐ and solution‐phase approaches. Our approach led to the first MC analogue with submicromolar inhibitory potency that is strongly selective for PP2A over PP1 and does not require the complex lipophilic Adda group. Through mutational and structural analyses, we identified a new key element for binding, as well as reasons for the selectivity. This work gives unprecedented insight into how selectivity between these phosphatases can be achieved with MC analogues. PMID:27723199

  2. Production and Characterization of Alkaline Protease from a High Yielding and Moderately Halophilic Strain of SD11 Marine Bacteria

    Directory of Open Access Journals (Sweden)

    Hongxia Cui

    2015-01-01

    Full Text Available A marine bacterium SD11, which was isolated from sea muds (Geziwo Qinhuangdao Sea area, China, was used to produce thermostable alkaline serine nonmetal protease in the skim milk agar plate medium with 10% NaCl. The optimal temperature about the manufacture of the extracellular protease was ~60°C. The crude enzyme was stable at 20–50°C. The activity was retained to 60% and 45% after heating for 1 h at 60 and 70°C, respectively. The protease was highly active in a wide pH scope (8.0–10.0 and maximum protease activity exhibited at pH 10.0. The activity was restrained by phenylmethylsulfonyl fluoride (PMSF but mildly increased (~107% in the presence of ethylenediaminetetraacetic acid (EDTA, indicating that the production contains serine-protease(s and nonmetal protease(s. Moreover, the crude alkaline protease was active with the 5 mM Ca2+, Mn2+, Zn2+, Cu2+, Na+, and K+ that existed separately. In addition, the protease showed superduper stability when exposed to an anionic surfactant (5 mM SDS, an oxidizing agent (1% H2O2, and several organic solvents (methanol, isopropanol, and acetone. These results suggest that the marine bacterium SD11 is significant in the industry from the prospects of its ability to produce thermally stable alkaline protease.

  3. Validation of a high-throughput in vitro alkaline elution/rat hepatocyte assay for DNA damage.

    Science.gov (United States)

    Gealy, Robert; Wright-Bourque, Jennifer L; Kraynak, Andrew R; McKelvey, Troy W; Barnum, John E; Storer, Richard D

    2007-04-20

    In vitro alkaline elution is a sensitive and specific short term assay which measures DNA strand breakage in a mammalian test system (primary rat hepatocytes). This lab has previously demonstrated the performance of the assay with known genotoxic and non-genotoxic compounds. The methodology employed has relatively low sample throughput and is labor-intensive, requiring a great deal of manual processing of samples in a format that is not amenable to automation. Here, we present an automated version of the assay. This high-throughput alkaline elution assay (HT-AE) was made possible through 3 key developments: (1) DNA quantitation using PicoGreen and OliGreen fluorescent DNA binding dyes; (2) design and implementation of a custom automation system; and (3) reducing the assay to a 96-well plate format. The assay can now be run with 5-50mg of test compound. HT-AE was validated in a similar manner as the original assay, including assessment of non-genotoxic and non-carcinogenic compounds and evaluation of cytotoxicity to avoid confounding effects of toxicity-associated DNA degradation. The validation test results from compounds of known genotoxic potential were used to set appropriate criteria to classify alkaline elution results for genotoxicity.

  4. A facile synthesis of highly stable multiblock poly(arylene ether)s based alkaline membranes for fuel cells

    Science.gov (United States)

    Jasti, Amaranadh; Shahi, Vinod K.

    2014-12-01

    Herein, we are disclosing simple route for the preparation of alkaline membranes (AMs) based on aminated multiblock poly(arylene ether)s (AMPEs) synthesized by nucleophilic substitution-poly condensation followed by quaternization and alkalization reactions. In this procedure, four quaternary ammonium groups are successfully introduced without use of carcinogenic reagents such as chloromethylmethyl ether (CMME). Hydrophilic/hydrophobic phase separation is responsible for their high hydroxide conductivity (∼150 mS cm-1 at 80 °C) due to development of interconnected ion transport pathway. AMs are exhibiting good alkaline stability due to the presence of two vicinal quaternary ammonium groups and avoid degradation such as Sommelet-Hauser rearrangement and Hofmann elimination. Vicinal quaternary ammonium groups also resist nucleophilic (OH-) attack and suppress the Stevens rearrangement as well as SN2 substitution reaction due to stearic hindrance. Optimized AM (AMPE-M20N15 (55% DCM)) exhibits about 0.95 V open circuit voltage (OCV) and 48.8 mW cm-2 power density at 65 °C in alkaline direct methanol fuel cell (ADMFC) operation. These results suggest promising begin for the preparation of stable and conductive AMs for ADMFC applications and useful for developing hydroxide conductive materials.

  5. Facile Alkaline Lysis of Escherichia coli Cells in High-Throughput Mode for Screening Enzyme Mutants: Arylsulfatase as an Example.

    Science.gov (United States)

    Yuan, Mei; Yang, Xiaolan; Li, Yuwei; Liu, Hongbo; Pu, Jun; Zhan, Chang-Guo; Liao, Fei

    2016-06-01

    Facile alkaline lysis of Escherichia coli cells in high-throughput (HTP) mode for screening enzyme mutants was tested with Pseudomonas aeruginosa arylsulfatase (PAAS). The alkaline lysis buffer was 1.0 M Tris-HCl at pH 9.0 plus 0.1 % Tween-20 and 2.0 mM 4-aminobenzamidine, mixed with cell suspension at 8:1 to 12:1 ratio for continuous agitation of mixtures in 96-well plates under room temperature; enzymatic activity in lysates was measured with 96-well microplate. PAAS activity tolerated final 0.1 % Tween-20. Individual clones were amplified for 12 h in 0.50 mL TB medium with 48-well plates to enhance the repeatability of induced expression. During continuous agitation of the mixture of cells and the lysis buffer, PAAS activities in lysates were steady from 3 to 9 h and comparable to sonication treatment but better than freezing-thawing. Coefficients of variation of activities of PAAS/mutants in lysates after treatment for 7 h reached ∼22 %. The mutant M72Q had specific activity 2-fold of G138S. By HTP lysis of cells, M72Q was recognized as a positive mutant over G138S with the area under the curve of 0.873. Therefore, for enzymes tolerating concentrated alkaline buffers, the proposed alkaline lysis approach may be generally applicable for HTP lysis of host cells during directed evolution.

  6. B56alpha/protein phosphatase 2A inhibits adipose lipolysis in high-fat diet-induced obese mice.

    Science.gov (United States)

    Kinney, Brice P; Qiao, Liping; Levaugh, Justin M; Shao, Jianhua

    2010-08-01

    Lipolysis and lipogenesis are two opposite processes that control lipid storage in adipocytes. Impaired adipose lipolysis has been observed in both obese human subjects and animal models. This study investigated the mechanisms underlying impaired adipose lipolysis in a high-fat diet-induced obese (DIO) mouse model. DIO models were created using male C57BL/6 mice. Our results show that beta3 adrenergic receptor-specific agonist BRL37344 induced adipose lipolysis was significantly blunted in DIO mice. The levels of Ser660 phosphorylation of hormone-sensitive lipase (HSL) were significantly decreased in the epididymal fat of DIO mice. However, protein levels of HSL, adipose triglyceride lipase and its coactivator comparative gene identification-58 were similar between DIO and control mice. It is known that upon lipolytic hormone stimulation, protein kinase A phosphorylates HSL Ser660 and activates HSL, whereas protein phosphatase 2A (PP2A) dephosphorylates and inactivates HSL. Interestingly, our study shows that high-fat feeding did not alter epididymal fat cAMP and protein kinase A protein levels but significantly increased the expression of the alpha-isoform of PP2A regulatory subunit B' (B56alpha). To study the role of B56alpha in obesity-associated lipolytic defect, B56alpha was overexpressed or knocked down by adenovirus-mediated gene transduction in cultured 3T3-L1CARDelta1 adipocytes. Overexpression of B56alpha significantly decreased HSL Ser660 phosphorylation. In contrast, knocking down B56alpha increased hormone-stimulated HSL activation and lipolysis in mature 3T3-L1CARDelta1 adipocytes. These results strongly suggest that elevated B56alpha/PP2A inhibits HSL and lipolysis in white adipose tissue of DIO mice.

  7. High Uric Acid (UA Negatively Affects Serum Tartrate-Resistant Acid Phosphatase 5b (TRACP 5b Immunoassay.

    Directory of Open Access Journals (Sweden)

    Zhi-Qi Wu

    Full Text Available Bone metastases often occur in the majority of patients with advanced cancer, such as prostate cancer, lung cancer and breast cancer. Serum tartrate-resistant acid phosphatase 5b (TRACP 5b, a novel bone resorption marker, has been used gradually in the clinics as a specific and sensitive marker of bone resorption for the early diagnosis of cancer patients with bone metastasis. Here, we reported that high concentrations of uric acid (UA lead to decrease of TRACP 5b levels and determined whether TRACP 5b level was associated with UA in interference experiment.A total of 77 patients with high concentrations of UA and 77 healthy subjects were tested to evaluate the differences in their TRACP 5b levels. Serial dilutions of UA were respectively spiked with a known concentration of TRACP 5b standard sample, then Serum TRACP 5b was detected by using bone TRAP® Assay. A correction equation was set to eliminate UA-derived TRACP 5b false-decrease. The effect of this correction was evaluated in high-UA individuals.The average TRACP level of the high-UA individuals (1.47 ± 0.62 U/L was significantly lower than that of the healthy subjects (2.62 ± 0.63 U/L (t-test, p < 0.0001. The UA correction equation derived: ΔTRACP 5b = -1.9751lgΔUA + 3.7365 with an R2 = 0.98899. Application of the UA correction equation resulted in a statistically non-significant difference in TRACP 5b values between the healthy subjects and high-UA individuals (p = 0.24.High UA concentrations can falsely decrease TRACP 5b levels due to a method-related systematic error. To avoid misdiagnoses or inappropriate therapeutic decisions, increased attention should be paid to UA interference, when TRACP 5b is used for early diagnosis of cancer patients with bone metastasis, evaluation of the aggressiveness of osteosarcoma or prediction of survival in prostate cancer and breast cancer with bone metastases.

  8. Phosphatase activity in the rhizosphere and root of mycorrhizal teak seedlings with three levels of NPK fertilization

    Directory of Open Access Journals (Sweden)

    CORRYANTI

    2007-07-01

    Full Text Available To examine the phosphatase alkaline activity of VA mycorrhizal fungi in the rizhosphere and in root, teak seedlings inoculated spores of VA mycorrhizal fungi were grown in sterilized soils. Teak seedlings were fertilized with NPK fertilizer consisting three levels, i.e. 0; 0.0625; 0.125 g per seedling. Phosphatase alkaline in rizhosphere was measured in terms of pNP on soil dry weight basis, meanwhile alkaline phosphatase activity in roots were quantified in using method developed by Tisserant. The results showed that alkaline phosphatase activity increased on inoculated seedlings compare to with uninoculated. NPK fertilization of 0.0625 g per seedling and inoculation on teak seedlings showed alkaline phosphatase activity in range 90-201 EU, and in roots indicated in range 14-72%. Gigaspora sp inoculation on teak seedlings was showing the highest of alkaline phosphatase activity. Increasing phosphatase alkaline activity relevant to hyphae growth, and increasing of root infection decreased alkaline phosphatase activity. Arbuscular mycorrhizal inoculation increased seedling dry weight.

  9. High power density alkaline fuel cell technology for MMW space burst power

    Science.gov (United States)

    Preston, J. Lawrence, Jr.; Trocciola, John C.; Wertheim, Ronald J.

    The use of advanced alkaline regenerative fuel cell energy storage systems to provide 10's to 100's of MWe of sprint (burst) power for 100's of seconds per orbit of SDI weapons platform was studied. Recharge power is supplied by a multimegawatt space based nuclear power system. Regenerative fuel cell energy storage systems offer the potential for significant platform mass reduction by reducing the size and mass of the nuclear power source required. This is because the reactor can be sized for the smaller average power level for the energy storage system, rather than the sprint power level. The regenerative fuel cell is a particularly attractive energy storage device because the fuel cell is essentially a static power conversion device, which results in excellent platform stability for weapon pointing and tracking. Based upon the detailed point design and conceptual layout, the alkaline regenerative fuel cell energy storage system is an attractive choice for integration with a nuclear thermionic system for providing multimegawatt burst power and multi orbit capability.

  10. Bioprecipitation of uranium from alkaline waste solutions using recombinant Deinococcus radiodurans

    Energy Technology Data Exchange (ETDEWEB)

    Kulkarni, Sayali; Ballal, Anand; Apte, Shree Kumar, E-mail: aptesk@barc.gov.in

    2013-11-15

    Highlights: • Deinococcus radiodurans was genetically engineered to overexpress alkaline phosphatase (PhoK). • Deino-PhoK bioprecipitated U efficiently over a wide range of input U concentration. • A maximal loading of 10.7 g U/g of biomass at 10 mM input U was observed. • Radioresistance and U precipitation by Deino-PhoK remained unaffected by γ radiation. • Immobilization of Deino-PhoK facilitated easy separation of precipitated U. -- Abstract: Bioremediation of uranium (U) from alkaline waste solutions remains inadequately explored. We engineered the phoK gene (encoding a novel alkaline phosphatase, PhoK) from Sphingomonas sp. for overexpression in the radioresistant bacterium Deinococcus radiodurans. The recombinant strain thus obtained (Deino-PhoK) exhibited remarkably high alkaline phosphatase activity as evidenced by zymographic and enzyme activity assays. Deino-PhoK cells could efficiently precipitate uranium over a wide range of input U concentrations. At low uranyl concentrations (1 mM), the strain precipitated >90% of uranium within 2 h while a high loading capacity of around 10.7 g U/g of dry weight of cells was achieved at 10 mM U concentration. Uranium bioprecipitation by Deino-PhoK cells was not affected in the presence of Cs and Sr, commonly present in intermediate and low level liquid radioactive waste, or after exposure to very high doses of ionizing radiation. Transmission electron micrographs revealed the extracellular nature of bioprecipitated U, while X-ray diffraction and fluorescence analysis identified the precipitated uranyl phosphate species as chernikovite. When immobilized into calcium alginate beads, Deino-PhoK cells efficiently removed uranium, which remained trapped in beads, thus accomplishing physical separation of precipitated uranyl phosphate from solutions. The data demonstrate superior ability of Deino-PhoK, over earlier reported strains, in removal of uranium from alkaline solutions and its potential use in

  11. Changes in the bacterial populations of the highly alkaline saline soil of the former lake Texcoco (Mexico) following flooding.

    Science.gov (United States)

    Valenzuela-Encinas, César; Neria-González, Isabel; Alcántara-Hernández, Rocio J; Estrada-Alvarado, Isabel; Zavala-Díaz de la Serna, Francisco Javier; Dendooven, Luc; Marsch, Rodolfo

    2009-07-01

    Flooding an extreme alkaline-saline soil decreased alkalinity and salinity, which will change the bacterial populations. Bacterial 16S rDNA libraries were generated of three soils with different electrolytic conductivity (EC), i.e. soil with EC 1.7 dS m(-1) and pH 7.80 (LOW soil), with EC 56 dS m(-1) and pH 10.11 (MEDIUM soil) and with EC 159 dS m(-1) and pH 10.02 (HIGH soil), using universal bacterial oligonucleotide primers, and 463 clone 16S rDNA sequences were analyzed phylogenetically. Library proportions and clone identification of the phyla Proteobacteria, Actinobacteria, Acidobacteria, Cyanobacteria, Bacteroidetes, Firmicutes and Cloroflexi showed that the bacterial communities were different. Species and genera of the Rhizobiales, Rhodobacterales and Xanthomonadales orders of the alpha- and gamma-subdivision of Proteobacteria were found at the three sites. Species and genera of the Rhodospirillales, Sphingobacteriales, Clostridiales, Oscillatoriales and Caldilineales were found only in the HIGH soil, Sphingomonadales, Burkholderiales and Pseudomonadales in the MEDIUM soil, Myxococcales in the LOW soil, and Actinomycetales in the MEDIUM and LOW soils. It was found that the largest diversity at the order and species level was found in the MEDIUM soil as bacteria of both the HIGH and LOW soils were found in it.

  12. Ultra-high performance supercritical fluid chromatography of lignin-derived phenols from alkaline cupric oxide oxidation.

    Science.gov (United States)

    Sun, Mingzhe; Lidén, Gunnar; Sandahl, Margareta; Turner, Charlotta

    2016-08-01

    Traditional chromatographic methods for the analysis of lignin-derived phenolic compounds in environmental samples are generally time consuming. In this work, an ultra-high performance supercritical fluid chromatography method with a diode array detector for the analysis of major lignin-derived phenolic compounds produced by alkaline cupric oxide oxidation was developed. In an analysis of a collection of 11 representative monomeric lignin phenolic compounds, all compounds were clearly separated within 6 min with excellent peak shapes, with a limit of detection of 0.5-2.5 μM, a limit of quantification of 2.5-5.0 μM, and a dynamic range of 5.0-2.0 mM (R(2) > 0.997). The new ultra-high performance supercritical fluid chromatography method was also applied for the qualitative and quantitative analysis of lignin-derived phenolic compounds obtained upon alkaline cupric oxide oxidation of a commercial humic acid. Ten out of the previous eleven model compounds could be quantified in the oxidized humic acid sample. The high separation power and short analysis time obtained demonstrate for the first time that supercritical fluid chromatography is a fast and reliable technique for the analysis of lignin-derived phenols in complex environmental samples.

  13. Highly Dispersed Palladium Nanoparticles on Functional MWNT Surfaces for Methanol Oxidation in Alkaline Solutions

    Institute of Scientific and Technical Information of China (English)

    WANG zhe; ZHU Zan-Zan; LI You-Xiang; LI Hu-Lin

    2008-01-01

    Palladium nanoparticles were crystallized on 4-aminobenzoic acid monolayer-grafted multi-walled carbon nanotubes (MWNT) by diazotization. The structure and nature of the resulting Pd/MWNT composite were characterized by transmission electron microscopy and X-ray diffraction, the results show that the chemically synthesized Pd nanoparticles were homogeneously dispersed and well-separated from one another on the modified MWNT surfaces. Cyclic voltammogram showed that the Pd-MWNT composite materials performed higher electrocatalytic activity and better long-term stability toward methanol oxidation in alkaline solution than Pd-C. The results imply that the Pd-MWNT composite materials as a promising support material improve the excellent electrocatalytic activity for methanol oxidation greatly. So the Pd/MWNT composites have a good application potential to fuel cells.

  14. A binary palladium-bismuth nanocatalyst with high activity and stability for alkaline glucose electrooxidation

    Science.gov (United States)

    Chen, Cheng-Chuan; Lin, Cheng-Lan; Chen, Lin-Chi

    2015-08-01

    Binary palladium-bismuth nanocatalysts supported on functionalized multi-walled carbon nanotubes (Pd-Bi/C) are synthesized using a one-pot polyol method. The prepared Pd-Bi/C catalysts have a metal particle range from 5.25 to 12.98 nm and are investigated for alkaline electrocatalytic glucose oxidation reaction (GOR). The physical properties of the catalysts are characterized by X-ray diffraction (XRD), scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The electrochemical activities are determined by cyclic voltammetry (CV), linear sweep voltammetry (LSV), Tafel analysis and chronoamperomtry (CA) for comparing the electrochemical active surface area (ECSA), GOR onset potential, GOR peak current density, Tafel slope, poisoning rate and cycling stability of the Pd-Bi/C catalysts. It is found that Pd-Bi/C (1:0.14) can significantly enhance the electrocatalytic activity on GOR about 40% times higher than Pd/C and as well as has a 3.7-fold lower poisoning rate. The in-use stability of Pd-Bi/C (1:0.14) is also remarkably improved, according to the results of the 200 cycling CV test. The effects of the operating temperature and the concentration of glucose and NaOH electrolyte on Pd-Bi/C (1:0.14) are further studied in this work. The highest Pd-Bi/C catalyzed GOR current density of 29.5 mA cm-2 is attained in alkaline medium.

  15. Chronic high glucose inhibits albumin reabsorption by lysosomal alkalinization in cultured porcine proximal tubular epithelial cells (LLC-PK1).

    Science.gov (United States)

    Ishibashi, Fukashi

    2006-06-01

    Lysosomal acidification is a key step of albumin reabsorption in proximal tubular epithelial cells (PTECs). This study was performed to examine the influence of chronic high glucose on lysosomal acidification in cultured PTECs. Porcine PTECs (LLC-PK(1) cells) were cultured in 16.7 mM (300 mg/dl) glucose (HG) alone or with 0.5 mM phlorizin for 24 weeks and subsequently for 12 weeks in 5.5 mM (100 mg/dl) glucose (NG). Chronic HG inhibited the fluorescein isothiocyanate (FITC)-albumin (A) uptake progressively, while phlorizin reversed the inhibition. NG for 12 weeks after HG normalized the uptake. The time-dependent uptake of FITC-A was inhibited by HG and bafilomycin A(1) (BafA(1)) after 15 min and by 4,4'-diisothiocyanato-2,2'-disulfonic acid (DIDS) and N-ethyl-N-isopropyl-amiloride (EIPA) after 3 min. Cellular ATP was depleted by HG and restored by NG. Lysosomal pH, assessed by an acidotropic fluorescent probe, was alkalinized (pH 4.5-7.8) with 5.5-27.8 mM glucose and normalized by subsequent NG. BafA(1) alkalinized lysosomes, and the concentration required to 50% change for the pH and 50% inhibition of FITC-A uptake was similar. EIPA inhibited FITC-A uptake, but did not influence lysosomal pH. DIDS inhibited FITC-A uptake, and unexpectedly lowered lysosomal pH. Real time PCR showed that HG reduced the mRNA level for vacuolar H(+)-ATPase, but did not alter those of chloride channel-5 and Na(+)-H(+)-exchanger-3. In conclusion, the chronic HG inhibits albumin reabsorption by lysosomal alkalinization in PTECs, probably due to ATP depletion and down-regulation of vacuolar H(+)-ATPase.

  16. Microbial impacts on (99m)Tc migration through sandstone under highly alkaline conditions relevant to radioactive waste disposal.

    Science.gov (United States)

    Smith, Sarah L; Boothman, Christopher; Williams, Heather A; Ellis, Beverly L; Wragg, Joanna; West, Julia M; Lloyd, Jonathan R

    2017-01-01

    Geological disposal of intermediate level radioactive waste in the UK is planned to involve the use of cementitious materials, facilitating the formation of an alkali-disturbed zone within the host rock. The biogeochemical processes that will occur in this environment, and the extent to which they will impact on radionuclide migration, are currently poorly understood. This study investigates the impact of biogeochemical processes on the mobility of the radionuclide technetium, in column experiments designed to be representative of aspects of the alkali-disturbed zone. Results indicate that microbial processes were capable of inhibiting (99m)Tc migration through columns, and X-ray radiography demonstrated that extensive physical changes had occurred to the material within columns where microbiological activity had been stimulated. The utilisation of organic acids under highly alkaline conditions, generating H2 and CO2, may represent a mechanism by which microbial processes may alter the hydraulic conductivity of a geological environment. Column sediments were dominated by obligately alkaliphilic H2-oxidising bacteria, suggesting that the enrichment of these bacteria may have occurred as a result of H2 generation during organic acid metabolism. The results from these experiments show that microorganisms are able to carry out a number of processes under highly alkaline conditions that could potentially impact on the properties of the host rock surrounding a geological disposal facility for intermediate level radioactive waste. Copyright © 2016. Published by Elsevier B.V.

  17. Optimized coagulation of high alkalinity, low temperature and particle water: pH adjustment and polyelectrolytes as coagulant aids.

    Science.gov (United States)

    Yu, Jianfeng; Wang, Dongsheng; Yan, Mingquan; Ye, Changqing; Yang, Min; Ge, Xiaopeng

    2007-08-01

    The Yellow River in winter as source water is characterized as high alkalinity, low temperature and low particle concentrations, which have brought many difficulties to water treatment plants. This study fully examines the optimized coagulation process of the Yellow River by conventional and pre-polymerized metal coagulants, pH adjustment and polyelectrolytes as the primary coagulants or coagulant aids. For all the metal coagulants, polyaluminum chlorides are superior to traditional metal coagulants due to their stable polymeric species and low consumption of alkalinity. The removal of natural organic matter by monomeric metal coagulants can be improved through pH adjustment, which is in accordance with the higher concentration of polymeric species formed at corresponding pH value. With the addition of polyelectrolytes as coagulant aids, the coagulation performance is significantly improved. The effective removal of dissolved organic matter is consistent with high charge density, while molecular weight is relatively important for removing particles, which is consistent with polyelectrolytes as primary coagulants. These results suggest that the coagulation mechanisms in the removal of dissolved organic matter and particles are different, which may be exploited for optimized coagulation for the typical source water in practice.

  18. Effects of Metal Ions and Organic Solvents on Alkaline Phosphatase from Rice-field Eel%金属离子·有机溶剂对黄鳝碱性磷酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    黄毅; 唐云明

    2008-01-01

    [Objective]The mechanism of alkaline phcephatase(ALP) was studied to promote rice-field eel aquaculture industry. [Method] The effects of effectors such as multiple metal ions and organic solvents on ALP in viscera of rice-field eel. [ Result] Na+ and K+ didn't generate big influences on en-zyme activity ;Mg2+ and Ca2+ could promote ALP while Li+,Cu2+ and Zn2+ could restrain ALP enzyme activity. Both KPO2-4 and WO3-4 generated by en-zyme catalyzing disodium phenyl phosphate possessed strong inhibitory effects on emzyme, and 9.5 mmol/L HPO2-4 would make enzyme activity decline by 13% while 9.5 mmol/L WO3-4 would make enzyme decline by 34%. The inhibition types of them were both competitive inhibition on enzyme activity. The organic solvents such as methanol,ethanol,ethylene glycol,isopropanol all generated influences on ALP and the order according to their inhibitory effects was isopropanol > ethanol > methanol > ethylene glycol. [Conclusion] The influences of various effectors on ALP activity of rice-field eel were studied from dy-namics perspective to provide theoretical basis for further clarifying ALP mechanism.

  19. Insights from the genome of a high alkaline cellulase producing Aspergillus fumigatus strain obtained from Peruvian Amazon rainforest.

    Science.gov (United States)

    Paul, Sujay; Zhang, Angel; Ludeña, Yvette; Villena, Gretty K; Yu, Fengan; Sherman, David H; Gutiérrez-Correa, Marcel

    2017-06-10

    Here, we report the complete genome sequence of a high alkaline cellulase producing Aspergillus fumigatus strain LMB-35Aa isolated from soil of Peruvian Amazon rainforest. The genome is ∼27.5mb in size, comprises of 228 scaffolds with an average GC content of 50%, and is predicted to contain a total of 8660 protein-coding genes. Of which, 6156 are with known function; it codes for 607 putative CAZymes families potentially involved in carbohydrate metabolism. Several important cellulose degrading genes, such as endoglucanase A, endoglucanase B, endoglucanase D and beta-glucosidase, are also identified. The genome of A. fumigatus strain LMB-35Aa represents the first whole sequenced genome of non-clinical, high cellulase producing A. fumigatus strain isolated from forest soil. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Biomineralization of Uranium by PhoY Phosphatase Activity Aids Cell Survival in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Yung, M C [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Jiao, Y [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2014-07-22

    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  1. Biomineralization of uranium by PhoY phosphatase activity aids cell survival in Caulobacter crescentus.

    Science.gov (United States)

    Yung, Mimi C; Jiao, Yongqin

    2014-08-01

    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  2. Novel Glucose-1-Phosphatase with High Phytase Activity and Unusual Metal Ion Activation from Soil Bacterium Pantoea sp. Strain 3.5.1.

    Science.gov (United States)

    Suleimanova, Aliya D; Beinhauer, Astrid; Valeeva, Liia R; Chastukhina, Inna B; Balaban, Nelly P; Shakirov, Eugene V; Greiner, Ralf; Sharipova, Margarita R

    2015-10-01

    Phosphorus is an important macronutrient, but its availability in soil is limited. Many soil microorganisms improve the bioavailability of phosphate by releasing it from various organic compounds, including phytate. To investigate the diversity of phytate-hydrolyzing bacteria in soil, we sampled soils of various ecological habitats, including forest, private homesteads, large agricultural complexes, and urban landscapes. Bacterial isolate Pantoea sp. strain 3.5.1 with the highest level of phytase activity was isolated from forest soil and investigated further. The Pantoea sp. 3.5.1 agpP gene encoding a novel glucose-1-phosphatase with high phytase activity was identified, and the corresponding protein was purified to apparent homogeneity, sequenced by mass spectroscopy, and biochemically characterized. The AgpP enzyme exhibits maximum activity and stability at pH 4.5 and at 37°C. The enzyme belongs to a group of histidine acid phosphatases and has the lowest Km values toward phytate, glucose-6-phosphate, and glucose-1-phosphate. Unexpectedly, stimulation of enzymatic activity by several divalent metal ions was observed for the AgpP enzyme. High-performance liquid chromatography (HPLC) and high-performance ion chromatography (HPIC) analyses of phytate hydrolysis products identify dl-myo-inositol 1,2,4,5,6-pentakisphosphate as the final product of the reaction, indicating that the Pantoea sp. AgpP glucose-1-phosphatase can be classified as a 3-phytase. The identification of the Pantoea sp. AgpP phytase and its unusual regulation by metal ions highlight the remarkable diversity of phosphorus metabolism regulation in soil bacteria. Furthermore, our data indicate that natural forest soils harbor rich reservoirs of novel phytate-hydrolyzing enzymes with unique biochemical features.

  3. High-T sub c thin films on low microwave loss alkaline-rare-earth-aluminate crystals

    Energy Technology Data Exchange (ETDEWEB)

    Sobolewski, R.; Gierlowski, P.; Kula, W.; Zarembinski, S.; Lewandowski, S.J.; Berkowski, M.; Pajaczkowska, A. (Instytut Fizyki, Polska Akatlemia Nauk, Al. Lotnikow 32/46, PL-02668 Warszawa (PL)); Gorshunov, B.P.; Lyudmirsky, D.B.; Sirotinsky, O.I. (Institute of General Physics, USSR Academy of Sciences, 38 Vavilova Street, SU-117924 Moscow (SU))

    1991-03-01

    This paper reports on the alkaline-rare-earth aluminates (K{sub 2}NiF{sub 4}-type perovskites) which are an excellent choice as the substrate material for the growth of high-T{sub c} thin films suitable for microwave and far-infrared applications. The CaNdAlO{sub 4}, and SrLaAlO{sub 4} single crystals have been grown by Czochralski pulling and fabricated into the form of (001) oriented wafers. The Y-Ba-Cu-O and Bi-Sr-Ca-Cu-O films deposited on these substrates by a single-target magnetron sputtering exhibited very good superconducting and structural properties.

  4. In-situ potentiostatic activation to optimize electrodeposited cobalt-phosphide electrocatalyst for highly efficient hydrogen evolution in alkaline media

    Science.gov (United States)

    Wei, Mengmeng; Yang, Liming; Wang, Longlu; Liu, Tian; Liu, Chengbin; Tang, Yanhong; Luo, Shenglian

    2017-08-01

    We first report a novel cobalt-phosphide (Co-P) hybrid with flake-like structure by a facile one-step electrodeposition combined with in-situ potentiostatic activation technique. Exotic microstructure transformation of Co-P hybrid from microspheres to nanosheets has been noted during the activation process. The Co-P catalyst exhibits striking kinetic metrics with an overpotential of 85 mV (at 10 mA cm-2) and Tafel slope of 37 mV dec-1, performing among the best of all the HER catalysts in strong alkaline media (at pH 14). This study offers a new in-situ approach to optimize catalytic materials for high-performance electrocatalysts towards energy-related applications.

  5. Diversity and food web structure of nematode communities under high soil salinity and alkaline pH.

    Science.gov (United States)

    Salamún, Peter; Kucanová, Eva; Brázová, Tímea; Miklisová, Dana; Renčo, Marek; Hanzelová, Vladimíra

    2014-10-01

    A long-term and intensive magnesium (Mg) ore processing in Slovenské Magnezitové Závody a.s. in Jelšava has resulted in a high Mg content and alkaline pH of the soil environment, noticeable mainly in the close vicinity of the smelter. Nematode communities strongly reacted to the contamination mostly by a decrease in abundance of the sensitive groups. Nematodes from c-p 1 group and bacterivores, tolerant to pollution played a significant role in establishing the dominance at all sites. With increasing distance from the pollution source, the nematode communities were more structured and complex, with an increase in proportion of sensitive c-p 4 and 5 nematodes, composed mainly of carnivores and omnivores. Various ecological indices (e.g. MI2-5, SI, H') indicated similar improvement of farther soil ecosystems.

  6. CASE REPORT: ALKALINE PHOSPHATASE, ?-GLUTAMYLTRANSFERASE, UREA AND CREATININE SERUM CONCENTRATION IN RABBITS (Oryctolagus cuniculus CONCENTRAÇÃO SÉRICA DE FOSFATASE ALCALINA, GAMA-GLUTAMIL TRANSFERASE, URÉIA E CREATININA EM COELHOS (Oryctolagus cuniculus

    Directory of Open Access Journals (Sweden)

    Mauren Picada Emanuelli

    2008-04-01

    Full Text Available

    The enzymes γ-Glutamyltransferase (GGT and Akaline phosphatase (AP are serum markers of cholestasis process, becoming an important way of diagnosis in hepatic diseases.  Urea and creatinine are eliminated by urine. When these metabolic elements are higher then normal, it is subside to diagnosis mammals’ renal dysfunction. This present study aimed to establish reference values for these enzymes in rabbits (laboratory animals, to obtain data for their use in scientific experiments. Thus, it was collected blood samples in 45 animals; three samples each one, making a total of 135 blood samples. Serum was separated by immediate centrifugation. Colorimetric method was realized and values from 36.44+/-10.66mg/dl for urea, minimum at 9.24mg/dl and a maximum at 66.06mg/dl; 0.94+/-0.22 mg/dl for creatinine, minimum at 0.51mg/dl and a maximum at 1.53mg/dl; and 72.41+/-29.68UI for AP, minimum at 10.66UI and a maximum at 167.39UI, were determinate. The GGT was determined for kinetic method and values from 6.85+/-3.31, minimum at 2UI and a maximum at 15UI.

    KEY WORDS: Hepatic function, rabbits, renal function, serum biochemistry

    As enzimas gama-glutamil transferase e fosfatase alcalina são marcadores séricos de processos colestásicos, sendo importantes no diagn

  7. Quantification of oxidative stress phenotypes based on high-throughput growth profiling of protein kinase and phosphatase knockouts

    DEFF Research Database (Denmark)

    Altıntaş, Ali; Martini, Jacopo; Mortensen, Uffe Hasbro;

    2016-01-01

    Cellular responses to oxidative stress are important for restoring redox balance and ensuring cell survival. Genetic defects in response factors can lead to impaired response to oxidative damage and contribute to disease and aging. In single cell organisms, such as yeasts, the integrity of the ox......Cellular responses to oxidative stress are important for restoring redox balance and ensuring cell survival. Genetic defects in response factors can lead to impaired response to oxidative damage and contribute to disease and aging. In single cell organisms, such as yeasts, the integrity...... different protein kinases and phosphatase mutants were selected for their known activities in oxidative stress or other stress response pathways and were investigated for their dosage-dependent response to hydrogen peroxide. Detailed growth profiles were analyzed after the induction of stress for growth...

  8. Alkaline Phosphatase Activity and Ⅰ Collagen Expression of Rat Dental Papilla Cells in Virto%体外诱导培养大鼠牙乳头细胞碱性磷酸酶和Ⅰ型胶原的表达

    Institute of Scientific and Technical Information of China (English)

    曹莹; 方厂云; 吕亚林; 姚志刚

    2012-01-01

    目的:观察体外诱导培养的大鼠牙乳头细胞碱性磷酸酶和Ⅰ型胶原蛋白合成的变化.方法:原代培养法获得大鼠牙乳头细胞,在条件培养基中进行诱导培养,在不同时间进行钙结节染色,ALP染色和定量测定,ELISA法和RT-PCR法测定细胞Ⅰ型胶原蛋白的量和mRNA表达水平.结果:体外诱导培养的大鼠牙乳头细胞形成含钙化物的细胞结节,体外培养至12 d的诱导组细胞ALP高于对照组(P< 0.05),细胞内1型胶原表达量高于对照组(P<0.05),且Ⅰ型胶原mRNA表达水平上调(P<0.05);培养15d后诱导组细胞分泌的Ⅰ型胶原高于对照组(P<0.05).结论:大鼠牙乳头细胞在体外分化的过程中碱性磷酸酶活性明显增高,矿化能力增强,并不断合成和分泌Ⅰ型胶原形成胞外基质.%Objective: To investigate the alkaline phosphatase (ALP ) activity and type I collagen content of rat dental papillae cells (rDPC) cultured in odontoblast differentiation-inducing medium in vitro. Methods: Neonatal rat dental papillae cells were obtained from molar tooth germs and expanded in vitro. rDPC were cultured in the medium containing dexamethasone, ascorbic acid and β-glycerophosphate. Cell cultures were analysed for morphology, mineralization potential, alkaline phosphatase and Type I collagen using immunocytochemistry, ELISA and reverse transcriptase-polymerase chain reaction. Results: Cell mineral nodules of rDPC cultured in odontoblast differentiation- inducing medium were detect by Alizarin red S staining after day 7. ALP activity, type I collagen in cell culture supernatants and type I collagen mRNA expression of rDPC were all increased significantly compared with control group after day 12. Type I collagen concentrations in intracellular protein of differentiation-inducing cultured rDPC were significantly enhanced compared to rDPC in control group after day 15. Conclusion: Odontoblast differentiation-inducing treatment promoted the

  9. Effect of alendronate sodium on serum alkaline phosphatase, osteocalcin and bone mineral density in patients with postmenopausal osteoporosis%阿仑膦酸钠对绝经后骨质疏松症患者骨密度及血清碱性磷酸酶、骨钙素的影响

    Institute of Scientific and Technical Information of China (English)

    白孟海; 白洁; 郑蓉; 陈克明; 刘茗露

    2012-01-01

    目的:探讨阿仑膦酸钠对绝经后骨质疏松症患者骨密度(BMD)及血清碱性磷酸酶(ALP)、骨钙素(BGP)的影响.方法:62例绝经后骨质疏松症患者随机分为2组,治疗组32例,对照组30例.治疗组口服阿仑膦酸钠、阿法迪三;对照组口服阿法迪三.分别于治疗前、治疗6个月、治疗12个月双能X线骨密度仪测定患者正位腰椎(L2~4)、髋部股骨颈(Neck)、大转子(Troch)和Ward’s三角区BMD,全自动生物分析仪测定血清ALP和BGP含量.结果;治疗组治疗6个月、12个月血清ALP和BGP含量与对照组比较有明显差异(P<0.01).治疗组,治疗12个月后L1_4和髋部(Neck,Troch.Ward's)BMD值与治疗前比较显著增加(P<0.01).治疗组与对照组比较L1-4和全髋部BMD值明显差异(P<0.01).结论:阿仑膦酸钠能有效阻止绝经后骨矿丢失,降低血清ALP含量和提高血清BGP含量.%Objective: To investigate the effect of alendronate sodium on serum alkaline phosphatase (ALP), osleocalein and bone mineral density (BMD) in patienis with postmenopausal osteoporosis. Methods,- A iota! Of 62 postmenopausal women with osteoporosis were randomly divided into 2 groups: 32 treatment patients received alendronate sodium and Alpha D3 treatment, and 30 control patients received Alpha D3 only. Before treatment, 6 and 12 months after treatment, the postmenopausal BMD of anterior - posterior lumbar spines and proximal femur were determined by dual - energy X - ray abaorptiometry, and the serum alkaline phosphatase and osteocalein were detected. Results-. Compared with control group, the serum ALP and osteocalcin levels were significantly decreased 6 and 12 months after treatment (P<0.01). Compared with that before treatment, the BMD value of the lumbar spine and proximal femur wag significantly increased 12 months after treatment (P <0.05 ). There was also obvious difference in BMD level of the lumbar spine and proximal femur 12 months afiei treatment between

  10. Measurement and analysis of alkaline phosphatase and aspartate aminotransferase activity levels in gingival cervical fluid after treatment of telescope retained fixed bridge%套筒冠式固定桥基牙龈沟液中碱性磷酸酶和天冬氨酸转氨酶的检测分析

    Institute of Scientific and Technical Information of China (English)

    武峰; 赵彬; 姚蔚

    2011-01-01

    Objective To explore the role of telescopic crown retainers fixed bridge in periodontal maintenance. Methods A total of 64 abutment teeth were selected from 16 patients and divided into experimental and control groups (n=32 each). The gingival index and sulcus bleeding index of abutments in the both two groups were detected at 6 and 12 months before and after repair. The alkaline phosphatase (ALP) and aspartate aminotransferase (AST) activity levels in gingival cervical fluid were measured using automatic biochemistry analyzer. Results There were significant differences in Gl and SBI between the experimental and control groups, as well as in ALP and AST activity levels (P<0.05). Conclusion Telescope retained fixed bridge might help protect the periodontal tissue of abutment teeth.%目的 探索套筒冠固位式固定桥对于牙周维护的作用.方法 选择16例患者共64颗基牙,分为实验组和对照组,每组32颗基牙.在修复前以及修复后6 、12个月,对基牙进行牙龈指数(GI)和龈沟出血指数(SBI)的检测,利用全自动生化分析仪检测基牙龈沟液中碱性磷酸酶(ALP)和天冬氨酸转氨酶(AST)活性水平.结果 在修复前与修复后12 个月实验组与对照组之间GI、SBI比较差异有统计学意义(P<0.05),ALP、AST活性水平差异也有统计学意义(P<0.05).结论 套筒冠式固定桥有利于基牙牙周组织的保护.

  11. Pyridoxine-responsive seizures as the first symptom of infantile hypophosphatasia caused by two novel missense mutations (c.677T>C, p.M226T; c.1112C>T, p.T371I) of the tissue-nonspecific alkaline phosphatase gene.

    Science.gov (United States)

    Baumgartner-Sigl, Sara; Haberlandt, Edda; Mumm, Steven; Scholl-Bürgi, Sabine; Sergi, Consolato; Ryan, Lawrence; Ericson, Karen L; Whyte, Michael P; Högler, Wolfgang

    2007-06-01

    Pyridoxine-responsive seizures (PRS) and the role of pyridoxine (PN, vitamin B(6)) in hypophosphatasia (HPP) are incompletely understood. Typically, PRS and HPP are rare, independent, metabolic disorders. In PRS, seizures resist standard anticonvulsants apart from PN, yet have a good prognosis. In HPP, inactivation of the tissue nonspecific isoenzyme of alkaline phosphatase (TNSALP) impairs skeletal mineralization and causes rickets in infants that can be fatal. Here, we report a 7-month-old girl, newly diagnosed with infantile HPP, who presented as a neonate with PRS but without bony abnormalities. Analysis of biogenic amines in cerebrospinal fluid (CSF) suggested brain pyridoxal 5'-phosphate (PLP) deficiency, although PLP in CSF was not decreased. She had normal cognitive milestones but failure to thrive and rickets. Nearly undetectable serum ALP activity, elevated plasma PLP and urinary phosphoethanolamine (PEA) and inorganic pyrophosphate (PPi) levels, hypercalcemia, hypercalciuria and nephrocalcinosis were consistent with infantile HPP. Only prednisolone reduced serum calcium levels. Despite improved growth and weight gain, she developed rib fractures and died from respiratory failure at age 9 months. Sequence analysis of the TNSALP gene revealed novel missense mutations in exon 7 (c.677T>C, p.M226T) and exon 10 (c.1112C>T, p.T371I). Our patient demonstrated that PRS in neonates may not necessarily be "idiopathic"; instead, such seizures can be caused by severe HPP that becomes clinically apparent later in infancy. The pathophysiology of PRS in HPP differs from the three other genetic defects known to cause PRS, but all may lead to brain PLP deficiency reducing seizure thresholds. All reported HPP patients with neonatal seizures died within 18 months of birth, suggesting that PRS is an indicator of HPP severity and lethal prognosis. We recommend that assessment of any neonate with PRS should include measurement of serum ALP activity.

  12. High electrocatalytic performance of nitrogen-doped carbon nanofiber-supported nickel oxide nanocomposite for methanol oxidation in alkaline medium

    Science.gov (United States)

    Al-Enizi, Abdullah M.; Elzatahry, Ahmed A.; Abdullah, Aboubakr M.; Vinu, Ajayan; Iwai, Hideo; Al-Deyab, Salem S.

    2017-04-01

    Nitrogen-Doped Carbon Nanofiber (N-CNF)-supported NiO composite was prepared by electrospinning a sol-gel mixture of graphene and polyaniline (PANi) with aqueous solutions of Polyvinylpyrrolidone (PVP) followed by a high-temperature annealing process. The electrospun was stabilized for 2 h at 280 °C, carbonized for 5 h at 1200 °C then loaded by 10% NiO. The electrocatalytic activities of the produced nanocomposite have been studied using cyclic voltammetry, and chronoamperometry. Also, N-CNF was characterized by X-ray diffraction (XRD), thermogravimetric analysis (TGA), surface area (BET), X-ray photoelectron spectroscopy (XPS), transmission electron microscope (TEM), and scanning-electron microscopy (SEM). The obtained N-doped carbon nanofiber was found to have a nitrogen content of 2.6 atomic% with a diameter range of (140-160) nm, and a surface area (393.3 m2 g-1). In addition, it showed a high electrocatalytic behavior towards methanol oxidation reaction in alkaline medium and high stability and resistivity to the adsorption of intermediates.

  13. Effect of Heat Stress on the Intestinal Flora Structure and Alkaline Phosphatase Activities and mRNA Expression of Amino Acid Transporters of Layer%热应激对蛋鸡肠道菌群结构、碱性磷酸酶活性及氨基酸转运载体mRNA表达丰度的影响

    Institute of Scientific and Technical Information of China (English)

    李永洙; 陈常秀; Yongquan Cui

    2013-01-01

    [目的]揭示热应激环境下蛋鸡肠道菌群结构、肠黏膜碱性磷酸酶活性和氨基酸转运载体mRNA表达量的变化机理。[方法]试验选择16周龄济宁百日鸡96只,随机分成对照组((24±1)℃;Ⅰ)和热应激((38±1)℃)组,各组设6个重复,每个重复8只,试验持续14 d。采用16S rDNA的PCR-DGGE技术和实时荧光定量PCR等手段,分析热应激2(Ⅱ)、7(Ⅲ)、14 d(Ⅳ)时,对十二指肠、空肠及回肠内容物菌群多样性和肠黏膜碱性磷酸酶活性以及氨基酸转运载体rBAT、y+LAT 1、CAT l mRNA基因表达的相对丰度变化规律。[结果]热应激7 d开始各肠段菌群多样性较为丰富,热应激7、14 d时空肠和回肠部位敏感乳杆菌(Lactobacillus agilis),回肠部位约氏乳杆菌(Lactobacillus johnsonii)、不可培养细菌(uncultured bacterium)和不可培养的拟杆菌属(uncultured Bacteroidalesbacterium)均末检测到;而热应激不同时间段空肠和回肠部位可检测到不可培养细菌、溃疡拟杆菌(Bacteroides helcogenes)、卵形拟杆菌(Bacteroides ovatus)、索氏志贺氏菌(Shigella sonnei);空肠和回肠部位黏膜上皮细胞表面的碱性磷酸酶活性与Ⅰ组比较显著下降(P<0.05);而空肠和回肠Ⅲ组的rBAT、y+LAT 1 mRNA表达丰度均最低,空肠在各热应激时段表达丰度变化幅度最大(P<0.05),回肠的CAT 1 mRNA表达丰度在Ⅲ、Ⅳ组与Ⅰ组比较影响更明显(P<0.01)。[结论]热应激对空肠和回肠部位微生物菌群影响较为明显,肠道微生物群落改变可导致肠道的消化吸收功能发生改变。%[Objective] The objective of this study is to reveal the influence mechanisms of heat stress affecting the intestinal flora structure of layer, the alkaline phosphatase activities of intestinal mucosa and the mRNA expression of amino acid transporters.[Method]A total of 96 Jining Bairi

  14. Alkaline Exchange Membrane (AEM) for High-Efficiency Fuel Cells, Electrolyzers and Regenerative Fuel Cell Systems Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Develop an alkaline exchange membrane (AEM)for use as a polymer electrolyte in both fuel cell and electrolyzer systems.  The ultimate goal in AEM development is...

  15. Highly sensitive detection and discrimination of LR and YR microcystins based on protein phosphatases and an artificial neural network.

    Science.gov (United States)

    Covaci, O I; Sassolas, A; Alonso, G A; Muñoz, R; Radu, G L; Bucur, B; Marty, J-L

    2012-08-01

    The inhibition characteristics of three different protein phosphatases by three microcystin (MC) variants--LR, YR, and RR--were studied. The corresponding K (I) for each enzyme-MC couple was calculated. The toxicity of MC varies in the following order: MC-LR > MC-YR > MC-RR. The sensitivity of the enzymes increased in the following order: mutant PP2A < mutant PP1 < natural PP2A. The best limit of detection obtained was 21.2 pM MC-LR using the most sensible enzyme. Methanol, ethanol, and acetonitrile up to 2 % (v/v) may be used in inhibition measurements. An artificial neural network (ANN) was used to discriminate two MC variants--LR and YR--using the differences in inhibition percentages measured with mutant PP1 and natural PP2A. The ANN is able to analyze mixtures with concentrations ranging from 8 to 98 pM MC-LR and 31 to 373 pM MC-YR.

  16. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Science.gov (United States)

    Ivanov, Delyan P; Parker, Terry L; Walker, David A; Alexander, Cameron; Ashford, Marianne B; Gellert, Paul R; Garnett, Martin C

    2014-01-01

    Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity) were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  17. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Directory of Open Access Journals (Sweden)

    Delyan P Ivanov

    Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  18. Dephosphorylation of Centrins by Protein Phosphatase 2C and

    Directory of Open Access Journals (Sweden)

    Marie-Christin Thissen

    2009-01-01

    Full Text Available In the present study, we identified protein phosphatases dephosphorylating centrins previously phosphorylated by protein kinase CK2. The following phosphatases known to be present in the retina were tested: PP1, PP2A, PP2B, PP2C, PP5, and alkaline phosphatase. PP2C and were capable of dephosphorylating P-Thr138-centrin1 most efficiently. PP2C was inactive and the other retinal phosphatases also had much less or no effect. Similar results were observed for centrins 2 and 4. Centrin3 was not a substrate for CK2. The results suggest PP2C and to play a significant role in regulating the phosphorylation status of centrins in vivo.

  19. High Temperature and Pressure Alkaline Electrochemical Reactor for Conversion of Power to Chemicals

    DEFF Research Database (Denmark)

    Chatzichristodoulou, Christodoulos

    2016-01-01

    Moving away from fossil fuels requires harvesting more and more intermittent renewable energy resources and establishing a sustainable system for the production of chemicals. This brings forward the need for efficient large scale energy storage technologies 1-3 and technologies for the conversion...... of renewable electricity to chemicals. Electrochemical reactors can play a crucial role in this endeavor, since they can efficiently and reversibly transform electricity to high-value chemicals, and thus serve as energy storage and recovery devices for balancing the grid, while offering a means...

  20. 基于微孔板化学发光法检测碱性磷酸酶的研究%Research of Chemiluminescent Assay of Alkaline Phosphatase Based on Microplate

    Institute of Scientific and Technical Information of China (English)

    王宇; 王宗良; 崔立国; 庄秀丽; 陈学思; 章培标

    2012-01-01

    CDP-Star能被ALP迅速水解,且水解产物能够持续发光.本研究对化学发光法检测ALP的反应条件进行了优化.优化后的反应条件为:125 μtmol/L的CDP- Star在pH 9.5缓冲液中,37℃下与ALP反应10min.在此条件下,△RLU与ALP的活力呈线性关系.因此,建立了一种基于微孔板化学发光法检测ALP的实验方法.结果表明,本方法检测ALP的检测线性范围是0.05~10 U/L和10~1000 U/L,相对于光吸收法具有速度快、灵敏度高、重复性好等优点.因此,本方法可以被开发成试剂盒,应用于高通量全自动生化分析仪的测试分析.%Enzymatic dephosphorylation of CDP-Star by ALP leads to CDP-Star decompose and continuously emit light. Optimal conditions for the determination of ALP by chemiluminescent method were investigated in this research. 125 μmol/L CDP-Star reacted with ALP in the buffer solution of pH = 9. 5 and incubated for 10 min at 37℃ were selected as the optimal conditions for the determination of ALP. The amount of ARLU is proportional to ALP activity. Therefore, the method of chemiluminescent assay of ALP based on microplate was established. The results showed that the linear range for the determination of ALP was 0. 05 -10 U/L and 10-1000 U/L. This method was more rapid, sensitive and reproducible than that of traditional colorimetric method. Therefore, this method can be developed into a kit used in the high-throughput automatic biochemical analyzer.

  1. The Effect of Two African Mineral Dyes on the Activity of Alkaline ...

    African Journals Online (AJOL)

    DR. MIKE HORSFALL

    activity of alkaline phosphatase (ALP) in the skin and serum of albino rats was ... analyses of the dyes were first carried out using solubility test, pH determination and X-ray fluorescence (XRF) ..... measurement to detect renal damage.

  2. A multi-scale model of the oxygen reduction reaction on highly active graphene nanosheets in alkaline conditions

    Science.gov (United States)

    Vazquez-Arenas, Jorge; Ramos-Sanchez, Guadalupe; Franco, Alejandro A.

    2016-10-01

    A multi-scale model based on a mean field approach, is proposed to describe the ORR mechanism on N-GN catalysts in alkaline media. The model implements activation energies calculated with Density Functional Theory (DFT) at the atomistic level, and scales up them into a continuum framework describing the cathode/electrolyte interface at the mesoscale level. The model also considers mass and momentum transports arising in the region next to the rotating electrode for all ionic species and O2; correction of potential drop and electrochemical double-layer capacitance. Most fitted parameters describing the kinetics of ORR elementary reactions are sensitive in the multi-scale model, which results from the incorporation of activation energies using the mean field method, unlike single-scale modelling Errors in the deviations from activation energies are found to be moderate, except for the elementary step (2) related to the formation of O2ads, which can be assigned to the inherent DFT limitations. The consumption of O2ads to form OOHads is determined as the rate-determining step as a result of its highest energy barrier (163.10 kJ mol-1) in the system, the largest error obtained for the deviation from activation energy (28.15%), and high sensitivity. This finding is confirmed with the calculated surface concentration and coverage of electroactive species.

  3. Microprofiles of oxygen, redox potential, and pH, and microbial fermentation products in the highly alkaline gut of the saprophagous larva of Penthetria holosericea (Diptera: Bibionidae).

    Science.gov (United States)

    Šustr, Vladimír; Stingl, Ulrich; Brune, Andreas

    2014-08-01

    The saprophagous larvae of bibionid flies harbor bacteria in their alkaline intestinal tracts, but little is known about the contribution of the gut microbiota to the digestion of their recalcitrant diet. In this study, we measured oxygen and hydrogen partial pressure, redox potential and pH in the midgut, gastric caeca and hindgut of larvae of the bibionid fly Penthetria holosericea with Clark-type O2 and H2 microsensors, platinum redox microelectrodes, and LIX-type pH microelectrodes. The center of the midgut lumen was anoxic, whereas gastric caeca and hindgut were hypoxic. However, redox potential profiles indicated oxidizing conditions throughout the gut, with lowest values in the midgut (+20 to +60mV). Hydrogen production was not detected. The midgut was extremely alkaline (pH around 11), whereas hindgut and gastric caeca were neutral to slightly alkaline. While HPLC analysis showed high concentrations of glucose in the midgut (15mM) and gastric caeca (27mM), the concentrations of microbial fermentation products such as lactate (2-4mM), acetate (larva of P. holosericea, which feeds selectively on decomposed leaves and its own microbe-rich faeces, differs fundamentally from those of detritivorous and humivorous insects, which host a highly active, fermentative microbiota in their alkaline midgut or hindgut compartments.

  4. Technetium in alkaline, high-salt, radioactive tank waste supernate: Preliminary characterization and removal

    Energy Technology Data Exchange (ETDEWEB)

    Blanchard, D.L. Jr.; Brown, G.N.; Conradson, S.D. [and others

    1997-01-01

    This report describes the initial work conducted at Pacific Northwest National Laboratory to study technetium (Tc) removal from Hanford tank waste supernates and Tc oxidation state in the supernates. Filtered supernate samples from four tanks were studied: a composite double shell slurry feed (DSSF) consisting of 70% from Tank AW-101, 20% from AP-106, and 10% from AP-102; and three complexant concentrate (CC) wastes (Tanks AN-107, SY-101, ANS SY-103) that are distinguished by having a high concentration of organic complexants. The work included batch contacts of these waste samples with Reillex{trademark}-HPQ (anion exchanger from Reilly Industries) and ABEC 5000 (a sorbent from Eichrom Industries), materials designed to effectively remove Tc as pertechnetate from tank wastes. A short study of Tc analysis methods was completed. A preliminary identification of the oxidation state of non-pertechnetate species in the supernates was made by analyzing the technetium x-ray absorption spectra of four CC waste samples. Molybdenum (Mo) and rhenium (Re) spiked test solutions and simulants were tested with electrospray ionization-mass spectrometry to evaluate the feasibility of the technique for identifying Tc species in waste samples.

  5. Alkaline phosphatases. Newer concepts in isoenzymes and clinical applications.

    Science.gov (United States)

    Griffiths, J

    1989-12-01

    ALP is a profoundly ubiquitous enzyme that originates in many, if not all, mammalian tissues. The development of a simple ISEF technique has allowed reflection of at least 12 cellular components. Improvements in technique may add further zones of information. The aura of darkness surrounding ALP is being slowly dissipated.

  6. Alginate-hydroxypropylcellulose hydrogel microbeads for alkaline phosphatase encapsulation

    NARCIS (Netherlands)

    Karewicz, A.; Zasada, K.; Bielska, D.; Douglas, T.E.L.; Jansen, J.A.; Leeuwenburgh, S.C.G.; Nowakowska, M.

    2014-01-01

    There is a growing interest in using proteins as therapeutics agents. Unfortunately, they suffer from limited stability and bioavailability. We aimed to develop a new delivery system for proteins. ALP, a model protein, was successfully encapsulated in the physically cross-linked sodium

  7. Kinetic studies of alkaline phosphatase extracted from rabbit (Lepus ...

    African Journals Online (AJOL)

    user

    activity, and the kinetic constants-maximum enzyme velocity (Vmax) and Michealis-Menten ... at temperature greater than 44.95°C with corresponding catalytic energy of ..... potential biomarker for the intergrity of the hepatic drainage system.

  8. Activation of Rat Intestinal Alkaline Phosphatase by Taurine May be ...

    African Journals Online (AJOL)

    Dr. K.J. Umar

    intestinal ALP activity is higher (12x10-3nmol-1min-1mg protein) in the presence of taurine and LPS when compared with the ... are essential for the toxic activity of lipid A. Removal of ... gram-negative bacteria and lipopolysaccaride (LPS) and.

  9. Kinetic Studies of Alkaline Phosphatase from the Liver of Agama ...

    African Journals Online (AJOL)

    Spectrophotometric method was used to assay the enzyme, and the kinetic constants: Maximum velocity (Vmax) ... was monitored by monitoring rate of hydrolysis of para – ... tubes to start the reaction at interval of 5 min for 30 min. The reaction ...

  10. Microprofiles of oxygen, redox potential, and pH, and microbial fermentation products in the highly alkaline gut of the saprophagous larva of Penthetria holosericea (Diptera: Bibionidae)

    KAUST Repository

    Šustr, Vladimír

    2014-08-01

    The saprophagous larvae of bibionid flies harbor bacteria in their alkaline intestinal tracts, but little is known about the contribution of the gut microbiota to the digestion of their recalcitrant diet. In this study, we measured oxygen and hydrogen partial pressure, redox potential and pH in the midgut, gastric caeca and hindgut of larvae of the bibionid fly Penthetria holosericea with Clark-type O2 and H2 microsensors, platinum redox microelectrodes, and LIX-type pH microelectrodes. The center of the midgut lumen was anoxic, whereas gastric caeca and hindgut were hypoxic. However, redox potential profiles indicated oxidizing conditions throughout the gut, with lowest values in the midgut (+20 to +60mV). Hydrogen production was not detected. The midgut was extremely alkaline (pH around 11), whereas hindgut and gastric caeca were neutral to slightly alkaline. While HPLC analysis showed high concentrations of glucose in the midgut (15mM) and gastric caeca (27mM), the concentrations of microbial fermentation products such as lactate (2-4mM), acetate (<1mM) and succinate (<0.5mM) were low in all gut regions, suggesting that the contribution of microorganisms to the digestive process, particularly in the alkaline midgut, is only of minor importance. We conclude that the digestive strategy of the saprophytic larva of P. holosericea, which feeds selectively on decomposed leaves and its own microbe-rich faeces, differs fundamentally from those of detritivorous and humivorous insects, which host a highly active, fermentative microbiota in their alkaline midgut or hindgut compartments. © 2014 Elsevier Ltd.

  11. FeS anchored reduced graphene oxide nanosheets as advanced anode material with superior high-rate performance for alkaline secondary batteries

    Science.gov (United States)

    Shangguan, Enbo; Guo, Litan; Li, Fei; Wang, Qin; Li, Jing; Li, Quanmin; Chang, Zhaorong; Yuan, Xiao-Zi

    2016-09-01

    A new nanocomposite formulation of the iron-based anode for alkaline secondary batteries is proposed. For the first time, FeS nanoparticles anchored on reduced graphene oxide (RGO) nanosheets are synthesized via a facile, environmentally friendly direct-precipitation approach. In this nanocomposite, FeS nanoparticles are anchored uniformly and tightly on the surface of RGO nanosheets. As an alkaline battery anode, the FeS@RGO electrode delivers a superior high-rate charge/discharge capability and outstanding cycling stability, even at a condition without any conductive additives and a high electrode loading of ∼40 mg cm-2. At high charge/discharge rates of 5C, 10C and 20C (6000 mA g-1), the FeS@RGO electrode presents a specific capacity of ∼288, 258 and 220 mAh g-1, respectively. Moreover, the FeS@RGO electrode exhibits an admirable long cycling stability with a superior capacity retention of 87.6% for 300 cycles at a charge/discharge rate of 2C. The excellent electrochemical properties of the FeS@RGO electrode can be stemmed from the high specific surface area, peculiar electric conductivity and robust sheet-anchored structure of the FeS@RGO nanocomposite. By virtue of its superior fast charge/discharge properties, the FeS@RGO nanocomposite is suitable as an advanced anode material for high-performance alkaline secondary batteries.

  12. Expression of bacterial alkaline phosphatase-steroid receptor coactivator-1 fusion protein and its application in regulation of drug metabolism enzymes%BAP-SRC-1融合蛋白的表达及其在药物代谢酶调控研究中的应用

    Institute of Scientific and Technical Information of China (English)

    陈亚坤; 陈枢青; 曾苏

    2005-01-01

    目的获得活性表达的细菌碱性磷酸酶(bacterial alkaline phosphatase,BAP)基因和人甾体受体辅活化因子-1(stero1d receptor coactivator-1,SRC-1)的融合蛋白,应用于辅活化因子与核受体的结合研究.方法从Escherichia coli JM83基因组和人肝总RNA中分别扩增获得BAP基因和SRC-1的186个氨基酸对应的基因序列(简称SRC186).用重组技术,构建BAP-SRC186-pET28a融合基因表达载体,在Escherichia coli Rosetta(DE3)中,以IPTG低温诱导表达.以对硝基苯磷酸盐(p-nitrophenyl-phos-phate,PNPP)为底物进行活性测定.活性表达的BAP-SRC186融合蛋白被应用于辅活化因子与核受体的结合研究.结果获得可溶性融合蛋白BAP-SRC186.该融合蛋白的BAP比活为(0.176±0.013 4)μmol·min-1·mg(pro).在利福平存在的情况下,BAP-SRC186能与孕烷X受体配体结合域(pregnane X receptor ligand binding domain,PXRLBD)发生利福平剂量依赖性的相互作用,作用强度通过BAP的显色反应可方便地检测.结论 BAP融合蛋白与核受体的结合研究为药物代谢酶调控的体外研究开辟了新的思路.

  13. Evaluation of gardenia yellow using crocetin from alkaline hydrolysis based on ultra high performance liquid chromatography and high-speed countercurrent chromatography.

    Science.gov (United States)

    Inoue, Koichi; Tanada, Chihiro; Nishikawa, Hiroaki; Matsuda, Satoru; Tada, Atsuko; Ito, Yusai; Min, Jun Zhe; Todoroki, Kenichiro; Sugimoto, Naoki; Toyo'oka, Toshimasa; Akiyama, Hiroshi

    2014-12-01

    Gardenia yellow is globally the most valuable spice and food color. It is generally a mixture of water-soluble carotenoid glycosyl esters which consist of crocetin bis(gentiobiosyl) ester as the main component. Crocetin is a natural carotenoid dicarboxylic acid that may be a candidate drug for pharmaceutical development, however, it is either present in trace amounts or is absent in natural gardenia yellow products. We here propose that crocetin produced by alkaline hydrolysis can be used to qualitatively evaluate gardenia yellow products using an ultra high performance liquid chromatographic assay. A useful and efficient isolation technique for isolating high-purity crocetin from gardenia yellow using high-speed countercurrent chromatography is described. High-speed countercurrent chromatographic fractionation followed by an ultra high performance liquid chromatographic assay showed that trans-crocetin is easily converted to about 15% cis-crocetin (85% trans-crocetin). Crocetin in gardenia yellow was quantitatively evaluated. Our approach is based on the hydrolysis process for converting crocetin glycosyl esters to crocetin before evaluation and isolation using the ultra high performance liquid chromatographic and high-speed countercurrent chromatographic methods. The combination of hydrolysis and chromatographic methods allows evaluation of the purity and quantity of crocetin in gardenia yellow.

  14. The effect of high glucose levels on the hypermethylation of protein phosphatase 1 regulatory subunit 3C (PPP1R3C) gene in colorectal cancer

    Indian Academy of Sciences (India)

    Soo Kyung Lee; Ji Wook Moon; Yong Woo Lee; Jung Ok Lee; Su Jin Kim; Nami Kim; Jin Kim; Hyeon Soo Kim; Sun-Hwa Park

    2015-03-01

    DNA methylation is an epigenetic event that occurs frequently in colorectal cancer (CRC). Increased glucose level is a strong risk factor for CRC. Protein phosphatase 1 regulatory subunit 3C (PPP1R3C) modulates glycogen metabolism, particularly glycogen synthesis. The aim of this study was to investigate the effect of high glucose levels on DNA methylation of PPP1R3C in CRC. PPP1R3C was significantly hypermethylated in CRC tissues (76/105, 72.38%, < 0.05) and colon cancer cell lines ( < 0.05). CRC tissues obtained from patients with high glucose levels showed that the methylation of PPP1R3C was lower than in patients who had normal levels of glucose. When DLD-1 cells were cultured under conditions of high glucose, the methylation of PPP1R3C was repressed. The expression of PPP1R3C was inversely related to methylation status. In addition, a promoter luciferase assay showed that the transcriptional activity of PPP1R3C was increased in high glucose culture conditions. The number of cells decreased when PPP1R3C was silenced in DLD-1 cells. These results suggest that PPP1R3C, a novel hypermethylated gene in CRC, may play a critical role in cancer cell growth in association with glucose levels.

  15. High-Mg# andesitic lavas of the Shisheisky Complex, Northern Kamchatka: implications for primitive calc-alkaline magmatism

    Science.gov (United States)

    Bryant, J. A.; Yogodzinski, G. M.; Churikova, T. G.

    2011-05-01

    Primitive arc magmatism and mantle wedge processes are investigated through a petrologic and geochemical study of high-Mg# (Mg/Mg + Fe > 0.65) basalts, basaltic andesites and andesites from the Kurile-Kamchatka subduction system. Primitive andesitic samples are from the Shisheisky Complex, a field of Quaternary-age, monogenetic cones located in the Aleutian-Kamchatka junction, north of Shiveluch Volcano, the northernmost active composite volcano in Kamchatka. The Shisheisky lavas have Mg# of 0.66-0.73 at intermediate SiO2 (54-58 wt%) with low CaO (3.0 wt%) and K2O (>1.0 wt%). Olivine phenocryst core compositions of Fo90 appear