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Sample records for alkaline phosphatase produced

  1. ALP (Alkaline Phosphatase) Test

    Science.gov (United States)

    ... Also known as: ALK PHOS; Alkp Formal name: Alkaline Phosphatase Related tests: AST ; ALT ; GGT ; Bilirubin ; Liver Panel ; Bone Markers ; Alkaline Phosphatase Isoenzymes; Bone Specific ALP All content on ...

  2. Alkaline Phosphatase in Stem Cells

    Directory of Open Access Journals (Sweden)

    Kateřina Štefková

    2015-01-01

    Full Text Available Alkaline phosphatase is an enzyme commonly expressed in almost all living organisms. In humans and other mammals, determinations of the expression and activity of alkaline phosphatase have frequently been used for cell determination in developmental studies and/or within clinical trials. Alkaline phosphatase also seems to be one of the key markers in the identification of pluripotent embryonic stem as well as related cells. However, alkaline phosphatases exist in some isoenzymes and isoforms, which have tissue specific expressions and functions. Here, the role of alkaline phosphatase as a stem cell marker is discussed in detail. First, we briefly summarize contemporary knowledge of mammalian alkaline phosphatases in general. Second, we focus on the known facts of its role in and potential significance for the identification of stem cells.

  3. Modulators of intestinal alkaline phosphatase.

    Science.gov (United States)

    Bobkova, Ekaterina V; Kiffer-Moreira, Tina; Sergienko, Eduard A

    2013-01-01

    Small molecule modulators of phosphatases can lead to clinically useful drugs and serve as invaluable tools to study functional roles of various phosphatases in vivo. Here, we describe lead discovery strategies for identification of inhibitors and activators of intestinal alkaline phosphatases. To identify isozyme-selective inhibitors and activators of the human and mouse intestinal alkaline phosphatases, ultrahigh throughput chemiluminescent assays, utilizing CDP-Star as a substrate, were developed for murine intestinal alkaline phosphatase (mIAP), human intestinal alkaline phosphatase (hIAP), human placental alkaline phosphatase (PLAP), and human tissue-nonspecific alkaline phosphatase (TNAP) isozymes. Using these 1,536-well assays, concurrent HTS screens of the MLSMR library of 323,000 compounds were conducted for human and mouse IAP isozymes monitoring both inhibition and activation. This parallel screening approach led to identification of a novel inhibitory scaffold selective for murine intestinal alkaline phosphatase. SAR efforts based on parallel testing of analogs against different AP isozymes generated a potent inhibitor of the murine IAP with IC50 of 540 nM, at least 65-fold selectivity against human TNAP, and >185 selectivity against human PLAP. PMID:23860652

  4. 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 apparent...... phosphatase activity could be demonstrated....

  5. 21 CFR 864.7660 - Leukocyte alkaline phosphatase test.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Leukocyte alkaline phosphatase test. 864.7660... Leukocyte alkaline phosphatase test. (a) Identification. A leukocyte alkaline phosphatase test is a device used to identify the enzyme leukocyte alkaline phosphatase in neutrophilic granulocytes...

  6. 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

  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. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    International Nuclear Information System (INIS)

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either 3H-fatty acids or [3H]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 3H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of [3H]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 3H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the 3H-fatty acid and the [3H]ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the [3H]ethanolamine label from the purified alkaline phosphatase. The 3H radioactivity in alkaline phosphatase purified from [3H]ethanolamine-labeled cells comigrated with authentic [3H]ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the 3H-fatty acid and [3H]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

  9. 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.

  10. Francisella DnaK Inhibits Tissue-nonspecific Alkaline Phosphatase*

    Science.gov (United States)

    Arulanandam, Bernard P.; Chetty, Senthilnath Lakshmana; Yu, Jieh-Juen; Leonard, Sean; Klose, Karl; Seshu, Janakiram; Cap, Andrew; Valdes, James J.; Chambers, James P.

    2012-01-01

    Following pulmonary infection with Francisella tularensis, we observed an unexpected but significant reduction of alkaline phosphatase, an enzyme normally up-regulated following inflammation. However, no reduction was observed in mice infected with a closely related Gram-negative pneumonic organism (Klebsiella pneumoniae) suggesting the inhibition may be Francisella-specific. In similar fashion to in vivo observations, addition of Francisella lysate to exogenous alkaline phosphatase (tissue-nonspecific isozyme) was inhibitory. Partial purification and subsequent proteomic analysis indicated the inhibitory factor to be the heat shock protein DnaK. Incubation with increasing amounts of anti-DnaK antibody reduced the inhibitory effect in a dose-dependent manner. Furthermore, DnaK contains an adenosine triphosphate binding domain at its N terminus, and addition of adenosine triphosphate enhances dissociation of DnaK with its target protein, e.g. alkaline phosphatase. Addition of adenosine triphosphate resulted in decreased DnaK co-immunoprecipitated with alkaline phosphatase as well as reduction of Francisella-mediated alkaline phosphatase inhibition further supporting the binding of Francisella DnaK to alkaline phosphatase. Release of DnaK via secretion and/or bacterial cell lysis into the extracellular milieu and inhibition of plasma alkaline phosphatase could promote an orchestrated, inflammatory response advantageous to Francisella. PMID:22923614

  11. 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....

  12. Nucleotide and amino acid sequences of human intestinal alkaline phosphatase: close homology to placental alkaline phosphatase

    International Nuclear Information System (INIS)

    A cDNA clone for human adult intestinal alkaline phosphatase (ALP) [orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1] was isolated from a λgt11 expression library. The cDNA insert of this clone is 2513 base pairs in length and contains an open reading frame that encodes a 528-amino acid polypeptide. This deduced polypeptide contains the first 40 amino acids of human intestinal ALP, as determined by direct protein sequencing. Intestinal ALP shows 86.5% amino acid identity to placental (type 1) ALP and 56.6% amino acid identity to liver/bone/kidney ALP. In the 3'-untranslated regions, intestinal and placental ALP cDNAs are 73.5% identical (excluding gaps). The evolution of this multigene enzyme family is discussed

  13. Placental-type alkaline phosphatase in cervical neoplasia.

    OpenAIRE

    McLaughlin, P. J.; Warne, P H; Hutchinson, G. E.; Johnson, P. M.; Tucker, D. F.

    1987-01-01

    Monoclonal antibodies reactive with placental-type alkaline phosphatase have formed the basis of methods for detection of this oncodevelopmental antigen in patients with pre-invasive and invasive cervical neoplasia, with or without evidence of papilloma virus infection. Disease-related elevations of placental-type alkaline phosphatase were not observed in patients' sera. Solubilised cervical smears or biopsy material, and cervical mucus swabs, often contained substantial amounts of this isoen...

  14. Characterization of the phosphatidylinositol-glycan membrane anchor of human placental alkaline phosphatase

    International Nuclear Information System (INIS)

    Placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] is a member of a diverse group of membrane proteins whose attachment to the lipid bilayer is mediated by a phosphatidylinositol-glycan. To investigate structural aspects of the glycolipid anchor, cultured WISH cells were used because, they produce the enzyme in abundant quantities. When cell suspensions were incubated with purified phosphatidylinositol-specific phospholipase C, most of the placental alkaline phosphatase was released from membranes in a hydrophilic form. On incubation of the cells with [14C]ethanolamine, [14C]myristic acid, or myo[3H]inositol, each was incorporated into the phosphatase near the carboxyl terminus, showing that these components, which are found in other phosphatidylinositol membrane-linked proteins, are also present in placental alkaline phosphatase

  15. 21 CFR 862.1050 - Alkaline phosphatase or isoenzymes test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alkaline phosphatase or isoenzymes test system... Test Systems § 862.1050 Alkaline phosphatase or isoenzymes test system. (a) Identification. An alkaline phosphatase or isoenzymes test system is a device intended to measure alkaline phosphatase or its...

  16. Serum alkaline phosphatase screening for vitamin D deficiency states

    International Nuclear Information System (INIS)

    Objective: To determine whether serum vitamin D levels are correlated with serum levels of alkaline phosphatase or not. Study Design: Cross-sectional, observational study. Place and Duration of Study: Multi-centre study, conducted at Liaquat National Hospital and Medical College, National Medical Centre and Medicare Hospital, Karachi, from January to October 2009. Methodology: Patients attending the Orthopaedic OPDs with complaints of pain in different body regions and serum vitamin D/sub 3/ levels of greater or equal to 30 ng/ml were included in the study. Patients with vitamin D deficiency were further categorized into mild deficiency or insufficiency (vit. D/sub 3/ = 20-29 ng/ml), moderate deficiency (vit. D/sub 3/ = 5 - 19 ng/ml) and severe deficiency forms (vit. D/sub 3/ < 5 ng/ml). Pearson correlation was applied to test the correlation of serum alkaline phosphatase levels with serum vitamin D/sub 3/ levels. P-value < 0.05 was considered to be significant. Results: Out of 110 samples, 26 had mild (23%), 61 had moderate (55%) and 21 had severe (19.1%) vitamin D deficiencies. All of the patients in the three groups had alkaline phosphatase with in normal limits and the total mean value of the enzyme was 135.97 +- 68.14I U/L. The inter group comparison showed highest values of alkaline phosphatase in the moderate vitamin D deficiency group. The correlation coefficient of alkaline phosphatase and serum vitamin D/sub 3/ levels was r =0.05 (p =0.593). Conclusion: Serum vitamin D/sub 3/ levels may not be correlated with increased serum alkaline phosphatase levels. Therefore, alkaline phosphatase may not be used as a screening test to rule out vitamin D deficiency. (author)

  17. 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

  18. 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.

  19. 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

  20. A physiologic function for alkaline phosphatase : Endotoxin detoxification

    NARCIS (Netherlands)

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

    1997-01-01

    Alkaline phosphatase (AP), a common enzyme present in many species including humans, has been studied extensively. Although the enzyme is routinely applied as a marker for liver function, its biologic relevance is poorly understood. The reason for this is obvious: the pH optimum of AP in vitro, as m

  1. 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

  2. Purification and properties of alkaline phosphatase of silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    TANG Yunming; CEN Liang; CHU Bo; LI Changchun; XU Min; LUO Ying; LU Cheng

    2006-01-01

    Alkaline phosphatase(AKP),from the succus entericus of silkworm,was purified using 10%-50% ammonium sulfate fractions,ion exchange chromatography Of DEAE-Sepharose,and size exclusion chromatography of Sephacryl S-200.The purification fold was 464 times and specified activity was 3936 U/mg.Optimum pH value of the phosphatase was 10.5,and was stable between pH 7.5 and 11.The optimum temperature of the phosphatase was 40℃ and it was unstable over 50℃.Km value of the phosphatase was 1.25 mmol/L.In a given condition,the phosphatase was selectively modified by PCMB,NBS,PMSE TNBS,SUAN,DTT,BrAc,and IAc,the results indicate that PMSF,SUA,BrAc,IAc,and TNBS could Obviously inhibit the activity of the phosphatase,and the degree of inhibition depended on the concentration of these reagents.There was little effect on the activity of phosphatase after treatment by PMSF,DTT,and NBT.We primarily conclude that mercapto and imidazole are essential for AKP from silkworm.Also,Lys residue and disulfide bands are necessary to protect the catalysis of the AKP.

  3. Alkaline phosphatase for immunocytochemical labelling: problems with endogenous enzyme activity.

    OpenAIRE

    Bulman, A. S.; Heyderman, E

    1981-01-01

    Alkaline phosphatase may be used as a label for immunocytochemistry and can be demonstrated in tissue sections using the single step naphthol phosphate method. Endogenous enzyme activity may not be destroyed by fixation in formalin, formol alcohol, Carnoy's or Baker's solutions and should be inhibited before results are assessed. Either Bouin's solution or periodic acid followed by potassium borohydride are satisfactory inhibitor and do not adversely affect immunocytochemical results.

  4. 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.

  5. Sensitive optical detection of alkaline phosphatase activity with quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Xiangling [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China); The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096 (China); Chen, Zhenzhen; Chen, Xiaoying; Liu, Jing [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China); Tang, Fangqiong, E-mail: tangfq@mail.ipc.ac.cn [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China)

    2014-01-15

    A simple method has been developed to detect the activity of alkaline phosphatase (ALP) by the changing of fluorescence intensities of the quantum dots (QDs). In this system, the fluorescence intensities of the QDs were quenched by p-nitrophenol (pNP) which was produced in the process of ALP catalytic reaction. A series of linear calibration curves of the activity of ALP were obtained in different pH buffer solutions. The wide linear range was 3–1000 U L{sup −1} and the detection limit was 3 U L{sup −1} (S/N=3). Furthermore, the experimental conditions of biosensor were optimized, and anti-interference ability was presented. The activity of ALP was also detected in serum and the recovery of ALP in serum samples was more than 95%. The excellent performance of this biosensor indicates that it can be used in practice detection of ALP. -- Highlights: • A sensitive ALP biosensor is constructed based on QDs without complex processes. • The analysis processing is very convenient, simple and rapid. • The detection mechanism of the ALP biosensor is studied by XPS. • The paper proposes a feasible approach for some substrates or enzymes detecting.

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

    OpenAIRE

    Eaman A. S. AL-Rubaee; Suha T. Abd; Nadia M. Kadim

    2015-01-01

    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 w...

  7. A description of alkaline phosphatases from marine organisms

    Science.gov (United States)

    Tian, Jiyuan; Jia, Hongbing; Yu, Juan

    2016-07-01

    Alkaline phosphatases (APs) are non-specific phosphohydrolases, and they are widely used in clinical diagnostics and biological studies. APs are widespread in nature and exhibit different structural formulations. Based on the diversity of biogenetic sources, APs exhibit temperature-propensity traits, and they are classified as psychrophilic, mesophilic, and thermophilic. In this article, the characteristics of psychrophilic APs from marine organisms were described, accompanied by a simple description of APs from other organisms. This review will facilitate better utilization of marine APs in the biotechnology field.

  8. 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.

  9. Alkaline Phosphatase Activity in San Francisco and Monterey Bays

    Science.gov (United States)

    Nicholson, D. P.

    2002-12-01

    Phosphorus (P) is an essential nutrient utilized by all living organisms, and has been recognized as a limiting nutrient in some oceanic systems (Cotner et al., 1997; Karl et al., 1995; Michaels et al., 1996; Wu et al., 2000). However, relatively little is known about the extent of P limitation in natural environments, how P limitation varies spatially and temporally, and what determines how and when P becomes limiting (Benitez-Nelson, 2000). A more direct estimate of the degree of P limitation in a variety of oceanic systems is needed to better understand P cycling and dynamics within the ocean and how these have and will change in response to global climate and environmental perturbation. Accordingly, the objective this study is to assess the P-status of marine planktonic communities in Monterey and San Francisco Bays using the activity of alkaline phosphatase in the water column. Alkaline phosphatase (AP) is the most widely used enzyme that marine organisms use to hydrolize organic P compounds to biologically available orthophosphate. Accordingly it is expected that in areas where P is a limiting nutrient organisms will produce and release more AP to seawater so they can utilize the dissolved and particulate organic P compounds. Indeed it has been suggested that the AP activity is a reliable indicator of P-availability to planktonic communities (Ammerman and Azam, 1985; Cotner and Wetzel, 1991; Hong et al., 1998). High enzyme activities indicate low dissolved inorganic phosphate (DIP) availability while low levels suggest that DIP supply satisfies the community P-demand. This study examines AP activity in San Francisco and Monterey Bays over a 12 month period, from November, 2001 through November, 2002 using two enzyme assays. The study encompasses data from a three-station transect in Monterey Bay, at depths ranging from 0-60 meters. The stations range from coastal waters to open ocean depths of several thousand meters. In San Francisco Bay, surface water from

  10. Improved double immunohistochemical staining method for cryostat and paraffin wax sections, combining alkaline phosphatase anti-alkaline phosphatase and indirect immunofluorescence

    OpenAIRE

    Tao, Q.; Srivastava, G; Loke, S L; Chan, E. Y.; Ho, F C

    1994-01-01

    Aims - To develop an immunohistochemical staining method for cryostat and paraffin wax sections so that two different antigens in the same section of tissues could be detected by combining immunoenzyme and immunofluorescence techniques. Methods - This double immunohistochemical staining method combines alkaline phosphatase-anti-alkaline phosphatase (APAAP) using New Fuchsin as a chromogen and indirect immunofluorescence. Results - APAAP staining for one antigen of this double immunohistochemi...

  11. 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

  12. 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.

  13. Characterization of the alkaline phosphatase expressed on the surface of a Hodgkin's lymphoma cell line

    NARCIS (Netherlands)

    Belland, L; Visser, L; Poppema, S; Stinson, R A

    1993-01-01

    Alkaline phosphatase solubilized from a human Hodgkin's lymphoma cell line (L428) was compared with purified amphiphilic and hydrophilic forms of the enzyme from human liver, and with the enzyme solubilized from a cultured osteosarcoma cell line (Saos-2). Purified hydrophilic alkaline phosphatases f

  14. 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.

  15. A Novel Bifunctional Hybrid with Marine Bacterium Alkaline Phosphatase and Far Eastern Holothurian Mannan-Binding Lectin Activities

    OpenAIRE

    Larissa Balabanova; Vasily Golotin; Svetlana Kovalchuk; Alexander Bulgakov; Galina Likhatskaya; Oksana Son; Valery Rasskazov

    2014-01-01

    A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa...

  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 alkaline...

  17. 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.

  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. How Should an Increase in Alkaline Phosphatase Activity Be Interpreted?

    International Nuclear Information System (INIS)

    Low-level laser therapy, commonly known as LLLT, is the application of low power, monochromatic, and coherent light to injuries and lesions to stimulate healing and give pain relief. There are conflicting reports in the literature regarding the role of ALP. Objective: this study aimed to compare the cellular responses of wounded human skin fibroblasts exposed to doses of 0.5 J/cm2, 2.5 J/cm2, 5 J/cm2, or 16 J/cm2 using LLLT with a Helium-Neon laser (632.8 nm, 18.8 mW power output, 2.07 mW/cm2 power density, and 3.4 cm diameter spot size or area 9.1?cm2) to elucidate the role of alkaline phosphatase (ALP) in cell proliferation. Methods: cellular responses to laser irradiation were evaluated using ALP enzyme activity, LDH membrane integrity, neutral red for cell proliferation, optical density at 540?nm, and basic fibroblast growth factor (bFGF) expression. Results: results suggest that an increase in ALP is negatively correlated with cell growth depending on the concentration of growth factors in the medium. Results also indicate that an increase in ALP may be related to cellular damage. Conclusion: since the exact role of ALP is unknown, the ALP enzyme activity assay should be considered in conjunction with other cell proliferation assays such as neutral red, optical density, or more specifically bFGF expression.

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

    OpenAIRE

    Chung-Chiun Liu; Kevin Wang; Wang, Joanne H.; Brandon Bartling

    2009-01-01

    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 line...

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

    OpenAIRE

    Faerk, J; Peitersen, B; Petersen, S; Michaelsen, K

    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.

  2. Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro

    OpenAIRE

    Katarzyna Bednarska; Magdalena Klink; Zofia Sulowska

    2006-01-01

    We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (104–106). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using...

  3. Optical Algal Biosensor using Alkaline Phosphatase for Determination of Heavy Metals

    OpenAIRE

    Durrieu, Claude; Tran-Minh, Canh

    2002-01-01

    International audience A biosensor is constructed to detect heavy metals from inhibition of alkaline phosphatase (AP) present on the external membrane of Chlorella vulgaris microalgae. The microalgal cells are immobilized on removable membranes placed in front of the tip of an optical fiber bundle inside a homemade microcell. C. vulgaris was cultivated in the laboratory and its alkaline phosphatase activity is strongly inhibited in the presence of heavy metals. This property has been used ...

  4. 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.

  5. 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.

  6. Electrochemical biosensor for detection of DNA hydroxymethylation based on glycosylation and alkaline phosphatase catalytic signal amplification

    International Nuclear Information System (INIS)

    Highlights: • DNA Hydroxymethylation was detected by electrochemical method. • 5-Hydroxymethylation cytosine in target DNA was chemically modified with glucose group. • Alkaline phosphatase catalytic signal amplification strategy was used. • The developed method also showed excellent reproducibility and stability. - Abstract: DNA hydroxymethylation (5-hydroxymethylcytosine, 5hmC) is a kind of new epigenetic modification, which plays key roles in nuclear reprogramming, regulates the gene activity, and initiates the DNA demethylation in mammals. For further understanding the functions of 5hmC and the correlation with tumour, it is essential to develop sensitive and selective methods for detecting and sequencing 5hmC. Herein, a kind of electrochemical biosensor was fabricated for 5hmC detection based on the glycosylation modification of 5hmC and enzymatic signal amplification. Under the catalytic effect of T4 β-glucosyltransferase, the 5hmC in target DNA was chemically modified with glucose. Then with the bridge connection of 1,4-phenyldiboronic acid, alkaline phosphatase was further captured on the electrode surface to catalyze the hydrolysis of p-nitrophenyl phosphate disodium salt to produce p-nitrophenol. Based on the relationship between the electrochemical oxidation signal of p-nitrophenol and the concentration of target DNA, the 5hmC level can be detected with high sensitivity and selectivity. The developed method also showed excellent reproducibility and stability

  7. 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.

  8. [Inhibition of alkaline phosphatase I of Pichia guilliermondii yeast in vitro and in vivo].

    Science.gov (United States)

    Sibirnyi, A A; Shavlovskii, G M

    1978-01-01

    The rate of p-nitrophenyl phosphate and flavin mononucleotide (FMN) hydrolysis by the partially purified preparation of alkaline phosphatase I of Pichia guilliermondii flavinogenic yeast was studied as affected by different substrates and inorganic ions. Their Km was established to be 2.0 X 10(-4) m and 2.5 X 10(-4) M, respectively. Dephosphorylation of p-nitrophenylphosphate and FMN was inhibited competitively by beta-glycerophosphate (Ki = 3.1 X 10(-3) M, respectively). The presence of inorganic phosphate ions in the reaction mixture decreases or removes inhibition of these compounds hydrolysis by other substrates of alkaline phosphatase I. The activity of alkaline phosphatase I increases in the presence of Mg2+ and was strongly inhibited in the presence of Be2+, Cu2+, Zn2+, Cd2+ and inorganic phosphate, the mixture of Be2+ and F- being the most effective. This mixture inhibited the phosphatase activity of the partially purified preparation of alkaline phosphatase I of the cell-free extract as well as of intact cells in both the alkaline and acid zones of pH (8.6 and 5.5, respectively). Incubation of the washed iron-deficient P. guilliermondii cells in the presence of Be2+ and F- did not result in accumulation of FMN in the yeast culture. A possible role of nonspecific phosphomonoesterases in hydrolysis of FMN in vivo is discussed. PMID:208203

  9. Application of intracellular alkaline phosphatase activity measurement in detection of neutrophil adherence in vitro.

    Science.gov (United States)

    Bednarska, Katarzyna; Klink, Magdalena; Sulowska, Zofia

    2006-01-01

    We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (10(4)-10(6)). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using the MTT reduction assay. The fluorimetric NAP activity test may be applied for resting as well as activated neutrophils without the risk of the activators interferences into the test. The alkaline phosphatase survey with the use of 4-MUP substrate is recommended herein as a sensitive, repeatable, simple, and reliable method of the neutrophil adherence determination in vitro. PMID:17047286

  10. 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.

  11. Electrochemical Determination of Alkaline Phosphatase in Human Serum by Differential Pulse Voltammetry

    Institute of Scientific and Technical Information of China (English)

    SUN Wei; JIAO Kui; WANG Hai-yu; LU Lu-de

    2004-01-01

    Differential pulse voltammetry(DPV) was applied to the determination of alkaline phosphatase(ALP) activity in human serum with phenyl phosphate as the substrate. Phenyl phosphate can enzymatically be hydrolyzed to produce phenol which is quantified by DPV at +660 mV(vs.Ag/AgCl) in the concentration range of 2.0-100 μmol/L. The standard curve for ALP is linear over the range from 0.06 to 1000 U/L with a relative standard deviation of 3.0%. The conditions for the enzymatic reaction and voltammetric detection were optimized and the kinetic constants were also examined.The human serum samples were tested by this method and the results were in good agreement with those obtained by the routine p-nitrophenyl phosphate spectrophotometric method.

  12. 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.

  13. Tubulin polymerization by paclitaxel (taxol) phosphate prodrugs after metabolic activation with alkaline phosphatase.

    Science.gov (United States)

    Mamber, S W; Mikkilineni, A B; Pack, E J; Rosser, M P; Wong, H; Ueda, Y; Forenza, S

    1995-08-01

    Paclitaxel (taxol) phosphate derivatives BMY46366, BMY-46489, BMS180661 and BMS180820 were used to determine the ability of alkaline phosphatase to convert these water-soluble potential prodrugs to tubulin-polymerizing metabolites (i.e., paclitaxel). Compounds were treated up to 180 min with an in vitro metabolic activation system composed of 10% bovine alkaline phosphatase in 0.2 M tris, pH 7.4, or in 0.2 M glycine, pH 8.8, plus 0.05 M MgCl2. Samples were tested (either by direct addition or after methylene chloride extraction/dimethyl-sulfoxide resuspension) in spectrophotometric tubulin polymerization assays utilizing bovine-derived microtubule protein. Pretreatment of 2'- and 7-phosphonoxyphenylpropionate prodrugs BMS180661 and BMS180820 with alkaline phosphatase for 30 to 120 min yielded relative initial slopes of about 20 to 100% at test concentrations equimolar to paclitaxel. High-performance liquid chromatography/mass spectrometry of BMS180661 treated with alkaline phosphatase confirmed the production of paclitaxel from the prodrug. In contrast, 2'- and 7-phosphate analogs BMY46366 and BMY46489 treated with alkaline phosphatase were not active in tubulin assays. None of the paclitaxel phosphate prodrugs polymerized tubulin in the absence of metabolic activation. The differences in tubulin polymerization with metabolic activation may be related both to accessibility of the phosphate group to the enzyme and to anionic charge effects. These results demonstrate that certain paclitaxel phosphate prodrugs can be metabolized by alkaline phosphatase to yield effective tubulin polymerization. PMID:7636751

  14. Identification of a macro-alkaline phosphatase complex in a patient with inflammatory bowel disease.

    Science.gov (United States)

    McTaggart, Malcolm P; Rawson, Catherine; Lawrence, David; Raney, Barbara S; Jaundrill, Linnet; Miller, Lorna A; Murtinho-Braga, Joseph; Kearney, Edward M

    2012-07-01

    We report the rare finding of a macro-alkaline phosphatase (macroALP) complex in a patient with a previously unexplained raised alkaline phosphatase activity. The clinical symptoms were persistent, daily diarrhoea for two months with blood in the stool. The patient was subsequently diagnosed with inflammatory bowel disease, specifically ulcerative colitis, following a rectal biopsy and colonoscopy. Two cases of macroALP associated with ulcerative colitis have been reported before, suggesting there could be an increased prevalence of macroALP in these patients. PMID:22454544

  15. Inhibition of acid, alkaline, and tyrosine (PTP1B) phosphatases by novel vanadium complexes.

    Science.gov (United States)

    McLauchlan, Craig C; Hooker, Jaqueline D; Jones, Marjorie A; Dymon, Zaneta; Backhus, Emily A; Greiner, Bradley A; Dorner, Nicole A; Youkhana, Mary A; Manus, Lisa M

    2010-03-01

    In the course of our investigations of vanadium-containing complexes for use as insulin-enhancing agents, we have generated a series of novel vanadium coordination complexes with bidentate ligands. Specifically we have focused on two ligands: anthranilate (anc(-)), a natural metabolite of tryptophan, and imidizole-4-carboxylate (imc(-)), meant to mimic naturally occurring N-donor ligands. For each ligand, we have generated a series of complexes containing the V(III), V(IV), and V(V) oxidation states. Each complex was investigated using phosphatase inhibition studies of three different phosphatases (acid, alkaline, and tyrosine (PTP1B) phosphatase) as prima facia evidence for potential use as an insulin-enhancing agent. Using p-nitrophenyl phosphate as an artificial phosphatase substrate, the levels of inhibition were determined by measuring the absorbance of the product at 405nm using UV/vis spectroscopy. Under our experimental conditions, for instance, V(imc)(3) appears to be as potent an inhibitor of alkaline phosphatase as sodium orthovanadate when comparing the K(cat)/K(m) term. VO(anc)(2) is as potent an inhibitor of acid phosphatase and tyrosine phosphatase as the Na(3)VO(4). Thus, use of these complexes can increase our mechanistic understanding of the effects of vanadium in vivo. PMID:20071031

  16. X-Ray Structure Reveals a New Class and Provides Insight into Evolution of Alkaline Phosphatases

    OpenAIRE

    Bihani, Subhash C.; Das, Amit; Nilgiriwala, Kayzad S.; Prashar, Vishal; Pirocchi, Michel; Apte, Shree Kumar; Ferrer, Jean-Luc; Hosur, Madhusoodan V.

    2011-01-01

    The alkaline phosphatase (AP) is a bi-metalloenzyme of potential applications in biotechnology and bioremediation, in which phosphate monoesters are nonspecifically hydrolysed under alkaline conditions to yield inorganic phosphate. The hydrolysis occurs through an enzyme intermediate in which the catalytic residue is phosphorylated. The reaction, which also requires a third metal ion, is proposed to proceed through a mechanism of in-line displacement involving a trigonal bipyramidal transitio...

  17. Extreme Elevation of Alkaline Phosphatase in a Pregnancy Complicated by Gestational Diabetes and Infant with Neonatal Alloimmune Thrombocytopenia.

    Science.gov (United States)

    Lozo, Svjetlana; Atabeygi, Amir; Healey, Michael

    2016-01-01

    There have been few case reports of isolated elevation of alkaline phosphatase beyond the normal physiologic amount with subsequent return to baseline after delivery. Here we present a similar case of extreme elevation of alkaline phosphatase in a pregnancy complicated by gestational diabetes and subsequently by neonatal alloimmune thrombocytopenia (NAIT). PMID:27610256

  18. A Novel Bifunctional Hybrid with Marine Bacterium Alkaline Phosphatase and Far Eastern Holothurian Mannan-Binding Lectin Activities

    Science.gov (United States)

    Balabanova, Larissa; Golotin, Vasily; Kovalchuk, Svetlana; Bulgakov, Alexander; Likhatskaya, Galina; Son, Oksana; Rasskazov, Valery

    2014-01-01

    A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25±5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens. PMID:25397876

  19. A novel bifunctional hybrid with marine bacterium alkaline phosphatase and Far Eastern holothurian mannan-binding lectin activities.

    Directory of Open Access Journals (Sweden)

    Larissa Balabanova

    Full Text Available A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25 ± 5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens.

  20. Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase

    NARCIS (Netherlands)

    Douglas, T.E.L.; Skwarczynska, A.; Modrzejewska, Z.; Balcaen, L.; Schaubroeck, D.; Lycke, S.; Vanhaecke, F.; Vandenabeele, P.; Dubruel, P.; Jansen, J.A.; Leeuwenburgh, S.C.G.

    2013-01-01

    Thermosensitive chitosan hydrogels containing sodium beta-glycerophosphate (beta-GP), whose gelation is induced by increasing temperature to body temperature, were functionalized by incorporation of alkaline phosphatase (ALP), an enzyme involved in mineralization of bone. ALP incorporation led to ac

  1. Protection against an Escherichia Coli-induced sepsis by alkaline phosphatase in mice

    NARCIS (Netherlands)

    Verweij, Willem; Bentala, Hafida; Huizenga-Van der Vlag, A; van Loenen - Weemaes, Anne-miek; Kooi, K.; Meijer, Dick; Poelstra, Klaas

    2004-01-01

    Alkaline phosphatase (AP) is a phosphate transferase present in bacteria and eukaryotes. In previous studies, we have shown that AP is able to dephosphorylate lipopolysaccharide (LPS) at physiological pH levels. Because LPS is the causative agent of gram-negative sepsis, we hypothesize that AP might

  2. 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

  3. Biological Apatite Formed from Polyphosphate and Alkaline Phosphatase May Exchange Oxygen Isotopes from Water through Carbonate

    Science.gov (United States)

    Omelon, S. J.; Stanley, S. Y.; Gorelikov, I.; Matsuura, N.

    2011-12-01

    The oxygen isotopic composition in bone mineral phosphate is known to reflect the local water composition, environmental humidity, and diet1. Once ingested, biochemical processes presumably equilibrate PO43- with "body water" by the many biochemical reactions involving PO43- 2. Blake et al. demonstrated that enzymatic release of PO43- from organophosphorus compounds, and microbial metabolism of dissolved orthophosphate, significantly exchange the oxygen in precipitated apatite within environmental water3,4, which otherwise does not exchange with water at low temperatures. One of the enzymes that can cleave phosphates from organic substrates is alkaline phosphastase5, the enzyme also associated with bone mineralization. The literature often states that the mineral in bone in hydroxylapatite, however the mineral in bone is carbonated apatite that also contains some fluoride6. Deprotonation of HPO32- occurs at pH 12, which is impossibly high for biological system, and the predominate carbonate species in solution at neutral pH is HCO3-. To produce an apatite mineral without a significant hydroxyl content, it is possible that apatite biomineralization occurs through a polyphosphate pathway, where the oxygen atom required to transform polyphosphate into individual phosphate ions is from carbonate: [PO3-]n + CO32- -> [PO3-]n-1 + PO43- + CO2. Alkaline phosphatase can depolymerise polyphosphate into orthophosphate5. If alkaline phosphatase cleaves an oxygen atom from a calcium-carbonate complex, then there is no requirement for removing a hydrogen atom from the HCO3- or HPO43- ions of body water to form bioapatite. A mix of 1 mL of 1 M calcium polyphosphate hydogel, or nano-particles of calcium polyphosphate, and amorphous calcium carbonate were reacted with alkaline phosphatase, and maintained at neutral to basic pH. After two weeks, carbonated apatite and other calcium phosphate minerals were identified by powder x-ray diffraction. Orthophosphate and unreacted

  4. Effects of sodium nitroprusside activity of acid and alkaline invertases and alkaline phosphatase in lemongrass (Cymbopogon flexuosus Steud Wats

    Directory of Open Access Journals (Sweden)

    Deepak Ganjewala

    2010-01-01

    Full Text Available Here we report the effects of SNP, a nitric oxide donor on sucrose metabolizing enzymes, acid and alkaline invertase (EC 3.2.1.26 and 3.2.1.153 and ubiquitous alkaline phosphatase (EC 3.1.3.1 in four lemongrass varieties viz., Krishna, Cauveri, Nima and Cheerharit. For the study, two 15 d lemongrass tillers were cut and immediately dipped into the test tubes containing SNP solution (5 mL of variable strength (1 to 5 mM and one without SNP (as control; kept for 4 h under mild sunlight. The results revealed that moderate SNP concentration (2 mM was most effective, caused drastic reduction (40% in protein content in var. Nima followed by Krishna (33%, Cauveri (17% and Cheerharit (12%. In contrast, SNP (1 mM has impressively enhanced protein content in all the lemongrass varieties. The SNP (2 mM markedly inhibited the activity of acid invertase by 38% in Cheerharit, 35% Nima and 28% Cauveri whereas and alkaline invertase by 21, 28 and 24% respectively in var. Cheerharit, Nima and Krishna. Similarly, SNP (5 mM severely inhibited (~ 63% the activity of the ALP in lemongrass var. Cauveri and Nima, 50% in Krishna and relatively less 23% in Cheerharit as compared to the control. However, in var. Nima, 50% loss in ALP activity had already been occurred after 2 mM SNP treatment. These results primarily suggests that NO interferes sucrose metabolism by anonymously hindering the activity of acid and alkaline invertase and ubiquitous alkaline phosphatase in lemongrasses.

  5. 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.

  6. 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

  7. Correlation Between Down Syndrome and the Level of Placental Alkaline Phosphatase in Amniotic Fluid

    OpenAIRE

    UÇAR, Canan; Balci, Sevim; TUNÇBİLEK, Ergül; ÜNSAL, İbrahim

    2005-01-01

    The aim of this study was to investigate the correlation between Down syndrome and the level of placental alkaline phosphatase (PLAP) in amniotic fluid. A total of 279 amniotic fluid samples taken between 14 and 24 gestational weeks were investigated in this study. Karyotype analysis was made in all samples. In 10 samples trisomy 21 was determined, in one sample trisomy 18, in two samples 47,XXY, in six samples various structural chromosomal anomalies and in 260 samples normal karyotype. PLA...

  8. Analytical validation of serum bone alkaline phosphatase (BAP OSTASE) on Liaison

    OpenAIRE

    Cavalier, Etienne; Rozet, Eric; Carlisi, Agnès; Bekaert, Anne-Catherine; ROUSSELLE, Olivier; Hubert, Philippe; Chapelle, Jean-Paul; Delanaye, Pierre

    2009-01-01

    Background: The goal of this study was to validate the DiaSorin Liaison BAP OSTASE, a new method for measurement of bone alkaline phosphatase (BAP), and to compare this method with the Beckman-Coulter Access Ostase. We also wanted to establish the reference range for BAP in adults and children. Methods: We determined the precision, functional sensitivity, recovery, linearity and measurement uncertainty, accuracy profile and β-expectation limits. We defined an adult reference interval using i...

  9. 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.

  10. 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 alkaline phosphatase in 18% and 7%. Neither the activity of gamma-glutamyltransferase, aspartate aminotransferase nor alkaline phosphatase showed any significant (P greater than 0.05) correlation with the history of alcohol consumption. The activities of gamma-glutamyltransferase and aspartate...... aminotransferase were raised significantly more often in patients with recent alcohol consumption than in patients who had abstained for more than 9 days. The concentration of alkaline phosphatase was not significantly (P greater than 0.05) different in these groups. The predictive value of raised and normal...

  11. Structural studies of human alkaline phosphatase in complex with strontium: Implication for its secondary effect in bones

    OpenAIRE

    Llinas, Paola; Masella, Michel; Stigbrand, Torgny; Ménez, André; Stura, Enrico A.; Le Du, Marie Hélène

    2006-01-01

    Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). Human alkaline phosphatases have four m...

  12. Genetic optimization of a bacteriophage-delivered alkaline phosphatase reporter to detect Escherichia coli.

    Science.gov (United States)

    Jackson, Angelyca A; Hinkley, Troy C; Talbert, Joey N; Nugen, Sam R; Sela, David A

    2016-10-01

    A large fraction of foodborne illnesses are linked to (∼46%) leafy green vegetables contaminated by pathogens harbored in agricultural water. To prevent this, accurate point-of-production detection tools are required to identify and quantify bacterial contaminants in produce before consumers are impacted. In this study, a proof-of-concept model was engineered for a phage-based Escherichia coli detection system. We engineered the coliphage T7 to express alkaline phosphatase (ALP) to serve as the signal for E. coli detection. Wild type phoA (T7ALP) and a dominant-active allele, phoA D153G D330N (T7ALP*) was inserted into the T7 genome, with engineered constructs selected by CRISPR-mediated cleavage of unaltered chromosomes and confirmed by PCR. Engineered phages and E. coli target cells were co-incubated for 16 hours to produce lysates with liberated ALP correlated with input cell concentrations. A colorimetric assay used p-nitrophenyl phosphate (pNPP) to demonstrate significant ALP production by T7ALP and T7ALP* compared to the vector control (T7EV) (p≤ 0.05). Furthermore, T7ALP* produced 2.5-fold more signal than T7ALP (p≤ 0.05) at pH 10. Due to the increase in signal for the modified ALP* allele, we assessed T7ALP* sensitivity in a dose-responsive manner. We observed 3-fold higher signal for target cell populations as low as ∼2 × 10(5) CFU mL(-1) (p≤ 0.05 vs. no-phage control). PMID:27412402

  13. 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.

  14. Diagnostic Utility of Heat Stable Alkaline Phosphatase in Hypertensive Disorders of Pregnancy

    Science.gov (United States)

    Abu Raghavan, Srinivasan; Ghosh, Seethesh; Basu, Sharbari; Ramasamy, Ramesh; Murugaiyan, Sathish Babu

    2014-01-01

    Background: Hypertensive disorders in pregnancy (HDP) complicate 3-10% of all pregnancies. Though there are several biochemical parameters which aid in predicting hypertension of pregnancy, human placental alkaline phosphatase (PLAP), synthesized in placenta during pregnancy by placental syncytiotrophoblast, assumes diagnostic relevance. The purpose of this study was to compare the total alkaline phosphatase (ALP) and heat stable placental alkaline phosphatase (PLAP) levels in the serum of normotensive and hypertensive disorders of pregnancy and to evaluate the clinical utility of ALP and PLAP as a reliable, sensitive, specific and economical biochemical marker of HDP. Materials and Methods: This was a case control study, carried out on pregnant women with hypertension, of south Indian population. Study included pregnant women, 60 patients with hypertension and 60 controls. Biochemical assays were carried out by the IFCC approved procedures based on spectrophotometric method and using fully automated random access chemistry analyser. Data was compared by using student t-test. ROC was drawn to find out optimum cut off for ALP, PLAP and PLAP/ALP ratio in HDP. Pearson’s correlation was performed to ascertain the association among markers. Results: Serum total ALP, PLAP and PLAP/ALP ratio levels were significantly higher in hypertensive pregnant women when compared to controls (p<0.05). There was significant correlation among ALP, PLAP and DBP. ROC analysis of ALP (169.5), PLAP (69) and PLAP/ALP (0.44) ratios showed optimum cut-offs in diagnosis of hypertension in pregnancy. Conclusion: Serum heat stable ALP isoenzyme and PLAP/ALP ratio could be useful adjuvant markers in diagnosis of HDP in association with other relevant and economically viable biochemical tests. PMID:25584211

  15. 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. PMID:26895537

  16. Effects of vasectomy on seminal plasma alkaline phosphatase in male alpacas (Vicugña pacos).

    Science.gov (United States)

    Pearson, L K; Campbell, A J; Sandoval, S; Tibary, A

    2013-12-01

    Azoospermia is a common finding in male alpacas which present for infertility. The challenge is to differentiate azoospermia of testicular origin from non-testicular origin. In several species, alkaline phosphatase (AP) concentrations in seminal plasma have been used as a diagnostic marker of contributions of the testis and epididymis to the ejaculate. The purpose of this study was to determine whether AP assay could differentiate testicular from non-testicular azoospermia in male alpacas. An experimental model of bilateral outflow obstruction (pre-scrotal vasectomy) was used in 22 male alpacas, aged 2-9 years. No reproductive history was available. Animals were submitted for electroejaculation (EE) under general anaesthesia and vasectomy performed. Five weeks later, animals were submitted for EE. Vasectomy was not successful in one animal, which was removed from analysis. AP levels were compared in seminal plasma in the pre- and post-vasectomy samples. The mean ± SEM concentration of AP in pre-vasectomy seminal plasma was 504.29 ± 166.45 U/l (range 10-2910); the post-vasectomy levels were 252.48 ± 81.77 U/l (range 0-1640; p = 0.06). In 71.4% of animals, AP levels decreased, varying from 18% to 100% reduction. Results of this study suggest that AP is not produced exclusively by the testis and epididymis in alpacas and that AP assay is not a valid diagnostic test for determination of origin of azoospermia; the gold standard for diagnosis of origin of azoospermia remains testicular biopsy. PMID:23790090

  17. Alkaline phosphatase activity in the subtropical ocean: insights from nutrient, dust and trace metal addition experiments

    Directory of Open Access Journals (Sweden)

    Claire eMahaffey

    2014-12-01

    Full Text Available Phosphorus is an essential nutrient for all life on earth. In the ocean, the most bioavailable form of phosphorus is inorganic phosphate, but in the extensive subtropical gyres, phosphate concentrations can be chronically low and limit primary productivity and nitrogen fixation. In these regions, organisms produce hydrolytic enzymes, such as alkaline phosphatase (AP, that enable them to utilize the more replete dissolved organic phosphorus (DOP pool to meet their cellular phosphorus demands. In this study, we synthesized data from 14 published studies and present our own findings from two research cruises (D326 and D361 in the eastern subtropical Atlantic to explore the relationship between AP activity (APA and nutrients, Saharan dust and trace metals. We found that below a threshold phosphate concentration of ~ 30 nM, APA increased with an inverse hyperbolic relationship with phosphate concentration. Meanwhile, DOP concentrations decreased with enhanced APA, indicating utilization of the DOP pool. We found APA rates were significantly higher in the subtropical Atlantic compared to the subtropical Pacific Ocean, even over the same low phosphate concentration range (0 to 50 nM. While the phosphate concentration may have a first order control on the APA rates, we speculate that other factors influence this basin scale contrast. Using bioassay experiments, we show that the addition of Saharan dust and zinc significantly increased the rate of APA. To our knowledge, our results are the first direct field-based evidence that APA is limited by zinc in the subtropical ocean. Further work is required to explore the relationship between trace metals such as iron and zinc, which are co-factors of phosphohydrolytic enzymes, specifically PhoX and PhoA, respectively, and APA in the ocean.

  18. 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.

  19. Evolution of osteocalcin, alkaline phosphatase and parathyroid hormone after parathyroidectomy in patients receiving chronic maintenance hemodialysis.

    Science.gov (United States)

    Peretz, A M; Dhaene, M M; Drowart, A; Kinnaert, P; Vertongen, F M; Bourdoux, P P

    1992-03-01

    Parathyroid hormone (PTH), osteocalcin and alkaline phosphatase (AP) were investigated before and after parathyroidectomy in 12 patients receiving hemodialysis. Early post-parathyroidectomy, PTH decreased (p less than 0.001), AP increased (p less than 0.05), but osteocalcin remained unchanged. At 3 months, osteocalcin and AP declined. A negative correlation was observed between aluminum staining and post-parathyroidectomy osteocalcin. In conclusion, early post-parathyroidectomy, osteocalcin and AP reflect persistent osteoblastic activity, which declined after 3 months. In patients receiving hemodialysis both variables may represent different aspects of osteoblastic activity and osteocalcin allows mixed uremic osteodystrophy after parathyroidectomy. PMID:1578455

  20. 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.

  1. Serial serum alkaline phosphatase as an early biomarker for osteopenia of prematurity.

    Science.gov (United States)

    Abdallah, Enas A A; Said, Reem N; Mosallam, Dalia S; Moawad, Eman M I; Kamal, Naglaa M; Fathallah, Mohammed G E-D

    2016-09-01

    Metabolic bone disease of prematurity is a condition characterized by reduction in bone mineral content (osteopenia). It is a problem faced by very low birth weight (VLBW) infants because of lack of fetal mineralization during the last trimester. Our aim was to assess serum alkaline phosphatase (ALP) level as an early biomarker for osteopenia in premature infants and to estimate an optimal cutoff value of serum ALP at which osteopenia is detected radiologically in premature newborns.This prospective study was conducted on a cohort of 120 newborn infants of both sex of ≤34 weeks' gestational age and premature infants particularly those <1000-g birth weight and <32 weeks' gestation. PMID:27631238

  2. Properties of a constitutive alkaline phosphatase from strain 74A of the mold Neurospora crassa

    OpenAIRE

    Morales A.C.; Nozawa S.R.; Thedei Jr. G.; Maccheroni Jr. W.; De Rossi A.

    2000-01-01

    A constitutive alkaline phosphatase was purified to apparent homogeneity as determined by polyacrylamide gel electrophoresis from mycelia of the wild strain 74A of the mold Neurospora crassa, after growth on acetate and in the presence of saturating amounts of inorganic phosphate (Pi) for 72 h at 30ºC. The molecular mass was 58 kDa and 56 kDa as determined by exclusion chromatography and SDS-PAGE, respectively. This monomeric enzyme shows an apparent optimum pH ranging from 9.5 to 10.5 and Mi...

  3. 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.

  4. 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.

  5. Temperature dependence of the absorbance of alkaline solutions of 4-nitrophenyl phosphate--a potential source of error in the measurement of alkaline phosphatase activity.

    Science.gov (United States)

    Burtis, C A; Seibert, L E; Baird, M A; Sampson, E J

    1977-09-01

    The absorbance of an alkaline solution of 4-nitrophenyl phosphate is a function of temperature. Quantitative evaluation of this phenomenon indicates that it (a) depends on the concentration of the compound and is independent of source, buffer concentration, and pH above 9.0; (b) is reversible; (c) is not a result of alkaline hydrolysis or 4-nitrophenol contamination; and (d) correlates with a temperature-induced shift of its absorbance spectrum. The phenomenon may represent a potential analytical problem in methods for alkaline phosphatase in which this compound is the substrate. If thermal equilibrium is not reached and maintained during an alkaline phosphatase assay, the thermochromic response will be included in the measured rate. The magnitude of this error depends on the thermal response and control characteristics of each particular instrument and the reaction conditions under which such an analysis is performed. PMID:19164

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

    Institute of Scientific and Technical Information of China (English)

    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.

  7. Alkaline phosphatase in osteoblasts is down-regulated by pulsatile fluid flow

    Science.gov (United States)

    Hillsley, M. V.; Frangos, J. A.

    1997-01-01

    It is our hypothesis that interstitial fluid flow plays a role in the bone remodeling response to mechanical loading. The fluid flow-induced expression of three proteins (collagen, osteopontin, and alkaline phosphatase) involved in bone remodeling was investigated. Rat calvarial osteoblasts subjected to pulsatile fluid flow at an average shear stress of 5 dyne/cm2 showed decreased alkaline phosphatase (AP) mRNA expression after only 1 hour of flow. After 3 hours of flow, AP mRNA levels had decreased to 30% of stationary control levels and remained at this level for an additional 5 hours of flow. Steady flow (4 dyne/cm2 fluid shear stress), in contrast, resulted in a delayed and less dramatic decrease in AP mRNA expression to 63% of control levels after 8 hours of flow. The reduced AP mRNA expression under pulsatile flow conditions was followed by reduced AP enzyme activity after 24 hours. No changes in collagen or osteopontin mRNA expression were detected over 8 hours of pulsatile flow. This is the first time fluid flow has been shown to affect gene expression in osteoblasts.

  8. Expression of a human placental alkaline phosphatase gene in transfected cells: Use as a reporter for studies of gene expression

    International Nuclear Information System (INIS)

    The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is appropriate for use as a reporter in studies of gene regulation since its expression is dependent on the presence of exogenous transcription control elements. The overall assay to detect the expression of the gene is quantitative, very rapid, and inexpensive. Cotransfections of cells with pSV2Apap and a related plasmid carrying the bacterial chloramphenicol acetyltransferase gene (pSV2Acat) indicate that transcription of these two genes is detected with roughly the same sensitivity

  9. X-ray structure reveals a new class and provides insight into evolution of alkaline phosphatases.

    Directory of Open Access Journals (Sweden)

    Subhash C Bihani

    Full Text Available The alkaline phosphatase (AP is a bi-metalloenzyme of potential applications in biotechnology and bioremediation, in which phosphate monoesters are nonspecifically hydrolysed under alkaline conditions to yield inorganic phosphate. The hydrolysis occurs through an enzyme intermediate in which the catalytic residue is phosphorylated. The reaction, which also requires a third metal ion, is proposed to proceed through a mechanism of in-line displacement involving a trigonal bipyramidal transition state. Stabilizing the transition state by bidentate hydrogen bonding has been suggested to be the reason for conservation of an arginine residue in the active site. We report here the first crystal structure of alkaline phosphatase purified from the bacterium Sphingomonas. sp. Strain BSAR-1 (SPAP. The crystal structure reveals many differences from other APs: 1 the catalytic residue is a threonine instead of serine, 2 there is no third metal ion binding pocket, and 3 the arginine residue forming bidentate hydrogen bonding is deleted in SPAP. A lysine and an aspargine residue, recruited together for the first time into the active site, bind the substrate phosphoryl group in a manner not observed before in any other AP. These and other structural features suggest that SPAP represents a new class of APs. Because of its direct contact with the substrate phosphoryl group, the lysine residue is proposed to play a significant role in catalysis. The structure is consistent with a mechanism of in-line displacement via a trigonal bipyramidal transition state. The structure provides important insights into evolutionary relationships between members of AP superfamily.

  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. 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+}.

  12. 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.

  13. 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

  14. 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.``

  15. Dietary free fatty acids form alkaline phosphatase-enriched microdomains in the intestinal brush border membrane

    DEFF Research Database (Denmark)

    Hansen, Gert H; Rasmussen, Karina; Niels-Christiansen, Lise-Lotte;

    2011-01-01

    mimicking a physiological solution of dietary mixed micelles, rearranged the lipid raft microdomain organization of the membranes. Thus, the fat mixture generated a low-density subpopulation of microvillar detergent resistant membranes (DRMs) highly enriched in alkaline phosphatase (AP). Since this GPI......-linked enzyme is the membrane protein in the brush border with the highest affinity for lipid rafts, this implies that free fatty acids selectively insert stably into these membrane microdomains. We have previously shown that absorption of dietary lipids transiently induce a selective endocytosis of AP from...... the brush border, and from work by others it is known that fat absorption is accompanied by a rise in serum AP and secretion of surfactant-like particles from enterocytes. We propose that these physiological processes may be triggered by the sequestering of dietary free fatty acids in lipid raft...

  16. Ionizing Groups at the Active Site of the Alkaline Phosphatase from Ostrea cucullate

    Institute of Scientific and Technical Information of China (English)

    CHEN Qiao; WANG Qin; LIAO Jinhua; CHEN Qingxi

    2006-01-01

    The ionizing groups at the active site of alkaline phosphatase (ALP, EC 3.1.3.1) from Ostrea cucullate were studied using kinetic methods. The ionization constant, pKe, of the ionizing groups at the active site of the enzyme was found to be 10.11 at 37.0℃ and the organic solvent, dioxin, had no effect on the pKe. The standard dissociation enthalpy (△Ho) was determined to be 10.57 kcal/mol (1 cal=4.18 J). The results show that the dissociation group of the enzyme active center is the з-NH3+ of lysine. Chemical modification of the enzyme by acetic anhydride and succinic anhydride demonstrates that the amino group is one of the enzyme's functional groups.

  17. Fluoride stimulates [3H]thymidine incorporation and alkaline phosphatase production by human osteoblasts

    International Nuclear Information System (INIS)

    The effect of sodium fluoride on alkaline phosphatase (ALP) release and [3H]thymidine uptake by human osteoblasts in culture was investigated. Sodium fluoride stimulated both ALP release and [3H]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. Diagnostic evaluation of canine serum alkaline phosphatase by immunochemical means and interpretation of results.

    Science.gov (United States)

    Saini, P K; Peavy, G M; Hauser, D E; Saini, S K

    1978-09-01

    Sera of several canine patients contained an isoenzyme of alkaline phosphatase (ALP) that resembled intestinal ALP with respect to heat inactivation, L-phenylalanine inhibition, and sensitivity to anti-canine intestinal ALP antibody, but differed with regard to the electrophoretic migration. The electrophoretic mobility of the isoenzyme was slightly cathodal than that of hepatic ALP, and its migration was reduced, similar to that of hepatic isoenzyme after neuraminidase treatment. This isoenzyme, which could be corticosteroid induced, was in the sera of numerous dogs with hepatobiliary disorders and was different from the hepatic isoenzyme that appeared in the sera of dogs with acute hepatitis, based on anti-canine intestinal ALP antibody interaction, heat inactivation, and electrophoretic migration. PMID:358873

  19. 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.

  20. 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…

  1. The Effects of Two Species of Daphne, Betulin and Betulinic Acid on Alkaline Phosphatase Activity in Two Human Cancer Cell lines, K562 and MCF-7

    Directory of Open Access Journals (Sweden)

    E Panahi Kokhdan

    2014-02-01

    Full Text Available Abstract Background & aim: Changes of alkaline phosphatase activity is one of the symptoms of many diseases. The aim of this study was to evaluate the effect of two types of Daphne, Betulin and Betulinic acid, on alkaline phosphatase activity in K562 and MCF-7 cell lines, respectively. Methods: In this study, 106 cancer cell lines of K562 and MCF-7 were cultured in presence of 5% carbon dioxide at 37 ° C. at doses near the IC50. The viability of cells, inside and outside alkaline phosphatase activity and the amount of total protein in each treatment were studied. The collected data was analyzed with a multivariate analysis of variance (Nested Design and Dunnett test. Results: The intracellular alkaline phosphatase activity of the cells showed different behavior compared to the extracellular alkaline phosphatase activity (p< 0.01. The highest increase of alkaline phosphatase activity in two cell lines (K562 and MCF-7 were 339% and 236% which was related to the treatment by macronata daphne. Conclusion: Unexpected increase in intracellular alkaline phosphatase activity in D. mucronata, D. oleides, Betulin, and Betulinic acid treatment may be due to changes in the composition of plasma membrane component and an increase the non-connected membrane of the protein which is due to the creation of more active proteins. Keywords: Daphne mucronata, Daphne oleoides, Alkaline Phosphatase, Betulinic Acid, Betulin

  2. 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.

  3. 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…

  4. Treatment with bortezomib in multiple myeloma is associated with only a transient and brief increase of bone specific alkaline phosphatase

    DEFF Research Database (Denmark)

    Haidl, Felix; Plesner, Torben; Lund, Thomas

    2012-01-01

    There are indications of a bone anabolic effect associated with bortezomib treatment. We present a study with long follow up, measuring bone specific alkaline phosphatase (bALP) for a year during and after treatment in an unselected cohort of myeloma patients treated with bortezomib, and assess...

  5. Production and characterization of a single-chain variable fragment linked alkaline phosphatase fusion protein for detection of O,O-diethyl organophosphorus pesticides in a one-step enzyme-linked immunosorbent assay

    Science.gov (United States)

    A single-chain variable fragment (scFv) and alkaline phosphatase (AP) fusion protein for detection of O, O-diethyl organophosphorus pesticides (OPs) was produced and characterized. The scFv gene was prepared by cloning VL and VH genes from a hybridoma cell secreting monoclonal antibody with broad-s...

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. Salivary alkaline phosphatase levels speak about association of smoking, diabetes and potentially malignant diseases???

    Directory of Open Access Journals (Sweden)

    A Ravi Prakash

    2016-01-01

    Full Text Available Background: Alkaline phosphatase (ALP is a hydrolase intracellular enzyme participating in the metabolic processes of cells. Rise in salivary ALP (S-ALP levels reflects inflammation and destruction of healthy tissues suggesting it as a clinical biomarker. S-ALP is used in analyzing the severity of the disease occurrence in smokers and nonsmokers who are diabetic and nondiabetic. S-ALP levels are analyzed using autoanalyzer in 40 patients who visited our department. Aims and Objectives: To determine the levels of S-ALP in diagnosing potentially malignant conditions and debilitating diseases in early stages of inflammation and altered cellular metabolism. Materials and Methods: The study groups include: (1 Group A - 10 smokers who are diabetic. (2 Group B - 10 smokers who are nondiabetic. (3 Group C - 10 nonsmokers who are diabetic. (4 Group D - 10 nonsmokers and nondiabetic as control. Unstimulated saliva samples are collected and run in auto-analyzer with ALP enzyme reagent to analyze ALP levels. Comparison is made between all the four groups. Results: Results were statistically significant with increased activity of ALP levels in saliva from Group A when compared to Group D. The results are Group A > Group B > Group C > Group D. The results also revealed significant raise in levels of ALP levels in saliva from smokers when compared to diabetes. Thus explaining adverse effects of smoking. Conclusion: S-ALP can be considered to be the biomarker for evaluating adverse effects of smoking, diabetes and other debilitating diseases in early stages.

  11. Correlation of alkaline phosphatase activity to clinical parameters of inflammation in smokers suffering from chronic periodontitis

    Science.gov (United States)

    Grover, Vishakha; Malhotra, Ranjan; Kapoor, Anoop; Bither, Rupika; Sachdeva, Sonia

    2016-01-01

    Context: Current clinical periodontal diagnostic techniques emphasize the assessment of clinical and radiographic signs of periodontal diseases which can provide a measure of history of disease. Hence, new methodologies for early identification and determination of periodontal disease activity need to be explored which will eventually result in expedited treatment. Aim: To evaluate the correlation of alkaline phosphatase (ALP) activity in gingival crevicular fluid (GCF) to clinical parameters of periodontal inflammation in smokers with chronic periodontitis. Materials and Methods: Study population included 15 smoker male patients in the age group of 35–55 years suffering from moderate generalized chronic periodontitis with history of smoking present. Following parameters were evaluated at baseline, 1 month and 3 months after scaling and root planing: plaque index, bleeding index, probing pocket depth (PD), relative attachment level (RAL), and GCF ALP activity. Statistical Analysis Used: Independent variables for measurements over time were analyzed by using Wilcoxon signed rank test. Results: A statistically significant reduction in all the clinical parameters and GCF ALP activity was observed from baseline to 1 month and 3 months. A correlation was observed between change in GCF ALP activity and PD reduction as well as gain in RAL at 3 months. Conclusion: The present study emphasizes that total ALP activity could be used as a marker for periodontal disease activity in smokers. Estimation of changes in the levels of this enzyme has a potential to aid in the detection of progression of periodontal disease and monitoring the response to periodontal therapy. PMID:27563197

  12. 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.

  13. 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.

  14. Biodistribution and translational pharmacokinetic modeling of a human recombinant alkaline phosphatase.

    Science.gov (United States)

    Peters, Esther; Stevens, Jasper; Arend, Jacques; Guan, Zheng; Raaben, Willem; Laverman, Peter; van Elsas, Andrea; Masereeuw, Rosalinde; Pickkers, Peter

    2015-11-10

    Clinical trials showed renal protective effects of bovine intestinal alkaline phosphatase (AP) in patients with sepsis-associated acute kidney injury (AKI). Subsequently, a human recombinant chimeric AP (recAP) was developed as a pharmaceutically acceptable alternative. Here, we investigated the biodistribution and pharmacokinetics (PK) of recAP and developed a translational population PK model. Biodistribution was studied during LPS-induced AKI in rats. Iodine-125-labeled recAP was primarily taken up by liver, spleen, adrenals, heart, lungs and kidneys followed by the gastro-intestinal tract and thyroid. Tissue distribution was not critically affected by endotoxemia. PK parameters were determined in rats and minipigs during IV bolus injections of recAP, administered once, or once daily during seven consecutive days. Plasma concentrations of recAP increased with increasing dose and disappeared in a biphasic manner. Exposure to recAP, estimated by AUC and Cmax, was similar on days 1 and 7. Subsequently, population approach nonlinear mixed effects modeling was performed with recAP rat and minipig and biAP phase I PK data. Concentration versus time data was accurately described in all species by a two-compartmental model with allometric scaling based on body weight. This model provides a solid foundation for determining the optimal dose and duration of first-in-man recAP studies. PMID:26325308

  15. Invited review: The application of alkaline phosphatase assays for the validation of milk product pasteurization.

    Science.gov (United States)

    Rankin, S A; Christiansen, A; Lee, W; Banavara, D S; Lopez-Hernandez, A

    2010-12-01

    Standard practices for indirectly assessing the pasteurization status of milk products are primarily based on the thermal inactivation kinetics of the endogenous milk enzyme, alkaline phosphatase (ALP). This assessment provides an invaluable, if not required, tool for both regulatory and in-house process control and validation. Endogenous milk ALP manifests a slightly higher heat resistance than the pathogenic microflora upon which pasteurization time and temperature requirements are based. Hence, ALP activity is recognized and accepted as the method of choice for the rapid validation of milk product pasteurization. However, ALP assays have notable limitations that must be understood if they are to be administered and interpreted correctly and the results are to be applied judiciously. Issues such as the reactivation of heat-denatured ALP and the presence of both heat-stable and -labile microbial ALP are addressed. A discussion of ALP in the milk of nonbovine species is presented based on the limited literature available. Some discussion of research involving alternative pasteurization indicators also is presented. This article is intended to summarize the pertinent details of the ALP assay for dairy products (noting the basis and limitations of various methods) and the processing, handling, and known compositional factors that influence the assay results.

  16. 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.

  17. 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.

  18. Correlation of alkaline phosphatase activity to clinical parameters of inflammation in smokers suffering from chronic periodontitis

    Directory of Open Access Journals (Sweden)

    Vishakha Grover

    2016-01-01

    Full Text Available Context: Current clinical periodontal diagnostic techniques emphasize the assessment of clinical and radiographic signs of periodontal diseases which can provide a measure of history of disease. Hence, new methodologies for early identification and determination of periodontal disease activity need to be explored which will eventually result in expedited treatment. Aim: To evaluate the correlation of alkaline phosphatase (ALP activity in gingival crevicular fluid (GCF to clinical parameters of periodontal inflammation in smokers with chronic periodontitis. Materials and Methods: Study population included 15 smoker male patients in the age group of 35–55 years suffering from moderate generalized chronic periodontitis with history of smoking present. Following parameters were evaluated at baseline, 1 month and 3 months after scaling and root planing: plaque index, bleeding index, probing pocket depth (PD, relative attachment level (RAL, and GCF ALP activity. Statistical Analysis Used: Independent variables for measurements over time were analyzed by using Wilcoxon signed rank test. Results: A statistically significant reduction in all the clinical parameters and GCF ALP activity was observed from baseline to 1 month and 3 months. A correlation was observed between change in GCF ALP activity and PD reduction as well as gain in RAL at 3 months. Conclusion: The present study emphasizes that total ALP activity could be used as a marker for periodontal disease activity in smokers. Estimation of changes in the levels of this enzyme has a potential to aid in the detection of progression of periodontal disease and monitoring the response to periodontal therapy.

  19. Kinetics of gene expression of alkaline phosphatase during healing of alveolar bone in rats.

    Science.gov (United States)

    Rodrigues, Willian Caetano; Fabris, André Luís da Silva; Hassumi, Jaqueline Suemi; Gonçalves, Alaíde; Sonoda, Celso Koogi; Okamoto, Roberta

    2016-06-01

    Immunohistochemical studies and molecular biology have enabled us to identify numerous proteins that are involved in the metabolism of bone, and their encoding genes. Among these is alkaline phosphatase (ALP), an enzyme that is responsible for the initiation of mineralisation of the extracellular matrix during alveolar bone repair. To evaluate the gene expression of ALP during this process, we studied nine healthy adult male rats, which had their maxillary central incisors extracted from the right side and were randomly divided into three groups. During three experimental periods, 7 days, 14 days, and 28 days, the alveoli were curetted, the rats killed, and samples analysed by real-time reverse transcription polymerase chain reaction (qRT-PCR). The RNAm that encodes the gene for the synthesis of ALP was expressed during the three periods analysed, but its concentration was significantly increased at 14 and 28 days compared with at 7 days. There was no significant difference between 14 and 28 days (p=0.0005). We conclude that genes related to ALP are expressed throughout the healing process and more intensively during the later periods (14 and 28 days), which coincides with the increased formation of mineralised bone. PMID:26935214

  20. 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.

  1. 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

    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 the en......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 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...

  2. Advances on Characteristic and Application of Alkaline Phosphatase%碱性磷酸酶特性及其应用的研究进展

    Institute of Scientific and Technical Information of China (English)

    王秋颖

    2011-01-01

    Alkaline phosphatase(AP) is a kind of hydrolase,which catalyzes the nonspecific hydrolysis of phosphate monoester to produce phosphate or transfer phosphate. AP exists widely in microbial and animal kingdom. AP plays a vital important role in phosphate cycle of world creatures, and has been used widely in the fields of diagnostics, biochemistry and molecular biology. In this paper,the history, the reaction mechanism and application of alkaline phosphatase was reviewed.%碱性磷酸酶(alkaline pllosplaatase,AP)是一种非特异性磷酸单酯酶,能催化磷酸单酯的水解反应,产生无机磷酸和相应的醇、酚或糖,也能催化磷酸基团的转移反应.AP广泛存在于微生物和动物体内,在磷生物地球化学循环过程中有重要作用,并广泛应用于诊断学、生物化学及分子生物学等领域.作者对碱性磷酸酶的研究历史、催化作用机制及其应用进行了综述.

  3. Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.

    OpenAIRE

    Temelli Ozlem; Ozhavzali MĂźzeyyen; Atay Meral; OÄuztĂźzĂźn Serpil; Yirtici Umit; TĂźrk Mustafa; Atay Ziya

    2009-01-01

    Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP), Cytokeratin7 (CK7) Cytokeratin8 (CK8) to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinom...

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

    OpenAIRE

    Malo, Madhu S.

    2015-01-01

    Highlights • Diabetes patients have very low levels of intestinal alkaline phosphatase (IAP) in their stool. • Obese people with high IAP do not develop diabetes. • A low level of IAP in a healthy person is indicative of having the incipient (latent) metabolic syndrome (diabetes). • Approximately 65% of the healthy population have the incipient metabolic syndrome including incipient diabetes. • ‘Temporal IAP profiling’ should diagnose the incipient metabolic syndrome (diabetes). Currently, di...

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

    OpenAIRE

    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 (A...

  6. Structural studies of human alkaline phosphatase in complex with strontium: Implication for its secondary effect in bones

    Science.gov (United States)

    Llinas, Paola; Masella, Michel; Stigbrand, Torgny; Ménez, André; Stura, Enrico A.; Le Du, Marie Hélène

    2006-01-01

    Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). Human alkaline phosphatases have four metal binding sites—two for zinc, one for magnesium, and one for calcium ion—that can be substituted by strontium. Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. Since calcium helps stabilize a large area that includes loops 210–228 and 250–297, its substitution by strontium could affect the stability of this region. Energy calculations suggest that only at high doses of strontium, comparable to those found for calcium, can strontium substitute for calcium. Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. PMID:16815919

  7. A Toxin-Binding Alkaline Phosphatase Fragment Synergizes Bt Toxin Cry1Ac against Susceptible and Resistant Helicoverpa armigera

    OpenAIRE

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

    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 p...

  8. Alkaline phosphatase and OmpA protein can be translocated posttranslationally into membrane vesicles of Escherichia coli.

    OpenAIRE

    Chen, L.; Rhoads, D; Tai, P C

    1985-01-01

    We previously described a system for translocating the periplasmic enzyme alkaline phosphatase and the outer membrane protein OmpA into inverted membrane vesicles of Escherichia coli. We have now optimized and substantially improved the translocation system by including polyamines and by reducing the amount of membrane used. Under these conditions, efficient translocation was seen even posttranslationally, i.e., when vesicles were not added until after protein synthesis was stopped. This was ...

  9. Energy-requiring translocation of the OmpA protein and alkaline phosphatase of Escherichia coli into inner membrane vesicles.

    OpenAIRE

    Rhoads, D B; Tai, P C; Davis, B D

    1984-01-01

    In developing a reliable in vitro system for translocating bacterial proteins, we found that the least dense subfraction of the membrane of Escherichia coli was superior to the total inner membrane, both for a secreted protein (alkaline phosphatase) and for an outer membrane protein (OmpA). Compounds that eliminated the proton motive force inhibited translocation, as already observed in cells; since protein synthesis continued, the energy for translocation appears to be derived from the energ...

  10. 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.

  11. 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. PMID:25885820

  12. 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]...

  13. 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.

  14. 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.

  15. [Influences of uncommon isoenzymes on determination of alkaline phosphatase activity by dry-chemistry analyzers].

    Science.gov (United States)

    Tozawa, T; Hashimoto, M

    2001-04-01

    Dry-chemistry(DC) analysis may be influenced by some matrix effects for measuring uncommon isoenzyme forms. Placental and intestinal alkaline phosphatase(AP) are overestimated by the VITROS DC, compared with results obtained with the wet-chemistry(WC) method of Bretaudiere, et al. using 2-amino-2-methyl-1-propanol (AMP) buffer, however, no such discrepancy between AP results in any DC method and that with a routine WC method recommended by Japanese Society of Clinical Chemistry in that 2-ethylaminoethanol(EAE) buffer is used, has been demonstrated. The type of buffer used affects differently the rates of AP isoenzymes activities. We therefore examined whether the presence of uncommon AP isoenzyme forms in serum caused aberrant DC results for AP in comparison with a routine WC method using EAE buffer. Here, serum samples with only liver AP and bone AP(n : 32); high-molecular-mass AP(n : 11); placental AP(n : 12); intestinal AP(n : 13) and immunoglobulin (Ig) bound AP(n : 12) were analyzed for total AP activity on three different DC analyzers: VITROS 700XR, FUJIDRYCHEM 5000, SPOTCHEM 4410 and a WC analyzer: HITACHI 7350. Values obtained in all of the DCs for sera containing only liver/bone AP agreed with those with the WC method. For sera containing placental AP, the VITROS values were higher than those with the WC method, while the FUJIDRYCHEM values and the SPOTCHEM values were lower. The VITROS values and the FUJIDRYCHEM values for sera containing intestinal AP were lower than those with the WC method, while the SPOTCHEM values were higher. All of the DCs did not affect high-molecular-mass AP and Ig bound liver/bone AP types of macro AP, but underestimated Ig bound intestinal type. Ig bound intestinal AP may be sieved by DC multilayer elements. PMID:11391954

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. Properties of a constitutive alkaline phosphatase from strain 74A of the mold Neurospora crassa

    Directory of Open Access Journals (Sweden)

    Morales A.C.

    2000-01-01

    Full Text Available A constitutive alkaline phosphatase was purified to apparent homogeneity as determined by polyacrylamide gel electrophoresis from mycelia of the wild strain 74A of the mold Neurospora crassa, after growth on acetate and in the presence of saturating amounts of inorganic phosphate (Pi for 72 h at 30ºC. The molecular mass was 58 kDa and 56 kDa as determined by exclusion chromatography and SDS-PAGE, respectively. This monomeric enzyme shows an apparent optimum pH ranging from 9.5 to 10.5 and Michaelis kinetics for the hydrolysis of p-nitrophenyl phosphate (the Km and Hill coefficient values were 0.35 mM and 1.01, respectively, alpha-naphthyl phosphate (the Km and Hill coefficient values were 0.44 mM and 0.97, respectively, ß-glycerol phosphate (the Km and Hill coefficient values were 2.46 mM and 1.01, respectively and L-histidinol phosphate (the Km and Hill coefficient values were 0.47 mM and 0.94, respectively at pH 8.9. The purified enzyme is activated by Mg2+, Zn2+ and Tris-HCl buffer, and is inhibited by Be2+, histidine and EDTA. Also, 0.3 M Tris-HCl buffer protected the purified enzyme against heat inactivation at 70ºC(half-life of 19.0 min, k = 0.036 min-1 as compared to 0.3 M CHES (half-life of 2.3 min, k = 0.392 min-1 in the same experiment.

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

    International Nuclear Information System (INIS)

    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

  2. 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.

  3. Kidney bone disease and mortality in CKD: revisiting the role of vitamin D, calcimimetics, alkaline phosphatase, and minerals.

    Science.gov (United States)

    Kalantar-Zadeh, Kamyar; Shah, Anuja; Duong, Uyen; Hechter, Rulin C; Dukkipati, Ramanath; Kovesdy, Csaba P

    2010-08-01

    Recent evidence suggests that the traditional syndromes known as renal osteodystrophy, secondary hyperparathyroidism, and vitamin D deficiency are related to mortality in persons with moderate to advanced chronic kidney disease (CKD). The so-called 'kidney bone disease', also known as 'mineral and bone disorders', is defined to include bone disorders, mineral disarrays, and vascular calcification. We have identified 14 common and clinically relevant conditions of contemporary nature that are related to the kidney bone disease, including calcitriol (active vitamin D) deficiency, 25(OH)-vitamin D deficiency, biochemical hyperparathyroidism, relatively low parathyroid hormone (PTH) level, increased serum alkaline phosphatase (hyperphosphatasemia), elevated fibroblast growth factor (FGF)-23, high turnover bone disease, adynamic bone disease, uremic osteoporosis, vascular calcification, hyper- and hypophosphatemia, and hyper- and hypocalcemia. We present a critical review of these 14 conditions with emphasis on patient survival and other pertinent clinical outcomes. We also review unresolved controversies surrounding the management of these conditions by administration of nutritional vitamin D (ergocalciferol and cholecalciferol), vitamin D receptor activators (calcitriol, alphacalcidiol, doxercalciferol), D-mimetics (paricalcitol, maxacalcitol), calcimimetics (cinacalcet), recombinant PTH (teriparatide), and receptor activator of nuclear factor-kappaB ligand modulators (denosumab); compare mortality predictability of PTH and alkaline phosphatase; and examine potential risks of bone disorders and mineral disarrays in CKD patients. PMID:20671739

  4. Ultrasensitive electrochemical DNAzyme sensor for lead ion based on cleavage-induced template-independent polymerization and alkaline phosphatase amplification.

    Science.gov (United States)

    Liu, Shufeng; Wei, Wenji; Sun, Xinya; Wang, Li

    2016-09-15

    In this article, a simple, highly sensitive and selective electrochemical DNAzyme sensor for Pb(2+) was developed on the basis of a 8-17 DNAzyme cleavage-induced template-independent polymerization and alkaline phosphatase amplification strategy. The hairpin-like substrate strand (HP DNA) of 8-17 DNAzyme was firstly immobilized onto the electrode. In the presence of Pb(2+) and the catalytic strand of 8-17 DNAzyme, the HP DNA could be cleaved to expose the free 3'-OH terminal, which could be then utilized for the cascade operation by terminal deoxynucleotidyl transferase (TdTase) for the base extension to incorporate biotinylated dUTP (dUTP-biotin). The further conjugated streptavidin-labeled alkaline phosphatase (SA-ALP) then catalyzed conversion of electrochemically inactive 1-naphthyl phosphate (1-NP) for the generation of electrochemical response signal. The currently fabricated Pb(2+) sensor effectively combines triply cascade amplification effects including cyclic Pb(2+)-dependent DNAzyme cleavage, TdTase-mediated base extension and enzymatic catalysis of ALP. An impressive detection limit of 0.043nM toward Pb(2+) with an excellent selectivity could be ultimately obtained, which was superior than most of the electrochemical methods. Thus, the developed amplification strategy opens a promising avenue for the detection of metal ions and may extend for the detection of other nucleic acid-related analytes. PMID:27093488

  5. Ultrasensitive detection of amifostine and alkaline phosphatase based on the growth of CdS quantum dots.

    Science.gov (United States)

    Na, Weidan; Liu, Siyu; Liu, Xiaotong; Su, Xingguang

    2015-11-01

    In this study, we reported a simple and sensitive fluorescence nanosensor for rapid detection of amifostine and alkaline phosphatase (ALP). The novel nanosensor was based on the fluorescence "turn on-off" of CdS quantum dots (QDs). Firstly, Cd(2+) cation could react with S(2-) anion to generate fluorescent CdS QDs in the presence of amifostine. The fluorescence (FL) intensity of amifostine-capped CdS QDs (Amifostine-CdS QDs) was increased with the increasing amounts of amifostine, and could be used for amifostine detection. However, amifostine could be converted to 2-(3-aminopropylamino) ethanethiol (WR1065) in the presence of ALP based on the dephosphorylation of ALP. Under the optimum conditions, the affinity of WR1065 to CdS QDs was weaker than that of amifostine. Therefore the new generation of WR1065-CdS QDs would reduce the FL intensity with the increase of ALP concentration, and the fluorescence of CdS QDs was turn off. The metabolic process of amifostine in the presence of alkaline phosphatase could be also studied via the change of FL intensity of CdS QDs. The present method was cost-effective, convenient, and does not require any complicated synthetic procedures. PMID:26452927

  6. Reduced L/B/K alkaline phosphatase gene expression in renal cell carcinoma: plausible role in tumorigenesis.

    Science.gov (United States)

    Sharma, Ujjawal; Pal, Deeksha; Singh, Shrawan Kumar; Kakkar, Nandita; Prasad, Rajendra

    2014-09-01

    Renal cell carcinoma (RCC) is the most common kidney cancer in adults. Although several genes have been found to be involved in carcinogenesis of RCC, more great efforts are needed to identify new genes which are responsible for the process. Clear cell RCC, originates from proximal tubule cells, is the most common pathological type of RCC. Alkaline phosphatase (ALP) is a marker enzyme of brush border membrane of proximal tubular cells. Our previous studies showed a significant decreased activity of Liver/Bone/Kidney (L/B/K) alkaline phosphatase in RCC. In the present study, we explored the molecular basis of the decreased activity of ALP in RCC. Immunohistochemistry, immunofluorescence and flow cytometry analysis showed decreased ALP protein in RCC. Additionally, real time PCR documented significantly reduced ALP gene expression (P = 0.009). Moreover, RCC cell lines (ACHN and A498) transfected with full length L/B/K cDNA showed decreased migratory property as well as viability of these cells as compared with controls (P = 0.000). Further, L/B/K ALP cDNA transfected cells (ACHN and A498) showed significant increased apoptosis as compared to control (P = 0.000). These findings suggest the new role of ALP in cell viability and apoptosis and involvement in RCC tumorigenesis. However, further studies are needed to explore the exact molecular mechanism. PMID:24909115

  7. Crystallization and preliminary X-ray crystallographic analysis of PhoK, an extracellular alkaline phosphatase from Sphingomonas sp. BSAR-1

    International Nuclear Information System (INIS)

    A new alkaline phosphatase enzyme from Sphingomonas sp. strain BSAR-1, termed PhoK, has been shown to be useful in uranium bioprecipitation. PhoK has been expressed, purified and crystallized. Alkaline phosphatases (APs) are widely distributed from microbes to humans and are involved in several important biological processes such as phosphate nutrition, signal transduction and pathogenesis. Alkaline phosphatases are also useful in various industrial applications and in recombinant DNA technology. A new AP enzyme from Sphingomonas sp. strain BSAR-1, termed PhoK, has been shown to be useful in uranium bioprecipitation. PhoK was expressed, purified and crystallized. The crystals belonged to space group P43212 or P41212, with unit-cell parameters a = b = 87.37, c = 168.16 Å, and contained one enzyme molecule in the asymmetric unit. Native diffraction data have been collected to 1.95 Å resolution at the ESRF

  8. 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

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

    DEFF Research Database (Denmark)

    Lisse, I M; Whittle, H; Aaby, P;

    1990-01-01

    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 trans......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...

  10. 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.

  11. 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. PMID:27343614

  12. A Further Investigation of the Effects of Extremely Low Frequency Magnetic Fields on Alkaline Phosphatase and Acetylcholinesterase.

    Directory of Open Access Journals (Sweden)

    Gary Silkstone

    Full Text Available Using a custom build spectrophotometer equipped with Helmholtz coils and designed to study the effects of magnetic fields on enzyme reactions in real-time we have investigated the influence of fields, from 100 μT to 10 mT and at a variety of field frequencies, on the membrane bound enzymes alkaline phosphatase and acetylcholinesterase. We have also employed other methods to apply a magnetic field, e.g. Biostim. In contrast to earlier reports we have been unable to detect any field effects on these enzymes under any field/frequency regime. We discuss possible reasons for the discrepancy between this and earlier work and note the particularly complex influence of small temperature changes that may confound analysis.

  13. 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.

  14. Significant Association Between Bone-Specific Alkaline Phosphatase and Vascular Calcification of the Hand Arteries in Male Hemodialysis Patients

    Directory of Open Access Journals (Sweden)

    Eiji Ishimura

    2014-09-01

    Full Text Available Background/Aims: Bone-specific alkaline phosphatase (BAP hydrolyzes pyrophosphate, which inhibits vascular calcification. We examined association between serum BAP and vascular calcification of male hemodialysis patients. Methods: Hand roentgenography of 167 male maintenance hemodialysis patients was conducted, and visible vascular calcification of the hand arteries was evaluated. Serum levels of 3 bone formation markers (BAP, osteocalcin, and N-terminal propeptide of type I collagen and 2 bone resorption markers (C-terminal telopeptide of type I collagen, and cross-linked N-telopeptide of type I collagen were measured, along with serum intact parathyroid hormone (PTH. Results: Of 167 patients, visible vascular calcification was seen in 37 patients. Among the bone formation and resorption markers, serum BAP was significantly higher in patients with vascular calcification than in those without (pConclusions: Higher serum BAP, but not other bone markers, is significantly associated with the presence of vascular calcification in male hemodialysis patients.

  15. Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins.

    Science.gov (United States)

    Noh, Soodong; Ha, Dat Thinh; Yang, Haesik; Kim, Moon-Soo

    2015-06-21

    Direct detection of double-stranded DNA (dsDNA) using zinc finger proteins (ZFPs) is of great importance in biomedical applications such as identifying pathogens and circulating DNAs. However, its sensitivity is still not sufficiently high because limited signalling labels can be conjugated or fused. Herein, we report sensitive and direct detection of dsDNA using (i) alkaline phosphatase (ALP) as a fast catalytic label conjugated to ZFPs along with (ii) electrochemical measurement of an ALP product (l-ascorbic acid) at the indium-tin oxide electrode with a high signal-to-background ratio. ALP is simply conjugated to a ZFP through lysine residues in a ZFP purification tag, a maltose binding protein (MBP). Sandwich-type electrochemical detection of dsDNA allows a detection limit of ca. 100 fM without using DNA amplification. PMID:25969923

  16. Comparison of the expression, activity, and fecal concentration of intestinal alkaline phosphatase between healthy dogs and dogs with chronic enteropathy.

    Science.gov (United States)

    Ide, Kaori; Kato, Kazuki; Sawa, Yuki; Hayashi, Akiko; Takizawa, Rei; Nishifuji, Koji

    2016-07-01

    OBJECTIVE To compare expression, activity, and fecal concentration of intestinal alkaline phosphatase (IAP) between healthy dogs and dogs with chronic enteropathy (CE). ANIMALS 9 healthy university-owned Beagles and 109 healthy client-owned dogs (controls) and 28 dogs with CE (cases). PROCEDURES Cases were defined as dogs with persistent (> 3 weeks) gastrointestinal signs that failed to respond to antimicrobials and anti-inflammatory doses of prednisolone or dietary trials, did not have mechanical gastrointestinal abnormalities as determined by abdominal radiography and ultrasonography, and had a diagnosis of lymphoplasmacytic enteritis or eosinophilic gastroenteritis on histologic examination of biopsy specimens. Duodenal and colonic mucosa biopsy specimens were obtained from the 9 university-owned Beagles and all cases for histologic examination and determination of IAP expression (by real-time quantitative PCR assay) and activity (by enzyme histochemical analysis). Fecal samples were obtained from all dogs for determination of fecal IAP concentration by a quantitative enzyme reaction assay. RESULTS For dogs evaluated, IAP expression and activity were localized at the luminal side of epithelial cells in the mucosa and intestinal crypts, although both were greater in the duodenum than in the colon. Active IAP was detected in the feces of all dogs. Intestinal alkaline phosphatase expression and activity were lower for cases than for controls, and fecal IAP concentration for dogs with moderate and severe CE was lower than that for dogs with mild CE. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that dogs with CE had impaired IAP expression and activity. Additional research is necessary to elucidate the role of IAP in the pathogenesis of CE. PMID:27347825

  17. 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.

  18. 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.

  19. CONTROL OF ALKALINE PHOSPHATASE ACTIVITY IN C3H10T1/2 CELLS: ROLE OF RETINOIC ACID AND CELL DENSITY

    Science.gov (United States)

    The enzyme alkaline phosphatase (AP) has been shown to be lost or inappropriately expressed during carcinogenesis in some tissues. ecause retinoic acid (RA) appears to play a role in the normal regulation of the enzyme (RA up-regulates AP in a variety of cell types) we have sugge...

  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. 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

    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 of GH treatment (p alkaline phosphatase increased significantly from 38.6 to 92.9 U/l during GH therapy in male patients (p ...-derived alkaline phosphatase was unaltered by GH. In the females, the increase in bone alkaline phosphatase did not reach statistical significance (19.1 vs 40.0 U/l, p = 0.06). The GH-induced increase in bone alkaline phosphatase correlated significantly with the increase in serum IGFBP-3 (r = 0.46, p = 0...

  2. 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.

  3. Screening of Alkaline Protease-Producing Streptomyces diastaticus and Optimization of Enzyme Production

    Directory of Open Access Journals (Sweden)

    Elham Dawoodi

    2014-12-01

    Full Text Available Background and Aim: Alkaline proteases are used in pharmaceutical, film and photography, silk production and food, leather and detergent industries. Actinomycetes are gram positive bacteria that produce different enzymes such as proteases. The aims of this research were isolation of native alkaline protease-producing Actinomycete spp. from different soil samples as well as optimizing the conditions for enzyme production. Materials and Methods: The different soil samples were collected from different locations of the provinces of Khouzestan, Chahar Mahalo Bakhtiari and Isfahan, Iran. After determining of the best alkaline protease producing species using Lowry method, the optimization of alkaline protease was performed. Results: The alkaline protease producing Actinomycete spp. was isolated from soil. The most enzyme activity was measured in S.diastaticus. The best concentration of sucrose as the carbon source for the highest production of alkaline protease was 10 g/l. The optimum pH and temperature for the alkaline protease production by S. diastaticus were 10 and 30°C respectively. The maximum activity of alkaline protease was measured at 200 rpm as the best aeration speed. Conclusions: This is the first report of alkaline protease production by Streptomyces diastaticus in Iran. The accomplished examinations in this research confirmed the previous theories of alkaline protease production by Actinomycetes relatively. Regarding the immense applications of alkaline proteases in several industries and isolation of a native alkaline protease producing Actinomycete, The production potential of this enzyme in our country could be accessible in the near future.

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

    International Nuclear Information System (INIS)

    Highlights: • Alkaline phosphatase from eggs of the sea urchin (StAP) proved to be active in seawater. • Activity of StAP is inhibited by very low concentrations of heavy metal. • A test to assess sea and fresh water quality has been developed basing on StAP. • For the first time a salt resistant alkaline phosphatase has been found in eukaryote. - Abstract: 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

  5. Origin and production of phosphatases in the acid Lake Gardsjoen

    Energy Technology Data Exchange (ETDEWEB)

    Olsson, H.

    1983-01-01

    The activity of acid phosphatases was followed for one year in Lake Gardsjoen as well as in the inlet and the outlet of the lake. A budget of the phosphatases was calculated, including an estimation of the production of phosphatases. The phosphatase activity was also measured in two basins upstream of L. Gardsjoen: the north basin and the south basin of L. Stora Haestevatten. The acid phosphatase activity was very high compared with reported alkaline phosphatase activities in other lakes. About 95% of the phosphatases in L. Gardsjoen was produced in the lake, and the production was highest in early summer. Small Chrysophyceae (< 10 ..mu..m) probably produced the majority of the acid phosphatases in the investigated lakes, and accordingly could be favoured in environments with low phosphorus supply due to their ability to produce large amounts of phosphatases. 10 references, 8 figures, 2 tables.

  6. 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.

  7. Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.

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    Temelli Ozlem

    2009-05-01

    Full Text Available Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP, Cytokeratin7 (CK7 Cytokeratin8 (CK8 to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinomas, demonstrated CK8 reactivity in 106 (72% of 148 cases. 95 primary lung carcinoma samples (65% were positive for CK7. AP was expressed in 81% of primary lung adenocarcinomas. Positive immunoreactivity for AP was characterized by a red, diffusely apical cytoplasmic staining in tumor cells that ocurred singly or in groups. There was a significant difference between AP, CK 7 and CK 8 expressions in primary lung adenocarcinomas (P=0.02; Chi-squared test. The sensitivity of AP, CK8, CK7 as a marker for primary lung adenocarcinomas were 82%, 72%, 64%, respectively. Thus the AP positive staining largely confirmed the cytologic diagnosis of lung adenocarcinoma.

  8. High-throughput screening of tissue-nonspecific alkaline phosphatase for identification of effectors with diverse modes of action.

    Science.gov (United States)

    Sergienko, Eduard A; Millán, José Luis

    2010-08-01

    Here we describe a protocol for the identification of effectors of tissue-nonspecific alkaline phosphatase (TNAP). It is based on a highly sensitive method for detecting TNAP activity. After dephosphorylation by TNAP, a dioxetane-based substrate undergoes a series of chemical transformations resulting in light production. Light intensity serves as a quantitative measure of the velocity of the TNAP-catalyzed reaction in the steady state. This protocol includes guidelines for optimizing the assay and for high-throughput screening in multiwell plates. The assay is sensitive to the influence of diverse effectors of TNAP as long as the assay optimization steps are repeated for each new batch of the enzyme; full optimization is accomplished in under 2 d. Depending on the available equipment, 10,000-100,000 compounds can be screened in an 8-h period. This protocol provides a method of screening TNAP that is 1,000-fold more sensitive and 10-fold faster than a conventional colorimetric assay with p-nitrophenyl phosphate. PMID:20671726

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

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    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.

  10. Monitoring the activity and inhibition of alkaline phosphatase via quenching and restoration of the fluorescence of carbon dots

    International Nuclear Information System (INIS)

    We report that the fluorescence of carbon dots (C-dots) in water is quenched by the addition of Cu2+ ions, and that the subsequent addition of pyrophosphate (PPi) restores fluorescence. This is likely to be due to the coordination of Cu2+ by PPi. This effect forms the basis for a method to determine the activity and inhibition of the enzyme alkaline phosphatase (ALP). If ALP is added to a system composed of C-dots, Cu2+ and PPi, fluorescence will decrease over time because ALP catalyzes the hydrolysis of PPi to form orthophosphate (Pi). This results in a release of the quencher Cu2+. The decrease in fluorescence is related to the activity of ALP. The method is simple and displays good sensitivity (with a limit of detection of 1 units per L) and selectivity. The method was successfully applied to the determination of ALP in serum samples. We also have studied the inhibitory effect of Pi on the activity of ALP. We presume that this method holds a large potential in terms of diagnosis of ALP-related diseases, to evaluate the function of ALP in biological systems and in screening for potential inhibitors of ALP. (author)

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

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    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.

  13. 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.

  14. 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. PMID:17268916

  15. 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

  16. Relation of fatty acid composition in lead-exposed mallards to fat mobilization, lipid peroxidation and alkaline phosphatase activity

    Science.gov (United States)

    Mateo, R.; Beyer, W.N.; Spann, J.W.; Hoffman, D.J.

    2003-01-01

    The increase of n-6 polyunsaturated fatty acids (PUFA) in animal tissues has been proposed as a mechanism of Pb poisoning through lipid peroxidation or altered eicosanoids metabolism. We have studied fatty acid (FA) composition in liver and brain of mallards (Anas platyrhynchos) feeding for three weeks on diets containing combinations of low or high levels of vitamin E (20 or 200 UI/kg) and Pb (0 or 2 g/kg). Saturated FA, n-6 PUFA and total concentrations of FA were higher in livers of Pb-exposed mallards, but not in their brains. The percentage of n-6 PUFA in liver and brain was slightly higher in Pb-exposed mallards. The increase of n-6 PUFA in liver was associated with increased triglycerides and cholesterol in plasma, thus could be in part attributed to feed refusal and fat mobilization. The hepatic ratios between adrenic acid (22:4 n-6) and arachidonic acid (20:4 n-6) or between adrenic acid and linoleic acid (18:2 n-6) were higher in Pb exposed birds, supporting the existing hypothesis of increased fatty acid elongation by Pb. Among the possible consequences of increased n-6 PUFA concentration in tissues, we found increased lipid peroxidation in liver without important histopathological changes, and decreased plasma alkaline phosphatase activity that may reflect altered bone metabolism in birds.

  17. A single-molecule digital enzyme assay using alkaline phosphatase with a cumarin-based fluorogenic substrate.

    Science.gov (United States)

    Obayashi, Yusuke; Iino, Ryota; Noji, Hiroyuki

    2015-08-01

    Digitalization of fluorogenic enzymatic assays through the use of femtoliter chamber array technology is an emerging approach to realizing highly quantitative bioassays with single-molecule sensitivity. However, only a few digital fluorogenic enzyme assays have been reported, and the variations of the digital enzyme assays are basically limited to fluorescein- and resorufin-based fluorogenic assays. This limitation hampers the realization of a multiplex digital enzyme assay such as a digital enzyme-linked immunosorbent assay (ELISA). In this study, after optimization of buffer conditions, we achieved a single-molecule digital enzyme alkaline phosphatase (ALP) assay with a cumarin-based fluorogenic substrate, 4-methylunbelliferyl phosphate (4-MUP). When ALP molecules were encapsulated in a 44-femtoliter chamber array at a low ratio of less than 1 molecule per chamber, each chamber showed a discrete fluorescence signal in an all-or-none manner, allowing the digital counting of the number of active enzyme molecules. The fraction of fluorescent chambers linearly decreased with the enzyme concentration, obeying the Poisson distribution as expected. We also demonstrated a dual-color digital enzyme assay with a ALP/4-MUP and β-galactosidase (β-gal)/resorufin-β-d-galactopyranoside combination. The activities of single ALP and β-gal molecules were clearly detected simultaneously. The method developed in this study will enable us to carry out a parallelized, multiplex digital ELISA. PMID:26101788

  18. Steric hindrance regulated supramolecular assembly between β-cyclodextrin polymer and pyrene for alkaline phosphatase fluorescent sensing

    Science.gov (United States)

    Song, Chunxia; Yang, Xiaohai; Wang, Kemin; Wang, Qing; Liu, Jianbo; Huang, Jin; Zhou, Maogui; Guo, Xiaochen

    2016-03-01

    We herein report a strategy for sensitive alkaline phosphatase (ALP) fluorescent sensing based on steric hindrance regulated supramolecular assembly between β-cyclodextrin polymer (polyβ-CD) and pyrene. The fluorescence of pyrene was enhanced more than 10 times through supramolecular assembly with polyβ-CD. The 5‧-phosphorylated dsDNA probe with pyrene attached on the 3‧-terminal could be cleaved by λ exonuclease (λ exo), yielding pyrene attached on mononucleotides. Pyrene attached on mononucleotides could easily enter the cavity of polyβ-CD, resulting in fluorescence enhancement. When ALP was introduced, it could remove 5‧-phosphate groups from dsDNA and then prevented the cleavage of dsDNA. Pyrene attached on dsDNA was difficult to enter the cavity of polyβ-CD because of steric hindrance, resulting in an inconspicuous fluorescence enhancement. Owing to the excellent fluorescence enhancement during steric hindrance regulated supramolecular assembly, excellent performance of the assay method was achieved for ALP with a detection limit of 0.04 U mL- 1. The detection limit was superior or comparable with the reported methods. Besides, this method was simple in design, avoiding double-labeling of probe.

  19. Transient species produced in irradiated alkaline aqueous solutions containing oxygen

    International Nuclear Information System (INIS)

    The spectra and decay kinetics of the optical absorption in the UV region and at 430nm (maximum of the ozonide ion absorption), were investigated in oxygenated neutral, slightly and strongly alkaline aqueous solutions. It is assumed that the initial absorption after the pulse in oxygenated alkaline solutions is due to the ozonide ion O3-, superoxide radical ion O2- and ozone O3. The long-living absorption in these solutions cannot be excluded as to be probably caused by the ozone O3 or some product formed from O2- or an alkaline stabilized form of this radical-ion. The advantage of applying additional non-optical measurements of the system is shown on the example of H2O2 role played in the mechanism. (author)

  20. Evaluation of Milk Trace Elements, Lactate Dehydrogenase, Alkaline Phosphatase and Aspartate Aminotransferase Activity of Subclinical Mastitis as and Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis)

    OpenAIRE

    Guha, Anirban; Gera, Sandeep; Sharma, Anshu

    2012-01-01

    Mastitis is a highly morbid disease that requires detection at the subclinical stage. Tropical countries like India mainly depend on milch buffaloes for milk. The present study was conducted to investigate whether the trace minerals viz. copper (Cu), iron (Fe), zinc (Zn), cobalt (Co) and manganese (Mn) and enzyme activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in riverine buffalo milk can be used as an indicator of subclinical mastitis ...

  1. Can Serum Levels of Alkaline Phosphatase and Phosphate Predict Cardiovascular Diseases and Total Mortality in Individuals with Preserved Renal Function? A Systemic Review and Meta-Analysis

    OpenAIRE

    Jing-Wei Li; Cui Xu; Ye Fan; Yong Wang; Ying-Bin Xiao

    2014-01-01

    Background It is demonstrated that elevated serum levels of alkaline phosphatase (ALP) and phosphate indicate a higher risks of cardiovascular disease (CVD) and total mortality in population with chronic kidney disease (CKD), but it remains unclear whether this association exists in people with normal or preserved renal function. Method Clinical trials were searched from Embase and PubMed from inception to 2013 December using the keywords “ALP”, “phosphate”, “CVD”, “mortality” and so on, and ...

  2. The Characteristic Absorption Peak Analysis of Alkaline Phosphatase Kit%对碱性磷酸酶试剂盒特征吸收峰的分析

    Institute of Scientific and Technical Information of China (English)

    何乐春

    2012-01-01

      The paper analysis the reasonableness of enterprise standards about detection wavelength according to the test result of reagent blank absorbance value of alkaline phosphatase ( ALP ) kit.%  从检测一厂家生产的碱性磷酸酶(ALP)试剂盒的试剂空白吸光度所得的值,分析企业标准中所涉及的检测波长是否合理。

  3. Improvement of the skeletal and dental hypophosphatasia phenotype in Alpl−/− mice by administration of soluble (non-targeted) chimeric alkaline phosphatase

    OpenAIRE

    Gasque, Kellen; Foster, Brian L.; Kuss, Pia; Yadav, Manisha C; Jin LIU; Kiffer-Moreira, Tina; van Elsas, Andrea; Hatch, Nan; Somerman, Martha J.; Millán, JoséLuis

    2014-01-01

    Hypophosphatasia (HPP) results from ALPL gene mutations, which lead to a deficiency of tissue-nonspecific alkaline phosphatase (TNAP), and accumulation of inorganic pyrophosphate, a potent inhibitor of mineralization that is also a natural substrate of TNAP, in the extracellular space. HPP causes mineralization disorders including soft bones (rickets or osteomalacia) and defects in teeth and periodontal tissues. Enzyme replacement therapy using mineral-targeting recombinant TNAP has proven ef...

  4. 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.

  5. 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 vivostudy

    Directory of Open Access Journals (Sweden)

    Jimly James Kunjappu

    2012-01-01

    Full Text Available 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.

  6. AN INTRA - ABDOMINAL FOREI GN BOCORRELATION OF SERUM LIPID PROFILE, SERUM CALCIUM, ALKALINE PHOSPHATASE AND SERUM PROTEIN WITH HISTOPATHOLOGICAL GRADING AND STAGING IN HEAD AND NECK CANCER

    Directory of Open Access Journals (Sweden)

    Neha

    2014-02-01

    Full Text Available Lipids have a key role in the maintenance of cell integrity. Hypolipidemia is related with increased risk of cancer and mortality. Elevated calcium and alkaline phosphatase has also been seen in cancer patients having m etastasis. Alteration in serum prote ins also has been reported. Hence present study was undertaken in 100 histopathologically confirmed head and neck cancer patients, in Department of Pathology Gandhi Medical College Bhopal, after taking informed written c onsent and ethical clearance. Lipid Profile, Serum Calcium, Alkaline Phosphatase, Serum Protein with Albumin were estimated and compared with age and sex matched healthy controls. Statistical analysis was done using chi square test. Total cholesterol (TC, high density lipoprotein - cholesterol (HDLC, low density lipoprotein - cholesterol (LDLC, Very low density lipoprotein - cholesterol (VLDLC all decreased in head & neck cancer patients. Hypercalcemia was seen in 35% of patients. Raised Alkaline phosphatase (ALP was seen in 8 % cancer patients. Serum calcium was found to be increasing with advancing stage. Further studies should be carried out in large number of patients to confirm the role of these parameters with special attention to modifiable ones and their relation with staging and grading of cancer, which can be used as prognostic markers.

  7. Estimation and Comparison of Salivary Calcium, Phosphorous, Alkaline Phosphatase and pH Levels in Periodontal Health and Disease: A Cross-sectional Biochemical Study

    Science.gov (United States)

    Patel, Rufi Murad; Suragimath, Girish; Zope, Sameer

    2016-01-01

    Introduction In oral diagnostics there is a great challenge to determine biomarkers for screening and evaluating the disease activity. Biomarkers can also serve as a useful tool to measure the efficacy of the therapy. Aim To evaluate and compare the levels of salivary calcium, phosphorous, alkaline phosphatase and pH levels in periodontally healthy subjects and patients with gingivitis and periodontitis. Materials and Methods The present study consisted of 150 subjects aged between 20-45 years who were divided into three groups; periodontally healthy, gingivitis and chronic periodontitis. Prior to the clinical examination the demographic details, relevant information of the subject, gingival index, plaque index, Oral Hygiene Index (OHI) and pH were recorded. Biochemical assay of saliva i.e., inorganic calcium, phosphorous and alkaline phosphatase were estimated by colorimetric method. ANOVA and Tukey’s test were applied for statistical analysis. Results The mean levels of biomarkers studied were; inorganic calcium (12.55μg/dl), phosphorous (14.50μg/dl), alkaline phosphatase (49.62μg/dl) and pH (11.65). There was a gradual increase in these levels as the condition progressed from health to gingivitis or periodontitis which was statistically significant at pperiodontal tissues in disease and health. PMID:27630955

  8. 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. PMID:27132001

  9. 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. PMID:22115786

  10. 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

  11. Orientation of mineral crystallites and mineral density during skeletal development in mice deficient in tissue nonspecific alkaline phosphatase.

    Science.gov (United States)

    Tesch, W; Vandenbos, T; Roschgr, P; Fratzl-Zelman, N; Klaushofer, K; Beertsen, W; Fratzl, P

    2003-01-01

    Tissue nonspecific alkaline phosphatase (TNALP) is thought to play an important role in mineralization processes, although its exact working mechanism is not known. In the present investigation we have studied mineral crystal characteristics in the developing skeleton of TNALP-deficient mice. Null mutants (n = 7) and their wild-type littermates (n = 7) were bred and killed between 8 and 22 days after birth. Skeletal tissues were processed to assess mineral characteristics (small angle X-ray scattering, quantitative backscattered electron imaging), and to analyze bone by light microscopy and immunolabeling. The results showed a reduced longitudinal growth and a strongly delayed epiphyseal ossification in the null mutants. This was accompanied by disturbances in mineralization pattern, in that crystallites were not orderly aligned with respect to the longitudinal axis of the cortical bone. Among the null mutants, a great variability in the mineralization parameters was noticed. Also, immunolabeling of osteopontin (OPN) revealed an abnormal distribution pattern of the protein within the bone matrix. Whereas in the wild-type animals OPN was predominantly observed in cement and reversal lines, in the null mutants, OPN was also randomly dispersed throughout the nonmineralized matrix, with focal densities. In contrast, the distribution pattern of osteocalcin (OC) was comparable in both types of animals. It is concluded that ablation of TNALP results not only in hypomineralization of the skeleton, but also in a severe disorder of the mineral crystal alignment pattern in the corticalis of growing long bone in association with a disordered matrix architecture, presumably as a result of impaired bone remodeling and maturation. PMID:12510812

  12. 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.

  13. 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.

  14. One-step immunoassay for tetrabromobisphenol a using a camelid single domain antibody-alkaline phosphatase fusion protein.

    Science.gov (United States)

    Wang, Jia; Majkova, Zuzana; Bever, Candace R S; Yang, Jun; Gee, Shirley J; Li, Ji; Xu, Ting; Hammock, Bruce D

    2015-01-01

    Tetrabromobisphenol A (TBBPA) is a ubiquitous brominated flame retardant, showing widespread environmental and human exposures. A variable domain of the heavy chain antibody (VHH), naturally occurring in camelids, approaches the lower size limit of functional antigen-binding entities. The ease of genetic manipulation makes such VHHs a superior choice to use as an immunoreagent. In this study, a highly selective anti-TBBPA VHH T3-15 fused with alkaline phosphatase (AP) from E. coli was expressed, showing both an integrated TBBPA-binding capacity and enzymatic activity. A one-step immunoassay based on the fusion protein T3-15-AP was developed for TBBPA in 5% dimethyl sulfoxide (DMSO)/phosphate buffered saline (PBS, pH 7.4), with a half-maximum signal inhibition concentration (IC50) of 0.20 ng mL(-1). Compared to the parental VHH T3-15, T3-15-AP was able to bind to a wider variety of coating antigens and the assay sensitivity was slightly improved. Cross-reactivity of T3-15-AP with a set of important brominated analogues was negligible (<0.1%). Although T3-15-AP was susceptible to extreme heat (90 °C), much higher binding stability at ambient temperature was observed in the T3-15-AP-based assay for at least 70 days. A simple pretreatment method of diluting urine samples with DMSO was developed for a one-step assay. The recoveries of TBBPA from urine samples via this one-step assay ranged from 96.7% to 109.9% and correlated well with a high-performance liquid chromatography-tandem mass spectroscopy (HPLC-MS/MS) method. It is expected that the dimerized fusion protein, VHH-AP, will show promising applications in human exposure and environmental monitoring.

  15. 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

  16. 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

  17. Versatile and Amplified Biosensing through Enzymatic Cascade Reaction by Coupling Alkaline Phosphatase in Situ Generation of Photoresponsive Nanozyme.

    Science.gov (United States)

    Jin, Lu-Yi; Dong, Yu-Ming; Wu, Xiu-Ming; Cao, Gen-Xia; Wang, Guang-Li

    2015-10-20

    The alkaline phosphatase (ALP) biocatalysis followed by the in situ enzymatic generation of a visible light responsive nanozyme is coupled to elucidate a novel amplification strategy by enzymatic cascade reaction for versatile biosensing. The enzymatic hydrolysis of o-phosphonoxyphenol (OPP) to catechol (CA) by ALP is allowed to coordinate on the surface of TiO2 nanoparticles (NPs) due to the specificity and high affinity of enediol ligands to Ti(IV). Upon the stimuli by CA generated from ALP, the inert TiO2 NPs is activated, which demonstrates highly efficient oxidase mimicking activity for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light (λ ≥ 400 nm) irradiation utilizing dissolved oxygen as an electron acceptor. On the basis of the cascade reaction of ALP and the nanozyme of CA coordinated TiO2 (TiO2-CA) NPs, we design exquisitely colorimetric biosensors for probing ALP activity and its inhibitor of 2, 4-dichlorophenoxyacetic acid (2,4-DA). Quantitative probing of ALP activity in a wide linear range from 0.01 to 150 U/L with the detection limit of 0.002 U/L is realized, which endows the methodology with sufficiently high sensitivity for potentially practical applications in real samples of human serum (ALP level of 40-190 U/L for adults). In addition, a novel immunoassay protocol by taking mouse IgG as an example is validated using the ALP/nanozyme cascade amplification reaction as the signal transducer. A low detection limit of 2.0 pg/mL is attained for mouse IgG, which is 4500-fold lower than that of the standard enzyme-linked immuno-sorbent assay (ELISA) kit. Although only mouse IgG is used as a proof-of-concept in our experiment, we believe that this approach is generalizable to be readily extended to other ELISA systems. This methodology opens a new horizon for amplified and versatile biosensing including probing ALP activity and following ALP-based ELISA immunoassays. PMID:26419907

  18. 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

  19. Characterization of Six Missense Mutations in the Tissue-Nonspecific Alkaline Phosphatase (TNSALP Gene in Chinese Children with Hypophosphatasia

    Directory of Open Access Journals (Sweden)

    Haiou Yang

    2013-09-01

    Full Text Available Aims: Hypophosphatasia, a rare inherited disease characterized by defective mineralization of bone and teeth, is caused by various mutations in the tissue-nonspecific isoenzyme of alkaline phosphatase (TNSALP gene. Our aim was to determine the mutations on TNSALP gene in three Chinese children diagnosed as having hypophosphatasia. Methods: Genomic DNA was extracted from whole blood samples of patients and their parents. The TNSALP coding regions were then sequenced. Plasmids expressing wild-type or various mutants were built and in vitro studies were performed in order to determine whether these amino acid replacements could affect the TNSALP enzymatic activity. Results: Six missense mutations were identified from three independent pedigrees. Of the six missense mutations, four were novel and two had been previously reported. The Y28D, A111T and T389N mutants displayed only negligible ALP activity in vitro compared to the wild-type (WT TNSALP. The defect was mainly due to the significantly decreased protein expression in the 66 KD immature forms and the nearly undetectable protein expression in the 80 KD mature forms. Moreover, all three mutants had a dominant negative effect on the WT protein when co-transfected with TNSALP (WT. M219V and R136L mutants both exhibited partial enzymatic activities which were consistent with reduced protein expression in both forms of TNSALP which further exhibited moderate dominant-negative effect. In addition, Y388H mutant showed weak ALP activity. Western blot analysis indicated that the extreme reduction in signal from the mature forms of TNSALP could be the main cause of decreased enzymatic activity, since a strong signal was observed in the immature forms. Conclusion: Six missense mutations were identified in three Chinese hypophosphatasia pedigrees with subnormal serum ALP activity. Our results show that the low activity of serum ALP in the three patients is due mainly to a defect in the protein expression

  20. Versatile and Amplified Biosensing through Enzymatic Cascade Reaction by Coupling Alkaline Phosphatase in Situ Generation of Photoresponsive Nanozyme.

    Science.gov (United States)

    Jin, Lu-Yi; Dong, Yu-Ming; Wu, Xiu-Ming; Cao, Gen-Xia; Wang, Guang-Li

    2015-10-20

    The alkaline phosphatase (ALP) biocatalysis followed by the in situ enzymatic generation of a visible light responsive nanozyme is coupled to elucidate a novel amplification strategy by enzymatic cascade reaction for versatile biosensing. The enzymatic hydrolysis of o-phosphonoxyphenol (OPP) to catechol (CA) by ALP is allowed to coordinate on the surface of TiO2 nanoparticles (NPs) due to the specificity and high affinity of enediol ligands to Ti(IV). Upon the stimuli by CA generated from ALP, the inert TiO2 NPs is activated, which demonstrates highly efficient oxidase mimicking activity for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light (λ ≥ 400 nm) irradiation utilizing dissolved oxygen as an electron acceptor. On the basis of the cascade reaction of ALP and the nanozyme of CA coordinated TiO2 (TiO2-CA) NPs, we design exquisitely colorimetric biosensors for probing ALP activity and its inhibitor of 2, 4-dichlorophenoxyacetic acid (2,4-DA). Quantitative probing of ALP activity in a wide linear range from 0.01 to 150 U/L with the detection limit of 0.002 U/L is realized, which endows the methodology with sufficiently high sensitivity for potentially practical applications in real samples of human serum (ALP level of 40-190 U/L for adults). In addition, a novel immunoassay protocol by taking mouse IgG as an example is validated using the ALP/nanozyme cascade amplification reaction as the signal transducer. A low detection limit of 2.0 pg/mL is attained for mouse IgG, which is 4500-fold lower than that of the standard enzyme-linked immuno-sorbent assay (ELISA) kit. Although only mouse IgG is used as a proof-of-concept in our experiment, we believe that this approach is generalizable to be readily extended to other ELISA systems. This methodology opens a new horizon for amplified and versatile biosensing including probing ALP activity and following ALP-based ELISA immunoassays.

  1. 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

  2. Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application.

    Science.gov (United States)

    Souza, Amanda Araújo; Leitão, Vanessa Oliveira; Ramada, Marcelo Henrique; Mehdad, Azadeh; Georg, Raphaela de Castro; Ulhôa, Cirano José; de Freitas, Sonia Maria

    2016-01-01

    Acid phosphatases (ACPases) are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65 °C, respectively, and is stable at 55 °C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0 was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments. PMID:26938873

  3. Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application.

    Directory of Open Access Journals (Sweden)

    Amanda Araújo Souza

    Full Text Available Acid phosphatases (ACPases are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65 °C, respectively, and is stable at 55 °C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0 was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments.

  4. 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.

  5. 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...

  6. 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

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

    International Nuclear Information System (INIS)

    Research highlights: → Incubating PCR products at a high temperature causes smears in gel electrophoresis. → Smears interfere with the interpretation of methylation analysis using COBRA. → Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. → 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 oC or 65 oC, 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.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. Identification and Biochemical Characterization of Protein Phosphatase 5 from the Cantharidin-Producing Blister Beetle, Epicauta chinensis

    Directory of Open Access Journals (Sweden)

    Xi'en Chen

    2013-12-01

    Full Text Available Protein phosphatase 5 (PP5 is a unique member of serine/threonine phosphatases which has been recognized in regulation of diverse cellular processes. A cDNA fragment encoding PP5 (EcPP5 was cloned and characterized from the cantharidin-producing blister beetle, E. chinensis. EcPP5 contains an open reading frame of 1500 bp that encodes a protein of 56.89 kDa. The deduced amino acid sequence shares 88% and 68% identities to the PP5 of Tribolium castaneum and humans, respectively. Analysis of the primary sequence shows that EcPP5 has three TPR (tetratricopeptide repeat motifs at its N-terminal region and contains a highly conserved C-terminal catalytic domain. RT-PCR reveals that EcPP5 is expressed in all developmental stages and in different tissues. The recombinant EcPP5 (rEcPP5 was produced in Escherichia coli and purified to homogeneity. The purified protein exhibited phosphatase activity towards pNPP (p-nitrophenyl phosphate and phosphopeptides, and its activity can be enhanced by arachidonic acid. In vitro inhibition study revealed that protein phosphatase inhibitors, okadaic acid, cantharidin, norcantharidin and endothall, inhibited its activity. Further, protein phosphatase activity of total soluble protein extract from E. chinensis adults could be impeded by these inhibitors suggesting there might be some mechanism to protect this beetle from being damaged by its self-produced cantharidin.

  12. PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR ALKALINE PROTEASE PRODUCED FROM AN ISOLATED BACILLUS SUBTILIS

    Directory of Open Access Journals (Sweden)

    Vijaya Bundela

    2013-03-01

    Full Text Available This paper describes the studies on the purification and partial characterization of serine alkaline protease produced through submerged fermentation process from a locally isolated Bacillus subtilis. This strain, grown in a highly alkaline medium (pH 10, produces an extracellular proteolytic enzyme. The alkaline protease was purified in a simple two-step procedure involving ammonium sulphate precipitation and gel filtration. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE analysis of the purified alkaline protease indicated an estimated molecular mass of 30KDa. It was more active in the range of 20-60ºC and had an optimum activity at 55ºC with optimum pH of 10.5. Characterization of the protease showed that it required certain cations such as Mg++, Mn++ and Ca++ for maximal activity. The serine nature of the alkaline protease was confirmed by PMSF inhibition. The temperature and pH stability of this Alkaline Protease from Bacillus Subtilismakes it potentially useful forindustrial applications.

  13. 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

  14. Screening of Strains Producing Alkaline Protease from Soil and Study on the Conditions for Enzyme Production

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    [Objective] The aim was to screen strains producing alkaline protease from soil and study the conditions for enzyme production.[Method] Eight strains producing alkaline protease were isolated from soil through plate isolation,and the ability of enzyme production was measured by filter paper and Folin-phenol method.The strain with the strongest ability of enzyme production was screened as a candidate strain,then the factors influencing the ability of enzyme production was studied,finally the conditions for e...

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

    International Nuclear Information System (INIS)

    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 capacity to

  16. 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

  17. Growth energetics of an alkaline serine protease-producing strain of Bacillus clausii during continuous cultivation

    DEFF Research Database (Denmark)

    Christiansen, Torben; Nielsen, Jens

    2002-01-01

    Glucose-limited chemostats were used to determine the growth yields of biomass of Bacillus clausii PP 473-8 producing an alkaline serine protease Savinase (Novozymes A/S, Bagsvaerd, Denmark) and a low yield of biomass on oxygen was observed. The energy metabolism was investigated further by setti...

  18. 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.

  19. 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.

  20. Comparison of measurements of canine plasma creatinine, glucose, proteins, urea, alanine aminotransferase, and alkaline phosphatase obtained with Spotchem SP 4430 and Vitros 250 analyzers.

    Science.gov (United States)

    Trumel, C; Diquélou, A; Germain, C; Palanché, F; Braun, J P

    2005-12-01

    The suitability of the Spotchem 4430 benchtop biochemistry analyzer for canine blood samples was tested for creatinine, glucose, proteins, urea, alkaline phosphatases and alanine aminotransferase. Results obtained from whole blood and corresponding heparin plasma were identical except for proteins which were higher in plasma (n=10). Between series imprecision (n=10) was 0.93). The slopes of the Passing-Bablock's regression ranged from 0.90 to 1.20 and intercepts were low. The mean biases were low, except for creatinine for which the results obtained by Spotchem (Jaffe reaction) were about 20 micromol/L higher than with the Vitros (enzymatic reaction). The results of this study show that the Spotchem analyzer is suitable for use in canine whole blood or plasma when small numbers of tests are to be performed and large analyzers are not available. PMID:16054888

  1. 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. PMID:26350570

  2. Multiwalled carbon nanotube modified screen-printed electrodes for the detection of p-aminophenol: Optimisation and application in alkaline phosphatase-based assays

    Energy Technology Data Exchange (ETDEWEB)

    Lamas-Ardisana, Pedro Jose [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain); Queipo, Paula [Departamento de Fisica, Universidad de Oviedo, 33007 Oviedo, Asturias (Spain); Fanjul-Bolado, Pablo [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain); Costa-Garcia, Agustin [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain)], E-mail: costa@fq.uniovi.es

    2008-05-12

    Carboxylated multiwalled carbon nanotubes (MWCNT-COOH) were used to modify the working electrode surface of different screen-printed electrodes. The effect of this modification on the electrodic characteristics (double layer capacitance, electroactive area and heterogeneous rate constants for the electron transfer) was evaluated and optimized for the cyclic voltammetric determination of p-aminophenol. The enzymatic hydrolysis of p-aminophenylphosphate was employed for the quantification of alkaline phosphatase, one of the most important label enzymes in immunoassays. Finally, ELISA assays were carried out to quantify pneumolysin using this enzymatic system. Results obtained indicated that low superficial densities of MWCNT-COOH (0.03-0.06 {mu}g mm{sup -2}) yielded the same electrodic improvements but with better analytical properties.

  3. 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.

  4. Cloning of the gene and characterization of the enzymatic properties of the monomeric alkaline phosphatase (PhoX) from Pasteurella multocida strain X-73.

    Science.gov (United States)

    Wu, Jin-Ru; Shien, Jui-Hung; Shieh, Happy K; Hu, Chung-Chi; Gong, Shuen-Rong; Chen, Ling-Yun; Chang, Poa-Chun

    2007-02-01

    We have identified a new phoX gene encoding the monomeric alkaline phosphatase from Pasteurella multocida X-73. This gene was not found in the published genome sequence of Pasteurella multocida pm70. Characterization of the recombinant PhoX of Pasteurella multocida X-73 showed that it is a monomeric enzyme, activated by Ca(2+) and possibly secreted by the Tat pathway. These features distinguish phosphatases of the PhoX family from those of the PhoA family. All proteins of the PhoX family were found to contain a conserved motif that shares significant sequence homology with the calcium-binding site of a phosphotriesterase known as diisopropylfluorophosphatase. Site-directed mutagenesis revealed that D527 of PhoX might be the ligand bound to the catalytic calcium. This is the first report on identification of homologous sequences between PhoX and the phosphotriesterase and on the potential calcium-binding site of PhoX. PMID:17156125

  5. 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.

  6. ZZ亲和肽-碱性磷酸酶融合蛋白的制备与初步应用%Preparation of fusion protein between ZZ affinity peptide and alkaline phosphatase and its primary application

    Institute of Scientific and Technical Information of China (English)

    鲍如梦; 赵爽佳; 薛敏; 杨洪鸣; 唐金宝

    2015-01-01

    目的:构建、表达ZZ亲和肽与碱性磷酸酶( AP)融合蛋白,并研究其生物学活性。方法:将ZZ亲和肽基因与AP基因重组,构建原核表达载体pEZZ-AP并利用大肠杆菌表达,金属离子亲和层析纯化目的蛋白,Western blot鉴定ZZ-AP生物学活性并初步应用于免疫细胞化学。结果:所构建重组质粒pEZZ-AP 经E.coli DH5α表达,SDS-PAGE结果显示目的蛋白相对分子量为62 kD,与理论值相近。 Western blot结果表明ZZ-AP融合蛋白既具有兔IgG抗体结合活性,又具有碱性磷酸酶活性,在细胞免疫组化应用中与AP标记的羊抗兔二抗显色模式与效果相似。结论:ZZ-AP融合蛋白构建成功,该融合蛋白具有兔IgG 抗体结合活性和碱性磷酸酶活性,细胞免疫组化应用结果表明该融合蛋白在免疫分析中具有潜在的应用价值。%Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic

  7. Multiple myeloma: Changes in serum C-terminal telopeptide of collagen type I and bone-specific alkaline phosphatase can be used in daily practice to detect imminent osteolysis

    DEFF Research Database (Denmark)

    Lund, Thomas; Abildgaard, Niels; Andersen, Thomas L;

    2010-01-01

    of collagen type-I (CTX-I), C-terminal crosslinked telopeptide of type-I collagen generated by MMPs (ICTP), N-terminal crosslinked telopeptide of type-I collagen (NTX-I), and the bone formation marker bone-specific alkaline phosphatase (bALP) monthly for two years. Retrospectively, we identified 40 cases...

  8. Application of Alcohol Soluble Fixing Solution in Alkaline Phosphatase Staining%醇溶性固定液在碱性磷酸酶染色中的应用

    Institute of Scientific and Technical Information of China (English)

    陈敬文; 张伟; 崔华娟; 陈晓东; 赖续文

    2011-01-01

    目的 探讨醇溶性固定液在碱性磷酸酶 (alkaline phosphatase,ALP)染色中的应用.方法 将60例新鲜标本各取2块组织设为A、B两组,A组冷丙酮固定,B组醇溶性固定液固定,两组均常规石蜡切片,同时设立对照组,进行碱性磷酸酶钙-钴法染色,并对染色结果 进行观察.结果 两组切片均可见黑色碱性磷酸酶存在,但B组的阳性强度明显高于A组,对照组呈阴性反应.结论 醇溶性固定液在对碱性磷酸酶的固定效果和显示方面,均优于冷丙酮固定液.%Objective To explore the application of alcohol soluble fixing solution in alkaline phosphatase staining.Methods Two tissues in each of 60 fresh samples were taken and assigned as group A and group B fixed by cold acetone and alcohol soluble fixing solution respectively.Conventional paraffin slice were obtained for the two groups and the corresponding control group was set.All groups underwent alkaline phosphatase Ca-Co staining.Results Black alkaline phosphatase were observed in groups A and B, the positive intensity in group B was higher, and the control group was negative.Conclusion Alcohol soluble fixing solution is better than cold acetone in fixing and staining alkaline phosphatase.

  9. Proposing and evaluating applications for products obtained during chromium chip alkaline hydrolysis produced during leather tanning

    Directory of Open Access Journals (Sweden)

    Andrea Díaz

    2010-04-01

    Full Text Available Some applications for products obtained by chromium chip alkaline hydrolysis produced during leather tanning were evaluated in this work, considering the concept of maximising tanneries’ solid residue reuse for different industrial applications and minimising the environmental impact so produced. When Cr(OH is transformed into Cr (OH(SO it can be used in tanning leather (i.e. as tanning salt. When compared to commercial salts, 2 4 it was determined that it could be applied to mixtures containing this salt, replacing it by up to 40%. Chromium content reduction was evaluated for collagen hydrolyzate by pH control after alkaline hydrolysis of the chips and by applying adsorbent materials such as bentonite, alfalfa and sorghum biomass and activated charcoal, a maximum 55% Cr removal being obtained when the first two adsorbent materials were used.

  10. 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

  11. 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

  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. 骨源性碱性磷酸酶与血钙测定结果比较及临床探讨%Bone alkaline phosphatase compared with the serum calcium measurement results and clinical

    Institute of Scientific and Technical Information of China (English)

    黎智彪

    2013-01-01

      Objective: To investigate the to study bone alkaline phosphatase and serum calcium measurement results, analysis of bone alkaline phosphatase test value, in order to improve the clinical diagnosis and treatment of calcium deficiency patients. Methods: August 2009 to October 2012 infants undergo a medical examination of 150 patients for the study, given the determination of bone alkaline phosphatase and serum calcium al samples caught. Comparative observation test results, analysis of the significance of the detection of the two indicators of lack of calcium. Results: 150 cases of infants and young children, 37 cases of abnormalities of bone alkaline phosphatase, calcium abnormalities in 15 cases, two methods difference was significantly (p <0.05), the difference was statisticaly significant. Conclusion: bone alkaline phosphatase and serum calcium response to a lack of calcium infants, abnormal bone alkaline phosphatase detection rate higher, but also help the reaction of calcium nutrition of patients, doctors are required to pay close attention.%  目的探讨研究骨源性碱性磷酸酶与血钙的测定结果,分析骨源性碱性磷酸酶的检验价值,以提高临床对缺钙患者的诊治效果。方法选择2009年8月~2012年10月我院进行体检的婴幼儿150例为研究对象,对所有被检者均给予骨源性碱性磷酸酶和血钙的测定。对比观察检查结果,分析两种指标对钙离子缺乏的检测意义。结果150例婴幼儿中,骨源性碱性磷酸酶异常者37例,血钙异常15例,两种方法比较差异明显,p <0.05,差异有统计学意义。结论骨源性碱性磷酸酶与血钙均可以反应婴幼儿的钙缺乏,其中骨源性碱性磷酸酶异常的检出率更高,更有助于反应患者的钙营养情况,需要医生引起高度重视。

  14. 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.

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. Fluorescence detection of adenosine-5'-triphosphate and alkaline phosphatase based on the generation of CdS quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Siyu; Wang, Xinyan; Pang, Shu; Na, Weidan; Yan, Xu; Su, Xingguang, E-mail: suxg@jlu.edu.cn

    2014-05-01

    Highlights: • Cd²⁺ reacts with S²⁻ to generate fluorescent CdS QDs with ATP. • ATP can be hydrolyzed by the enzymatic dephosphorylation of ALP. • Fluorescent CdS QDs could not be generated in the presence of ALP. • The analysis system was successfully applied to assay ATP and ALP. Abstract: We have developed an analytical method to detect adenosine-5'-triphosphate (ATP) and alkaline phosphatase (ALP) based on the generation of CdS quantum dots (QDs). We demonstrated that Cd²⁺ cation reacts with S²⁻anion to generate fluorescent CdS QDs in the presence of some certain amount of ATP. With increase in the ATP concentration, the fluorescence intensity of CdS QDs was also enhanced. ATP can be converted into adenosine by the dephosphorylation of ALP, so that the generation of CdS QDs would be inhibited in the presence of ALP. Therefore, this novel analysis system could be applied to assay ATP and ALP based on the growth of fluorescent CdS QDs.

  20. Evaluation of Milk Trace Elements, Lactate Dehydrogenase, Alkaline Phosphatase and Aspartate Aminotransferase Activity of Subclinical Mastitis as and Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis).

    Science.gov (United States)

    Guha, Anirban; Gera, Sandeep; Sharma, Anshu

    2012-03-01

    Mastitis is a highly morbid disease that requires detection at the subclinical stage. Tropical countries like India mainly depend on milch buffaloes for milk. The present study was conducted to investigate whether the trace minerals viz. copper (Cu), iron (Fe), zinc (Zn), cobalt (Co) and manganese (Mn) and enzyme activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in riverine buffalo milk can be used as an indicator of subclinical mastitis (SCM) with the aim of developing suitable diagnostic kit for SCM. Trace elements and enzyme activity in milk were estimated with Atomic absorption Spectrophotometer, GBC 932 plus and biochemical methods, respectively. Somatic cell count (SCC) was done microscopically. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. A statistically significant (pbenchmark. Only ALP and Zn, the former being superior, were found to be suitable for diagnosis of SCM irrespective of etiological agents. LDH, Co and Fe can be introduced in the screening programs where Gram positive bacteria are omnipresent. It is recommended that both ALP and Zn be measured together in milk to diagnose buffalo SCM, irrespective of etiology. PMID:25049573

  1. 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.

  2. Alkaline Phosphatase Tagged Antibodies on Gold Nanoparticles/TiO2 Nanotubes Electrode: A Plasmonic Strategy for Label-Free and Amplified Photoelectrochemical Immunoassay.

    Science.gov (United States)

    Zhu, Yuan-Cheng; Zhang, Nan; Ruan, Yi-Fan; Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan

    2016-06-01

    This work reports a plasmonic strategy capable of label-free yet amplified photoelectrochemical (PEC) immunoassay for the sensitive and specific detection of model protein p53, an important transcription factor that regulates the cell cycle and functions as a tumor suppressor. Specifically, on the basis of Au nanoparticles (NPs) deposited on hierarchically ordered TiO2 nanotubes (NTs), a protein G molecular membrane was used for immobilization of alkaline phosphatase (ALP) conjugated anti-p53 (ALP-a-p53). Due to the immunological recognition between the receptor and target, the plasmonic charge separation from Au NPs to the conduction band of TiO2 NTs could be influenced greatly that originated from multiple factors. The degree of signal suppression is directly associated with the target concentration, so by monitoring the changes of the plasmonic photocurrent responding after the specific binding, a new plasmonic PEC immunoassay could be tailored for label-free and amplified detection. The operating principle of this study could be extended as a general protocol for numerous other targets of interest. PMID:27150939

  3. 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.

  4. 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. PMID:26663676

  5. Development of a nanobody-alkaline phosphatase fusion protein and its application in a highly sensitive direct competitive fluorescence enzyme immunoassay for detection of ochratoxin A in cereal.

    Science.gov (United States)

    Liu, Xing; Xu, Yang; Wan, De-bin; Xiong, Yong-hua; He, Zhen-yun; Wang, Xian-xian; Gee, Shirley J; Ryu, Dojin; Hammock, Bruce D

    2015-01-20

    A rapid and sensitive direct competitive fluorescence enzyme immunoassay (dc-FEIA) for ochratoxin A (OTA) based on a nanobody (Nb)-alkaline phosphatase (AP) fusion protein was developed. The VHH (variable domain of heavy chain antibody) gene of Nb28 was subcloned into the expression vector pecan45 containing the AP double-mutant gene. The Nb28-AP construct was transformed into Escherichia coli BL21(DE3)plysS, and soluble expression in bacteria was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. Both the Nb properties and AP enzymatic activity were validated by colorimetric and fluorometric analysis. The 50% inhibitory concentration and the detection limit of the dc-FEIA were 0.13 and 0.04 ng/mL, respectively, with a linear range of 0.06-0.43 ng/mL. This assay was compared with LC-MS/MS, and the results indicated the reliability of Nb-AP fusion protein-based dc-FEIA for monitoring OTA contamination in cereal.

  6. 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

  7. Immunocytochemical Phenotyping of Disseminated Tumor Cells in Bone Marrow by uPA Receptor and CK18: Investigation of Sensitivity and Specificity of an Immunogold/Alkaline Phosphatase Double Staining Protocol

    OpenAIRE

    Allgayer, Heike; Heiss, Markus Maria; Riesenberg, Rainer; Babic, Rudolf; Jauch, Karl Walter; Schildberg, Friedrich Wilhelm

    1997-01-01

    Phenotyping of cytokeratin (CK) 18-positive cells in bone marrow is gaining increasing importance for future prognostic screening of carcinoma patients. Urokinase-type plasminogen activator receptor (uPA-R) is one example of a potential aggressive marker for those cells. However, a valid and reliable double staining method is needed. Using monoclonal antibodies against uPA-R and CK18, we modified an immunogold/alkaline phosphatase double staining protocol. UPA-R/CK18-positive tumor cell contr...

  8. 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

  9. 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。

  10. 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

  11. Effects of sediment and turbulence on alkaline phosphatase activity and photosynthetic activity of phytoplankton in the shallow hyper-eutrophic Lake Taihu, China.

    Science.gov (United States)

    Ding, Yanqing; Qin, Boqiang; Xu, Hai; Wang, Xiaodong

    2016-08-01

    Sediments play important roles, as nutrient reservoir, especially in shallow lake ecosystem. The water column of large shallow lakes is often stable but also disturbed by turbulence causing resuspension of sediments. While considerable research has been carried out to investigate the influence of sediment resuspension on nutrient release, fewer studies have been done to understand the contribution of alkaline phosphatase activity (APA) in water as a response to the two conditions (turbulence and stability). Also, effects of the two lake conditions on photosynthetic efficiency of phytoplankton are still poorly understood. This study will evaluate the effect of these two conditions on photosynthetic efficiency and APA. Sediments used in the indoor experiments were collected from Zhushan Bay in Lake Taihu. Turbulence was generated by rotors to simulate the strong wind-induced disturbance in Lake Taihu. Results of the experiments showed that TN and TP in the stable and episodically turbulent conditions were not significantly different, with TN ranging from 1.34 to 1.90 mg/L and TP from 0.08 to 0.18 mg/L. Whereas, the soluble reactive phosphorus in the episodically turbulent condition was significantly higher than in the stable condition. Episodic turbulence could enhance P cycling by resuspending sediment-associated P, which alleviated algal P limitation. In stable conditions, P deficiency induced the production of high APA, which enhanced the availability of P. Although episodic turbulence could also cause increased algal biomass, photosynthetic efficiency of the algae was also affected not only by the nutrients but also by many other factors, especially light availability. Our results suggest that episodic turbulence is an important driver of biogeochemical cycling in large shallow hypertrophic lake ecosystem. PMID:27151245

  12. 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.

  13. A New 4.0-Generation Dendrimer Phosphorescence Labeling Reagent and Its Application to Determination of Trace Alkaline Phosphatase by Affinity Adsorption Solid Substrate-room Temperature Phosphorimetry

    Institute of Scientific and Technical Information of China (English)

    LI, Zhi-Ming; LIU, Jia-Ming; CHEN, Xiao-Hua; YANG, Min-Lan; CHEN, Xin-Hua; SHI, Xiu-Mei

    2007-01-01

    A Triton X-100-4.0G-D (4.0G-D refers to a 4.0-generation dendrimer) was brought forward as a new phosphorescence labeling reagent. Two types of specific affinity adsorption (AA) reactions (direct method and sandwich method) were carried out between the labeling product of Triton X-100-4.0G-D-Wheat germ agglutinin (WGA) and alkaline phosphatase (ALP), the product of AA reaction preserved the good characteristics of room temperature phosphorescence (RTP) of 4.0G-D and △Ip of the product was proportional to the content of ALP. According to the fact stated above, a new method for the determination of trace ALP by affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) was established on the basis of WGA labeled with the Triton X-100-4.0G-D. The detection limits were 0.20 ag·spot-1 (corresponding concentration: 5.0 × 10-16 g·mL-1, namely 5.0× 10-18 mol·L-1) for a direct method and 0.14 ag·spot-1 (corresponding concentration: 3.5× 10-16 g·mL-1,namely 3.5 × 10-18 mol·L-1) for a sandwich method, respectively. For their high sensitivity, good repeatability and high accuracy, the direct method and sandwich method have been successfully applied to determine the content of ALP in human serum, and the results were coincided with the clinical detection results of the enzyme-linked immunosorbent assay method by the Zhangzhou Hospital of Traditional Chinese Medicine. Meanwhile, the mechanism for the determination of trace ALP by AA-SS-RTP was discussed.

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

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    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.

  15. 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.

  16. Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease

    OpenAIRE

    Hongxia Cui; Muyang Yang; Liping Wang; Xian, Cory J.

    2015-01-01

    While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated....

  17. 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

  18. 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

  19. Study on Temporal and Spatial Variation of Alkaline Phosphatase Activity in the Surface Sediments of Erhai Lake%洱海表层沉积物碱性磷酸酶活性时空变化

    Institute of Scientific and Technical Information of China (English)

    黄睿智; 王圣瑞; 赵海超; 朱乐辉

    2013-01-01

    The effects of physicochemical parameters in the overlying water and sediments on the alkaline phosphatase activity in Erhai Lake were discussed by studying the temporal and spatial variation of this activity from November 2010 to August 2011. The results indicated that; ①The activity of alkaline phosphatase of the surface sediments changed from 45.63 to 144.67 μmol/(g·h) , and the decreasing order of the activities in different regions were the north, the south and the central. The regions where the exogenous pollutant input had a significant impact had high activity levels of nitrogen, phosphorus and the alkaline phosphatase. ② The alkaline phosphatase activity in the surface sediment showed a obvious seasonal variation in general and its seasonal increasing order were autumn ( November) , winter (February), spring (May) and summer (August). Its seasonal variation showed significant differences in the western coastal region. ③ The alkaline phosphatase activity had significant positive correlations with concentrations of OM, TN, OP, TP and C/N in the sediments and with concentrations of OP, TN and CODMn in the overlying water. The physicochemical parameters in the sediments and overlying water were the internal factors and the exogenous pollutants input were the external factors that are important to affect alkaline phosphatase activity.%研究了2010-2011年洱海表层沉积物中碱性磷酸酶活性的时空变化,并探讨了沉积物及水体理化因子对碱性磷酸酶活性的影响.结果表明:①洱海表层沉积物中碱性磷酸酶活性为45.63 ~ 144.67 μmol/(g·h),空间分布总体呈北部>南部>中部,受外源污染物输入影响较大的区域,其沉积物中w(TN)、w(TP)较高,碱性磷酸酶活性也较高;②洱海表层沉积物中碱性磷酸酶活性呈明显的季节性变化特征,秋季(11月)<冬季(2月)<春季(5月)<夏季(8月),并且西部沿岸区域的季节性变化差异显著;③碱性磷酸酶

  20. 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.

  1. 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

    2016-02-01

    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. PMID:26637593

  2. Bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase levels in patients with cleidocranial dysplasia: one case report%颅骨锁骨发育不良患者骨特异性碱性磷酸酶和抗酒石酸酸性磷酸酶的水平:1例报告

    Institute of Scientific and Technical Information of China (English)

    李阳飞; 秦晗; 徐宏志

    2011-01-01

    BACKGROUND: Bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase levels are closely related to skeletal development. OBJECTIVE: To observe bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase levels in patients with cleidocranial dysplasia (CCD). METHODS: Clinical data of one case of CCD were collected. The osseous malformation over the entire body was examined by panoramic radiography. A CCD phenotype was defined by the presence of hypoplastic clavicles and delayed closure of the anterior fontanels in addition to the presence of classic eraniofaeial features. Laboratory examination was used to detect the change of bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase levels. RESULTS AND CONCLUSION: The patients had obvious clinical characteristics of CCD, such as clavicle rudiment or absence, delayed closure of cranial fontanels and sutures, supernumerary and late erupting teeth, wide pubic symphysis, and other skeletal anomalies. The skeletal abnormalities as well as oral manifestations of the syndrome were variable within the affected patient. But abnormal chromosome was not found in this patient. Bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase levels were increased. These findings suggest that CCD patients may accompany with the genetic variation of tartrate-resistant acid phosphatase.%背景:骨特异性碱性磷酸酶和抗酒石酸酸性磷酸酶与骨骼的发育密切相关.目的:观察颅骨锁骨发育不良患者骨特异性碱性磷酸酶和抗酒石酸酸性磷酸酶的变化.方法:收集1 例颅骨锁骨发育不良患者临床资料,利用全颌曲面断层片检查全面了解患者的颌骨发育情况;详细进行口腔检查,记录恒牙萌出及乳牙迟萌以及埋伏牙、多生牙等;综合临床资料和实验室检查确定临床诊断,并检测患者骨特异性碱性磷酸酶和抗酒石酸酸性磷酸酶的变化.结果与结论:该患者有

  3. Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease.

    Directory of Open Access Journals (Sweden)

    Hongxia Cui

    Full Text Available While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production.

  4. Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease.

    Science.gov (United States)

    Cui, Hongxia; Yang, Muyang; Wang, Liping; Xian, Cory J

    2015-01-01

    While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth) was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production. PMID:26716833

  5. 热灭活法与干化学法检测骨碱性磷酸酶技术的对照分析%Comparison between heat-inactivity method and dry chemistry method on assaying bone alkaline phosphatase

    Institute of Scientific and Technical Information of China (English)

    李常林; 张艳; 马胜; 林虹梅; 卢秀芬

    2009-01-01

    Objective Assaying the content of bone alkaline phosphatase in childen's large batch samples by the method of heat-inactivity in compensation for the deficiency of dry chemistry.Methods Collecting finger blood and examinging alkaline phosphatase by biochemistry analyzer before and after heat-inactivity of the samples,then calculating the ratio of results between after and before heat-inactivity to filtrate suspect positive cases.Detecting the suspect positive cases by dry chemistry method,comparing the results of the two methods.Results The positive results of this method are in accord with the results of dry chemistry.Conclusion In contrast with dry chemistry method, the method of heat-inactivity is in possession of the characteristics of lower cost,quick analysis,quantity,fitting for large batch samples asssaying,etc.%目的 应用热灭活法检测大批量儿童骨碱性磷酸酶(bone alkaline phosphatase,BALP)项目以弥补干化学法测定BALP之不足.方法 采末梢血于生化分析仪上测定灭活前后的碱性磷酸酶(alkaline phosphatase,ALP)值,计算灭活后与灭活前ALP的百分比以筛出疑似阳性病例.热灭活法筛查出的BALP阳性病例,再用干化学法测定其BALP,之后将二种方法的测定结果进行对照.结果 热灭活法筛查出的BALP阳性结果同干化学法测定的BALP结果基本一致.结论 热灭活法检测骨碱性磷酸酶技术具有成本低、检测速度快、可定量分析、适合大批量BALP检测的优点.

  6. 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 mycotoxins concentrations used in the study, AP from farmed L. 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. PMID:27040818

  7. 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

  8. 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

  9. 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.

  10. Research on Phosphatases of Belladona Leaves and Their Purification

    Directory of Open Access Journals (Sweden)

    M. Khorsand

    1957-01-01

    Full Text Available Through experimentation with several leaves it has been possible for us to point out the existance of two different acid phosphatases. We have studied in more detail the phosphatases of belldon a leaves (Atropa Belladona L. Solanacees. The great part of the phosphatase activity is water extractable. We have compared the activity of the soluble fraction with that not directly extractable by means of water. The insoluble fraction could not be solubilized in a satisfaetC'fY m.anner.The digestion by papaine produced a slight solubilizing effect; on the other hand salt solutions, neutral or alkaline, or water glycerol mixtures had no solubilizing effect on the enzyme, It has been possible to demonstrate the existence of two different phosphatases in the insoluble fraction: the first of the type II,

  11. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes

    OpenAIRE

    Eman Zakaria Gomaa

    2013-01-01

    The potentiality of 23 bacterial isolates to produce alkaline protease and carboxymethyl-cellulase (CMCase) on Ficus nitida wastes was investigated. Bacillus pumillus ATCC7061 was selected as the most potent bacterial strain for the production of both enzymes. It was found that the optimum production of protease and CMCase were recorded at 30 °C, 5% Ficus nitida leaves and incubation period of 72 h. The best nitrogen sources for protease and CMCase production were yeast extract and casein, re...

  12. 966 cases of infants with bone alkaline phosphatase detection analysis%婴幼儿骨源性碱性磷酸酶966例检测分析

    Institute of Scientific and Technical Information of China (English)

    张炬光

    2013-01-01

    Objective to understand the incidence of infant rickets in Yulin city. Methods 966 cases of infants with bone alkaline phosphatase detection results of a retrospective analysis. Results the detection of bone alkaline phosphatase in 966 cases, 132 cases were rickets, a 13.66%detection rate, Among them, the boy was detected in 617 patients, the detection of rickets in 70 cases, the detection rate of 11.35%; the girl was detected in 349 patients, 62 cases were rickets, a 17.77%detection rate. Conclusion The detection of bone alkaline phosphatase in early diagnosis of rickets in infants and young children with sensitivity and specificity, the detection method is simple, to provide basis for prevention and treatment of rickets, is worth popularizing.%  目的评价骨源性碱性磷酸酶检测对婴幼儿佝偻病的早期诊断价值。方法对966例婴幼儿行骨源性碱性磷酸酶检测。结果966例婴幼儿中检出佝偻病治疗水平132例,阳性检出率为13.66%。其中,男孩检测617例,检出佝偻病治疗水平70例,阳性检出率为11.35%;女孩检测349例,检出佝偻病治疗水平62例,阳性检出率为17.77%。结论骨源性碱性磷酸酶检测对婴幼儿佝偻病的早期诊断具有一定的灵敏性和特异性,检测方法简单,能够为佝偻病的防治提供参考依据,值得推广应用。

  13. 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

  14. The selection of alkaline protease-producing yeasts from marine environments and evaluation of their bioactive peptide production

    Institute of Scientific and Technical Information of China (English)

    LI Jing; CHI Zhenming; WANG Xianghong; PENG Ying; CHI Zhe

    2009-01-01

    A total of 400 yeast strains from seawater, sediments, saltern mud, marine fish guts, and marine algae were obtained. The protease activity of the yeast cultures was estimated, after which four strains (HN3.11, N11b, YF04C and HN4.9) capable of secreting extracellular alkaline protease were isolated. The isolated strains were identified as Aureobasidium pullulans, Yarrowia lipolytica, Issatchenkia orientalis and Cryptococcus cf. aureus. The optimal pH of the protease activity produced by strains HN3.11, YF04C, and HN4.9 was 9.0, while that of the protease produced by strain N11b was 10.0. The optimal temperature for protease activity was 45°C for strains HN3.11, N11b, and YF04C, and 50°C for strain HN4.9. After digestion of shrimp (Penaeus vannamei) protein and spirulina (Arthospira platensis) protein with the four crude alkaline proteases, the filtrate from spirulina (Arthrospira platensis) powder digested by the crude alkaline protease of strain HN3.11 was found to have the highest antioxidant activity (61.4%) and the highest angiotensin I converting enzyme (ACE)-inhibitory activities (68.4%). The other filtrates had much lower antioxidant activity and ACE-inhibitory activities.

  15. 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

  16. Purification and characterization of alkaline protease produced by a mutant strain of bacillus subtilis

    International Nuclear Information System (INIS)

    The present study describes the production, purification and characterization of alkaline protease from mutant strain of Bacillus subtilis EMS-8. The enzyme was purified using ammonium sulphate precipitation which gave 2.64 fold purification with 81.5% yield at 70% saturation. The molecular weight of the enzyme was determined using SDS-PAGE and it was found to be 25 KDa. The optimum pH of enzyme activity was 8.5; however the enzyme remained stable up to pH 10 after 24 hrs of incubation. Similarly, the optimum temperature for enzyme activity was 40 degree C, whereas it remained stable up to 90 degree C with greatly reduced activity. Alkaline protease showed highest specificity towards casein. Among different inhibitors, Phenylmethylsulphonyl fluoride (PMSF) completely inhibited the enzyme activity indicating the serine nature of protease. Similarly, the protease activity was greatly reduced in the presence of MnCl/sub 2/, whereas MgCl/sub 2/ enhanced its activity. The shelf life of the protease was also determined and it was found that the activity of the enzyme came to an end after second week, when the enzyme was stored at room temperature. (author)

  17. Three-photon process for producing a degenerate gas of metastable alkaline-earth-metal atoms

    Science.gov (United States)

    Barker, D. S.; Pisenti, N. C.; Reschovsky, B. J.; Campbell, G. K.

    2016-05-01

    We present a method for creating a quantum degenerate gas of metastable alkaline-earth-metal atoms. This has yet to be achieved due to inelastic collisions that limit evaporative cooling in the metastable states. Quantum degenerate samples prepared in the 1S0 ground state can be rapidly transferred to either the 3P2 or 3P0 state via a coherent three-photon process. Numerical integration of the density-matrix evolution for the fine structure of bosonic alkaline-earth-metal atoms shows that transfer efficiencies of ≃90 % can be achieved with experimentally feasible laser parameters in both Sr and Yb. Importantly, the three-photon process can be set up such that it imparts no net momentum to the degenerate gas during the excitation, which will allow for studies of metastable samples outside the Lamb-Dicke regime. We discuss several experimental challenges to successfully realizing our scheme, including the minimization of differential ac Stark shifts between the four states connected by the three-photon transition.

  18. A 3-photon process for producing degenerate gases of metastable alkaline-earth atoms

    Science.gov (United States)

    Barker, Daniel S.; Pisenti, Neal C.; Reschovsky, Benjamin J.; Campbell, Gretchen K.

    2016-05-01

    We present a method for creating quantum degenerate gases of metastable alkaline-earth atoms. A degenerate gas in any of the 3 P metastable states has not previously been obtained due to large inelastic collision rates, which are unfavorable for evaporative cooling. Samples prepared in the 1S0 ground state can be rapidly transferred to either the 3P2 or 3P0 state via a coherent 3-photon process. Numerical integration of the density matrix evolution for the fine structure of bosonic alkaline-earth atoms shows that transfer efficiencies of ~= 90 % can be achieved with experimentally feasible laser parameters in both Sr and Yb. Importantly, the 3-photon process does not impart momentum to the degenerate gas during excitation, which allows studies of these metastable samples outside the Lamb-Dicke regime. We discuss several experimental challenges to the successful realization of our scheme, including the minimization of differential AC Stark shifts between the four states connected by the 3-photon transition.

  19. Amylase produced by Bacillus sp. SI-136 isolated from sodic-alkaline soil for efficient starch desizing

    OpenAIRE

    Sarethy, Indira P; Yashi Saxena; Aditi Kapoor; Manisha Sharma; Rohan Seth; Harsh Sharma; Sharma, Sanjeev K; Sanjay Gupta

    2012-01-01

    Bacillus sp. SI-136, isolated from sodic-alkaline soil, showed 94% similarity to B. cereus group based on 16S rDNA sequence. It produced α-amylase of 26 kDa with maximum activity at pH 10.0, stable up to pH 12.0 and 80oC. Mn2+ enhanced its activity as also 10% NaCl in medium. Agricultural waste substrates supported growth and enzyme activity was enhanced by 30% with sugarcane bagasse. The partially purified enzyme showed efficient desizing of cotton fabric at 50oC (40-60 min) or 70oC (...

  20. Amylase produced by Bacillus sp. SI-136 isolated from sodic-alkaline soil for efficient starch desizing

    Directory of Open Access Journals (Sweden)

    Indira P. Sarethy

    2012-12-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Bacillus sp. SI-136, isolated from sodic-alkaline soil, showed 94% similarity to B. cereus group based on 16S rDNA sequence. It produced α-amylase of 26 kDa with maximum activity at pH 10.0, stable up to pH 12.0 and 80oC. Mn2+ enhanced its activity as also 10% NaCl in medium. Agricultural waste substrates supported growth and enzyme activity was enhanced by 30% with sugarcane bagasse. The partially purified enzyme showed efficient desizing of cotton fabric at 50oC (40-60 min or 70oC (60 min with Tegewa rating 7-8, and at 95oC (20 min with Tegewa rating 9, properties enabling utility in textile industries.

  1. Partial purification and properties of cellulase-free alkaline xylanase produced by Rhizopus stolonifer in solid-state fermentation

    OpenAIRE

    Antonio José Goulart; Eleonora Cano Carmona; Rubens Monti

    2005-01-01

    Rhizopus stolonifer was cultivated in wheat bran to produce a cellulase-free alkaline xylanase. The purified enzyme obtained after molecular exclusion chromatography in Sephacryl S-200 HR showed optimum temperature as 45º C and hydrolysis pHs optima as pH 6.0 and 9.0. Xylanase presented higher Vmax at pH 9.0 (0.87 µmol/mg protein) than at pH 6.0 and minor Km at pH 6.0 (7.42 mg/mL) than at pH 9.0.Rhizopus stolonifer foi cultivado em meio de farelo de trigo para produzir uma xilanase alcalina c...

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. The two neutrophil plasma membrane markers alkaline phosphatase and HLA class I antigen localize differently in granule-deficient cytoplasts. An ideal plasma membrane marker in human neutrophils is still lacking.

    Science.gov (United States)

    Pellmé, Sara; Dahlgren, Claes; Karlsson, Anna

    2007-08-31

    Neutrophil function relies largely on the ability of the cell to mobilize its different granules and vesicles to the cell surface and thereby expose and/or release effector molecules to the surrounding tissue. To properly identify these subcellular compartments is thus a prerequisite for studies of neutrophil physiology. A range of specific markers for the classical granules is available, but finding optimal markers for the secretory vesicles and plasma membrane has historically been more challenging. Latent and non-latent alkaline phosphatase activities are often used to distinguish these two light membrane structures, but the outcome using this technique depends on the level of cellular activation. Therefore, HLA-I was introduced some years ago as a specific, stimulation-independent marker for the plasma membrane. In this study we however report that detailed fractionation studies of neutrophil cytoplasts, lacking secretory vesicles, granules and other dense organelles, reveal that the HLA-I antigen is not only co-localizing with the plasma membrane marker ALP, but is also present in other, more dense organelles. Further, we found the mixed enzyme-linked immunosorbent assay (MELISA), detecting the beta(2)-microglobulin/HLA-I complex, to be negatively influenced by uncomplexed beta(2)-microglobulin present in the specific granules and secretory vesicles, making it difficult to use HLA-I as a plasma membrane marker during maturation of for example phagolysosomes. PMID:17673253

  7. The Correlation Analysis of Alkaline Phosphatase and Parathyroid Hormone in Chronic Kidney Disease%慢性肾脏病碱性磷酸酶与甲状旁腺素相关性分析

    Institute of Scientific and Technical Information of China (English)

    余娜

    2012-01-01

      Objective To explore the correlation analysis of alkaline phosphatase(ALP) and parathyroid hormone(PTH) in chronic kidney disease(CKD), and evaluate their significance in diagnosis and treatment of renal osteodystrophy(ROD). Methods 60 CKD3-5 stage patients were extracted of fasting venous blood, to determinate ALP and PTH and analysis their correlation. Results ALP and PHT showed a significant positive correlation when PTH>300pg/ml(r=0.865, P300pg/ml时,ALP与PTH相关性良好,且呈正相关,有统计学意义(r=0.865,P<0.05);当PTH≤300pm/dl时, ALP与PTH无相关性.结论 ALP与PTH相关性良好,随着PTH的增高,ALP增高越明显.两者结合,对诊断和治疗高转运骨病(HTO)有重要的临床价值.且ALP检测方法简单,花费少.

  8. 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 shrimp was inhibited when incubated with AFB1 and FB1. The greatest inhibition occurred when AP was incubated with a mixture of AFB1 and FB1. The IC50 for AFB1 on AP activity of wild and farmed shrimp hepatopancreases was 0.790 and 0.398 μg/mL, respectively. The IC50 of FB1 was 0.87 μg/mL for wild shrimp and 0.69 μg/mL for farmed shrimp. These results suggest that, at the mycotoxins concentrations used in the study, AP from farmed L. 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.

  9. Distribution and impact factor of alkaline phosphatase activity in the intertidal surface sediments of the Yangtze Estuary%长江口潮滩表层沉积物中碱性磷酸酶活性及其影响因素

    Institute of Scientific and Technical Information of China (English)

    姜经梅; 赵慧; 沈铭能; 丁平兴; 尹国宇; 赵迪; 侯立军

    2011-01-01

    以长江口潮滩作为研究区域,对表层沉积物中碱性磷酸酶活性、解磷微生物量、磷组分及理化性质的时空分布特征进行了分析.结果表明,长江口潮滩湿地沉积物中碱性磷酸酶活性具有显著的季节性、区域性差异,冬季(1月)碱性磷酸酶活性较低,介于0.65~1.59μg.g-.1h-1之间;夏季(8、9月份)最高,变化范围为1.80~5.64μg.g-.1h-1.相关性分析结果显示,碱性磷酸酶活性与解磷细菌数量呈显著的正相关关系(rOPB=0.91,rIPB=0.89,p〈0.01),说明解磷细菌是影响碱性磷酸酶活性的主要因素之一.同时,碱性磷酸酶活性与磷组分、C/N、盐分等因素显著相关,而与粒度、有机碳氮含量的相关性不显著.%Spatial and temporal distributions of alkaline phosphatase activity(APA) in the intertidal sediments of the Yangtze Estuary were investigated,together with abundances of phosphate solubilizing bacteria and the phosphorus forms.It was shown that there were significantly seasonal and regional variations of alkaline phosphatase activity in the intertidal sediments.The alkaline phosphatase activities were relatively lower in winter(January),with values of 0.65 ~ 1.59 μg.g-.1h-1.In contrast,higher alkaline phosphatase activities were detected in summer(August and September),with values of 1.80 ~ 5.64 μg.g-1.h-1.Statistical analyses indicated that the alkaline phosphatase activities were related significantly to the numbers of phosphate solubilizing bacteria(rOPB = 0.91,rIPB = 0.89,p 0.01).This implied that the numbers of phosphate solubilizing bacteria were the primary factor controlling the changes of alkaline phosphatase activities.In addition,the alkaline phosphatase activities in the intertidal sediments were also affected by phosphorus forms,C:N molar ratios and salinity.However,there were no significant correlations of the alkaline phosphatase activities with the contents of sand

  10. Effects of TNF-α on the proliferation and alkaline phosphatase of the human dental follicle cells%TNF-α对体外培养的人牙囊细胞增殖特性及碱性磷酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    毕迎春; 李芸; 金作林

    2012-01-01

    Objective To study the effects of different concentration TNF-a on the activity of proliferation and the activity of alkaline phosphatase of human dental follicle cells. Methods The 5th passage human dental follicle cells were seeded on 96-well plates and co-cultured with different concentration of TNF-a, the activity of proliferation and the activity of alkaline phosphatase was assayed. Results The 10~50ng/ml TNF-a enhanced the activity of proliferation profoundly and the effects were significant at 3 days after co-culture. 10~200ng/ml TNF-a decreased the activity of alkaline phosphatase. Conclusions TNF-a enhance the activity of proliferation and decreased the activity of alkaline phosphatase of human dental follicle cells at special concentration and time.%目的:检测不同浓度TNF-对体外培养人牙囊细胞增殖活性和碱性磷酸酶活性的影响.方法:第 5代人牙囊细胞接种于96孔培养板上,分别与不同浓度的TNF-共同孵育,检测TNF-对人牙囊细胞的增殖活性和碱性磷酸酶活性的影响. 结果:TNF-在浓度为10~ 50ng/ml,时间为3天时促进人牙囊细胞的增殖,浓度为10~ 200ng/ml时抑制碱性磷酸酶活性.结论:TNF-在特定的浓度和时间促进人牙囊细胞的增殖,抑制牙囊细胞的成骨特性.

  11. Quantitative Analysis of Protein Concentration by Absorption Peak Integration Method of UV Spectroscopy-Taking Alkaline Phosphatase as an Example%紫外吸收光谱积分法分析蛋白质浓度-以碱性磷酸酶为例

    Institute of Scientific and Technical Information of China (English)

    朱元荣; 吴丰昌; 林樱

    2013-01-01

    利用矿物(针铁矿,蒙脱石)和太湖沉积物吸附碱性磷酸酶(alkaline phosphatase,APase),测定吸附后上清液中剩余碱性磷酸酶浓度时发现其紫外吸收光谱发生了变化,利用传统280 nm处紫外吸收法无法直接准确测定其浓度值.基于对碱性磷酸酶252~305 nm处吸收峰面积积分方法可以消除影响,并准确分析测定碱性磷酸酶浓度.其测定结果与考马斯亮蓝法测定结果进行比较,表明了该方法可以方便,快速和准确地测定此类实验中碱性磷酸酶浓度.同时,该方法还可以扩展至其他蛋白质的定量分析,甚至其他类似实验中,一定程度上克服传统方法应用单波长进行定量分析中存在的易受干扰的缺点.%Adsorption of alkaline phosphatase (APase) on minerals (goethite,montmorillonite) and sediments from Taihu Lake was studied.However,the concentration of alkaline phosphatase in the supernatant cannot be analyzed by the adsorption at 280 nm due to that the UV spectroscopy was changed.Quantitative analysis of alkaline phosphatase in the supernatant by the absorption peak (252~305 nm) integration method of UV spectroscopy was developed.This method determined the concentrations of alkaline phosphatase well.Compared to the results of Bradford,the absorption peak integration method can determine the concentrations of supernatant fast,conveniently,and accurately.This method can also be applied to other protein solution analysis and similar experiments.The drawbacks of traditional single wavelength method (280nm) were overcome to some extent based on the method of absorption peak integration.

  12. Phylogenetic diversity of alkaline protease-producing psychrotrophic bacteria from glacier and cold environments of Lahaul and Spiti, India.

    Science.gov (United States)

    Salwan, Richa; Gulati, Arvind; Kasana, Ramesh Chand

    2010-04-01

    The diversity of proteolytic bacteria associated with a glacier and cold environment soils from three different locations in Lahaul and Spiti, India was investigated. Two hundred seventeen bacterial strains were isolated in pure culture. Subsequently these strains were screened for protease-production and one hundred nine showed protease production. From these protease producing psychrotrophic bacteria twenty showing high enzyme production at low temperature and alkaline pH were characterized and identified. The 16S rRNA phylogenetic analysis revealed that none of the strains showed 100% identity with the validly published species of various genera. Isolates belonged to three classes i.e. Actinobacteria, Gammaproteobacteria and Alphaproteobacteria, and were affiliated with the genera Acinetobacter, Arthrobacter, Mycoplana, Pseudomonas, Pseudoxanthomonas, Serratia and Stenotrophomonas. The optimal growth temperature ranged from 10 to 28 degrees C and interestingly, high levels of enzyme productions were measured at growth temperatures between 15 and 25 degrees C, for most of the isolates in plate assay. Most of the isolates were found to produce at least two other hydrolytic enzymes along with protease. The crude protease from one strain was active over broad range of temperature and pH with optima at 30 degrees C and 7.5, respectively. The protease activity was enhanced by Ca(2+), dithiothreitol and beta-mercaptoethanol. While Na(+), Hg(2+), Zn(2+), Mn(2+), phenylmethanesulfonyl fluoride and ethylenediaminetetraacetic acid did not showed much effect on protease activity. The results enrich our knowledge on the psychrotrophic bacterial diversity and biogeographic distribution of enzyme producing bacteria in western Himalaya. PMID:20082368

  13. 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.

  14. Sensitive and selective determining ascorbic acid and activity of alkaline phosphatase based on electrochemiluminescence of dual-stabilizers-capped CdSe quantum dots in carbon nanotube-nafion composite.

    Science.gov (United States)

    Ma, Xiaolong; Zhang, Xin; Guo, Xinli; Kang, Qi; Shen, Dazhong; Zou, Guizheng

    2016-07-01

    Sensitive and selective determining bio-related molecule and enzyme play an important role in designing novel procedure for biological sensing and clinical diagnosis. Herein, we found that dual-stabilizers-capped CdSe quantum dots (QDs) in composite film of multi-walled carbon nanotubes (CNTs) and Nafion, displaying eye-visible monochromatic electrochemiluminescence (ECL) with fwhm of 37nm, which offers promising ECL signal for detecting ascorbic acid (AA) as well as the activity of alkaline phosphatase (ALP) in biological samples. It was also shown that the dual-stabilizers-capped CdSe QDs can preserve their highly passivated surface states with prolonged lifetime of excited states in Nafion mixtures, and facilitate electron-transfer ability of Nafion film along with CNTs. Compared with the QDs/GCE, the ECL intensity is enhanced 1.8 times and triggering potential shifted to lower energy by 0.12V on the CdSe-CNTs-Nafion/GCE. The ECL quenching degree increases with increasing concentration of AA in the range of 0.01-30nM with a limit of detection (LOD) of 5pM. The activity of ALP was determined indirectly according to the concentration of AA, generated in the hydrolysis reaction of l-ascorbic acid 2-phosphate sesquimagnesium (AA-P) in the presence of ALP as a catalyst, with an LOD of 1μU/L. The proposed strategy is favorable for developing simple ECL sensor or device with high sensitivity, spectral resolution and less electrochemical interference. PMID:27154663

  15. 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

  16. Draft genome sequences of two protease-producing strains of Arsukibacterium, isolated from two cold and alkaline environments

    DEFF Research Database (Denmark)

    Lylloff, Jeanette Eva; Hansen, Lea Benedicte Skov; Jepsen, Morten;

    2015-01-01

    Arsukibacterium ikkense GCM72(T) and a close relative, Arsukibacterium sp. MJ3, were isolated from two cold and alkaline environments as producers of extracellular proteolytic enzymes active at high pH and low temperature. This report describes the two draft genome sequences, which may serve as...

  17. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes.

    Science.gov (United States)

    Gomaa, Eman Zakaria

    2013-01-01

    The potentiality of 23 bacterial isolates to produce alkaline protease and carboxymethyl-cellulase (CMCase) on Ficus nitida wastes was investigated. Bacillus pumillus ATCC7061 was selected as the most potent bacterial strain for the production of both enzymes. It was found that the optimum production of protease and CMCase were recorded at 30 °C, 5% Ficus nitida leaves and incubation period of 72 h. The best nitrogen sources for protease and CMCase production were yeast extract and casein, respectively. Also maximum protease and CMCase production were reported at pH 9 and pH 10, respectively. The enzymes possessed a good stability over a pH range of 8-10, expressed their maximum activities at pH10 and temperature range of 30-50 °C, expressed their maximum activities at 50 °C. Ions of Hg(2+), Fe2+ and Ag(+) showed a stimulatory effect on protease activity and ions of Fe(2+), Mg(2+), Ca(2+), Cu(2+) and Ag(+) caused enhancement of CMCase activity. The enzymes were stable not only towards the nonionic surfactants like Triton X-100 and Tween 80 but also the strong anionic surfactant, SDS. Moreover, the enzymes were not significantly inhibited by EDTA or cystein. Concerning biotechnological applications, the enzymes retained (51-97%) of their initial activities upon incubation in the presence of commercials detergents for 1 h. The potential use of the produced enzymes in the degradation of human hair and cotton fabric samples were also assessed. PMID:24294252

  18. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes

    Directory of Open Access Journals (Sweden)

    Eman Zakaria Gomaa

    2013-01-01

    Full Text Available The potentiality of 23 bacterial isolates to produce alkaline protease and carboxymethyl-cellulase (CMCase on Ficus nitida wastes was investigated. Bacillus pumillus ATCC7061 was selected as the most potent bacterial strain for the production of both enzymes. It was found that the optimum production of protease and CMCase were recorded at 30 °C, 5% Ficus nitida leaves and incubation period of 72 h. The best nitrogen sources for protease and CMCase production were yeast extract and casein, respectively. Also maximum protease and CMCase production were reported at pH 9 and pH 10, respectively. The enzymes possessed a good stability over a pH range of 8-10, expressed their maximum activities at pH10 and temperature range of 30-50 °C, expressed their maximum activities at 50 °C. Ions of Hg2+, Fe2+ and Ag+ showed a stimulatory effect on protease activity and ions of Fe2+, Mg2+, Ca2+, Cu2+ and Ag+ caused enhancement of CMCase activity. The enzymes were stable not only towards the nonionic surfactants like Triton X-100 and Tween 80 but also the strong anionic surfactant, SDS. Moreover, the enzymes were not significantly inhibited by EDTA or cystein. Concerning biotechnological applications, the enzymes retained (51-97% of their initial activities upon incubation in the presence of commercials detergents for 1 h. The potential use of the produced enzymes in the degradation of human hair and cotton fabric samples were also assessed.

  19. 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

  20. Serum proteins, trace metals and phosphatases in psoriasis

    Directory of Open Access Journals (Sweden)

    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.

  1. 2143例0~12岁儿童血清碱性磷酸酶结果探讨%Clinical value of serum alkaline phosphatase in children aged 0 to 12 years

    Institute of Scientific and Technical Information of China (English)

    穆迎娟; 李忠信; 郑小玲

    2012-01-01

    目的 探讨2143例0~12岁儿童血清碱性磷酸酶(ALP)的结果.方法 应用SIEMENS DimensionRxL Max全自动生化分析仪检测2 143例来本院体检的0~12岁儿童血清ALP,对所得的数据进行统计学处理.结果 儿童血清ALP值在3~岁、9~岁和10~岁组男女性别间差异有统计学意义(P<0.05),其余组男女性别间差异均无统计学意义.儿童血清ALP值在1~岁、10~岁、11~12岁年龄组间差异有统计学意义(P<0.05),而其他年龄组之间差异无统计学意义(P>0.05).结论 0~12岁儿童血清ALP水平显著高于成人.儿童时期血清ALP水平与其年龄和性别相关,应根据不同性别、年龄的儿童相应的发育情况给予不同的参考范围.%Objective To analyze the clinical significance of serum alkaline phosphatase (ALP) in children. Methods Serum level of ALP from 2 143 children with the age of 0 to 12 years old children in our hospital attending for the physical examination were measured by SIEMENS Dimension RxL Max AUTOLAB, and the results were analyzed statistically. Results There was a significant difference in the level of serum ALP in children aged 3 to 4 years old, 9 to 10 years old and 10 to 11 years old. The level of ALP in children aged 0 to 1 year old and 2 to 10 years was similar in both sex. Conclusion The serum level of ALP in children was higher than the adult. The level of ALP is related to their age and sex. so children serum ALP values should be based on the development situation of children of different gender, age, given the different reference ranges.

  2. Partial purification and properties of cellulase-free alkaline xylanase produced by Rhizopus stolonifer in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Antonio José Goulart

    2005-05-01

    Full Text Available Rhizopus stolonifer was cultivated in wheat bran to produce a cellulase-free alkaline xylanase. The purified enzyme obtained after molecular exclusion chromatography in Sephacryl S-200 HR showed optimum temperature as 45º C and hydrolysis pHs optima as pH 6.0 and 9.0. Xylanase presented higher Vmax at pH 9.0 (0.87 µmol/mg protein than at pH 6.0 and minor Km at pH 6.0 (7.42 mg/mL than at pH 9.0.Rhizopus stolonifer foi cultivado em meio de farelo de trigo para produzir uma xilanase alcalina celulase-free. Uma amostra parcialmente purificada desta enzima foi obtida após cromatografia de exclusão molecular em Sephacryl S-200 HR. A temperatura ótima de hidrólise determinada (45º C está dentro do intervalo citado na literatura (45º C a 60º C para xilanases microbianas. Quanto ao pH ótimo, a amostra obtida apresentou atividades máximas em pH 6,0 e 9,0. Estes dados diferem da literatura, uma vez que o pH ótimo citado para a maioria das xilanases estudadas varia entre 4,0 e 5,5. De acordo com os estudos cinéticos realizados, a xilanase apresentou maior Vmax em pH 9,0 (0,87 µmol/mg proteína e menor Km em pH 6,0 (7,42 mg/mL. Os dois pHs ótimos determinados podem indicar a presença de isoformas desta enzima. Estes dados são interessantes pelo fato de que enzimas xilanolíticas alcalinas celulase-free podem ser utilizadas para o biobranqueamento da polpa na indústria de papel.

  3. Isolation of phosphatase-producing phosphate solubilizing bacteria from Loriya hot spring: Investigation of phosphate solubilizing in the presence of different parameters

    Directory of Open Access Journals (Sweden)

    Maryam Parhamfar

    2014-04-01

    Full Text Available Introduction: Biofertilizers are the microorganisms that can convert useless nutrient to usable compounds. Unlike fertilizer, cost of biofertilizer production is low and doesn’t produce ecosystem pollution. Phosphate fertilizers can be replaced by phosphate biofertilizer to produce improvement. So, it is necessary to screen the climate-compatible phosphate solubilizing bacteria. Materials and methods: In this project samples were picked up from Loriya hot spring, which are located in Jiroft. Samples were incubated in PKV medium for 3 days. Screening of phosphate solubilizing bacteria was performed on the specific media, based on clear area diameter. The best bacterium was identified based on 16s rDNA gene. Phosphate solubilizing activity of this strain was considered in different carbon, nitrogen, phosphate and pH sources. Results: Sequence alignment and phylogenetic tree results show that B. sp. LOR033 is closely related to Bacillus licheniformis, with 97% homology. In addition, results show that maximum enzyme production was performed after 2 days that incubation pH was decreased simultaneously when the time was increased. Carbon sources investigation show that glucose is the most appropriate in enzyme production and phosphate releasing. Furthermore, results show that the optimum initial pH for phytase production was pH5.0. Different phosphate sources show that tricalcium phosphate has the suitable effect on enzyme activity in three days of incubation. Discussion and conclusion: Phosphatase enzyme production capacity, growth in acidic pH and phosphate solubilizing potential in different salt and phosphate sources show that this strain has considerable importance as biofertilizers.

  4. Screening and characterization of alkaline protease produced by a pink pigmented facultative methylotrophic (PPFM) strain, MSF 46

    OpenAIRE

    Shanmugam Jayashree; Balumuri Annapurna; Renganathan Jayakumar; Tongmin Sa; Sundaram Seshadri

    2014-01-01

    Among the various bacterial isolates, the strain MSF 46 isolated from thorn forest soil samples, Tamil Nadu, India, was screened and characterized for its proteolytic activity. While the 16S rRNA sequencing and biochemical characterization revealed that the strain closely resembles Methylobacterium sp., methylotrophy of the strain was confirmed by the sequence homology of mxaF gene with other relative Methylobacterium sp. The alkaline protease was purified to homogeneity using DEAE cellulose ...

  5. 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

  6. 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.

  7. Co-cultivation of Aspergillus nidulans Recombinant Strains Produces an Enzymatic Cocktail as Alternative to Alkaline Sugarcane Bagasse Pretreatment

    Science.gov (United States)

    Lima, Matheus S.; Damasio, André R. de L.; Crnkovic, Paula M.; Pinto, Marcelo R.; da Silva, Ana M.; da Silva, Jean C. R.; Segato, Fernando; de Lucas, Rosymar C.; Jorge, João A.; Polizeli, Maria de L. T. de M.

    2016-01-01

    Plant materials represent a strategic energy source because they can give rise to sustainable biofuels through the fermentation of their carbohydrates. A clear example of a plant-derived biofuel resource is the sugar cane bagasse exhibiting 60–80% of fermentable sugars in its composition. However, the current methods of plant bioconversion employ severe and harmful chemical/physical pretreatments raising biofuel cost production and environmental degradation. Replacing these methods with co-cultivated enzymatic cocktails is an alternative. Here we propose a pretreatment for sugarcane bagasse using a multi-enzymatic cocktail from the co-cultivation of four Aspergillus nidulans recombinant strains. The co-cultivation resulted in the simultaneous production of GH51 arabinofuranosidase (AbfA), GH11 endo-1,4-xylanase (XlnA), GH43 endo-1,5-arabinanase (AbnA) and GH12 xyloglucan specific endo-β-1,4-glucanase (XegA). This core set of recombinant enzymes was more efficient than the alternative alkaline method in maintaining the cellulose integrity and exposing this cellulose to the following saccharification process. Thermogravimetric and differential thermal analysis revealed residual byproducts on the alkali pretreated biomass, which were not found in the enzymatic pretreatment. Therefore, the enzymatic pretreatment was residue-free and seemed to be more efficient than the applied alkaline method, which makes it suitable for bioethanol production. PMID:27199917

  8. Co-cultivation of Aspergillus nidulans recombinant strains produces an enzymatic cocktail as alternative to alkaline sugarcane bagasse pretreatment

    Directory of Open Access Journals (Sweden)

    Matheus Sanita Lima

    2016-04-01

    Full Text Available Plant materials represent a strategic energy source because they can give rise to sustainable biofuels through the fermentation of their carbohydrates. A clear example of a plant-derived biofuel resource is the sugar cane bagasse exhibiting 60 % - 80 % of fermentable sugars in its composition. However, the current methods of plant bioconversion employ severe and harmful chemical/physical pretreatments raising biofuel cost production and environmental degradation. Replacing these methods with co-cultivated enzymatic cocktails is an alternative. Here we propose a pretreatment for sugarcane bagasse using a multi-enzymatic cocktail from the co-cultivation of four Aspergillus nidulans recombinant strains. The co-cultivation resulted in the simultaneous production of GH51 arabinofuranosidase (AbfA, GH11 endo-1,4-xylanase (XlnA, GH43 endo-1,5-arabinanase (AbnA and GH12 xyloglucan specific endo-β-1,4-glucanase (XegA. This core set of recombinant enzymes was more efficient than the alternative alkaline method in maintaining the cellulose integrity and exposing this cellulose to the following saccharification process. Thermogravimetric and differential thermal analysis revealed residual byproducts on the alkali pretreated biomass, which were not found in the enzymatic pretreatment. Therefore, the enzymatic pretreatment was residue-free and seemed to be more efficient than the applied alkaline method, which makes it suitable for bioethanol production.

  9. Co-cultivation of Aspergillus nidulans Recombinant Strains Produces an Enzymatic Cocktail as Alternative to Alkaline Sugarcane Bagasse Pretreatment.

    Science.gov (United States)

    Lima, Matheus S; Damasio, André R de L; Crnkovic, Paula M; Pinto, Marcelo R; da Silva, Ana M; da Silva, Jean C R; Segato, Fernando; de Lucas, Rosymar C; Jorge, João A; Polizeli, Maria de L T de M

    2016-01-01

    Plant materials represent a strategic energy source because they can give rise to sustainable biofuels through the fermentation of their carbohydrates. A clear example of a plant-derived biofuel resource is the sugar cane bagasse exhibiting 60-80% of fermentable sugars in its composition. However, the current methods of plant bioconversion employ severe and harmful chemical/physical pretreatments raising biofuel cost production and environmental degradation. Replacing these methods with co-cultivated enzymatic cocktails is an alternative. Here we propose a pretreatment for sugarcane bagasse using a multi-enzymatic cocktail from the co-cultivation of four Aspergillus nidulans recombinant strains. The co-cultivation resulted in the simultaneous production of GH51 arabinofuranosidase (AbfA), GH11 endo-1,4-xylanase (XlnA), GH43 endo-1,5-arabinanase (AbnA) and GH12 xyloglucan specific endo-β-1,4-glucanase (XegA). This core set of recombinant enzymes was more efficient than the alternative alkaline method in maintaining the cellulose integrity and exposing this cellulose to the following saccharification process. Thermogravimetric and differential thermal analysis revealed residual byproducts on the alkali pretreated biomass, which were not found in the enzymatic pretreatment. Therefore, the enzymatic pretreatment was residue-free and seemed to be more efficient than the applied alkaline method, which makes it suitable for bioethanol production. PMID:27199917

  10. 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.

  11. Screening and characterization of alkaline protease produced by a pink pigmented facultative methylotrophic (PPFM strain, MSF 46

    Directory of Open Access Journals (Sweden)

    Shanmugam Jayashree

    2014-12-01

    Full Text Available Among the various bacterial isolates, the strain MSF 46 isolated from thorn forest soil samples, Tamil Nadu, India, was screened and characterized for its proteolytic activity. While the 16S rRNA sequencing and biochemical characterization revealed that the strain closely resembles Methylobacterium sp., methylotrophy of the strain was confirmed by the sequence homology of mxaF gene with other relative Methylobacterium sp. The alkaline protease was purified to homogeneity using DEAE cellulose ion exchange chromatography, with a 5.2-fold increase in specific activity and 34% recovery. The apparent molecular weight of the enzyme was determined as 40 kDa by SDS–PAGE study. The pH and temperature optima were 9.0 and 50 °C respectively with maximum protease activity of 1164 U/ml. Protease of MSF 46 was active in a broad pH range 7.0–11.0 with a maximum at pH 8.5 and exhibited thermostability at 50 °C. The enzyme activity was inhibited by PMSF but showed stability with Tween 20, Triton X-100 and hydrogen peroxide. Nearly 30% reduction in enzyme activity was observed in the presence of EDTA and DTT. The enzyme was effective in hydrolyzing gelatin, skimmed milk and blood clots and exhibited the potency for dehairing of goat skin and removing blood stain from cotton fabric. Significant morphological changes were observed under scanning electron microscope between cells grown in normal and casein amended medium. This first detailed report on the production of alkaline protease by a PPFM strain appears promising toward development of protocols for mass production, study of the molecular mechanism and other applications.

  12. The role of phosphatases in the initiation of skeletal mineralization.

    Science.gov (United States)

    Millán, José Luis

    2013-10-01

    Endochondral ossification is a carefully orchestrated process mediated by promoters and inhibitors of mineralization. Phosphatases are implicated, but their identities and functions remain unclear. Mutations in the tissue-nonspecific alkaline phosphatase (TNAP) gene cause hypophosphatasia, a heritable form of rickets and osteomalacia, caused by an arrest in the propagation of hydroxyapatite (HA) crystals onto the collagenous extracellular matrix due to accumulation of extracellular inorganic pyrophosphate (PPi), a physiological TNAP substrate and a potent calcification inhibitor. However, TNAP knockout (Alpl(-/-)) mice are born with a mineralized skeleton and have HA crystals in their chondrocyte- and osteoblast-derived matrix vesicles (MVs). We have shown that PHOSPHO1, a soluble phosphatase with specificity for two molecules present in MVs, phosphoethanolamine and phosphocholine, is responsible for initiating HA crystal formation inside MVs and that PHOSPHO1 and TNAP have nonredundant functional roles during endochondral ossification. Double ablation of PHOSPHO1 and TNAP function leads to the complete absence of skeletal mineralization and perinatal lethality, despite normal systemic phosphate and calcium levels. This strongly suggests that the Pi needed for initiation of MV-mediated mineralization is produced locally in the perivesicular space. As both TNAP and nucleoside pyrophosphohydrolase-1 (NPP1) behave as potent ATPases and pyrophosphatases in the MV compartment, our current model of the mechanisms of skeletal mineralization implicate intravesicular PHOSPHO1 function and Pi influx into MVs in the initiation of mineralization and the functions of TNAP and NPP1 in the extravesicular progression of mineralization.

  13. P depletion and activity of phosphatases in the rhizosphere of mycorrhizal and non-mycorrhizal cucumber (Cucumis Sativus L.)

    DEFF Research Database (Denmark)

    Joner, E.J.; Magid, J.; Gahoonia, T.S.;

    1995-01-01

    was sectioned in a freezing microtome and analyzed for extracellular acid (pH 5.2) and alkaline (pH 8.5) phosphatase activity as well as depletion of NaHCO-3-extractable inorganic P (P-i) and P-o. Roots and mycorrhizal hyphae depleted the soil of P-i but did not influence the concentration of P-o in spite......An experiment was set up to test the ability of arbuscular mycorrhizal (AM) roots and hyphae to produce extracellular phosphatases and to study the relationship between phosphatase activity and soil organic P (P-o). Non-mycorrhizal cucumber and cucumber in symbiosis with either of two mycorrhizal...... of increased phosphatase activity in soil influenced by roots. Phosphatase activity at both pH values was highest in soil influenced by uncolonized roots, but this was attributed to higher root length densities as compared to mycorrhizal roots. Mycorrhizal hyphae showed no influence on soil phosphatase...

  14. The change of serum alkaline phosphatase and fracture healing about the fracture with type 2 diabetes%骨折合并Ⅱ型糖尿病患者骨转换生化指标及骨折愈合的变化

    Institute of Scientific and Technical Information of China (English)

    杨东辉

    2012-01-01

    [ Objective ] To observe the change of serum alkaline phosphatase and fracture-healing about the fracture with type 2 diabetes. [ Method ] The patients with fracture were divided into two groups; fracture-patient with type 2 diabetes, and fracture-patient with normal blood glucose. The change of serum alkaline phosphatase( AKP)and fracture-healing were observed in different groups. [Result] AKP in the group of the fracture-patient with type 2 diabetes changed significantly as compared with the group of the fracture-patient with normal blood glucose. Type 2 diabetes would slow down the speed of fracture-healing (P<0.01). [ Conclusion ] Type 2 diabetes could influence the metabolism-speed of fracture-healing.%[目的]观察Ⅱ型糖尿病骨折患者骨转换生化指标-碱性磷酸酶及骨折愈合的变化.[方法]将在我科住院行手术治疗的骨折患者分为非糖尿病骨折组72例及Ⅱ型糖尿病骨折组79例,观察术前、术后两组患者血浆碱性磷酸酶(AKP)及骨折愈合情况.[结果]非糖尿病骨折组与Ⅱ型糖尿病骨折组比较,血AKP检测指标及骨折愈合情况差异具有显著性意义(P<0.01,P<0.05).Ⅱ型糖尿病可影响骨折后血AKP浓度的变化及骨折愈合速度.[结论]Ⅱ型糖尿病可影响骨质代谢,并易造成骨折延缓愈合.

  15. 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.

  16. Production and characterization of alkaline protease from hemoglobin-degrading Bacillus pumilus NJM4 to produce fermented blood meal

    OpenAIRE

    Yao, Dawei; Qu, Jiao; Chang, Peiwei; Tao, Yanhua; Yang, Deji

    2011-01-01

    The aim of the research was to isolate the hemoglobin-degrading bacterial strain to produce fermented blood meal and to characterize the protease produced by this strain. The strain NJM4, a kind of hemoglobin-degrading bacterial strain, was isolated by blood agar plates from slaughterhouse and identified as a Bacillus pumilus by physiological, biochemical, and morphological characteristics and by 16S rRNA gene sequencing. Bacillus pumilus NJM4 could degrade hemoglobin up to 85% in 36 h under ...

  17. [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. PMID:16350755

  18. 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.

  19. 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.

  20. Assessment the value of ascites alkaline phosphatase and lactate dehydrogenase in surgical timing decision making in patients with intestinal obstruction%腹水碱性磷酸酶及乳酸脱氢酶在判断肠梗阻手术治疗时机的应用价值

    Institute of Scientific and Technical Information of China (English)

    梁栋; 赖苏何

    2014-01-01

    Objective To investigate the value of alkaline phosphatase,lactate dehydrogenase,glucose, Na+ 、K+ 、Ca2 + in the ascites for assessment the timing of surgical treatment of intestinal obstruction. Meth-ods The clinical records of 94 patients with intestinal obstruction underwent surgery in our hospital were ret-rospectively analyzed.They were assigned to a necrosis group (n =50,with bowel necrosis)and a non- nec-rosis group (n =44,without bowel necrosis)according to the presence or absence of bowel necrosis,and the differences of ascites - related indicators between the two groups were compared. Results Necrotic group showed significantly higher levels of alkaline phosphatase and lactate dehydrogenase than non-necrosis group (158.25±37.85 vs.43.47±17.54;5087.25±1218.1 1 vs.389.58±154.22),and the group-pair comparison exhibited statistically significant differences (Z = 18.440,P 0.05). Conclusion Alkaline phosphatase and lactate dehydrogenase in ascites are important in evaluating the bowel with or without necrosis,and they could be used as indicators of making de-cision for timing of surgical treatment of intestinal obstruction.%目的:探讨腹水碱性磷酸酶、乳酸脱氢酶、葡萄糖、Na+、K+、Ca2+对判断肠梗阻手术治疗时机的价值。方法回顾性分析我院收治的94例肠梗阻手术治疗患者的临床资料,根据肠管有无坏死分为坏死组(50例)和未坏死组(44例),比较两组患者腹水相关指标的差异。结果坏死组碱性磷酸酶和乳酸脱氢酶含量均显著高于非坏死组[(158.25±37.85)比(43.47±17.54);(5087.25±1218.11)比(389.58±154.22)],且差异具有统计学意义(Z =18.440,P 0.05)。结论腹水碱性磷酸酶和乳酸脱氢酶对判断肠管有无坏死具有重要意义,可作为判断肠梗阻手术治疗时机的指标。

  1. 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.

  2. HbNIN2, a cytosolic alkaline/neutral-invertase, is responsible for sucrose catabolism in rubber-producing laticifers of Hevea brasiliensis (para rubber tree).

    Science.gov (United States)

    Liu, Shujin; Lan, Jixian; Zhou, Binhui; Qin, Yunxia; Zhou, Yihua; Xiao, Xiaohu; Yang, Jianghua; Gou, Jiqing; Qi, Jiyan; Huang, Yacheng; Tang, Chaorong

    2015-04-01

    In Hevea brasiliensis, an alkaline/neutral invertase (A/N-Inv) is responsible for sucrose catabolism in latex (essentially the cytoplasm of rubber-producing laticifers, the source of natural rubber) and implicated in rubber yield. However, neither the gene encoding this enzyme nor its molecular and biochemical properties have been well documented. Three Hevea A/N-Inv genes, namely HbNIN1, 2 and 3, were first cloned and characterized in planta and in Escherichia coli. Cellular localizations of HbNIN2 mRNA and protein were probed. From latex, active A/N-Inv proteins were purified, identified, and explored for enzymatic properties. HbNIN2 was identified as the major A/N-Inv gene functioning in latex based on its functionality in E. coli, its latex-predominant expression, the conspicuous localization of its mRNA and protein in the laticifers, and its expressional correlation with rubber yield. An active A/N-Inv protein was partially purified from latex, and determined as HbNIN2. The enhancement of HbNIN2 enzymatic activity by pyridoxal is peculiar to A/N-Invs in other plants. We conclude that HbNIN2, a cytosolic A/N-Inv, is responsible for sucrose catabolism in rubber laticifers. The results contribute to the studies of sucrose catabolism in plants as a whole and natural rubber synthesis in particular. PMID:25581169

  3. Isolation of phosphatase-producing phosphate solubilizing bacteria from Loriya hot spring: Investigation of phosphate solubilizing in the presence of different parameters

    OpenAIRE

    Maryam Parhamfar; Arastoo Badoei-Dalfard; Mouj Khaleghi; Mehdi Hassanshahian

    2014-01-01

    Introduction: Biofertilizers are the microorganisms that can convert useless nutrient to usable compounds. Unlike fertilizer, cost of biofertilizer production is low and doesn’t produce ecosystem pollution. Phosphate fertilizers can be replaced by phosphate biofertilizer to produce improvement. So, it is necessary to screen the climate-compatible phosphate solubilizing bacteria. Materials and methods: In this project samples were picked up from Loriya hot spring, which are located in Jiro...

  4. Growth and extracellular phosphatase activity of arbuscular mycorrhizal hyphae as influenced by soil organic matter

    DEFF Research Database (Denmark)

    Joner, E.J.; Jakobsen, I.

    1995-01-01

    length density was twice as high in soil with added straw compared to the two other treatments. Mycorrhizal colonization resulted in lower activity of acid phosphatase in the HC for two out of three treatments. Alkaline phosphatase activity was only decreased by mycorrhiza in soil without organic matter...... additions. In soil with added clover alkaline phosphatase activity increased due to the presence of mycorrhizal hyphae. We suggest that mycorrhizas may influence the exudation of acid phosphatase by roots. Hyphae of G. invermaium did apparently not excrete extracellular phosphatases, but their presence may......Two experiments were set up to investigate the influence of soil organic matter on growth of arbuscular mycorrhizal (AM) hyphae and concurrent changes in soil inorganic P, organic P and phosphatase activity. A sandy loam soil was kept for 14 months under two regimes (outdoor where surplus...

  5. Draft Genome Sequence of Halobacillus sp. Strain KGW1, a Moderately Halophilic and Alkaline Protease-Producing Bacterium Isolated from the Rhizospheric Region of Phragmites karka from Chilika Lake, Odisha, India.

    Science.gov (United States)

    Panda, Ananta Narayan; Mishra, Samir R; Ray, Lopamudra; Sahu, Neha; Acharya, Ankita; Jadhao, Sudhir; Suar, Mrutyunjay; Adhya, Tapan Kumar; Rastogi, Gurdeep; Pattnaik, Ajit Kumar; Raina, Vishakha

    2016-01-01

    Halobacillus sp. strain KGW1 is a moderately halophilic, rod shaped, Gram-positive, yellow pigmented, alkaline protease-producing bacterium isolated from a water sample from Chilika Lake, Odisha, India. Sequencing of bacterial DNA assembled a 3.68-Mb draft genome. The genome annotation analysis showed various gene clusters for tolerance to stress, such as elevated pH, salt concentration, and toxic metals. PMID:27365341

  6. Draft Genome Sequence of Halobacillus sp. Strain KGW1, a Moderately Halophilic and Alkaline Protease-Producing Bacterium Isolated from the Rhizospheric Region of Phragmites karka from Chilika Lake, Odisha, India

    Science.gov (United States)

    Panda, Ananta Narayan; Mishra, Samir R.; Ray, Lopamudra; Sahu, Neha; Acharya, Ankita; Jadhao, Sudhir; Suar, Mrutyunjay; Adhya, Tapan Kumar; Rastogi, Gurdeep; Pattnaik, Ajit Kumar

    2016-01-01

    Halobacillus sp. strain KGW1 is a moderately halophilic, rod shaped, Gram-positive, yellow pigmented, alkaline protease-producing bacterium isolated from a water sample from Chilika Lake, Odisha, India. Sequencing of bacterial DNA assembled a 3.68-Mb draft genome. The genome annotation analysis showed various gene clusters for tolerance to stress, such as elevated pH, salt concentration, and toxic metals. PMID:27365341

  7. Draft Genome Sequence of Halobacillus sp. Strain KGW1, a Moderately Halophilic and Alkaline Protease-Producing Bacterium Isolated from the Rhizospheric Region of Phragmites karka from Chilika Lake, Odisha, India

    OpenAIRE

    Panda, Ananta Narayan; Mishra, Samir R.; Ray, Lopamudra; Sahu, Neha; Acharya, Ankita; Jadhao, Sudhir; Suar, Mrutyunjay; Adhya, Tapan Kumar; Rastogi, Gurdeep; Pattnaik, Ajit Kumar; Raina, Vishakha

    2016-01-01

    Halobacillus sp. strain KGW1 is a moderately halophilic, rod shaped, Gram-positive, yellow pigmented, alkaline protease-producing bacterium isolated from a water sample from Chilika Lake, Odisha, India. Sequencing of bacterial DNA assembled a 3.68-Mb draft genome. The genome annotation analysis showed various gene clusters for tolerance to stress, such as elevated pH, salt concentration, and toxic metals.

  8. 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

  9. 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

  10. Study on oxidative stress and activity of alkaline phosphatase of rats exposed to different period of fluoride%投氟不同时间大鼠血清氧化应激和碱性磷酸酶活性的变化研究

    Institute of Scientific and Technical Information of China (English)

    徐辉; 范海清; 张金铭; 李广生

    2010-01-01

    Objective To observe the status of oxidative stress and activity of alkaline phosphatase(ALP) in rats exposed to high fluoride for the different periods and to analyze the effect of fluoride on the activity of ALP and oxidative stress in fluorosis rats. Methods Twenty-four Wistar rats were divided into control and high-fluoride groups according to their body mass, 12 rats in each group. The control group drank tap water(sodium fluoride concentrations 0.05); the level of uric acid in serum was significantly higher in high-fluoride group for 1,4 week[ (89.53 ± 13.21 ), (88.47 ± 19.78 )μmol/L] compared to the control [ (77.79 ± 11.43 ), (65.42 ± i 3.42) μ mol/L, all P 0.05);投氟1、4周大鼠血清尿酸[(89.53±13.21)、(88.47±19.78)μmol/L]较对照组[(77.79±11.43)、(65.42±13.42)μmol/L]明显增高(P均<0.05),而投氟8、12周大鼠尿酸[(67.21±9.44)、(73.95±9.52)μmol/L]低于对照组[(77.79±11.43)、(65.42±13.42)μmol/L,P均<0.05].结论 投与过量氟大鼠的ALP活性变化具有一定的时间依赖性;而投氟刺激大鼠氧化应激水平增强,这种刺激与投氟时间没有明显关联.

  11. 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

    目的:了解小剂量葛根素联合雌二醇对去卵巢大鼠的骨组织、血钙、磷和碱性磷酸酶的影响,为中西医结合治疗绝经后骨质疏松症提供实验依据.方法:5月龄健康雌性大白鼠120只,分成5个实验组(每组24只):①假手术组(sham);②去卵巢模型组(OVX);③葛根索组(Pr),皮下注射葛根素,50 mg/kg,1次/d;④雌二醇组(E2),皮下注射雌二醇200 μg/kg,2次/周;⑤小剂量葛根素+雌二醇组(Pr+ E2),皮下注射雌二醇100 μg/kg,2次/周和葛根素25 mg/kg,1次/d.各实验组在第4、8、12和20周,随机取6只大鼠取股骨切片观察骨组织,采血测量血钙、磷和碱性磷酸酶,数据进行统计学分析.结果:OVX组第4、8、12、20周的骨组织呈骨质疏松病理改变,血钙、磷明显低于sham组(P<0.01),OVX组的第4、8、12、20周血碱性磷酸酶均明显高于sham组(P<0.01).3个治疗组各时间的骨组织和血钙、磷和碱性磷酸酶与sham组无统计学意义(P>0.05).小剂量的葛根素联合雌二醇治疗能使去卵巢大鼠骨组织和血钙、磷和血碱性磷酸酶基本恢复正常(P>0.05),与较大剂量的葛根素组或较大剂量的雌二醇组相比无统计学意义(P>0.05).结论:小剂量的雌二醇与葛根素对去卵巢大鼠的骨质疏松症的治疗效果与单独使用较大剂量的葛根素或较大剂量的雌二醇相比治疗效果相近.%Objective: To understand the effect of puerarin combined with estradiol on bone tissue and serum calcium, phosphorus, and alkaline phosphatase in ovariectomized rats, and provide experimental basis for treating postmenopausal osteoporosis with traditional Chinese medicine combined with Western medicine. Methods: One hundred and twenty 5 - month - old female rats were selected and divided into five experimental groups, 24 rats in each group: sham - operation group, ovariectomized model group, puerarin group (the rats were treated with subcutaneous injection of puerarin

  12. Application of alkaline thermo-stable lipase(s) enzyme produced from irradiated microbial isolate in the field of detergent technology

    International Nuclear Information System (INIS)

    Due to continuous demand for manufacture of high quality, low coast industrial detergents containing lipolytic enzymes and due to continuous accumulation of enviro-agro-industrial wastes which are good and suitable conditions for growth and reproduction of pathogenic microorganisms, our study aims at isolating thermoalkalophilic lipase producer microorganisms from enviro-agro-industrial wastes and selection of the most potent isolate for studying physiological conditions controlling enzyme formation also purification characterization and some applications on purified and crude enzyme as bio-detergent. Some environmental and industrial wastes were collected from different places. The industrial wastes include, cotton seed, soyabean, sun flower, lin seed and olive oil wastes. Environmental wastes include poultry and fish wastes, all these wastes were dried at 70 degree C, grounded and used for isolation of microorganisms and lipase(s) production.Nine thermoalkalophilic bacterial isolates were isolated from enviro-agro-industrial wastes at ph 11.5 and 70 degree C. They were purified and screening for their ability of thermoalkalo-stable lipase(s) formation, this is followed by examining the effect of different nutritional media and exposure of bacterial isolates to different doses of gamma irradiation and the influence of these radiation on lipase(s) productivity by these isolates. From the results it was found that.1- The most potent lipase(s) forming bacterial isolates were isolates number B2 and B3 which cultivated on medium A amended with fish-wastes as being the best nutritional medium for enzyme formation. 2-Bacterial isolate B2 finally was selected as being the most potent lipase(s) forming bacterial isolate cultivated on fish-wastes and yeast extract (in tap water) and identified according to key's of Bergey Manual of Systematic Bacteriology (1984) as being Bacillus brevis B2.The optimum culture conditions for maximum biosynthesis of extracellular lipase

  13. Optimization of NaOH Molarity, LUSI Mud/Alkaline Activator, and Na2SiO3/NaOH Ratio to Produce Lightweight Aggregate-Based Geopolymer

    Directory of Open Access Journals (Sweden)

    Rafiza Abdul Razak

    2015-05-01

    Full Text Available This paper presents the mechanical function and characterization of an artificial lightweight geopolymer aggregate (ALGA using LUSI (Sidoarjo mud and alkaline activator as source materials. LUSI stands for LU-Lumpur and SI-Sidoarjo, meaning mud from Sidoarjo which erupted near the Banjarpanji-1 exploration well in Sidoarjo, East Java, Indonesia on 27 May 2006. The effect of NaOH molarity, LUSI mud/Alkaline activator (LM/AA ratio, and Na2SiO3/NaOH ratio to the ALGA are investigated at a sintering temperature of 950 °C. The results show that the optimum NaOH molarity found in this study is 12 M due to the highest strength (lowest AIV value of 15.79% with lower water absorption and specific gravity. The optimum LUSI mud/Alkaline activator (LM/AA ratio of 1.7 and the Na2SiO3/NaOH ratio of 0.4 gives the highest strength with AIV value of 15.42% with specific gravity of 1.10 g/cm3 and water absorption of 4.7%. The major synthesized crystalline phases were identified as sodalite, quartz and albite. Scanning Electron Microscope (SEM image showed more complete geopolymer matrix which contributes to highest strength of ALGA produced.

  14. Optimization of NaOH Molarity, LUSI Mud/Alkaline Activator, and Na2SiO3/NaOH Ratio to Produce Lightweight Aggregate-Based Geopolymer.

    Science.gov (United States)

    Razak, Rafiza Abdul; Abdullah, Mohd Mustafa Al Bakri; Hussin, Kamarudin; Ismail, Khairul Nizar; Hardjito, Djwantoro; Yahya, Zarina

    2015-05-21

    This paper presents the mechanical function and characterization of an artificial lightweight geopolymer aggregate (ALGA) using LUSI (Sidoarjo mud) and alkaline activator as source materials. LUSI stands for LU-Lumpur and SI-Sidoarjo, meaning mud from Sidoarjo which erupted near the Banjarpanji-1 exploration well in Sidoarjo, East Java, Indonesia on 27 May 2006. The effect of NaOH molarity, LUSI mud/Alkaline activator (LM/AA) ratio, and Na2SiO3/NaOH ratio to the ALGA are investigated at a sintering temperature of 950 °C. The results show that the optimum NaOH molarity found in this study is 12 M due to the highest strength (lowest AIV value) of 15.79% with lower water absorption and specific gravity. The optimum LUSI mud/Alkaline activator (LM/AA) ratio of 1.7 and the Na2SiO3/NaOH ratio of 0.4 gives the highest strength with AIV value of 15.42% with specific gravity of 1.10 g/cm3 and water absorption of 4.7%. The major synthesized crystalline phases were identified as sodalite, quartz and albite. Scanning Electron Microscope (SEM) image showed more complete geopolymer matrix which contributes to highest strength of ALGA produced.

  15. 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.

  16. 中性粒细胞碱性磷酸酶活性在红细胞增多症鉴别诊断中的临床意义%The significance of neutrophil alkaline phosphatase activity in the differential diagnosis of polycythemia

    Institute of Scientific and Technical Information of China (English)

    汤俊峰; 叶龙飞

    2016-01-01

    Objective To discuss clinical significance of the neutrophil alkaline phosphatase(NAP) activity in polycythemia pa‐tients .Methods 35 cases of patients with secondary polycythemia(secondary group) and 22 patients with polycythemia vera (poly‐cythemia vera group) were enrolled in the study ,whose peripheral blood smears were stained by using chemical method and the pos‐itive rate and scores of NAP were assessed .In addition to that ,30 cases of healthy people who had underwent physical examination were recruited as control group .Results NAP positive rate and scores of the polycythemia vera group were significantly higher than those of secondary and control group(P0 .05) .Conclusion NAP score is a good methord to diagnose polycythemia ,which could facilitate rapid ,accurate diagno‐sis and be used as auxiliary indicators in the differential diagnosis of polycythemia .%目的:探讨中性粒细胞碱性磷酸酶(NAP)活性在红细胞增多症鉴别诊断中的临床意义。方法对35例继发性红细胞增多症患者(继红组)、22例真性红细胞增多症患者(真红组)外周血涂片用化学法染色并对NAP阳性率和积分进行判断并以30例健康体检者作为对照组。结果真性组患者NAP阳性率及积分明显高于继红组及对照组,差异均有统计学意义(P<0.05);继红组NAP积分及阳性率与健康组比较差异均无统计学意义(P>0.05)。结论NAP积分应用于诊断红细胞增多症是一种有效的方法,有助于快速、准确的鉴别诊断,可作为红细胞增多症鉴别诊断的重要辅助指标。

  17. 僵蚕提取物对棉铃虫的胃毒作用和碱性磷酸酶活性的影响%Toxicity and Alkaline Phosphatase Activity Inhibition of the Extracts from Bombyx batryticatus Against Helicoverpa armigera

    Institute of Scientific and Technical Information of China (English)

    曹军; 于欢; 张瑞凤; 王沛; 张雅林; 王敦

    2012-01-01

    僵蚕提取物对棉铃虫有胃毒作用,对饲毒后不同时间棉铃虫3龄幼虫体内碱性磷酸酶的活性进行了研究.结果表明,在3龄棉铃虫人工饲料中添加僵蚕提取物浓度为3 mg·g-1时,饲毒第3天试虫的校正死亡率为66.73%;5 mg·g-1的剂量导致试虫死亡率可以高达83.33%,表明僵蚕提取物对棉铃虫具有很强的胃毒作用.试虫碱性磷酸酶活性分析表明,1~5 mg·g-1浓度处理组的试虫碱性磷酸酶活性与对照组相比均显著下降,并有明显的剂量相关性.推测侵蚕对棉铃虫幼虫的毒杀作用机制之一与其对碱性磷酸酶活性的抑制有关.%This paper reported the study results on feeding toxicity and inhibition effects on alkaline phos-phatase (ALP) of Helicoverpa armigera treated with the extracts of Bombyx batryticatus. The corrected mortality of H. armigera treated with the extracts of B. batryticatus at 3 mg · g-1 concentration in insect diet was 66. 7% after feeding on the toxic diet three days, while the corrected mortality was up to 83% to the larvae treated with a concentration of 5 mg · g-1, indicating that the B. batryticatus extracts was high toxic to the third instar larvae of H. armigera. The enzyme assay showed that the ALP activities decreased significantly while treated with the B. batryticatus extracts concentration ranged from 1 mg · g-1 to 5 mg · g-1, and the correlationship was responsible to the dosage of B. batryticatus extracts. Thus, the inhibition on ALP activity is a possible mechanism for the pesticidal activity of B. batryticatus extracts.

  18. Mechanisms of Action for Total Flavones of Hippophae Rhamnoides and Quercetin on Alkaline Phosphatase Activity in Chicken Osteoblasts%沙棘总黄酮、槲皮素对鸡成骨细胞碱性磷酸酶活性作用机制的体外研究

    Institute of Scientific and Technical Information of China (English)

    刘洪柱; 李垚; 陈鑫; 赵伟; 张丽欣; 欧阳文文; 刘红南

    2011-01-01

    The experiment was conducted to study the effects of total flavones of Hippophae rhamnoides (TFH) and quercetin on alkaline phosphatase (ALP) activity in osteoblasts of arbor acres (AA) broilers' embryo in vitro and its mechanisms of action. Cells were cultured in basal culture medium, basal culture medium supplemented with β-estradiol (E2) , TFH and quercetin for 24 h, respectively; after treated by estrogen receptor (ER) antagonist for 2 h, the cells were cultured for another 24 h in basal culture medium, basal culture medium supplemented with TFH and quercetin, respectively; after treated with extracellular signal-regulated ki-nase (ERK) inhibitor for 2 h, the cells were cultured for another 24 h in basal culture medium, basal culture medium supplemented with TFH and quercetin, respectively; the cells were collected for the analysis of ALP activity. Cells were cultured in basal culture medium supplemented with TFH or quercetin, and collected at six time points (0, 2, 5, 15, 30, 60 min) for the determination of phosphorylated extracellular-regulated kinase (p-ERK) concentration by ELASE. Cells were cultured in basal culture medium, basal culture medium supplemented with E2, TFH or quercetin for 30 min, respectively; after treated by ER antagonist for 15 min, the cells were cultured for 30 min in basal culture medium, basal culture medium supplemented with E2, TFH and quercetin, respectively; cells were collected for the determination of p-ERK concentration by ELASE. The results showed as follows:1) culture medium supplemented with TFH or quercetin significantly increased the ALP activity {P <0.05), and the effects of which were totally prevented by ER antagonist and MEK inhibitor (P<0.05);2)the phosphorylation level of ERK were significantly increased in culture medium supplemented with TFH after 5, 15 min incubation and with quercetin after 2, 5, 15, 30, 60 min incubation (P < 0.05); 3) culture medium supplemented with TFH or quercetin significantly increased

  19. Research on Phosphatases of Belladona Leaves and Their Purification (Part 1

    Directory of Open Access Journals (Sweden)

    M. Khorsand

    1956-12-01

    Full Text Available Belladona leaves as well as all other studied leaves contains two distinct phosphatase fractions belonging respectively to types II and IIIi the major parts of these enzymes is extraetible by water. It was not possible to extract the non soluble fraction which is solidly retained by the cellular constituents. Phosphatase II does not differ from other phosphatnses of the same type. Whereas phosphatase III is distinetely different from enzymes of the same type of vegetal or animal origins. It is activated by bivalent metallic ions which are specific activators of the alkaline phcspbatnses: Mg-Zn-Ni and Co.

  20. Research on Phosphatases of Belladona Leaves and Their Purification (Part 1

    Directory of Open Access Journals (Sweden)

    M. Khorsand

    1956-07-01

    Full Text Available Belladona leaves as well as all other studied leaves contains two distinct phosphatase fractions belonging respectively to types II and IIIi the major parts of these enzymes is extraetible by water. It was not possible to extract the non soluble fraction which is solidly retained by the cellular constituents. Phosphatase II does not differ from other phosphatnses of the same type. Whereas phosphatase III is distinetely different from enzymes of the same type of vegetal or animal origins. It is activated by bivalent metallic ions which are specific activators of the alkaline phcspbatnses: Mg-Zn-Ni and Co.

  1. Comparison of alkaline phosphatase in Bemisia tabaci B-biotype(Homoptera: Aleyrodidae) and Trialeurodes vaporariorum (Homoptera:Aleyrodidae) at different developmental stages%B型烟粉虱与温室白粉虱不同虫态的碱性磷酸酶性质比较

    Institute of Scientific and Technical Information of China (English)

    严盈; 刘万学; 万方浩

    2008-01-01

    为了探明B型烟粉虱,Bemisia tobaci B-biotype和温室白粉虱Trialeurodes vaporariorum体内的碱性磷酸酶(alkaline phosphatase,ALP)在两者竞争替代中所起的作用,以对硝基苯磷酸二钠(pNPP)为底物,采用个体测定和群体测定的方法,研究比较了2种粉虱不同虫态中该酶的性质.结果表明:2种粉虱的碱性磷酸酶比活力在整个发育历期均逐渐增加,成虫期达到最大.温室白粉虱2至4龄若虫(伪蛹)期的碱性磷酸酶比活力分别是B型烟粉虱对应龄期酶比活力的2.58、2.68和3.14倍;B型烟粉虱雌雄成虫的碱性磷酸酶比活力分别是温室白粉虱雌雄成虫酶比活力的1.24和1.26倍,且2种粉虱雌虫的酶比活力显著大于其雄虫.2种粉虱2龄若虫到成虫的碱性磷酸酶最适pH均为7.8,最适温度均为47 ℃;在1龄若虫中均未能检测到该酶活性.测定并比较2种粉虱不同虫态碱性磷酸酶动力学特征参数的结果显示,温室白粉虱碱性磷酸酶在3、4龄若虫的亲和力以及在2,3,4龄若虫的酶蛋白浓度均显著大于B型烟粉虱的对应值,而在成虫期2种粉虱的亲和力、酶蛋白浓度无差异,B型烟粉虱的活化能显著小于温室白粉虱.据此推测,B型烟粉虱利用碱性磷酸酶在若虫期进行组织骨化和生长发育不如温室白粉虱,但羽化为成虫后利用其进行解毒代谢则可能强于温室白粉虱.

  2. 全身垂直振动对去卵巢骨质疏松大鼠血清和骨组织碱性磷酸酶水平的影响%Effects of Whole-body Vertical Vibration on Serum and Bone Levels of Alkaline Phosphatase in Ovariectomized Rats with Osteoporosis

    Institute of Scientific and Technical Information of China (English)

    张洋; 杨少峰; 卜淑敏

    2013-01-01

    目的:观察全身垂直振动对去卵巢骨质疏松大鼠血清和骨组织碱性磷酸酶水平的影响.方法:将36只3月龄雌性SD大鼠按体重分层后随机分为假手术、去卵巢静止和去卵巢振动三组.去卵巢10周后,去卵巢振动组每天进行2次全身垂直振动治疗,振动频率90 Hz,时间15 min.治疗7周时,采用双能X线骨密度仪检测各组大鼠左侧离体股骨骨密度,用721分光光度计检测血清和右侧股骨远端碱性磷酸酶水平.结果:与假手术组比较,去卵巢静止组血清碱性磷酸酶水平显著增加,股骨远端碱性磷酸酶水平以及离体股骨近端和远端的骨密度显著下降,而离体股骨中段骨密度无显著变化.与去卵巢静止组比较,去卵巢振动组离体股骨远端碱性磷酸酶水平以及股骨近端和远端骨密度显著增加,而血清碱性磷酸酶水平和离体股骨中段骨密度无显著变化.结论:全身垂直振动可能通过增加骨组织碱性磷酸酶水平改善去卵巢骨质疏松大鼠的骨密度.%Objective This paper investigated the effects of whole-body vertical vibration on levels of alkaline phosphatase (ALP)in serum and bone of ovariectomized (OVX)rats with osteoporosis. Methods Thirty six 3-month-old female SD rats were randomly divided into following three groups according to their body weight:sham-operation group (Sham) ,OVX group,and OVX plus vibration group (V). After 10 weeks of ovariectomy.the group V was treated with whole-body vertical vibration for 15 min at a frequency of 90Hz twice a day. After 7 weeks of whole-body vertical vibration treatment,the bone mineral density (BMD)of left femur in vitro was detected by dual energy X-ray absorptiometry. The changes of serum and bone ALP levels were detected by 721 Photometer. Results Compared with the group Sham, the serum level of ALP in group OVX increased significantly, while the ALP level in distal femur and the BMD of proximal and distal femur in vitro

  3. Cd2+和Cr3+对崇明东滩湿地土壤碱性磷酸酶的低剂量兴奋效应%Low-Dose Hormetic Effects of Cd2+ and Cr3+on Alkaline Phosphatase in Wetland Soil in Dongtan of Chongming

    Institute of Scientific and Technical Information of China (English)

    范弟武; 徐莎; 周曼丽; 张倩楠; 朱咏莉; 韩建刚

    2016-01-01

    为了明确Cd2+、Cr3+与碱性磷酸酶(ALP)活性之间的低剂量兴奋效应关系,以崇明东滩湿地土壤为对象,通过添加不同剂量的外源Cd2+( CdCl2)和Cr3+( CrCl3),使土壤中w( Cd2+)分别为0、0�001、0�01、0�1、1、5、10、20、100和500 mg·kg-1,w(Cr3+)为0、0�5、5、50、100、500和5000 mg·kg-1,观测土壤ALP活性随时间(0、6、12、24、48、72和120 h)的变化特征。结果表明:(1)培养12 h后,Cd2+添加量为1 mg·kg-1时,ALP 活性比对照高8�6%(P<0�05);当Cd2+添加量大于10 mg·kg-1时,酶活性受到明显抑制。 Cr3+添加量为5 mg·kg-1时,ALP活性比对照显著升高22�8%( P<0�05);当Cr3+添加量大于100 mg·kg-1时,酶活性显著降低。这表明Cd2+和Cr3+与ALP之间存在典型的低剂量兴奋效应,但效应的表达与两者接触时间的长短密切相关。(2)以培养24 h的土壤样品为例, Cd2+添加量为1和5 mg·kg-1时,ALP的催化效率( Vmax/Km ),即最大反应速率( Vmax )与Michaelis常数( Km )的比值为1�7;当Cd2+添加量增加到20 mg·kg-1时,Vmax/Km比降至0�8,而Vmax和Km的值均低于对照。 Cr3+添加量为0�5和5 mg·kg-1时,Vmax/Km比为1�7;当Cr3+添加量增至100 mg·kg-1时,Vmax/Km比降为1�4,但Vmax和Km值均高于对照,这表明重金属与土壤酶之间的低剂量兴奋效应机理可能与其离子特性密切相关。%To explore low⁃dose hormetic effects ( a biphasic dose⁃response characterized by a low dose benefit and a high dose inhibition) of Cd2+ and Cr3+on activity of alkaline phosphatase in wetland soil, soil samples were collected from the wetland in Dongtan of Chongming, treated with different doses of Cd2+ and Cr3+, making the samples 0, 0�001, 0�01, 0�1, 1, 5, 10, 20, 100 and 500 mg·kg-1 in Cd2+ concentration and 0, 0�5, 5

  4. A new process for efficiently producing methane from waste activated sludge: alkaline pretreatment of sludge followed by treatment of fermentation liquid in an EGSB reactor.

    Science.gov (United States)

    Zhang, Dong; Chen, Yinguang; Zhao, Yuxiao; Ye, Zhengxiang

    2011-01-15

    In the literature the production of methane from waste activated sludge (WAS) was usually conducted in a continuous stirred tank reactor (CSTR) after sludge was pretreated. It was reported in our previous publication that compared with other pretreatment methods the methane production in CSTR could be significantly enhanced when sludge was pretreated by NaOH at pH 10 for 8 days. In order to further improve methane production, this study reported a new process for efficiently producing methane from sludge, that is, sludge was fermented at pH 10 for 8 days, which was adjusted by Ca(OH)(2), and then the fermentation liquid was treated in an expanded granular sludge bed (EGSB) for methane generation. First, for comparing the methane production observed in this study with that reported in the literature, the conventional operational model was applied to produce methane from the pH 10 pretreated sludge, that is, directly using the pH 10 pretreated sludge to produce methane in a CSTR. It was observed that the maximal methane production was only 0.61 m(3)CH(4)/m(3)-reactor/day. Then, the use of fermentation liquid of pH 10 pretreated sludge to produce methane in the reactors of up-flow anaerobic sludge bed (UASB), anaerobic sequencing batch reactor (ASBR) and EGSB was compared. The maximal methane production in UASB, ASBR, and EGSB reached 1.41, 3.01, and 12.43 m(3)CH(4)/m(3)-reactor/day, respectively. Finally, the mechanisms for EGSB exhibiting remarkably higher methane production were investigated by enzyme, adenosine-triphosphate (ATP), scanning electron microscope (SEM) and fluorescence in situ hybridization (FISH) analyses. It was found that the granular sludge in EGSB had the highest conversion efficiency of acetic acid to methane, and the greatest activity of hydrolysis and acidification enzymes and general physiology with much more Methanosarcinaceae.

  5. Radioprotective effect of Panax ginseng on the phosphatases and lipid peroxidation level in testes of Swiss albino mice

    Energy Technology Data Exchange (ETDEWEB)

    Kumar M.; Sharma M.K.; Saxena P.S.; Kumar A. [Rajasthan Univ., Jaipur (India)

    2003-03-01

    The Panax ginseng has been used as traditional medicine for past several years among oriental people. The present investigation has been made to assess the radioprotective efficacy of ginseng root extract in the testicular enzymes of Swiss albino mice. The Swiss albino mice were divided into different groups. Ginseng treated group: The animals were administered 10 mg/kg body weight ginseng root extract intraperitoneal (i.p.). Radiation treated group: The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 cm. Combination group: Animals were administered ginseng extract continuously for 4 d and on 4th day they were irradiated to 8 Gy gamma radiation after 30 min of extract administration. The animals from above groups were autopsied on day 1, 3, 7, 14 and 30. Biochemical estimations of acid and alkaline phosphatases and Lipid peroxidation (LPO) in testes were done. In ginseng treated group acid and alkaline phosphatases activity and LPO level did not show any significant alteration. In irradiated animals there was a significant increase in acid phosphatase activity and LPO level. However, significant decline in alkaline phosphatase activity was observed. The treatment of ginseng before irradiation causes significant decrease in acid phosphatase and LPO level and significant increase in alkaline phosphatase activity. One of the cause of radiation damage is lipid peroxidation. Due to lipid peroxidation, lysosomal membrane permeability alters and thus results in release of hydrolytic enzymes. So, an increase in acid phosphatase was noticed after radiation treatment. The alkaline phosphatase activity is associated with membrane permeability and different stages of spermatogenesis. Due to membrane damage and depletion of germ cells of testes after irradiation the enzyme activity was decreased. Ginseng markedly inhibits lipid peroxidation. It acts in indirect fashion to protect radical processes by inhibition of initiation of

  6. Carbon and Nitrogen Sources Influence Tricalcium Phosphate Solubilization and Extracellular Phosphatase Activity by Talaromyces flavus.

    Science.gov (United States)

    Stefanoni Rubio, P J; Godoy, M S; Della Mónica, I F; Pettinari, M J; Godeas, A M; Scervino, J M

    2016-01-01

    The aim of this work was to study phosphate (P) solubilization (and the processes involved in this event) by Talaromyces flavus (BAFC 3125) as a function of carbon and/or nitrogen sources. P solubilization was evaluated in NBRIP media supplemented with different carbon (glucose, sorbitol, sucrose, and fructose) and nitrogen (L-asparagine, urea, ammonium sulfate (AS), and ammonium nitrate (AN) combinations. The highest P solubilization was related to the highest organic acid production (especially gluconic acid) and pH drop for those treatments where glucose was present. Also P solubilization was higher when an inorganic nitrogen source was supplemented to the media when compared to an organic one. Although not being present an organic P source, phosphatase activity was observed. This shows that P mineralization and P solubilization can occur simultaneously, and that P mineralization is not induced by the enzyme substrate. The combination that showed highest P solubilization was for AN-glucose. The highest acid phosphatase activity was for AS-fructose, while for alkaline phosphatase were for AS-fructose and AN-fructose. Acid phosphatase activity was higher than alkaline. P solubilization and phosphatase activity (acid and alkaline) were influenced by the different carbon-nitrogen combinations. A better understanding of phosphate-solubilizing fungi could bring a better use of soil P.

  7. Phosphatase Activity of Microbial Populations in Different Milk Samples in Relation to Protein and Carbohydrate Content

    Directory of Open Access Journals (Sweden)

    Sosanka Protim SANDILYA

    2014-12-01

    Full Text Available Cattle milk is a rich source of protein, carbohydrate, vitamins, minerals and all other major and micro nutrients. At a moderate pH, milk is an excellent media for the growth of microbes and thus, intake of raw milk is precarious. In this study, attempt was made for a qualitative study of eight raw milk samples of different varieties of cow and goat milk, collected from Jorhat district of Assam, India, on the basis of nutritional value and microbial population. The highest microbial population was found in the milk collected from cross hybrid variety of cow, whereas microbial contamination was the least in Jersey cow milk. Samples of C1 (Jersey cow variety showed presence of the highest amount of protein and carbohydrate content as compared to the others. Almost all the milk samples showed positive acid and alkaline phosphatase activity. Maximum acid phosphatase activity was observed in cross hybrid cow milk, whereas local cow milk exhibited the highest alkaline phosphatase activity. Phosphatase activity did not show any co-relationship with microbial population of the milk samples. Similarly, the protein and carbohydrate content of the samples did not have any significant impact on both acid and alkaline phosphatase activity.

  8. 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.

  9. 饲料锌含量对方格星虫稚虫生长性能、体成分、体腔液中锌含量及碱性磷酸酶活性的影响%Effects of Dietary Zinc Content on Growth Performance, Body Composition, Coelomic Fluid Zinc Content and Alkaline Phosphatase Activity of Juvenile Peanut Worm, Sipunculus nudus Linnaeus

    Institute of Scientific and Technical Information of China (English)

    许明珠; 张琴; 童潼; 董兰芳; 杨家林; 蒋艳; 黄国强

    2016-01-01

    This trial was conducted to investigate the effects of dietary zinc content on growth performance, body composition, coelomic fluid zinc content and alkaline phosphatase ( AKP ) activity of juvenile peanut worm, Sipunculus nudus Linnaeus ( S. nudus ) . Juvenile peanut worm with the average body weight of (14.54±0.10) mg were fed six isonitrogenous and isoenergetic microparticle diets with different supplemental levels of zinc ( zinc methionine as zinc source) for 8 weeks, and the measured zinc contents in diets were 9.3, 31.7, 49.9, 90.1, 168.6 and 326.5 mg/kg, respectively. Each diet had three replicates and each replicate had 400 juvenile peanut worm. The results showed that the dietary zinc content had significant effects on weight gain rate ( WGR) , specific growth rate ( SGR) and survival ratio ( SR) of S. nudus ( P0.05). Dietary zinc content significantly affected coelomic fluid zinc content and AKP activity of S. nudus ( P<0.05) . With the increase of dietary zinc content, the coelomic fluid zinc content of S. nudus was sustained rise, when the dietary zinc content was 326.5 mg/kg, it produced the maximum value. Moreover, the coelomic fluid AKP activity was firstly increased and then tended to be stable, when the dietary zinc content was 49.9 mg/kg, it produced the maximum value. With WGR as the evaluating index, the suitable dietary zinc content of juvenile peanut worm is 41.93 mg/kg by regression analysis.%以蛋氨酸锌为锌源,在等氮等能的基础饲料中添加不同水平的锌,配制实际锌含量分别为9.3、31.7、49.9、90.1、168.6和326.5 mg/kg 的6种试验饲料,饲喂平均体重为(14.54±0.10) mg的方格星虫稚虫8周,用以研究饲料锌含量对方格星虫稚虫生长、体成分、体腔液中锌含量及碱性磷酸酶活性的影响。每种试验饲料设3个重复,每个重复饲喂400尾方格星虫稚虫。结果表明:饲料锌含量对方格星虫的成活率、增重率和特定

  10. Alkaline tolerant dextranase from streptomyces anulatus

    Science.gov (United States)

    Decker, Stephen R.; Adney, William S.; Vinzant, Todd B.; Himmel, Michael E.

    2003-01-01

    A process for production of an alkaline tolerant dextranase enzyme comprises culturing a dextran-producing microorganism Streptomyces anulatus having accession no. ATCC PTA-3866 to produce an alkaline tolerant dextranase, Dex 1 wherein the protein in said enzyme is characterized by a MW of 63.3 kDa and Dex 2 wherein its protein is characterized by a MW of 81.8 kDa.

  11. Serum alkaline phosphatases concentration in adult patients with skeleton class Ⅱ malocclusion:preliminary results%成人骨性Ⅱ类错牙合畸形患者血清碱性磷酸酶浓度的初步研究

    Institute of Scientific and Technical Information of China (English)

    马艳; 施优灵; 李金超; 刘继光

    2016-01-01

    Objective To explore the concentration and roles of serum alkaline phosphatases ( ALP) in patients with skeleton class Ⅱ malocclusion. Methods 45 adult patients with skeleton class Ⅱ malocclusion and 47 age-and sex-matched healthy controls were se-lected in this study, and the concentrations of serum ALP were collected. The difference between experimental and control groups was assessed by independent Student's t-test with SPSS 18. 0 software. Results The serum ALP level in patients and that in healthy con-trols were (41. 62±1. 97)U/L and (65. 83±2. 70)U/L, respectively. The serum ALP level in male patients and that in healthy con-trols were (52.07±4.63)U/L and (68.25±2.65)U/L, whereas that in female patients and that in healthy controls were (36.40± 0. 92)U/L and (64. 04±4. 28) U/L, respectively. The serum ALP level in patients25 years old vs. that in healthy controls was (41. 20±2. 45)U/L vs. (66. 00±3. 35)U/L, and (51. 29±18. 68)U/L vs. (66. 88±11. 76)U/L for males, while (37. 28±5. 56)U/L vs (65. 18±24. 68)U/L for females. The significant statistical difference was observed (P<0. 05). Conclusion The serum ALP level in adult patients with skeleton class Ⅱ malocclusion is lower than that in healthy controls, suggesting weaker bone metabolism in the patients, which may be resulted from mandibular hypodevelopment.%目的 研究骨性Ⅱ类错牙合畸形患者的血清碱性磷酸酶浓度及意义.方法 选择45例成人骨性Ⅱ类错牙合畸形患者和47例年龄和性别匹配的正常对照成人,检测其血清碱性磷酸酶浓度.应用SPSS 18.0软件对检测值进行成组t检验统计学分析.结果 成人骨性Ⅱ类错牙合畸形患者和正常对照成人的血清ALP浓度分别为(41.62±1.97)U/L和(65.83±2.70)U/L.其中男性骨性Ⅱ类错牙合畸形患者和对照成人的血清ALP浓度分别为(52.07±4.63)U/L和(68.25±2.65)U/L,女性分别为(36.40±0.92)U/L和(64.04±4.28)U/L.25岁以下骨性Ⅱ类错牙合畸形患

  12. Observation on the activities of alpha-L-fucosidase,alkaline phosphatase and adenosine deaminase in pregnant women%妊娠女性α-L-岩藻糖苷酶、碱性磷酸酶和腺苷脱氨酶活性观察

    Institute of Scientific and Technical Information of China (English)

    王玮玮; 孙伟才; 周惠玉

    2014-01-01

    目的:妊娠期女性观察血清α-L-岩藻糖苷酶(AFU)、碱性磷酸酶(ALP)及腺苷脱氨酶(ADA)活性的变化及其与妊娠周期的相关性。方法分别检测352例体检正常的不同妊娠期女性(妊娠组,其中早期妊娠210例、晚期妊娠142例)及322例因不孕就诊且尚未妊娠的体检正常的女性(非妊娠组)血清 AFU、ALP、ADA 活性。对所有对象均进行动态监测(早期妊娠组于妊娠第14、22、30及38周分别检测,晚期妊娠组于妊娠第38及40周检测,非妊娠组于就诊时、就诊后第8及24周检测)并做比较。结果妊娠组血清 ALP、AFU 活性明显高于非妊娠组(P =0.000),ADA 活性则低于非妊娠组(P =0.000)。晚期妊娠组血清 ALP、AFU 活性明显高于早期妊娠组(P =0.000),但 ADA 活性两组间差异无统计学意义(P >0.05)。对早期妊娠组的动态监测显示,随着妊娠周期的增加,AFU 及 ALP 活性逐渐升高,但 ADA 基本无变化。晚期妊娠组第38周及第40周血清 AFU、ALP、ADA 活性差异均无统计学意义(P >0.05)。对非妊娠组的动态监测显示,与就诊时比较,就诊后第8周 ALP、ADA 活性差异有统计学意义(P =0.000),就诊后第24周 ADA 活性差异有统计学意义;与就诊后第8周比较,就诊后第24周 ALP、AFU、ADA 活性差异均有统计学意义(P <0.05)。结论妊娠期女性血清 AFU 及 ALP 活性明显升高,且随孕周的增加,升高趋势越明显。%Objective To observe the activity changes of alpha-L-fucosidase (AFU),alkaline phosphatase (ALP) and adenosine deaminase (ADA)in pregnant women and their correlations with pregnant period.Methods The activities of AFU,ALP and ADA were determined in 352 pregnant women (210 cases with early-stage pregnancy and 142 cases with late-stage pregnancy)and 322 unpregnant women.Dynamic monitoring was performed (early-stage pregnancy

  13. Glucose-6-phosphatase deficiency

    Directory of Open Access Journals (Sweden)

    Labrune Philippe

    2011-05-01

    Full Text Available Abstract Glucose-6-phosphatase deficiency (G6P deficiency, or glycogen storage disease type I (GSDI, is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, between the ages of 3 to 4 months by symptoms of hypoglycemia (tremors, seizures, cyanosis, apnea. Patients have poor tolerance to fasting, marked hepatomegaly, growth retardation (small stature and delayed puberty, generally improved by an appropriate diet, osteopenia and sometimes osteoporosis, full-cheeked round face, enlarged kydneys and platelet dysfunctions leading to frequent epistaxis. In addition, in GSDIb, neutropenia and neutrophil dysfunction are responsible for tendency towards infections, relapsing aphtous gingivostomatitis, and inflammatory bowel disease. Late complications are hepatic (adenomas with rare but possible transformation into hepatocarcinoma and renal (glomerular hyperfiltration leading to proteinuria and sometimes to renal insufficiency. GSDI is caused by a dysfunction in the G6P system, a key step in the regulation of glycemia. The deficit concerns the catalytic subunit G6P-alpha (type Ia which is restricted to expression in the liver, kidney and intestine, or the ubiquitously expressed G6P transporter (type Ib. Mutations in the genes G6PC (17q21 and SLC37A4 (11q23 respectively cause GSDIa and Ib. Many mutations have been identified in both genes,. Transmission is autosomal recessive. Diagnosis is based on clinical presentation, on abnormal basal values and absence of hyperglycemic response to glucagon. It can be confirmed by demonstrating a deficient activity of a G6P system component in a liver biopsy. To date, the diagnosis is most

  14. Probing protein phosphatase substrate binding

    DEFF Research Database (Denmark)

    Højlys-Larsen, Kim B.; Sørensen, Kasper Kildegaard; Jensen, Knud Jørgen;

    2012-01-01

    profile of the integrin-linked kinase associated phosphatase (ILKAP), a member of the protein phosphatase 2C (PP2C) family. Phosphatases can potentially dephosphorylate these phosphopeptide substrates but, interestingly, performing the binding studies at 4 °C allowed efficient binding to phosphopeptides...... around the phosphorylated residue are important for the binding affinity of ILKAP. We conclude that solid-phase affinity pull-down of proteins from complex mixtures can be applied in phosphoproteomics and systems biology.......Proteomics and high throughput analysis for systems biology can benefit significantly from solid-phase chemical tools for affinity pull-down of proteins from complex mixtures. Here we report the application of solid-phase synthesis of phosphopeptides for pull-down and analysis of the affinity...

  15. Inorganic-organic separators for alkaline batteries

    Science.gov (United States)

    Sheibley, D. W. (Inventor)

    1978-01-01

    A flexible separator is reported for use between the electrodes of Ni-Cd and Ni-Zn batteries using alkaline electrolytes. The separator was made by coating a porous substrate with a battery separator composition. The coating material included a rubber-based resin copolymer, a plasticizer and inorganic and organic fillers which comprised 55% by volume or less of the coating as finally dried. One or more of the filler materials, whether organic or inorganic, is preferably active with the alkaline electrolyte to produce pores in the separator coating. The plasticizer was an organic material which is hydrolyzed by the alkaline electrolyte to improve conductivity of the separator coating.

  16. Bioengineered protein phosphatase 2A: Update on need

    OpenAIRE

    Rubiolo, Juan A.; López-Alonso, Henar; Alfonso, Amparo; Vega, Félix V.; Vieytes, Mercedes Rodríguez; Botana, Luis M

    2013-01-01

    Harmful algal blooms caused by phytoplankton can occur in all aquatic environments. Some of the algae present in these blooms are capable of producing extremely potent toxins. Due to climate change and eutrophication, harmful algal blooms are increasing on a global scale. One kind of toxin producing algae are those that produce okadaic acid, its derivatives (dinophysistoxin-1 and 2), and microcystins. These toxins are potent inhibitors of protein phosphatase 2A, so this protein is used to det...

  17. Anodes for alkaline electrolysis

    Science.gov (United States)

    Soloveichik, Grigorii Lev

    2011-02-01

    A method of making an anode for alkaline electrolysis cells includes adsorption of precursor material on a carbonaceous material, conversion of the precursor material to hydroxide form and conversion of precursor material from hydroxide form to oxy-hydroxide form within the alkaline electrolysis cell.

  18. Alkaline "Permanent" Paper.

    Science.gov (United States)

    Pacey, Antony

    1991-01-01

    Discussion of paper manufacturing processes and their effects on library materials focuses on the promotion of alkaline "permanent" paper, with less acid, by Canadian library preservation specialists. Standards for paper acidity are explained; advantages of alkaline paper are described, including decreased manufacturing costs; and recyclability is…

  19. The activity of some phosphatases in tissues of adult Hymenolepis nana Siebold (Csetoda).

    Science.gov (United States)

    Humiczewska, M

    1989-01-01

    Histochemical methods were used to study the localization and activity of acid and alkaline phosphatases, ATP-ase, 5-nucleotidase, and glucose-6-phosphatase in tissues of the mature form of Hymenolepis nana. Considerable differences in activity and localization of particular enzymes were observed in the organs of the parasite. The results obtained permit the statement that the integument is the most active enzymatically; in connection with the literature data, this gives grounds for the thesis that the integument of the cestodes functions as an absorbent-digestive organ. PMID:2558920

  20. Alkaline battery operational methodology

    Energy Technology Data Exchange (ETDEWEB)

    Sholklapper, Tal; Gallaway, Joshua; Steingart, Daniel; Ingale, Nilesh; Nyce, Michael

    2016-08-16

    Methods of using specific operational charge and discharge parameters to extend the life of alkaline batteries are disclosed. The methods can be used with any commercial primary or secondary alkaline battery, as well as with newer alkaline battery designs, including batteries with flowing electrolyte. The methods include cycling batteries within a narrow operating voltage window, with minimum and maximum cut-off voltages that are set based on battery characteristics and environmental conditions. The narrow voltage window decreases available capacity but allows the batteries to be cycled for hundreds or thousands of times.

  1. Optimization of NaOH Molarity, LUSI Mud/Alkaline Activator, and Na2SiO3/NaOH Ratio to Produce Lightweight Aggregate-Based Geopolymer

    OpenAIRE

    Rafiza Abdul Razak; Mohd Mustafa Al Bakri Abdullah; Kamarudin Hussin; Khairul Nizar Ismail; Djwantoro Hardjito; Zarina Yahya

    2015-01-01

    This paper presents the mechanical function and characterization of an artificial lightweight geopolymer aggregate (ALGA) using LUSI (Sidoarjo mud) and alkaline activator as source materials. LUSI stands for LU-Lumpur and SI-Sidoarjo, meaning mud from Sidoarjo which erupted near the Banjarpanji-1 exploration well in Sidoarjo, East Java, Indonesia on 27 May 2006. The effect of NaOH molarity, LUSI mud/Alkaline activator (LM/AA) ratio, and Na2SiO3/NaOH ratio to the ALGA are investigated at a sin...

  2. Effects of synthetic detergents on in vivo activity of tissue phosphatases and succinic dehydrogenase from Mystus vittatus

    Energy Technology Data Exchange (ETDEWEB)

    Mohan, D.; Verma, S.R.

    1981-05-01

    African catfish (Mystus vittatus) were exposed to three sub-lethal concentrations of Swascofix E45 (13.8, 9.2 and 4.6 mg/l) and Swascol 3L (69.3, 46.2 and 23.1 mg/l) for 15 and 30 days, and their effects on alkaline and acid phosphatase, and succinic dehydrogenase in liver, kidney and intestine were measured. The enzymes were found to be inhibited in all the tissues. Maximum inhibition (38.44%) was observed in liver alkaline phosphatase activity after 30 days with the highest concentration of Swascofix E45 and the lowest inhibition (0.118%) was found in kidney acid phosphatase activity with the lowest concentration of Swascol 3L after 15 days. Insignificant enzyme stimulation in some cases was also observed.

  3. Uranium in alkaline rocks

    International Nuclear Information System (INIS)

    Geologic and geochemical criteria were developed for the occurrence of economic uranium deposits in alkaline igneous rocks. A literature search, a limited chemical analytical program, and visits to three prominent alkaline-rock localities (Ilimaussaq, Greenland; Pocos de Caldas, Brazil; and Powderhorn, Colorado) were made to establish criteria to determine if a site had some uranium resource potential. From the literature, four alkaline-intrusive occurrences of differing character were identified as type-localities for uranium mineralization, and the important aspects of these localities were described. These characteristics were used to categorize and evaluate U.S. occurrences. The literature search disclosed 69 U.S. sites, encompassing nepheline syenite, alkaline granite, and carbonatite. It was possible to compare two-thirds of these sites to the type localities. A ranking system identified ten of the sites as most likely to have uranium resource potential

  4. Uranium in alkaline rocks

    Energy Technology Data Exchange (ETDEWEB)

    Murphy, M.; Wollenberg, H.; Strisower, B.; Bowman, H.; Flexser, S.; Carmichael, I.

    1978-04-01

    Geologic and geochemical criteria were developed for the occurrence of economic uranium deposits in alkaline igneous rocks. A literature search, a limited chemical analytical program, and visits to three prominent alkaline-rock localities (Ilimaussaq, Greenland; Pocos de Caldas, Brazil; and Powderhorn, Colorado) were made to establish criteria to determine if a site had some uranium resource potential. From the literature, four alkaline-intrusive occurrences of differing character were identified as type-localities for uranium mineralization, and the important aspects of these localities were described. These characteristics were used to categorize and evaluate U.S. occurrences. The literature search disclosed 69 U.S. sites, encompassing nepheline syenite, alkaline granite, and carbonatite. It was possible to compare two-thirds of these sites to the type localities. A ranking system identified ten of the sites as most likely to have uranium resource potential.

  5. Fluorescence labelling of phosphatase activity in digestive glands of carnivorous plants.

    Science.gov (United States)

    Płachno, B J; Adamec, L; Lichtscheidl, I K; Peroutka, M; Adlassnig, W; Vrba, J

    2006-11-01

    A new ELF (enzyme labelled fluorescence) assay was applied to detect phosphatase activity in glandular structures of 47 carnivorous plant species, especially Lentibulariaceae, in order to understand their digestive activities. We address the following questions: (1) Are phosphatases produced by the plants and/or by inhabitants of the traps? (2) Which type of hairs/glands is involved in the production of phosphatases? (3) Is this phosphatase production a common feature among carnivorous plants or is it restricted to evolutionarily advanced species? Our results showed activity of the phosphatases in glandular structures of the majority of the plants tested, both from the greenhouse and from sterile culture. In addition, extracellular phosphatases can also be produced by trap inhabitants. In Utricularia, activity of phosphatase was detected in internal glands of 27 species from both primitive and advanced sections and different ecological groups. Further positive reactions were found in Genlisea, Pinguicula, Aldrovanda, Dionaea, Drosera, Drosophyllum, Nepenthes, and Cephalotus. In Utricularia and Genlisea, enzymatic secretion was independent of stimulation by prey. Byblis and Roridula are usually considered as "proto-carnivores", lacking digestive enzymes. However, we found high activity of phosphatases in both species. Thus, they should be classified as true carnivores. We suggest that the inflorescence of Byblis and some Pinguicula species might also be an additional "carnivorous organ", which can trap a prey, digest it, and finally absorb available nutrients. PMID:16865659

  6. 髁突软骨细胞碱性磷酸酶活性及体外矿化能力的检测%Study on the activity of alkaline phosphatase and the calcified ability of mandibular condylar chondrocyte in vitro

    Institute of Scientific and Technical Information of China (English)

    毛勇; 段小红; 张玉梅

    2001-01-01

    @@0 引言髁突软骨细胞(mandibular condylar chondro cyte, MCC)体外培养时存在细胞形态变化、Ⅰ型和Ⅱ型胶原表达的更迭等表型变化特征[1],但还不清楚MCC碱性磷酸酶(alkaline phosphatase,ALP)活性的变化特点. 本实验采用ALP活性测定和Von Kossa染色法对兔MCC进行检测,以补充对MCC这方面生物学特征的认识.

  7. The involvement of glucose-6-phosphatase in mucilage secretion by root cap cells of Zea mays

    Science.gov (United States)

    Moore, R.; McClelen, C. E.

    1985-01-01

    In order to determine the involvement of glucose-6-phosphatase in mucilage secretion by root cap cells, we have cytochemically localized the enzyme in columella and peripheral cells of root caps of Zea mays. Glucose-6-phosphatase is associated with the plasmalemma and cell wall of columella cells. As columella cells differentiate into peripheral cells and begin to produce and secrete mucilage, glucose-6-phosphatase staining intensifies and becomes associated with the mucilage and, to a lesser extent, the cell wall. Cells being sloughed from the cap are characterized by glucose-6-phosphatase staining being associated with the vacuole and plasmalemma. These changes in enzyme localization during cellular differentiation in root caps suggest that glucose-6-phosphatase is involved in the production and/or secretion of mucilage by peripheral cells of Z. mays.

  8. Spectroscopic investigations of the chiral interactions of metolachlor and its (S)-isomer with lipase and phosphatase.

    Science.gov (United States)

    Wen, Yue Z; Yuan, Yu L; Chen, Hui; Wang, He L; Liu, Hui J; Kang, Xiao D; Fu, Liu S

    2010-04-01

    Metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide] is a chiral acetanilide herbicide. We investigated its enantioselective interactions, and that of its (S)-isomer, with Penicillium expansum alkaline lipase and phosphatase. UV differential spectroscopy and fluorescence spectrophotometry studies were conducted in phosphate buffered solution at pH 7. Chiral differences in the UV absorption and fluorescence spectra of lipase and phosphatase with metolachlor and its (S)-isomer were detected. The results showed that the interactions of metolachlor and its (S)-isomer with lipase and phosphatase occur statically through complex formation, and enantioselectivity was clearly observed. In addition, both UV absorption and fluorescence spectrophotometry showed that the (S)-isomer interacted more strongly with lipase and phosphatase than metolachlor. PMID:20390958

  9. Ecto-phosphatase activity on the external surface of Rhodnius prolixus salivary glands: modulation by carbohydrates and Trypanosoma rangeli.

    Science.gov (United States)

    Gomes, Suzete A O; Fonseca de Souza, André L; Kiffer-Moreira, Tina; Dick, Claudia F; dos Santos, André L A; Meyer-Fernandes, José R

    2008-05-01

    The salivary glands of insect's vectors are target organs to study the vectors-pathogens interactions. Rhodnius prolixus an important vector of Trypanosoma cruzi can also transmit Trypanosoma rangeli by bite. In the present study we have investigated ecto-phosphatase activity on the surface of R. prolixus salivary glands. Ecto-phosphatases are able to hydrolyze phosphorylated substrates in the extracellular medium. We characterized these ecto-enzyme activities on the salivary glands external surface and employed it to investigate R. prolixus-T. rangeli interaction. Salivary glands present a low level of hydrolytic activity (4.30+/-0.35 nmol p-nitrophenol (p-NP)xh(-1)xgland pair(-1)). The salivary glands ecto-phosphatase activity was not affected by pH variation; and it was insensitive to alkaline inhibitor levamisole and inhibited approximately 50% by inorganic phosphate (Pi). MgCl2, CaCl2 and SrCl2 enhanced significantly the ecto-phosphatase activity detected on the surface of salivary glands. The ecto-phosphatase from salivary glands surface efficiently releases phosphate groups from different phosphorylated amino acids, giving a higher rate of phosphate release when phospho-tyrosine is used as a substrate. This ecto-phosphatase activity was inhibited by carbohydrates as d-galactose and d-mannose. Living short epimastigotes of T. rangeli inhibited salivary glands ecto-phosphatase activity at 75%, while boiled parasites did not. Living long epimastigote forms induced a lower, but significant inhibitory effect on the salivary glands phosphatase activity. Interestingly, boiled long epimastigote forms did not loose the ability to modulate salivary glands phosphatase activity. Taken together, these data suggest a possible role for ecto-phosphatase on the R. prolixus salivary glands-T. rangeli interaction. PMID:18407240

  10. Grace DAKASEP alkaline battery separator

    Science.gov (United States)

    Giovannoni, R. T.; Lundquist, J. T.; Choi, W. M.

    1987-01-01

    The Grace DAKASEP separator was originally developed as a wicking layer for nickel-zinc alkaline batteries. The DAKASEP is a filled non-woven separator which is flexible and heat sealable. Through modification of formulation and processing variables, products with a variety of properties can be produced. Variations of DAKASEP were tested in Ni-H2, Ni-Zn, Ni-Cd, and primary alkaline batteries with good results. The properties of DAKASEP which are optimized for Hg-Zn primary batteries are shown in tabular form. This separator has high tensile strength, 12 micron average pore size, relatively low porosity at 46-48 percent, and consequently moderately high resistivity. Versions were produced with greater than 70 percent porosity and resistivities in 33 wt percent KOH as low as 3 ohm cm. Performance data for Hg-Zn E-1 size cells containing DAKASEP with the properties shown in tabular form, are more reproducible than data obtained with a competitive polypropylene non-woven separator. In addition, utilization of active material is in general considerably improved.

  11. Effects of Betaine Aldehyde Dehydrogenase-Transgenic Soybean on Phosphatase Activities and Rhizospheric Bacterial Community of the Saline-Alkali Soil

    Directory of Open Access Journals (Sweden)

    Ying Nie

    2016-01-01

    Full Text Available The development of transgenic soybean has produced numerous economic benefits; however the potential impact of root exudates upon soil ecological systems and rhizospheric soil microbial diversity has also received intensive attention. In the present study, the influence of saline-alkali tolerant transgenic soybean of betaine aldehyde dehydrogenase on bacterial community structure and soil phosphatase during growth stages was investigated. The results showed that, compared with nontransgenic soybean as a control, the rhizospheric soil pH of transgenic soybean significantly decreased at the seedling stage. Compared to HN35, organic P content was 13.5% and 25.4% greater at the pod-filling stage and maturity, respectively. The acid phosphatase activity of SRTS was significantly better than HN35 by 12.74% at seedling, 14.03% at flowering, and 59.29% at podding, while alkaline phosphatase achieved maximum activity in the flowering stage and was markedly lower than HN35 by 13.25% at pod-filling. The 454 pyrosequencing technique was employed to investigate bacterial diversity, with a total of 25,499 operational taxonomic units (OTUs obtained from the 10 samples. Notably, the effect of SRTS on microbial richness and diversity of rhizospheric soil was marked at the stage of podding and pod-filling. Proteobacteria, Acidobacteria, and Actinobacteria were the dominant phyla among all samples. Compared with HN35, the relative abundance of Proteobacteria was lower by 2.01%, 2.06%, and 5.28% at the stage of seedling, at pod-bearing, and at maturity. In genus level, the relative abundance of Gp6, Sphingomonas sp., and GP4 was significantly inhibited by SRTS at the stage of pod-bearing and pod-filling.

  12. Alkaline quinone flow battery.

    Science.gov (United States)

    Lin, Kaixiang; Chen, Qing; Gerhardt, Michael R; Tong, Liuchuan; Kim, Sang Bok; Eisenach, Louise; Valle, Alvaro W; Hardee, David; Gordon, Roy G; Aziz, Michael J; Marshak, Michael P

    2015-09-25

    Storage of photovoltaic and wind electricity in batteries could solve the mismatch problem between the intermittent supply of these renewable resources and variable demand. Flow batteries permit more economical long-duration discharge than solid-electrode batteries by using liquid electrolytes stored outside of the battery. We report an alkaline flow battery based on redox-active organic molecules that are composed entirely of Earth-abundant elements and are nontoxic, nonflammable, and safe for use in residential and commercial environments. The battery operates efficiently with high power density near room temperature. These results demonstrate the stability and performance of redox-active organic molecules in alkaline flow batteries, potentially enabling cost-effective stationary storage of renewable energy. PMID:26404834

  13. Alkaline broadening in Stars

    CERN Document Server

    De Kertanguy, A

    2015-01-01

    Giving new insight for line broadening theory for atoms with more structure than hydrogen in most stars. Using symbolic software to build precise wave functions corrected for ds;dp quantum defects. The profiles obtained with that approach, have peculiar trends, narrower than hydrogen, all quantum defects used are taken from atomic database topbase. Illustration of stronger effects of ions and electrons on the alkaline profiles, than neutral-neutral collision mechanism. Keywords : Stars: fundamental parameters - Atomic processes - Line: profiles.

  14. Use Alkalinity Monitoring to Optimize Bioreactor Performance.

    Science.gov (United States)

    Jones, Christopher S; Kult, Keegan J

    2016-05-01

    In recent years, the agricultural community has reduced flow of nitrogen from farmed landscapes to stream networks through the use of woodchip denitrification bioreactors. Although deployment of this practice is becoming more common to treat high-nitrate water from agricultural drainage pipes, information about bioreactor management strategies is sparse. This study focuses on the use of water monitoring, and especially the use of alkalinity monitoring, in five Iowa woodchip bioreactors to provide insights into and to help manage bioreactor chemistry in ways that will produce desirable outcomes. Results reported here for the five bioreactors show average annual nitrate load reductions between 50 and 80%, which is acceptable according to established practice standards. Alkalinity data, however, imply that nitrous oxide formation may have regularly occurred in at least three of the bioreactors that are considered to be closed systems. Nitrous oxide measurements of influent and effluent water provide evidence that alkalinity may be an important indicator of bioreactor performance. Bioreactor chemistry can be managed by manipulation of water throughput in ways that produce adequate nitrate removal while preventing undesirable side effects. We conclude that (i) water should be retained for longer periods of time in bioreactors where nitrous oxide formation is indicated, (ii) measuring only nitrate and sulfate concentrations is insufficient for proper bioreactor operation, and (iii) alkalinity monitoring should be implemented into protocols for bioreactor management.

  15. Reaction of dissolved ozone in hydrogen peroxide produced during ozonization of an alkaline medium in a bubble column - doi: 10.4025/actascitechnol.v36i1.11834

    Directory of Open Access Journals (Sweden)

    Marco Aurélio Cremasco

    2014-01-01

    Full Text Available The effect of a reaction of dissolved ozone in an alkaline water solution with hydrogen peroxide was evaluated. Assays were conducted in a bubble column with a gaseous ozone concentration (mixed with oxygen at 21 and 27 g m-3. Concentration curve of dissolved ozone in the liquid revealed an ozone concentration peak in the liquid phase, subsequent decreasing concentration and constant maintenance in the stationary phase. Highest concentration was not detected for the lowest ozone concentration in the gaseous phase. Peak was due to the self-decomposition of ozone in the liquid phase and the production of hydrogen peroxide. The latter reacted with molecular dissolved ozone and decreased ozone concentration after maximum concentration.

  16. Phosphatase-mediated heavy metal accumulation by a Citrobacter sp. and related enterobacteria.

    Science.gov (United States)

    Macaskie, L E; Bonthrone, K M; Rouch, D A

    1994-08-15

    A Citrobacter sp. was reported previously to accumulate heavy metals as cell-bound heavy metal phosphates. Metal uptake is mediated by the activity of a periplasmic acid-type phosphatase that liberates inorganic phosphate to provide the precipitant ligand for heavy metals presented to the cells. Amino acid sequencing of peptide fragments of the purified enzyme revealed significant homology to the phoN product (acid phosphatase) of some other enterobacteria. These organisms, together with Klebsiella pneumoniae, previously reported to produce acid phosphatase, were tested for their ability to remove uranium and lanthanum from challenge solutions supplemented with phosphatase substrate. The coupling of phosphate liberation to metal bioaccumulation was limited to the metal accumulating Citrobacter sp.; therefore the participation of species-specific additional factors in metal bioaccumulation was suggested.

  17. Development of electrochemical biosensor based on the immobilisation of alkaline fosfatase for the determination of 2,4-dichlorophenoxyacetic acid

    International Nuclear Information System (INIS)

    An electrochemical biosensor based on the immobilisation of alkaline phosphatase was developed for the determination of 2,4-dichlorophenoxyacetic acid (2,4-D). The biosensor was constructed from the immobilization of alkaline phosphatase enzyme onto a screen-printed electrode (SPE). The ascorbic acid 2-phosphate (AA2P) was used as substrate for the enzymic reaction. The enzyme was entrapped in a hybrid sol-gel/ chitosan material with certain fixed composition. The determination of toxicity of 2,4-D pesticides quantitatively and qualitatively could be carried out by the inhibition of the alkaline phosphatase. A potential of +600 mV was suitable to be used for the oxidation of the products from the enzyme-substrate reaction, where the reaction pH was at 8.5. The linear response range of the biosensor to the AA2P substrates was 10 μM - 80 μM. The inhibition of the alkaline phosphatase enzyme of the 2,4-D biosensor was maximum at 80 ppm 2,4-D (50 % inhibition). (author)

  18. High temperature and pressure alkaline electrolysis

    DEFF Research Database (Denmark)

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

    2013-01-01

    Alkaline electrolyzers have proven to operate reliable for decades on a large scale, but in order to become commercially attractive and compete against conventional technologies for hydrogen production, the production and investment costs have to be reduced. This may occur by increasing 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...... SrTiO3 was used 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...

  19. Effect of andrographolide on phosphatases activity and cytotoxicity against Spodoptera litura

    Directory of Open Access Journals (Sweden)

    E Edwin

    2016-05-01

    Full Text Available Andrographolide was isolated from ethanol extraction of Andrographis paniculata by column chromatography. Evaluation of larvicidal efficacy, enzymatic changes and cytotoxic activities against Spodoptera litura were conducted across a range of concentrations. The compound showed significant larvicidal activity between 5 - 25 ppm, post ingestion. Morphological deformities observed in larval-pupal stages. Enzymatic profiles were altered by reduction in acid phosphatase, ACP activity by 69.18 %, alkaline phosphatase, ALP activity 75.3 % and 74.9 % reduction in ATPase. Binding affinity to midgut epithelium cells suggests disintegration of cellular organelles observed was directly associated with ingestion of the compound. The results suggest that andrographolide has potential for development as a significant inhibitor of development against the pest Spodoptera litura.

  20. Alkaline fuel cells applications

    Science.gov (United States)

    Kordesch, Karl; Hacker, Viktor; Gsellmann, Josef; Cifrain, Martin; Faleschini, Gottfried; Enzinger, Peter; Fankhauser, Robert; Ortner, Markus; Muhr, Michael; Aronson, Robert R.

    On the world-wide automobile market technical developments are increasingly determined by the dramatic restriction on emissions as well as the regimentation of fuel consumption by legislation. Therefore there is an increasing chance of a completely new technology breakthrough if it offers new opportunities, meeting the requirements of resource preservation and emission restrictions. Fuel cell technology offers the possibility to excel in today's motive power techniques in terms of environmental compatibility, consumer's profit, costs of maintenance and efficiency. The key question is economy. This will be decided by the costs of fuel cell systems if they are to be used as power generators for future electric vehicles. The alkaline hydrogen-air fuel cell system with circulating KOH electrolyte and low-cost catalysed carbon electrodes could be a promising alternative. Based on the experiences of Kordesch [K. Kordesch, Brennstoffbatterien, Springer, Wien, 1984, ISBN 3-387-81819-7; K. Kordesch, City car with H 2-air fuel cell and lead-battery, SAE Paper No. 719015, 6th IECEC, 1971], who operated a city car hybrid vehicle on public roads for 3 years in the early 1970s, improved air electrodes plus new variations of the bipolar stack assembly developed in Graz are investigated. Primary fuel choice will be a major issue until such time as cost-effective, on-board hydrogen storage is developed. Ammonia is an interesting option. The whole system, ammonia dissociator plus alkaline fuel cell (AFC), is characterised by a simple design and high efficiency.

  1. Assessing the Biological Activity of the Glucan Phosphatase Laforin.

    Science.gov (United States)

    Romá-Mateo, Carlos; Raththagala, Madushi; Gentry, Mathew S; Sanz, Pascual

    2016-01-01

    Glucan phosphatases are a recently discovered family of enzymes that dephosphorylate either starch or glycogen and are essential for proper starch metabolism in plants and glycogen metabolism in humans. Mutations in the gene encoding the only human glucan phosphatase, laforin, result in the fatal, neurodegenerative, epilepsy known as Lafora disease. Here, we describe phosphatase assays to assess both generic laforin phosphatase activity and laforin's unique glycogen phosphatase activity. PMID:27514803

  2. The diagnostic value of alpha-fetoprotein, α-L-fucosidase,gamma-glutamyltransferase, alkaline phosphatase and carbohydrate antigen 19-9 in the diagnosis of primary hepatocellular carcinoma%甲胎蛋白、α-L-岩藻糖苷酶、γ-谷氨酰转移酶、碱性磷酸酶及糖类抗原19-9联合检测在原发性肝癌中的诊断价值

    Institute of Scientific and Technical Information of China (English)

    潘倩雄

    2014-01-01

    Objective To investigate the diagnostic value of alpha-fetoprotein (AFP),α-L-fucosidase(AFU),gamma-glutamyltransferase(GGT),alkaline phosphatase (ALP) and carbohydrate antigen 19-9(CA19-9) in the diagnosis of primary hepatocellular carcinoma(PHC).Methods Seventy-eight PHC patients as PHC group,76 patients with other liver diseases as other liver diseases group and 60 healthy volunteers as control group.The level of serum AFP,AFU,GGT,ALP,CA19-9 and positive rate in each group were compared.Results The level of AFP,AFU,GGT,ALP,CA19-9 in PHC group were (276.3 ±72.6)μg/L,(63.1 ± 11.7) U/L,(268.5 ±57.2) U/L,(293.4 ±61.7) U/L and (56.9 ± 12.5) μg/L,in other liver diseases group were (81.6 ± 12.1) μg/L,(28.3 ±7.3) U/L,(54.9 ±7.8) U/L,(116.5 ± 23.8) U/L and (27.8 7.1) μ g/L,in control group were (13.8 ± 2.7) μ g/L,(12.6 ± 3.9) U/L,(12.3 ± 3.2) U/L,(47.2 ± 11.3) U/L and (12.9 ± 3.4) μ g/L.The level of above index in PHC group and other liver diseases group were significantly higher than those in control group,while the level of above index in PHC group were significantly higher than those in other liver diseases group (P < 0.01 or < 0.05).The positive rate of AFP,AFU,GGT,ALP,CA19-9 in PHC group were 62.8%(49/78),79.5%(62/78),83.3% (65/78),85.9% (67/78),66.7% (52/78),and significantly higher than those in other liver diseases group[31.6%(24/76),32.9%(25/76),27.6%(21/76),22.4%(17/76),15.8 %(12/76)].The positive rate of AFP in control group was 0,the positive rate of AFU,GGT,ALP,CA19-9 in control group were 1.7% (1/60),3.3% (2/60),1.7% (1/60),8.3% (5/60),which significantly lower than those in other two groups.The positive rate of combined detection in PHC group was 100.0%(78/78),which significantly higher than that in other liver diseases group [40.8%(31/76)] and control group[8.3%(5/60)](P< 0.01).Conclusion The combined detection of AFP,AFU,GGT,ALP,CA19-9 in PHC diagnosis has a higher sensitivity,and thus to make

  3. Bifunctional alkaline oxygen electrodes

    Science.gov (United States)

    Swette, L.; Kackley, N.; Mccatty, S. A.

    1991-01-01

    The authors describe the identification and testing of electrocatalysts and supports for the positive electrode of moderate-temperature, single-unit, rechargeable alkaline fuel cells. Recent work on Na(x)Pt3O4, a potential bifunctional catalyst, is described, as well as the application of novel approaches to the development of more efficient bifunctional electrode structures. The three dual-character electrodes considered here showed similar superior performance; the Pt/RhO2 and Rh/RhO2 electrodes showed slightly better performance than the Pt/IrO2 electrode. It is concluded that Na(x)Pt3O4 continues to be a promising bifunctional oxygen electrode catalyst but requires further investigation and development.

  4. Silica in alkaline brines

    Science.gov (United States)

    Jones, B.F.; Rettig, S.L.; Eugster, H.P.

    1967-01-01

    Analysis of sodium carbonate-bicarbonate brines from closed basins in volcanic terranes of Oregon and Kenya reveals silica contents of up to 2700 parts per million at pH's higher than 10. These high concentrations of SiO 2 can be attributed to reaction of waters with silicates, and subsequent evaporative concentration accompanied by a rise in pH. Supersaturation with respect to amorphous silica may occur and persist for brines that are out of contact with silicate muds and undersaturated with respect to trona; correlation of SiO2 with concentration of Na and total CO2 support this interpretation. Addition of moredilute waters to alkaline brines may lower the pH and cause inorganic precipitation of substantial amounts of silica.

  5. An alkaline element

    Energy Technology Data Exchange (ETDEWEB)

    Arita, T.; Murakami, K.; Okha, K.

    1983-04-28

    A cathode with a dual layer active mass is installed in the disk shaped alkaline silver and zinc element. The first layer, which is turned towards the anode, contains 85 parts Ag2O, 5 parts electrolytic MnO2 and 10 parts graphite. The second layer, which contacts the bottom of the element, contains 35 parts Ag2O, 60 parts electrolytic MnO2 and 5 parts graphite. The electrical capacity of the first and second layers is 60 and 40, respectively. The first layer may be discharged with a high current density and the second layer with less current density. The element has high characteristics with comparatively low cost.

  6. 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.

  7. Phosphatase activity in sandy soil influenced by mycorrhizal and non-mycorrhizal cover crops

    Directory of Open Access Journals (Sweden)

    Alceu Kunze

    2011-06-01

    Full Text Available Cover crops may difffer in the way they affect rhizosphere microbiota nutrient dynamics. The purpose of this study was to evaluate the effect of mycorrhizal and non-mycorrhizal cover crops on soil phosphatase activity and its persistence in subsequent crops. A three-year experiment was carried out with a Typic Quartzipsamment. Treatments were winter species, either mycorrhizal black oat (Avena strigosa Schreb or the non-mycorrhizal species oilseed radish (Raphanus sativus L. var. oleiferus Metzg and corn spurry (Spergula arvensis L.. The control treatment consisted of resident vegetation (fallow in the winter season. In the summer, a mixture of pearl millet (Pennisetum americanum L. with sunnhemp (Crotalaria juncea L. or with soybean (Glycine max L. was sown in all plots. Soil cores (0-10 cm and root samples were collected in six growing seasons (winter and summer of each year. Microbial biomass P was determined by the fumigation-extraction method and phosphatase activity using p-nitrophenyl-phosphate as enzyme substrate. During the flowering stage of the winter cover crops, acid phosphatase activity was 30-35 % higher in soils with the non-mycorrhizal species oilseed radish, than in the control plots, regardless of the amount of P immobilized in microbial biomass. The values of enzyme activity were intermediate in the plots with corn spurry and black oat. Alkaline phosphatase activity was 10-fold lower and less sensitive to the treatments, despite the significant relationship between the two phosphatase activities. The effect of plant species on the soil enzyme profile continued in the subsequent periods, during the growth of mycorrhizal summer crops, after completion of the life cycle of the cover crops.

  8. Construction of single chain Fv antibody against transferrin receptor and its protein fusion with alkaline phosphatase

    Institute of Scientific and Technical Information of China (English)

    Dao-Feng Yang; Hui-Fen Zhu; Zhi-Hua Wang; Guan-Xin Shen; De-Ying Tian

    2005-01-01

    AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by SfiⅠ and NotⅠ, it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E. coli TG1. The positive colonies were screened by colony PCR and their expressions were induced by IPTG. ScFv gene was gained by digesting ScFv expression vector pUC19/119 with Sfi I and NotⅠ restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E. coli TG1. The positive colonies were selected by bacterial colony PCR. The expression of fusion protein (scFv-AP) was induced by IPTG. Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku. Immunofluorescent assay (IFA) demonstrated its reactivity with TfR. The molecular weight of scFv-AP was 75 ku. Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity.CONCLUSION: We have successfully prepared the antihuman TfR scFv and constructed the fusion protein of scFv and AP. It is promising for immunological experiments.

  9. 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.

    nutrient to all life, being a structural and functional component of organisms. Large quantities of elemental phosphorus are immobilized in living organism, and are locked up in the sediments as insoluble organic and inorganic compounds (De et. al.,2011... of phenol released. Suitable controls were maintained in all analyses and the results represent the average of replicates. Biological data was log transformed for statistical analysis. Inter- relationships between biological and environmental parameters...

  10. 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

    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....... A lipid analysis revealed that fat absorption caused a marked increase in the microvillar membrane contents of free fatty acids. In conclusion, fat absorption rapidly induces a transient clathrin-dependent endocytosis via coated pits from the enterocyte brush border. The process selectively internalizes...

  11. Alkaline phosphatase : An old enzyme newly discovered : Implications in cardiac surgery

    NARCIS (Netherlands)

    Kats, S.

    2011-01-01

    After myocardial infarction complicated by cardiogenic shock the inflammatory response is the result of an ischemic insult. Elevated procalcitonin levels, correlated to IL-6 levels, are triggered by bacterial toxins. In cardiac surgery with the use of cardiopulmonary bypass (CPB) hypoperfusion of th

  12. The potential of alkaline phosphatase as a treatment for sepsis-associated acute kidney injury

    NARCIS (Netherlands)

    Peters, Esther; Masereeuw, R.; Pickkers, Peter

    2014-01-01

    Sepsis-associated acute kidney injury (AKI) is associated with a high attributable mortality and an increased risk of developing chronic kidney failure in survivors. As a successful therapy is, as yet, unavailable, a pharmacological treatment option is clearly warranted. Recently, two small phase II

  13. Alkaline phosphatase : a possible treatment for sepsis-associated acute kidney injury in critically ill patients

    NARCIS (Netherlands)

    Peters, Esther; Heemskerk, Suzanne; Masereeuw, R.; Pickkers, Peter

    2014-01-01

    Acute kidney injury (AKI) is a common disease in the intensive care unit and accounts for high morbidity and mortality. Sepsis, the predominant cause of AKI in this setting, involves a complex pathogenesis in which renal inflammation and hypoxia are believed to play an important role. A new therapy

  14. Alkaline phosphatase predicts relapse in chronic hepatitis C patients with end-of-treatment response

    Institute of Scientific and Technical Information of China (English)

    Gerd; Bodlaj; Rainer; Hubmann; Karim; Saleh; Tatjana; Stojakovic; Georg; Biesenbach; Jrg; Berg

    2010-01-01

    AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genoty...

  15. 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. PMID:25242559

  16. The significance of amperometric detection of alkaline phosphatase in colorectal cancer diagnostics

    Science.gov (United States)

    Belkin, Anton; Freynd, Genrietta; Katsnelson, Mikhail

    2016-08-01

    Colorectal cancer (CRC) is one of the most common cancers in the world; it takes the second place in oncological morbidity. The ALP activity of intestinal epithelial differentiation marker (ALP) was investigated in surgical material and colon biopsies of 47 patients with the electrochemical method using biosensors (biochips). The average current obtained in the studies of colorectal cancer tissues was lower than in the studies of intact colon mucosa. Histology of tumors matched the differentiation types/stages of adenocarcinomas. The study of ALP activity in the surgical material and biopsies of colon tumors can become one of the most useful methods for evaluating the functional atypia in tumor tissue.

  17. 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

    -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...

  18. 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

    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...

  19. In situ hybridization of cytokine mRNA using alkaline phosphatase-labelled oligodeoxynucleotide probes

    DEFF Research Database (Denmark)

    Clausen, Bettina Hjelm; Fenger, Christina; Finsen, B.

    2013-01-01

    In situ hybridization is a powerful tool for visualizing cellular gene expression in morphologically preserved brain tissue giving precise information on the regional expression of specific mRNA sequences in cells of diverse phenotype. Here, we describe a sensitive, simple, and robust method using...

  20. 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

  1. Alkaline battery, separator therefore

    Science.gov (United States)

    Schmidt, George F. (Inventor)

    1980-01-01

    An improved battery separator for alkaline battery cells has low resistance to electrolyte ion transfer and high resistance to electrode ion transfer. The separator is formed by applying an improved coating to an electrolyte absorber. The absorber, preferably, is a flexible, fibrous, and porous substrate that is resistant to strong alkali and oxidation. The coating composition includes an admixture of a polymeric binder, a hydrolyzable polymeric ester and inert fillers. The coating composition is substantially free of reactive fillers and plasticizers commonly employed as porosity promoting agents in separator coatings. When the separator is immersed in electrolyte, the polymeric ester of the film coating reacts with the electrolyte forming a salt and an alcohol. The alcohol goes into solution with the electrolyte while the salt imbibes electrolyte into the coating composition. When the salt is formed, it expands the polymeric chains of the binder to provide a film coating substantially permeable to electrolyte ion transfer but relatively impermeable to electrode ion transfer during use.

  2. Stability and selectivity of alkaline proteases in hydrophilic solvents

    DEFF Research Database (Denmark)

    Pedersen, Lars Haastrup; Ritthitham, Sinthuwat; Pleissner, Daniel

    2008-01-01

    Hydrophilic, organic solvents can be used as co-solvents with water to produce one phase systems sustaining optimal mass transfer of substrates and products of mixed polarity in biocatalysed processes. At concentrations below 50 % hydrophilic solvents can even have a stabilising effect on alkaline...... proteases, but at higher concentrations and particularly in anhydrous systems most enzymes including alkaline proteases will denature and consequently loose activity [1]. However, partial denaturing and increased structural flexibility due to the interaction between hydrophilic solvents and alkaline...... proteases has been agued as the primary reasons for increasing activity, influencing regio-selectivity and improving the enantio-selectivity of these enzymes [2]. Alkaline proteases have been shown to be active not only on peptides, but on a wide range of renewable resources for synthesis of biologically...

  3. Activity of Follicular Fluid Phosphatases and Their Correlation with Levels of Serum Esteroidal Hormones and Gonadotropins

    Directory of Open Access Journals (Sweden)

    Sh Byranvand

    2006-10-01

    Full Text Available Introduction: The aim of this study was to evaluate the correlation between serum levels of ovarian and gonadotropin hormones, age and number of follicles with follicular alkaline and acid phosphatase activity in infertile women under controlled ovarian hyperstimulation. Methods: After collection of follicular fluid and calculation of the number of follicles, the specific activity of alkaline (ALP and acid phosphatase (ACP was determined according to the total protein in 19 women at the time of puncture. Also at that time, the levels of progesterone, estradiol, and follicle stimulating hormone (FSH and leuteinizing hormone (LH of their sera were measured. The correlation of follicular ALP and ACP with each serum hormone levels, women age and number of follicles was calculated using non-parametric analysis. Results: The ALP has a correlation with progesterone (P=0.01 levels but doesn’t have any correlation with the other factors. However, the ACP activity has a correlation not only with follicular number but also with estradiol and progesterone levels (P=0.05. Conclusion: Thus ACP activity is more affected by ovarian hormone than ALP and it can affect the ovarian microenvironment and oocyte development.

  4. Effect of Phosphatases Activity in the Hepatopancreas and Muscle of the Fresh Water Female Field Crab, Spiralothelphusa hydrodroma (Herbst Treated with Cypermethrin

    Directory of Open Access Journals (Sweden)

    R. S. Sreenivasan

    2011-04-01

    Full Text Available The fresh water field crab, Spiralothelphusa hydrodroma is an important human food source in parts of South India and the crab is constantly exposed to pesticides, which are used extensively to control agricultural pests. Evaluation of the toxic effect of cypermethrin on the experimental crab for the LC₅₀ value was carried out. Effect of cypermethrin on the biochemical changes in the hepatopancreas and muscle was observed. Quantitative study of biochemical changes of acid phosphatase and alkaline phosphatase were undertaken.

  5. Protein-tyrosine phosphatases in zebrafish gastrulation

    NARCIS (Netherlands)

    van Eekelen, M.J.L.

    2011-01-01

    Protein tyrosine phosphorylation plays a key role in relaying external stimuli and signals into the cell towards the appropriate responses. This process is mediated by protein-tyrosine kinases adding a phosphor group to a tyrosine residue and protein-tyrosine phosphatases removing a phosphor group f

  6. Defining Starch Binding by Glucan Phosphatases

    DEFF Research Database (Denmark)

    Auger, Kyle; Raththagala, Madushi; Wilkens, Casper;

    2015-01-01

    Starch is a vital energy molecule in plants that has a wide variety of uses in industry, such as feedstock for biomaterial processing and biofuel production. Plants employ a three enzyme cyclic process utilizing kinases, amylases, and phosphatases to degrade starch in a diurnal manner. Starch is ...

  7. Enzyme kinetic characterization of protein tyrosine phosphatases

    DEFF Research Database (Denmark)

    Peters, Günther H.J.; Branner, S.; Møller, K. B.;

    2003-01-01

    Protein tyrosine phosphatases (PTPs) play a central role in cellular signaling processes, resulting in an increased interest in modulating the activities of PTPs. We therefore decided to undertake a detailed enzyme kinetic evaluation of various transmembrane and cytosolic PTPs (PTPalpha, PTPbeta...

  8. Cathepsin D-mediated yolk protein degradation is blocked by acid phosphatase inhibitors.

    Science.gov (United States)

    Fialho, Eliane; Nakamura, Angelica; Juliano, Luiz; Masuda, Hatisaburo; Silva-Neto, Mário A C

    2005-04-15

    Vitellin (VT) is a lipoglycophosphoprotein stored inside the eggs of every oviparous organism during oogenesis. In the blood-sucking bug Rhodnius prolixus, VT is deposited inside growing oocytes together with two acid hydrolases: acid phosphatase (AP) and cathepsin D (CD). Egg fertilization triggers AP activity and VT proteolysis in vivo [Insect Biochem. Mol. Biol. 2002 (32) 847]. Here, we show that CD is the main protease targeting VT proteolysis during egg development. CD activity in total egg homogenates is blocked by the classical aspartyl protease inhibitor, pepstatin A. Surprisingly, AP inhibitors such as NaF, Na+/K+ tartrate, and inorganic phosphate also block VT proteolysis, whereas this effect is not observed when tyrosine phosphatase inhibitors such as vanadate and phenylarsine oxide or an inhibitor of alkaline phosphatases such as levamisole are used in a VT proteolysis assay. NaF concentrations that block isolated AP activity do not affect the activity of partially purified CD. Therefore, a specific repressor of VT proteolysis must be dephosphorylated by AP in vivo. In conclusion, these results demonstrate for the first time that acid hydrolases act cooperatively to promote yolk degradation during egg development in arthropods. PMID:15797237

  9. Essential domain of receptor tyrosine phosphatase beta (RPTPbeta) for interaction with Helicobacter pylori vacuolating cytotoxin

    DEFF Research Database (Denmark)

    Yahiro, Kinnosuke; Wada, Akihiro; Yamasaki, Eiki;

    2004-01-01

    Helicobacter pylori produces a potent exotoxin, VacA, which causes progressive vacuolation as well as gastric injury. Although VacA was able to interact with two receptor-like protein tyrosine phosphatases, RPTPbeta and RPTPalpha, RPTPbeta was found to be responsible for gastric damage caused...

  10. Revisiting histidine-dependent acid phosphatases: a distinct group of tyrosine phosphatases

    OpenAIRE

    Veeramani, Suresh; Lee, Ming-Shyue; Lin, Ming-Fong

    2009-01-01

    Although classical protein tyrosine phosphatase (PTP) superfamily members are cysteine-dependent, emerging evidence shows that many acid phosphatases (AcPs) function as histidine-dependent PTPs in vivo. These AcPs dephosphorylate phospho-tyrosine substrates intracellularly and could have roles in development and disease. In contrast to cysteine-dependent PTPs, they utilize histidine, rather than cysteine, for substrate dephosphorylation. Structural analyses reveal that active site histidine, ...

  11. Research on Alkaline Filler Flame-Retarded Asphalt Pavement

    Institute of Scientific and Technical Information of China (English)

    HU Shuguang; ZHANG Houji; WANG Jiaolan

    2006-01-01

    Used as flame retardant of tunnel asphalt pavement, organic bromides produce a large amount of poisons and smoke in construction and flame retardation stage. The alkaline filler was found to replace mineral filler, and the flame-retarded asphalt mixtures were produced. Experimental results show that these asphalt mixtures are smoke restrained; the performances and construction technology of asphalt pavement are not influenced; also the alkaline filler is of low-price. So this kind of flame-retarded asphalt mixtures is suitable for tunnel pavement.

  12. Biochemical Properties and Inhibition Kinetics of Phosphatase from Wheat Thylakoid Membranes

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A phosphatase that hydrolyses phosphate monoesters has been isolated from wheat thylakoid membranes.Biochemical properties and inhibition kinetics of the phosphatase were investigated using several ions, organic solvents, and inhibitors. Wheat (Triticum aestivum L. cv. PH82-2-2) thylakoid membrane phosphatase activity was activated by Mg2+, Ca2+, and Fe2+ and was inhibited by Mn2+ and Cu2+. For example, enzyme activity was activated 34.81% by 2 mmol/L Mg2+, but was inhibited 22.3% and 8.5% by 2 and 1 mmol/L Cu2+, respectively.Methanol, ethanol and glycol were all able to activate enzyme activity. Enzyme activity was activated 58.5%, 48.2%,and 8.7% by 40% ethanol, methanol and glycol, respectively. From these results, it can be seen that the degree of activation of the phosphatase was greatest for ethanol and the type of activation was uncompetitive. Moreover,the activity of the thylakoid membrane phosphatase was inhibited by molybdate, vanadate, phosphate, and fluoride and the type of inhibition produced by these elements was uncompetitive, non-competitive, competitive and mixed, respectively.

  13. The alkaline and alkaline-carbonatite magmatism from Southern Brazil

    Science.gov (United States)

    Ruberti, E.; Gomes, C. D. B.; Comin-Chiaramonti, P.

    2015-12-01

    Early to Late Cretaceous lasting to Paleocene alkaline magmatism from southern Brazil is found associated with major extensional structural features in and around the Paraná Basin and grouped into various provinces on the basis of several data. Magmatism is variable in size, mode of occurrence and composition. The alkaline rocks are dominantly potassic, a few occurrences showing sodic affinity. The more abundant silicate rocks are evolved undersaturated to saturated in silica syenites, displaying large variation in igneous forms. Less evolved types are restricted to subvolcanic environments and outcrops of effusive suites occur rarely. Cumulatic mafic and ultramafic rock types are very common, particularly in the alkali-carbonatitic complexes. Carbonatite bodies are represented by Ca-carbonatites and Mg-carbonatites and more scarcely by Fe-carbonatites. Available radiometric ages for the alkaline rocks fit on three main chronological groups: around 130 Ma, subcoveal with the Early Cretaceous flood tholeiites of the Paraná Basin, 100-110 Ma and 80-90 Ma (Late Cretaceous). The alkaline magmatism also extends into Paleocene times, as indicated by ages from some volcanic lavas. Geochemically, alkaline potassic and sodic rock types are distinguished by their negative and positive Nb-Ta anomalies, respectively. Negative spikes in Nb-Ta are also a feature common to the associated tholeiitic rocks. Sr-Nd-Pb systematics confirm the contribution of both HIMU and EMI mantle components in the formation of the alkaline rocks. Notably, Early and Late Cretaceous carbonatites have the same isotopic Sr-Nd initial ratios of the associated alkaline rocks. C-O isotopic Sr-Nd isotopic ratios indicate typical mantle signature for some carbonatites and the influence of post-magmatic processes in others. Immiscibility of liquids of phonolitic composition, derived from mafic alkaline parental magmas, has been responsible for the origin of the carbonatites. Close association of alkaline

  14. Screening of Alkaline-Cellulose Producing Strain and Study on Its Enzymaic Characterization%产碱性纤维素酶菌株的筛选及酶学性质研究

    Institute of Scientific and Technical Information of China (English)

    卢金珍; 任俊; 杨文超; 熊汉国

    2011-01-01

    从采集的造纸厂土样中,筛选出l株产碱性纤维素酶酶活力较高的菌株H-1,经鉴定,该菌株为革兰氏阳性芽孢杆菌属.发酵产酶条件和酶学性质研究表明H-1菌株的适宜发酵务件为:接种量为6%,pH 9.0,温度为37℃,此条件下的酶活力可达8.69 U/mL,是初始酶活力(2.93 U/mL)的3倍.该酶最适反应温度为40℃,最适反应pH 9.0.在25~55℃,pH 7.0~ 11.0仍能保持60%以上的酶活力,能较好地满足日常洗涤的环境要求.%From soil samples collected from paper mills, one strain called H-l capable of degrading cellulose was screened and identified as Gram-positive Bacillus. The optimum enzyme-producing conditions were as follows, inoculum amount 6%, Ph 9.0 , temperature 37℃. The enzyme activity was up to 8.69 U/Ml, which was 3 times higher than the initial enzyme activity (2.93 U/Ml). The optimal conditions for the enzymatic reaction were about 40 ℃ and Ph 9.0. The enzyme activity could be maintained above 60% at temperature 25-55℃ and Ph 7.0~11.0, which meet the environmental requirements of the daily washing.

  15. Acid phosphatase production by recombinant Arxula adeninivorans.

    Science.gov (United States)

    Minocha, Neha; Kaur, Parvinder; Satyanarayana, T; Kunze, G

    2007-08-01

    Acid phosphatase production by recombinant Arxula adeninivorans was carried out in submerged fermentation. Using the Plackett-Burman design, three fermentation variables (pH, sucrose concentration, and peptone concentration) were identified to significantly affect acid phosphatase and biomass production, and these were optimized using response surface methodology of central composite design. The highest enzyme yields were attained in the medium with 3.9% sucrose and 1.6% peptone at pH 3.8. Because of optimization, 3.86- and 4.19-fold enhancement in enzyme production was achieved in shake flasks (17,054 U g(-1) DYB) and laboratory fermenter (18,465 U g(-1) DYB), respectively. PMID:17541580

  16. Electrolytic process for producing hydrogen peroxide

    International Nuclear Information System (INIS)

    An electrolytic process for producing hydrogen peroxide in an aqueous alkaline solution includes simultaneously passing an aqueous alkaline electrolyte and oxygen through a fluid permeable conductive cathode comprising reticulated vitreous carbon foam, separating the fluid permeable conductive cathode from an anode by a barrier and connecting the fluid permeable conductive electrode and the anode with an external power source to cause generation of hydrogen peroxide ion within the aqueous alkaline solution

  17. Assessment and kinetics of soil phosphatase in Brazilian Savanna systems.

    Science.gov (United States)

    Ferreira, Adão S; Espíndola, Suéllen P; Campos, Maria Rita C

    2016-05-31

    The activity and kinetics of soil phosphatases are important indicators to evaluate soil quality in specific sites such as the Cerrado (Brazilian Savanna). This study aimed to determine the activity and kinetic parameters of soil phosphatase in Cerrado systems. Soil phosphatase activity was assessed in samples of native Cerrado (NC), no-tillage (NT), conventional tillage (CT) and pasture with Brachiaria brizantha (PBb) and evaluated with acetate buffer (AB), tris-HCl buffer (TB), modified universal buffer (MUB) and low MUB. The Michaelis-Menten equation and Eadie-Hofstee model were applied to obtain the kinetic parameters of soil phosphatase using different concentrations of p-nitrophenol phosphate (p-NPP). MUB showed the lowest soil phosphatase activity in all soils whereas AB in NC and NT presented the highest. Low MUB decreased interferences in the assessment of soil phosphatase activity when compared to MUB, suggesting that organic acids interfere on the soil phosphatase activity. In NC and NT, soil phosphatase activity performed with TB was similar to AB and low MUB. Km values from the Michaels-Menten equation were higher in NC than in NT, which indicate a lower affinity of phosphatase activity for the substrate in NC. Vmax values were also higher in NC than in NT. The Eadie-Hofstee model suggests that NC had more phosphatase isoforms than NT. The study showed that buffer type is of fundamental importance when assessing soil phosphatase activity in Cerrado soils. PMID:27254453

  18. Acid Phosphatase Development during Ripening of Avocado.

    Science.gov (United States)

    Sacher, J A

    1975-02-01

    The activity and subcellular distribution of acid phosphatase were assayed during ethylene-induced ripening of whole fruit or thick slices of avocado (Persea americana Mill. var. Fuerte and Hass). The activity increased up to 30-fold during ripening in both the supernatant fraction and the Triton X-100 extract of the precipitate of a 30,000g centrifugation of tissue homogenates from whole fruit or slices ripening in moist air. Enzyme activity in the residual precipitate after Triton extraction remained constant. The development of acid phosphatase in thick slices ripened in moist air was similar to that in intact fruit, except that enzyme development and ripening were accelerated about 24 hours in the slices. The increase in enzyme activity that occurs in slices ripening in moist air was inhibited when tissue sections were infiltrated with solutions, by aspiration for 2 minutes or by soaking for 2 hours, anytime 22 hours or more after addition of ethylene. This inhibition was independent of the presence or absence of cycloheximide or sucrose (0.3-0.5m). However, the large decline in enzyme activity in the presence of cycloheximide, as compared with the controls, indicated that synthesis of acid phosphatase was occurring at all stages of ripening.

  19. Isolation, identification and optimization of alkaline protease production by Candida viswanathii

    Directory of Open Access Journals (Sweden)

    Mandana Lotfi

    2014-03-01

    Conclusion: Due to the high demand for industrial enzymes in the Country and the high activity of alkaline proteases produced by strain. It seems that the native strain can achieve high production of alkaline proteases.These native strains could be resulted in the independence of our country in industrial enzymes production.

  20. Production, purification and characterization of a thermotolerant alkaline serine protease from a novel species Bacillus caseinilyticus

    OpenAIRE

    Mothe, Thirumala; Sultanpuram, Vishnuvardhan Reddy

    2016-01-01

    Alkaline proteases are important enzymes in many industrial applications, especially as additives in laundry detergent industry. Though there are a number of Bacillus species which are reported to be producing proteases, the efficiency of a protease produced by a novel strain has to be studied in comparison to the others. Hence, in this study, an alkaline serine protease produced by a novel species Bacillus caseinilyticus was purified and characterized for its possible usage in detergent indu...

  1. Synthesis of functionalized fluorescent gold nanoclusters for acid phosphatase sensing

    Science.gov (United States)

    Sun, Jian; Yang, Fan; Yang, Xiurong

    2015-10-01

    A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by introducing an alkaline aqueous solution of MUA into the GSH-Au+ complexes or AuNC@GSH solution. Subsequently, a reliable AuNC@GSH/MUA-based real-time assay of acid phosphatase (ACP) is established for the first time, inspired by the selective coordination of Fe3+ with surface ligands of AuNCs, the higher binding affinity between the pyrophosphate ion (PPi) and Fe3+, and the hydrolysis of PPi into orthophosphate by ACP. Our fluorescent chemosensor can also be applied to assay ACP in a real biological sample and, furthermore, to screen the inhibitor of ACP. This report paves a new avenue for synthesizing AuNCs based on either the bottom-up reduction or top-down etching method, establishing real-time fluorescence assays for ACP by means of PPi as the substrate, and further exploring the sensing applications of fluorescent AuNCs.A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by

  2. Aluminum/alkaline earth metal composites and method for producing

    Science.gov (United States)

    Russell, Alan M; Anderson, Iver E; Kim, Hyong J; Freichs, Andrew E

    2014-02-11

    A composite is provided having an electrically conducting Al matrix and elongated filaments comprising Ca and/or Sr and/or Ba disposed in the matrix and extending along a longitudinal axis of the composite. The filaments initially comprise Ca and/or Sr and/or Ba metal or allow and then may be reacted with the Al matrix to form a strengthening intermetallic compound comprising Al and Ca and/or Sr and/or Ba. The composite is useful as a long-distance, high voltage power transmission conductor.

  3. Nucleotide sequences encoding a thermostable alkaline protease

    Science.gov (United States)

    Wilson, David B.; Lao, Guifang

    1998-01-01

    Nucleotide sequences, derived from a thermophilic actinomycete microorganism, which encode a thermostable alkaline protease are disclosed. Also disclosed are variants of the nucleotide sequences which encode a polypeptide having thermostable alkaline proteolytic activity. Recombinant thermostable alkaline protease or recombinant polypeptide may be obtained by culturing in a medium a host cell genetically engineered to contain and express a nucleotide sequence according to the present invention, and recovering the recombinant thermostable alkaline protease or recombinant polypeptide from the culture medium.

  4. Advanced inorganic separators for alkaline batteries

    Science.gov (United States)

    Sheibley, D. W. (Inventor)

    1982-01-01

    A flexible, porous battery separator comprising a coating applied to a porous, flexible substrate is described. The coating comprises: (1) a thermoplastic rubber-based resin which is insoluble and unreactive in the alkaline electrolyte; (2) a polar organic plasticizer which is reactive with the alkaline electrolyte to produce a reaction product which contains a hydroxyl group and/or a carboxylic acid group; and (3) a mixture of polar particulate filler materials which are unreactive with the electrolyte, the mixture comprising at least one first filler material having a surface area of greater than 25 meters sq/gram, at least one second filler material having a surface area of 10 to 25 sq meters/gram, wherein the volume of the mixture of filler materials is less than 45% of the total volume of the fillers and the binder, the filler surface area per gram of binder is about 20 to 60 sq meters/gram, and the amount of plasticizer is sufficient to coat each filler particle. A method of forming the battery separator is also described.

  5. Mannitol metabolism in brown algae involves a new phosphatase family.

    Science.gov (United States)

    Groisillier, Agnès; Shao, Zhanru; Michel, Gurvan; Goulitquer, Sophie; Bonin, Patricia; Krahulec, Stefan; Nidetzky, Bernd; Duan, Delin; Boyen, Catherine; Tonon, Thierry

    2014-02-01

    Brown algae belong to a phylogenetic lineage distantly related to green plants and animals, and are found predominantly in the intertidal zone, a harsh and frequently changing environment. Because of their unique evolutionary history and of their habitat, brown algae feature several peculiarities in their metabolism. One of these is the mannitol cycle, which plays a central role in their physiology, as mannitol acts as carbon storage, osmoprotectant, and antioxidant. This polyol is derived directly from the photoassimilate fructose-6-phosphate via the action of a mannitol-1-phosphate dehydrogenase and a mannitol-1-phosphatase (M1Pase). Genome analysis of the brown algal model Ectocarpus siliculosus allowed identification of genes potentially involved in the mannitol cycle. Among these, two genes coding for haloacid dehalogenase (HAD)-like enzymes were suggested to correspond to M1Pase activity, and thus were named EsM1Pase1 and EsM1Pase2, respectively. To test this hypothesis, both genes were expressed in Escherichia coli. Recombinant EsM1Pase2 was shown to hydrolyse the phosphate group from mannitol-1-phosphate to produce mannitol but was not active on the hexose monophosphates tested. Gene expression analysis showed that transcription of both E. siliculosus genes was under the influence of the diurnal cycle. Sequence analysis and three-dimensional homology modelling indicated that EsM1Pases, and their orthologues in Prasinophytes, should be seen as founding members of a new family of phosphatase with original substrate specificity within the HAD superfamily of proteins. This is the first report describing the characterization of a gene encoding M1Pase activity in photosynthetic organisms. PMID:24323504

  6. Development of alkaline fuel cells.

    Energy Technology Data Exchange (ETDEWEB)

    Hibbs, Michael R.; Jenkins, Janelle E.; Alam, Todd Michael; Janarthanan, Rajeswari [Colorado School of Mines, Golden, CO; Horan, James L. [Colorado School of Mines, Golden, CO; Caire, Benjamin R. [Colorado School of Mines, Golden, CO; Ziegler, Zachary C. [Colorado School of Mines, Golden, CO; Herring, Andrew M. [Colorado School of Mines, Golden, CO; Yang, Yuan [Colorado School of Mines, Golden, CO; Zuo, Xiaobing [Argonne National Laboratory, Argonne, IL; Robson, Michael H. [University of New Mexico, Albuquerque, NM; Artyushkova, Kateryna [University of New Mexico, Albuquerque, NM; Patterson, Wendy [University of New Mexico, Albuquerque, NM; Atanassov, Plamen Borissov [University of New Mexico, Albuquerque, NM

    2013-09-01

    This project focuses on the development and demonstration of anion exchange membrane (AEM) fuel cells for portable power applications. Novel polymeric anion exchange membranes and ionomers with high chemical stabilities were prepared characterized by researchers at Sandia National Laboratories. Durable, non-precious metal catalysts were prepared by Dr. Plamen Atanassov's research group at the University of New Mexico by utilizing an aerosol-based process to prepare templated nano-structures. Dr. Andy Herring's group at the Colorado School of Mines combined all of these materials to fabricate and test membrane electrode assemblies for single cell testing in a methanol-fueled alkaline system. The highest power density achieved in this study was 54 mW/cm2 which was 90% of the project target and the highest reported power density for a direct methanol alkaline fuel cell.

  7. Development of alkaline fuel cells.

    Energy Technology Data Exchange (ETDEWEB)

    Hibbs, Michael R.; Jenkins, Janelle E.; Alam, Todd Michael; Janarthanan, Rajeswari; Horan, James L.; Caire, Benjamin R.; Ziegler, Zachary C.; Herring, Andrew M.; Yang, Yuan; Zuo, Xiaobing; Robson, Michael H.; Artyushkova, Kateryna; Patterson, Wendy; Atanassov, Plamen Borissov

    2013-09-01

    This project focuses on the development and demonstration of anion exchange membrane (AEM) fuel cells for portable power applications. Novel polymeric anion exchange membranes and ionomers with high chemical stabilities were prepared characterized by researchers at Sandia National Laboratories. Durable, non-precious metal catalysts were prepared by Dr. Plamen Atanassovs research group at the University of New Mexico by utilizing an aerosol-based process to prepare templated nano-structures. Dr. Andy Herrings group at the Colorado School of Mines combined all of these materials to fabricate and test membrane electrode assemblies for single cell testing in a methanol-fueled alkaline system. The highest power density achieved in this study was 54 mW/cm2 which was 90% of the project target and the highest reported power density for a direct methanol alkaline fuel cell.

  8. Production of Alkaline Cellulase by Fungi Isolated from an Undisturbed Rain Forest of Peru

    OpenAIRE

    Karin Vega; Gretty K. Villena; Sarmiento, Victor H.; Yvette Ludeña; Nadia Vera; Marcel Gutiérrez-Correa

    2012-01-01

    Alkaline cellulase producing fungi were isolated from soils of an undisturbed rain forest of Peru. The soil dilution plate method was used for the enumeration and isolation of fast growing cellulolytic fungi on an enriched selective medium. Eleven out of 50 different morphological colonies were finally selected by using the plate clearing assay with CMC as substrate at different pH values. All 11 strains produced cellulases in liquid culture with activities at alkaline pH values without an ap...

  9. An electron microscopic study of the phosphatases in the ciliate Balantidium coli.

    Science.gov (United States)

    Skotarczak, B; Kołodziejczyk, L

    2005-11-01

    The localisation and activity of D glucose-6-phosphatase (G-6-Pase) and alkaline phosphatase (AlP) in the trophozoites of Balantidium coli isolated from pig intestine content were investigated using ultrastructural and cytochemical methods. The activity of G-6-Pase was demonstrated on the membranes of the endoplasmic reticulum, particularly in the cortical part of the trophozoites. In addition, the product of the reaction to G-6-Pase was concentrated in the vesicular structures, which were distributed along the reticular membranes. These structures were described as vesicles similar to glycosomes, containing enzymes of glycogenolysis. It is very likely that hydrolases in B. coli are formed on the rough reticular membranes without the involvement of cisterns of the Golgi complex. The ultrastructural deposits of the reaction to G-6-Pase and AlP in the trophozoites of B. coli described here indicate that some membranes of the rough endoplasmic reticulum and small vacuoles with a strong reaction to these enzymes can play a similar role to the Golgi complex. PMID:16425154

  10. Hydrogen in aluminum during alkaline corrosion

    Energy Technology Data Exchange (ETDEWEB)

    Adhikari, Saikat; Ai, Jiahe [Department of Chemical and Biological Engineering, Iowa State University, Ames, IA 50011 (United States); Hebert, Kurt R., E-mail: krhebert@iastate.ed [Department of Chemical and Biological Engineering, Iowa State University, Ames, IA 50011 (United States); Ho, K.M.; Wang, C.Z. [US DOE, Ames Laboratory, Ames, IA 50011 (United States)] [Department of Physics and Astronomy, Iowa State University, Ames, IA 50011 (United States)

    2010-07-30

    The thermodynamic state of hydrogen in aluminum during alkaline corrosion was investigated, using a two-compartment hydrogen permeation cell with an Al/Pd bilayer membrane. The open-circuit potential of the Pd layer in a pH 7.0 buffer solution was monitored to sense the hydrogen chemical potential, {mu}{sub H}. At pH 12.5-13.5, the measurements established a minimum {mu}{sub H} of 0.55 eV relative to the ideal gas reference, equivalent to a H{sub 2} gas pressure of 5.7 GPa. Statistical mechanics calculations show that vacancy-hydrogen defects are stable in Al at this condition. A dissolution mechanism was proposed in which H at very high {mu}{sub H} is produced by oxidation of interfacial aluminum hydride. The mechanism explains the observed rapid accumulation of H in the metal by extensive formation of vacancy-hydrogen defects.

  11. Counter-regulatory phosphatases TNAP and NPP1 temporally regulate tooth root cementogenesis

    Institute of Scientific and Technical Information of China (English)

    Laura E Zweifler; Mudita K Patel; Francisco H Nociti Jr; Helen F Wimer; Jose L Milla n; Martha J Somerman; Brian L Foster

    2015-01-01

    Cementum is critical for anchoring the insertion of periodontal ligament fibers to the tooth root. Several aspects of cementogenesis remain unclear, including differences between acellular cementum and cellular cementum, and between cementum and bone. Biomineralization is regulated by the ratio of inorganic phosphate (Pi) to mineral inhibitor pyrophosphate (PPi), where local Pi and PPi concentrations are controlled by phosphatases including tissue-nonspecific alkaline phosphatase (TNAP) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1). The focus of this study was to define the roles of these phosphatases in cementogenesis. TNAP was associated with earliest cementoblasts near forming acellular and cellular cementum. With loss of TNAP in the Alpl null mouse, acellular cementum was inhibited, while cellular cementum production increased, albeit as hypomineralized cementoid. In contrast, NPP1 was detected in cementoblasts after acellular cementum formation, and at low levels around cellular cementum. Loss of NPP1 in the Enpp1 null mouse increased acellular cementum, with little effect on cellular cementum. Developmental patterns were recapitulated in a mouse model for acellular cementum regeneration, with early TNAP expression and later NPP1 expression. In vitro, cementoblasts expressed Alpl gene/protein early, whereas Enpp1 gene/protein expression was significantly induced only under mineralization conditions. These patterns were confirmed in human teeth, including widespread TNAP, and NPP1 restricted to cementoblasts lining acellular cementum. These studies suggest that early TNAP expression creates a low PPi environment promoting acellular cementum initiation, while later NPP1 expression increases PPi, restricting acellular cementum apposition. Alterations in PPi have little effect on cellular cementum formation, though matrix mineralization is affected.

  12. Changes in Phosphorus Fractions, pH, and phosphatase Activity in rhizosphere of Two Rice Genotypes

    Institute of Scientific and Technical Information of China (English)

    LI Yong-Fu; LUO An-Cheng; WEI Xing-Hua; YAO Xu-Guo

    2008-01-01

    A rhizobox experiment with two phosphorus (P) treatments, zero-P (0 mg P kg-1) and plus-P (100 mg P kg-1) as Ca(H2PO4)2·H2O, was conducted to study the chemical and biochemical properties in the rhizosphere of two rice genotypes (cv. Zhongbu 51 and Pembe) different in P uptake ability and their relationship with the depletion of soil P fractions. Plant P uptake, pH, phosphatase activity, and soil P fractions in the rhizcephere were measured. Both total dry weight and total P uptake of Pembe were significantly (P < 0.05) higher than those of Zhongbu 51 in the zero-P and plns-P treatments. Significant depletions of resin-Pi, NaHCO3-Pi, NaHCO3-Po, and NaOH-Pi, where Pi stands for inorganic P and Po for organic P, were observed in the rhizosphere of both Zhongbu 51 and Pembe under both P treatments. Pembe showed a greater ability than Zhongbu 51 in depleting resin-Pi, NaHCO2-Pi, NaHCO3-Po, NaOH-Pi, and NaOH-Po in the rhizosphere. HCl-Pi and residual-P were not depleted in the rhizosphere of both genotypes, regardless of P treatments despite significant acidification in the rhizosphere of Pembe under zero-P treatment. Higher acid phosphatase (AcPME) activity and alkaline phosphatase (A1PME) activity were observed in the rhizosphere of both Zhongbu 51 and Pembe compared to the corresponding controls without plant. AcPME activity was negatively (P < 0.01) correlated to NaHCO3-Po concentration in the rhizosphere of both Zhongbu 51 and Pembe, suggesting that AcPME was associated with the mineralization of soil organic P.

  13. Enzymatic Hydrolysis of Alkaline Pretreated Coconut Coir

    Directory of Open Access Journals (Sweden)

    Akbarningrum Fatmawati

    2013-06-01

    Full Text Available The purpose of this research is to study the effect of concentration and temperature on the cellulose and lignin content, and the reducing sugars produced in the enzymatic hydrolysis of coconut coir. In this research, the coconut coir is pretreated using 3%, 7%, and 11% NaOH solution at 60oC, 80oC, and 100oC. The pretreated coir were assayed by measuring the amount of cellulose and lignin and then hydrolysed using Celluclast and Novozyme 188 under various temperature (30oC, 40oC, 50oC and pH (3, 4, 5. The hydrolysis results were assayed for the reducing sugar content. The results showed that the alkaline delignification was effective to reduce lignin and to increase the cellulose content of the coir. The best delignification condition was observed at 11% NaOH solution and 100oC which removed 14,53% of lignin and increased the cellulose content up to 50,23%. The best condition of the enzymatic hydrolysis was obtained at 50oC and pH 4 which produced 7,57 gr/L reducing sugar. © 2013 BCREC UNDIP. All rights reservedReceived: 2nd October 2012; Revised: 31st January 2013; Accepted: 6th February 2013[How to Cite: Fatmawati, A., Agustriyanto, R., Liasari, Y. (2013. Enzymatic Hydrolysis of Alkaline Pre-treated Coconut Coir. Bulletin of Chemical Reaction Engineering & Catalysis, 8 (1: 34-39 (doi:10.9767/bcrec.8.1.4048.34-39[Permalink/DOI: http://dx.doi.org/10.9767/bcrec.8.1.4048.34-39] | View in  |

  14. Structural mechanisms of plant glucan phosphatases in starch metabolism.

    Science.gov (United States)

    Meekins, David A; Vander Kooi, Craig W; Gentry, Matthew S

    2016-07-01

    Glucan phosphatases are a recently discovered class of enzymes that dephosphorylate starch and glycogen, thereby regulating energy metabolism. Plant genomes encode two glucan phosphatases, called Starch EXcess4 (SEX4) and Like Sex Four2 (LSF2), that regulate starch metabolism by selectively dephosphorylating glucose moieties within starch glucan chains. Recently, the structures of both SEX4 and LSF2 were determined, with and without phosphoglucan products bound, revealing the mechanism for their unique activities. This review explores the structural and enzymatic features of the plant glucan phosphatases, and outlines how they are uniquely adapted to perform their cellular functions. We outline the physical mechanisms used by SEX4 and LSF2 to interact with starch glucans: SEX4 binds glucan chains via a continuous glucan-binding platform comprising its dual-specificity phosphatase domain and carbohydrate-binding module, while LSF2 utilizes surface binding sites. SEX4 and LSF2 both contain a unique network of aromatic residues in their catalytic dual-specificity phosphatase domains that serve as glucan engagement platforms and are unique to the glucan phosphatases. We also discuss the phosphoglucan substrate specificities inherent to SEX4 and LSF2, and outline structural features within the active site that govern glucan orientation. This review defines the structural mechanism of the plant glucan phosphatases with respect to phosphatases, starch metabolism and protein-glucan interaction, thereby providing a framework for their application in both agricultural and industrial settings. PMID:26934589

  15. Serum total and bone alkaline phosphatase and tartrate-resistant acid phosphatase activities for the assessment of bone fracture healing in dogs

    Directory of Open Access Journals (Sweden)

    C. Sousa

    2011-08-01

    Full Text Available O objetivo deste trabalho foi estudar o padrão de variação da atividade sérica da fosfatase alcalina total (tALP, da isoenzima óssea da fosfatase alcalina (BALP e da fosfatase ácida resistente ao tartarato (TRAP, assim como a variação da concentração dos minerais séricos durante o processo de cicatrização de fraturas ósseas no cão. A variação sérica destes marcadores do metabolismo ósseo foi avaliada em nove cães com fraturas diafisárias fechadas de ossos longos, submetidas a tratamento cirúrgico para osteosíntese. Durante o período pós-operatório, sete animais evoluíram no sentido de uma normal união óssea, sendo que dois deles desenvolveram um processo de não união óssea. Foram observados, relativamente à BALP, valores de actividade sérica mais elevados e com diferença estatística (P<0,05 no grupo de animais que evoluiu no sentido de uma normal união óssea, comparativamente ao grupo de animais que evoluiu no sentido do processo de não união. No grupo de animais que evoluiu para a completa união óssea foram, adicionalmente, observados valores diminuidos (P<0,05 da atividade sérica da TRAP, até ao dia 60 do período pós-operatório seguido de uma elevação estatisticamente significativa após este período. Em conclusão, os biomarcadores do metabolismo ósseo poderão vir a constituir um método auxiliar de diagnóstico na monitorização do processo de cicatrização de fracturas ósseas, possibilitando, a detecção precoce de complicações pós-operatórias.

  16. The content of macro- and microelements and the phosphatase activity of soils under a varied plant cultivation technology

    Science.gov (United States)

    Bartkowiak, A.; Lemanowicz, J.; Kobierski, M.

    2015-12-01

    The paper presents the results of the analyses of selected physicochemical properties and the activity of alkaline and acid phosphatase in the soils which differed in terms of plant cultivation technology. Profile sI represented arable land in the crop rotation with cereals dominating (medium intensive technology), without irrigation, while profile sII—represented arable land with vegetable crops cultivation (intensive technology), intensively fertilized and irrigated. The content of available phosphorus in the two soil profiles investigated ranged from 6.6 to 69.1 mg/kg. The highest contents of phosphorus available to plants were reported in the plough horizon of both soils, while the abundance of potassium and magnesium was highest in the illuvial horizon of both soils. The soil profiles investigated showed a significant variation in terms of the cultivation technologies applied. The contents of plant-available Cu and Zn in soil were low and they resulted in the inhibition of neither alkaline nor acid phosphatase. The intensive vegetable crops cultivation technology decreased the content of organic matter and increased the content of the nutrients in soil. Using the Ward method, it was found that relatively similar physicochemical and chemical properties were reported for the genetic horizons of both soil profiles, especially Ap horizon of the soil representing arable land with intensive cultivation of vegetable crops.

  17. Effects icariin of on the alkline phosphatase activity of human periodontal ligament cells inhibited by LPS%淫羊藿苷对内毒素抑制牙周膜细胞ALP表达的影响

    Institute of Scientific and Technical Information of China (English)

    姜瑛; 王祥; 许华; 刘英群

    2013-01-01

    Objective: To investigate the effects of icariin (ICA) on proliferation of huaman periodontal ligament cells (human periodontal ligament cells, hPDLC) and the alkaline phosphatase (alkaline phosphatase, ALP) activity inhibited by LPS. Method: HPDLC were cultured in virto and stimulated with icariin with different concentrations (0, 10-5、 10-6、 10-7、 10-8、 10-9 mol / L) for 96 hours.The ability of proliferation of HPDLC was detected by MTT method.The activity of alkaline phosphatase and the expression of alkaline phosphatase mRNA was seperately determined by p-Nitrophnyl phosphate (pNPP) method and RT-PCR after being treated with icariin at the concentration of 10-6 mol / L inhibited by LPS. Result: icariin had a dose-dependent effect on the proliferation the hPDLC in a suitable concentration range from l0-6-10-8 mol / L. The alkaline phosphatase activity was remarkably inhibited by incubation of PDLC with 10 μg / mL LPS and was improved in the presence of icariin at the concentration of 10-6 mol / L. Conclusion: Icariin have the effect on the proliferation of PDLC and improve the ALP activity inhibited by LPS, which could help for Peri—apical tissue regeneration.%目的:探讨淫羊藿苷对人牙周膜细胞(periodontal ligament cells,PDLC)增殖及对内毒素(Lipopolysaccharide,LPS)干扰下碱性磷酸酶(alkaline phosphatase,ALP)的影响.方法:原代培养PDLC,MTT法检测不同浓度(0、10-5~10-9 mol/L)淫羊藿苷对PDLC增殖的影响;RT-PCR、对硝基苯酚法测定淫羊藿苷对LPS抑制PDLC的ALP mRNA表达和分泌的影响.结果:淫羊藿苷在一定浓度下(10-6~10-7 mol/L)可促进PDLC增殖;10 μg/mL LPS可抑制PDLC的ALP活性;加入10-6 mol/L淫羊藿苷干扰后,对LPS抑制PDLC的ALP有拮抗作用,可提高其mRNA表达和活性.结论:淫羊藿苷在一定浓度时可促进PDLC增殖;可能通过拮抗LPS抑制其ALP的活性.

  18. Voltage sensitive phosphatases: emerging kinship to protein tyrosine phosphatases from structure-function research

    Directory of Open Access Journals (Sweden)

    Kirstin eHobiger

    2015-02-01

    Full Text Available The transmembrane protein Ci-VSP from the ascidian Ciona intestinalis was described as first member of a fascinating family of enzymes, the voltage sensitive phosphatases (VSPs. Ci-VSP and its voltage-activated homologs from other species are stimulated by positive membrane potentials and dephosphorylate the head groups of negatively charged phosphoinositide phosphates (PIPs. In doing so, VSPs act as control centers at the cytosolic membrane surface, because they intervene in signaling cascades that are mediated by PIP lipids. The characteristic motif CX5RT/S in the active site classifies VSPs as members of the huge family of cysteine-based protein tyrosine phosphatases (PTPs. Although PTPs have already been well characterized regarding both, structure and function, their relationship to VSPs has drawn only limited attention so far. Therefore, the intention of this review is to give a short overview about the extensive knowledge about PTPs in relation to the facts known about VSPs. Here, we concentrate on the structural features of the catalytic domain which are similar between both classes of phosphatases and their consequences for the enzymatic function. By discussing results obtained from crystal structures, molecular dynamics simulations, and mutagenesis studies, a possible mechanism for the catalytic cycle of VSPs is presented based on that one proposed for PTPs. In this way, we want to link the knowledge about the catalytic activity of VSPs and PTPs.

  19. Different designs of kinase-phosphatase interactions and phosphatase sequestration shapes the robustness and signal flow in the MAPK cascade

    Directory of Open Access Journals (Sweden)

    Sarma Uddipan

    2012-07-01

    Full Text Available Abstract Background The three layer mitogen activated protein kinase (MAPK signaling cascade exhibits different designs of interactions between its kinases and phosphatases. While the sequential interactions between the three kinases of the cascade are tightly preserved, the phosphatases of the cascade, such as MKP3 and PP2A, exhibit relatively diverse interactions with their substrate kinases. Additionally, the kinases of the MAPK cascade can also sequester their phosphatases. Thus, each topologically distinct interaction design of kinases and phosphatases could exhibit unique signal processing characteristics, and the presence of phosphatase sequestration may lead to further fine tuning of the propagated signal. Results We have built four architecturally distinct types of models of the MAPK cascade, each model with identical kinase-kinase interactions but unique kinases-phosphatases interactions. Our simulations unravelled that MAPK cascade’s robustness to external perturbations is a function of nature of interaction between its kinases and phosphatases. The cascade’s output robustness was enhanced when phosphatases were sequestrated by their target kinases. We uncovered a novel implicit/hidden negative feedback loop from the phosphatase MKP3 to its upstream kinase Raf-1, in a cascade resembling the B cell MAPK cascade. Notably, strength of the feedback loop was reciprocal to the strength of phosphatases’ sequestration and stronger sequestration abolished the feedback loop completely. An experimental method to verify the presence of the feedback loop is also proposed. We further showed, when the models were activated by transient signal, memory (total time taken by the cascade output to reach its unstimulated level after removal of signal of a cascade was determined by the specific designs of interaction among its kinases and phosphatases. Conclusions Differences in interaction designs among the kinases and phosphatases can

  20. Alkaline fuel cell performance investigation

    Science.gov (United States)

    Martin, R. E.; Manzo, M. A.

    1988-01-01

    An exploratory experimental fuel cell test program was conducted to investigate the performance characteristics of alkaline laboratory research electrodes. The objective of this work was to establish the effect of temperature, pressure, and concentration upon performance and evaluate candidate cathode configurations having the potential for improved performance. The performance characterization tests provided data to empirically establish the effect of temperature, pressure, and concentration upon performance for cell temperatures up to 300 F and reactant pressures up to 200 psia. Evaluation of five gold alloy cathode catalysts revealed that three doped gold alloys had more that two times the surface areas of reference cathodes and therefore offered the best potential for improved performance.

  1. INFLUENCE OF LIMING AND WASTE ORGANIC MATERIALS ON THE ACTIVITY OF PHOSPHATASE IN SOIL CONTAMINATED WITH NICKEL

    Directory of Open Access Journals (Sweden)

    Beata Kuziemska

    2014-10-01

    Full Text Available A study was carried out on soil following a two-year pot experiment that was conducted in 2009–2010, in three repetitions in Siedlce. The experiment included the following factors: 1 – amount of Ni in soil (0, 75, 150 and 225 mg·kg-1 soil by applying an aqueous NiSO4·7H2O solution; 2 – liming (0 and Ca according to 1 Hh as CaCO3; 3 – organic waste products (rye straw at a dose of 4 t·ha-1 and brown coal at a dose of 40 t·ha-1. In each experimental year, orchard grass was the test plant and four swaths were harvested. The activities of acidic and alkaline phosphatase, pH and the content of carbon in organic compounds were determined in the soil samples collected after each grass swath and in each experimental year. It was found that Ni at 75 mg·kg-1 soil activated the enzymes under study, whereas higher doses caused their statistically-confirmed inactivation. The lowest activity of the investigated enzymes was detected in soil supplemented with 225 Ni·kg-1 soil. Liming caused an increase in the activity of alkaline phosphatase and a reduction in the activity of acidic phosphatase. Straw and brown coal induced a substantial increase in the activity of both enzymes in the tested soil samples. Both liming and straw and carbon eliminated the negative effect of higher nickel doses on the activity of the enzymes under study.

  2. When can ocean acidification impacts be detected from decadal alkalinity measurements?

    Science.gov (United States)

    Carter, B. R.; Frölicher, T. L.; Dunne, J. P.; Rodgers, K. B.; Slater, R. D.; Sarmiento, J. L.

    2016-04-01

    We use a large initial condition suite of simulations (30 runs) with an Earth system model to assess the detectability of biogeochemical impacts of ocean acidification (OA) on the marine alkalinity distribution from decadally repeated hydrographic measurements such as those produced by the Global Ship-Based Hydrographic Investigations Program (GO-SHIP). Detection of these impacts is complicated by alkalinity changes from variability and long-term trends in freshwater and organic matter cycling and ocean circulation. In our ensemble simulation, variability in freshwater cycling generates large changes in alkalinity that obscure the changes of interest and prevent the attribution of observed alkalinity redistribution to OA. These complications from freshwater cycling can be mostly avoided through salinity normalization of alkalinity. With the salinity-normalized alkalinity, modeled OA impacts are broadly detectable in the surface of the subtropical gyres by 2030. Discrepancies between this finding and the finding of an earlier analysis suggest that these estimates are strongly sensitive to the patterns of calcium carbonate export simulated by the model. OA impacts are detectable later in the subpolar and equatorial regions due to slower responses of alkalinity to OA in these regions and greater seasonal equatorial alkalinity variability. OA impacts are detectable later at depth despite lower variability due to smaller rates of change and consistent measurement uncertainty.

  3. Coupling the Alkaline-Surfactant-Polymer Technology and The Gelation Technology to Maximize Oil Production

    Energy Technology Data Exchange (ETDEWEB)

    Malcolm Pitts; Jie Qi; Dan Wilson; Phil Dowling; David Stewart; Bill Jones

    2005-12-01

    Performance and produced polymer evaluation of four alkaline-surfactant-polymer projects concluded that only one of the projects could have benefited from combining the alkaline-surfactant-polymer and gelation technologies. Cambridge, the 1993 Daqing, Mellott Ranch, and the Wardlaw alkaline-surfacant-polymer floods were studied. An initial gel treatment followed by an alkaline-surfactant-polymer flood in the Wardlaw field would have been a benefit due to reduction of fracture flow. Numerical simulation demonstrated that reducing the permeability of a high permeability zone of a reservoir with gel improved both waterflood and alkaline-surfactant-polymer flood oil recovery. A Minnelusa reservoir with both A and B sand production was simulated. A and B sands are separated by a shale layer. A sand and B sand waterflood oil recovery was improved by 196,000 bbls or 3.3% OOIP when a gel was placed in the B sand. Alkaline-surfactant-polymer flood oil recovery improvement over a waterflood was 392,000 bbls or 6.5% OOIP. Placing a gel into the B sand prior to an alkaline-surfactant-polymer flood resulted in 989,000 bbl or 16.4% OOIP more oil than only water injection. A sand and B sand alkaline-surfactant-polymer flood oil recovery was improved by 596,000 bbls or 9.9% OOIP when a gel was placed in the B sand.

  4. Crystallization and preliminary X-ray diffraction analysis of rat protein tyrosine phosphatase η

    Energy Technology Data Exchange (ETDEWEB)

    Matozo, Huita C.; Nascimento, Alessandro S.; Santos, Maria A. M. [Instituto de Física de São Carlos, Departamento de Física e Informática, Universidade de São Paulo, Avenida Trabalhador São Carlense 400, CEP 13566-590 São Carlos, SP (Brazil); Iuliano, Rodolfo [Dipartimento di Medicina Sperimentale e Clinica, Facoltà di Medicina e Chirurgia, Università di Catanzaro, 88100 Catanzaro (Italy); Fusco, Alfredo [Dipartimento di Biologia e Patologia Cellulare e Molecolare, c/o Instituto di Endocrinologia ed Oncologia Sperimentale del CNR, Facolta di Medicina e Chirurgia, Università degli Studi di Napoli ‘Federico II’, Via Pansini 5, 80131 Naples (Italy); NOGEC (Naples Oncogenomocs Center)-CEINGE, Biotecnologie Avanzate, Via Comunale Margherita 482, 80145 Naples (Italy); Polikarpov, Igor, E-mail: ipolikarpov@if.sc.usp.br [Instituto de Física de São Carlos, Departamento de Física e Informática, Universidade de São Paulo, Avenida Trabalhador São Carlense 400, CEP 13566-590 São Carlos, SP (Brazil); Laboratório Nacional de Luz Síncrotron, Campinas, SP (Brazil)

    2006-09-01

    In this study, the catalytic domain of rat protein tyrosine phosphatase η was produced in Escherichia coli in soluble form and purified to homogeneity. Crystals were obtained by the hanging-drop vapour-diffusion method. The rat protein tyrosine phosphatase η (rPTPη) is a cysteine-dependent phosphatase which hydrolyzes phosphoester bonds in proteins and other molecules. rPTPη and its human homologue DEP-1 are involved in neoplastic transformations. Thus, expression of the protein is reduced in all oncogene-transformed thyroid cell lines and is absent in highly malignant thyroid cells. Moreover, consistent with the suggested tumour suppression role of PTPη, inhibition of the tumorigenic process occurs after its exogenous reconstitution, suggesting that PTPη might be important for gene therapy of cancers. In this study, the catalytic domain of rPTPη was produced in Escherichia coli in soluble form and purified to homogeneity. Crystals were obtained by the hanging-drop vapour-diffusion method. Diffraction data were collected to 1.87 Å resolution. The crystal belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 46.46, b = 63.07, c = 111.64 Å, and contains one molecule per asymmetric unit.

  5. Alkaline Water and Longevity: A Murine Study.

    Science.gov (United States)

    Magro, Massimiliano; Corain, Livio; Ferro, Silvia; Baratella, Davide; Bonaiuto, Emanuela; Terzo, Milo; Corraducci, Vittorino; Salmaso, Luigi; Vianello, Fabio

    2016-01-01

    The biological effect of alkaline water consumption is object of controversy. The present paper presents a 3-year survival study on a population of 150 mice, and the data were analyzed with accelerated failure time (AFT) model. Starting from the second year of life, nonparametric survival plots suggest that mice watered with alkaline water showed a better survival than control mice. Interestingly, statistical analysis revealed that alkaline water provides higher longevity in terms of "deceleration aging factor" as it increases the survival functions when compared with control group; namely, animals belonging to the population treated with alkaline water resulted in a longer lifespan. Histological examination of mice kidneys, intestine, heart, liver, and brain revealed that no significant differences emerged among the three groups indicating that no specific pathology resulted correlated with the consumption of alkaline water. These results provide an informative and quantitative summary of survival data as a function of watering with alkaline water of long-lived mouse models.

  6. Purification and characterization of alkaline protease from Lysinibacillus fusiformis

    OpenAIRE

    Suppiah S*; Sendeshkannan K; Prabakaran P; Rajkumar G; Yasothkumar N

    2012-01-01

    A novel alkaline protease producing bacterium was isolated from the gut of an estuarine fish Etroplus suratensis. The strain was identified by sequencing the fragment of their bacterial 16s rRNA and its homology was 97% closest to the Lysinibacillus fusiformis. An extracellular protease from this organism was purified by acetone precipitation, ion exchange chromatography and gel filtration chromatography methods and the specific activity of the purified enzyme was found to be 20.39 U/mg, 169....

  7. On the variation of alkalinity during phytoplankton photosynthesis

    Directory of Open Access Journals (Sweden)

    2005-01-01

    Full Text Available The alkalinity of the organic constituents of marine phytoplankton and their participation in the total alkalinity (TA change of seawater during photosynthesis are carefully assessed. Quantification of the contribution of phytoplankton chlorophyll, proteins and phosphorus compounds to the hydrogen ion balance of seawater in terms of total inorganic nitrogen (∆[NT] = ∆[NH4 +] + ∆[N2] + ∆[NO2 –] + ∆[NO3 –] and total inorganic phosphorus (∆[PT] changes during photosynthesis yielded that the organic components of marine phytoplankton are alkaline by –0.06 × ∆[NT] – 0.49 × ∆[PT], and that the potential total alkalinity (TAP during photosynthesis is TAP = TA – [NH4 –] + 0.93 × [NO2 –] + [NO3 –] + 0.08 × [NT] + 0.23 × [PT] for unfiltered seawater samples and TAP = TA – [NH4 –] + 0.93 × [NO2 –] + [NO3 –] + 0.02 × [NT] + 0.26 × [PT] for filtered seawater samples. These equations correct the traditionally used expression TAP = TA + [NO3 –]. The TAP anomalies are produced, in order of increasing importance, by N2 fixation, DMSP production and CaCO3 fixation.

  8. Structure-Function Analysis of the 3' Phosphatase Component of T4 Polynucleotide Kinase/phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Zhu,H.; Smith, P.; Wang, L.; Shuman, S.

    2007-01-01

    T4 polynucleotide kinase/phosphatase (Pnkp) exemplifies a family of bifunctional enzymes with 5'-kinase and 3' phosphatase activities that function in nucleic acid repair. T4 Pnkp is a homotetramer of a 301-aa polypeptide, which consists of an N-terminal kinase domain of the P-loop phosphotransferase superfamily and a C-terminal phosphatase domain of the DxD acylphosphatase superfamily. The homotetramer is formed via pairs of phosphatase-phosphatase and kinase-kinase homodimer interfaces. Here we identify four side chains-Asp187, Ser211, Lys258, and Asp277-that are required for 3' phosphatase activity. Alanine mutations at these positions abolished phosphatase activity without affecting kinase function or tetramerization. Conservative substitutions of asparagine or glutamate for Asp187 did not revive the 3' phosphatase, nor did arginine or glutamine substitutions for Lys258. Threonine in lieu of Ser211 and glutamate in lieu of Asp277 restored full activity, whereas asparagine at position 277 had no salutary effect. We report a 3.0 A crystal structure of the Pnkp tetramer, in which a sulfate ion is coordinated between Arg246 and Arg279 in a position that we propose mimics one of the penultimate phosphodiesters (5'NpNpNp-3') of the polynucleotide 3'-PO(4) substrate. The amalgam of mutational and structural data engenders a plausible catalytic mechanism for the phosphatase that includes covalent catalysis (via Asp165), general acid-base catalysis (via Asp167), metal coordination (by Asp165, Asp277 and Asp278), and transition state stabilization (via Lys258, Ser211, backbone amides, and the divalent cation). Other critical side chains play architectural roles (Arg176, Asp187, Arg213, Asp254). To probe the role of oligomerization in phosphatase function, we introduced six double-alanine cluster mutations at the phosphatase-phosphatase domain interface, two of which (R297A-Q295A and E292A-D300A) converted Pnkp from a tetramer to a dimer

  9. Alkaline and alkaline earth metal phosphate halides and phosphors

    Science.gov (United States)

    Lyons, Robert Joseph; Setlur, Anant Achyut; Cleaver, Robert John

    2012-11-13

    Compounds, phosphor materials and apparatus related to nacaphite family of materials are presented. Potassium and rubidium based nacaphite family compounds and phosphors designed by doping divalent rare earth elements in the sites of alkaline earth metals in the nacaphite material families are descried. An apparatus comprising the phosphors based on the nacaphite family materials are presented herein. The compounds presented is of formula A.sub.2B.sub.1-yR.sub.yPO.sub.4X where the elements A, B, R, X and suffix y are defined such that A is potassium, rubidium, or a combination of potassium and rubidium and B is calcium, strontium, barium, or a combination of any of calcium, strontium and barium. X is fluorine, chlorine, or a combination of fluorine and chlorine, R is europium, samarium, ytterbium, or a combination of any of europium, samarium, and ytterbium, and y ranges from 0 to about 0.1.

  10. Progress of research on the influence of alkaline cation and alkaline solution on bentonite properties

    International Nuclear Information System (INIS)

    Based on the previous laboratory studies and numerical simulation on bentonite in alkaline environments, the effects of alkaline cation and alkaline solution on mineral composition, microstructure, swelling capacity and hydraulic properties of bentonite are emphasized in this paper, temperature, pH values and concentration are discussed as main affecting factors. When bentonite is exposed to alkaline cation or alkaline solution, microstructure of bentonite will be changed due to the dissolution of montmorillonite and the formation of secondary minerals, which results in the decrease of swelling pressure. The amount of the reduction of swelling pressure depends on the concentration of alkaline solution. Temperature, polyvalent cation, salinity and concentration are the main factors affecting hydraulic properties of bentonite under alkaline conditions. Therefore, future research should focus on the mechanism of coupling effects of weak alkaline solutions on the mineral composition, microstructure, swelling capacity and hydraulic properties of bentonite under different temperatures and different pH values. (authors)

  11. 产蛋白酶混合菌系对碱性剩余污泥水解酸化的影响%Effect of mixed microbial consortium capable of protease-producing on hydrolysis and acidification of excess sludge under alkaline condition

    Institute of Scientific and Technical Information of China (English)

    接伟光; 彭永臻

    2014-01-01

    为提高剩余污泥水解酸化过程中挥发性脂肪酸(VFAs)的累积,从剩余污泥中分离产蛋白酶活力较高的耐碱细菌,并构建产蛋白酶混合菌系.将其接种于碱性( pH 10.0)发酵剩余污泥的不同发酵时期,评价其对溶解性有机化合物和VFAs累积的影响,探讨利用剩余污泥生产VFAs的最佳条件.从剩余污泥中分离到2株产蛋白酶活力较高的耐碱细菌,并构建产蛋白酶混合菌系.在发酵初期接种混合菌系效果最显著,且可缩短发酵启动时间2 d.发酵初期接种混合菌系后,溶解性蛋白质和VFAs质量浓度在第8天均达到最高值,分别为未接种混合菌系样品中相应值的1.25和1.41倍,分别占溶解性化学需氧量( SCOD)总量的29.87%和44.54%.乙酸和丙酸为剩余污泥水解酸化过程中VFAs的主要组分,分别占VFAs总量的50.69%和18.19%.%To improve volatile fatty acids ( VFAs ) accumulation from hydrolysis and acidification of excess sludge ( ES) , alkali⁃tolerant bacteria capable of protease⁃producing were isolated from ES. A mixed microbial consortium capable of protease⁃producing was constructed by the isolated bacterial strains. The mixed microbial consortium was inoculated into the different fermentation periods of ES to investigate their effects on soluble organic compounds and VFAs accumulation from ES under alkaline conditions ( pH 10. 0 ) . The optimal condition for VFAs accumulation from ES was investigated. The results showed that two alkali⁃tolerant bacterial strains capable of protease⁃producing were isolated from ES and constructed as a mixed microbial consortium. The soluble organic compounds concentrations and VFAs accumulation were improved significantly after the mixed microbial consortium was inoculated at the initial fermentation, and the start⁃up phase was shortened by 2 days. On the 8th day of fermentation, the concentrations of

  12. The SHP-2 tyrosine phosphatase: Signaling mechanisms and biological functions

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals.Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction.SHP-2, a cytoplasmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation.

  13. Process for treating alkaline wastes for vitrification

    Science.gov (United States)

    Hsu, Chia-lin W.

    1994-01-01

    According to its major aspects and broadly stated, the present invention is a process for treating alkaline waste materials, including high level radioactive wastes, for vitrification. The process involves adjusting the pH of the wastes with nitric acid, adding formic acid (or a process stream containing formic acid) to reduce mercury compounds to elemental mercury and MnO{sub 2} to the Mn(II) ion, and mixing with class formers to produce a melter feed. The process minimizes production of hydrogen due to noble metal-catalyzed formic acid decomposition during, treatment, while producing a redox-balanced feed for effective melter operation and a quality glass product. An important feature of the present invention is the use of different acidifying and reducing, agents to treat the wastes. The nitric acid acidifies the wastes to improve yield stress and supplies acid for various reactions; then the formic acid reduces mercury compounds to elemental mercury and MnO{sub 2}) to the Mn(II) ion. When the pH of the waste is lower, reduction of mercury compounds and MnO{sub 2}) is faster and less formic acid is needed, and the production of hydrogen caused by catalytically-active noble metals is decreased.

  14. Pregnancy-secreted Acid phosphatase, uteroferrin, enhances fetal erythropoiesis.

    Science.gov (United States)

    Ying, Wei; Wang, Haiqing; Bazer, Fuller W; Zhou, Beiyan

    2014-11-01

    Uteroferrin (UF) is a progesterone-induced acid phosphatase produced by uterine glandular epithelia in mammals during pregnancy and targeted to sites of hematopoiesis throughout pregnancy. The expression pattern of UF is coordinated with early fetal hematopoietic development in the yolk sac and then liver, spleen, and bone to prevent anemia in fetuses. Our previous studies suggested that UF exerts stimulatory impacts on hematopoietic progenitor cells. However, the precise role and thereby the mechanism of action of UF on hematopoiesis have not been investigated previously. Here, we report that UF is a potent regulator that can greatly enhance fetal erythropoiesis. Using primary fetal liver hematopoietic cells, we observed a synergistic stimulatory effect of UF with erythropoietin and other growth factors on both burst-forming unit-erythroid and colony-forming unit-erythroid formation. Further, we demonstrated that UF enhanced erythropoiesis at terminal stages using an in vitro culture system. Surveying genes that are crucial for erythrocyte formation at various stages revealed that UF, along with erythropoietin, up-regulated transcription factors required for terminal erythrocyte differentiation and genes required for synthesis of hemoglobin. Collectively, our results demonstrate that UF is a cytokine secreted by uterine glands in response to progesterone that promotes fetal erythropoiesis at various stages of pregnancy, including burst-forming unit-erythroid and colony-forming unit-erythroid progenitor cells and terminal stages of differentiation of hematopoietic cells in the erythroid lineage. PMID:25093463

  15. Uranium Biomineralization By Natural Microbial Phosphatase Activities in the Subsurface

    Energy Technology Data Exchange (ETDEWEB)

    Taillefert, Martial [Georgia Tech Research Corporation, Atlanta, GA (United States)

    2015-04-01

    This project investigated the geochemical and microbial processes associated with the biomineralization of radionuclides in subsurface soils. During this study, it was determined that microbial communities from the Oak Ridge Field Research subsurface are able to express phosphatase activities that hydrolyze exogenous organophosphate compounds and result in the non-reductive bioimmobilization of U(VI) phosphate minerals in both aerobic and anaerobic conditions. The changes of the microbial community structure associated with the biomineralization of U(VI) was determined to identify the main organisms involved in the biomineralization process, and the complete genome of two isolates was sequenced. In addition, it was determined that both phytate, the main source of natural organophosphate compounds in natural environments, and polyphosphate accumulated in cells could also be hydrolyzed by native microbial population to liberate enough orthophosphate and precipitate uranium phosphate minerals. Finally, the minerals produced during this process are stable in low pH conditions or environments where the production of dissolved inorganic carbon is moderate. These findings suggest that the biomineralization of U(VI) phosphate minerals is an attractive bioremediation strategy to uranium bioreduction in low pH uranium-contaminated environments. These efforts support the goals of the SBR long-term performance measure by providing key information on "biological processes influencing the form and mobility of DOE contaminants in the subsurface".

  16. Alkaline azide mutagenicity in cowpea

    International Nuclear Information System (INIS)

    Sodium azide is known as a potent mutagen in cereals and legumes. It is very effective in acidic medium in barley. Here an attempt is made to measure the effectiveness of sodium azide in alkaline medium (pH 7.4) on cowpea (Vigna unguiculata (L.) Walp., variety FS-68). Seeds pre-soaked in distilled water for 5 hours were treated with different concentrations (10-6, 10-5, 10-4 and 10-3M) of sodium azide (NaN3) for 4 hours at 28± 2 deg. C. Bottles were intermittently shaken, then the seeds were thoroughly washed in running tap water and subsequently planted in pots. The treatment caused significant biological damage such as reduction in seed germination, length of root and shoot, number of nodules and pods per plant and morphological leaf variations. Morphological, as well as chlorophyll mutants, were detected in M2

  17. Energetics of Amino Acid Synthesis in Alkaline Hydrothermal Environments

    Science.gov (United States)

    Kitadai, Norio

    2015-12-01

    Alkaline hydrothermal systems have received considerable attention as candidates for the origin and evolution of life on the primitive Earth. Nevertheless, sufficient information has not yet been obtained for the thermodynamic properties of amino acids, which are necessary components for life, at high temperatures and alkaline pH. These properties were estimated using experimental high-temperature volume and heat capacity data reported in the literature for several amino acids, together with correlation algorithms and the revised Helgeson-Kirkham-Flowers (HKF) equations of state. This approach enabled determination of a complete set of the standard molal thermodynamic data and the revised HKF parameters for the 20 protein amino acids in their zwitterionic and ionization states. The obtained dataset was then used to evaluate the energetics of amino acid syntheses from simple inorganic precursors (CO2, H2, NH3 and H2S) in a simulated alkaline hydrothermal system on the Hadean Earth. Results show that mixing between CO2-rich seawater and the H2-rich hydrothermal fluid can produce energetically favorable conditions for amino acid syntheses, particularly in the lower-temperature region of such systems. Together with data related to the pH and temperature dependences of the energetics of amino acid polymerizations presented in earlier reports, these results suggest the following. Hadean alkaline hydrothermal settings, where steep pH and temperature gradients may have existed between cool, slightly acidic Hadean ocean water and hot, alkaline hydrothermal fluids at the vent-ocean interface, may be energetically the most suitable environment for the synthesis and polymerization of amino acids.

  18. Studies on the production of alkaline α-amylase from Bacillus subtilis CB-18

    OpenAIRE

    Ogbonnaya Nwokoro; Odiase Anthonia

    2015-01-01

    Background. Amylases are among the main enzymes used in food and other industries. They hydrolyse starch molecules into polymers composing glucose units. Amylases have potential applications in a number of industrial processes including foods and pharmaceutical industries. Alkaline α-amylase has the potential of hydrolysing starch under alkaline pH and is useful in the starch and textile industries and as an ingredient of detergents. Amylases are produced from plants, however, microbial...

  19. Effect of phosphoric fertilizer and starter rates of nitrogen fertilizers on the phosphatase activity in the rhizosphere soil and nonlignified soybean roots under drought conditions

    Science.gov (United States)

    Emnova, E. E.; Daraban, O. V.; Bizgan, I. V.; Toma, S. I.

    2014-02-01

    In a small-plot field experiment, two soybean ( Glycine max L.) cultivars were grown on a calcareous chernozem under the drought conditions of 2012 with the preplanting application of simple superphosphate (Ps) at 60 kg/ha, urea (Nu) at 10 and 20 kg/ha, and ammonium nitrate (Nan) at 20 kg/ha. The phosphatase activity was measured in the rhizosphere soil (0- to 20-cm layer) and the fine nonlignified roots of soybean plants at the blossoming and pod-formation stages (the soil water content was 19 and 33% of the total water capacity, respectively). The maximum content of available phosphorus in the rhizosphere of both soybean cultivars (4.3-4.8 mg/100 g dry soil) was found at the simultaneous application of Ps and Nu20. Higher activities of the predominant phosphatases (alkaline phosphatase in the rhizosphere and acid phosphatase in the roots) were observed in the root-inhabited zone of the soil under the Indra cultivar compared to the Aura cultivar, which correlated with the lower content of available phosphorus in the rhizosphere soil (especially at the simultaneous application of Ps and Nu20) and the higher productivity of this cultivar in this treatment.

  20. Protein Phosphatases Involved in Regulating Mitosis: Facts and Hypotheses.

    Science.gov (United States)

    Kim, Hyun-Soo; Fernandes, Gary; Lee, Chang-Woo

    2016-09-01

    Almost all eukaryotic proteins are subject to post-translational modifications during mitosis and cell cycle, and in particular, reversible phosphorylation being a key event. The recent use of high-throughput experimental analyses has revealed that more than 70% of all eukaryotic proteins are regulated by phosphorylation; however, the mechanism of dephosphorylation, counteracting phosphorylation, is relatively unknown. Recent discoveries have shown that many of the protein phosphatases are involved in the temporal and spatial control of mitotic events, such as mitotic entry, mitotic spindle assembly, chromosome architecture changes and cohesion, and mitotic exit. This implies that certain phosphatases are tightly regulated for timely dephosphorylation of key mitotic phosphoproteins and are essential for control of various mitotic processes. This review describes the physiological and pathological roles of mitotic phosphatases, as well as the versatile role of various protein phosphatases in several mitotic events.

  1. Protein Phosphatases Involved in Regulating Mitosis: Facts and Hypotheses.

    Science.gov (United States)

    Kim, Hyun-Soo; Fernandes, Gary; Lee, Chang-Woo

    2016-09-01

    Almost all eukaryotic proteins are subject to post-translational modifications during mitosis and cell cycle, and in particular, reversible phosphorylation being a key event. The recent use of high-throughput experimental analyses has revealed that more than 70% of all eukaryotic proteins are regulated by phosphorylation; however, the mechanism of dephosphorylation, counteracting phosphorylation, is relatively unknown. Recent discoveries have shown that many of the protein phosphatases are involved in the temporal and spatial control of mitotic events, such as mitotic entry, mitotic spindle assembly, chromosome architecture changes and cohesion, and mitotic exit. This implies that certain phosphatases are tightly regulated for timely dephosphorylation of key mitotic phosphoproteins and are essential for control of various mitotic processes. This review describes the physiological and pathological roles of mitotic phosphatases, as well as the versatile role of various protein phosphatases in several mitotic events. PMID:27669825

  2. Phosphatase of Regenerating Liver and Its Association with Tumors

    Institute of Scientific and Technical Information of China (English)

    Yuqiong Liu; Huixiang Li

    2007-01-01

    @@ INTRODUCTION Protein kinases and protein phosphatases play key roles in regulating functions of diverse proteins which control numerous essential events in eukaryotes, such as transcriptional regulation, apoptosis, cell cycle progression, protein degradation and protein trafficking[1-3].

  3. Acid phosphatase and protease activities in immobilized rat skeletal muscles

    Science.gov (United States)

    Witzmann, F. A.; Troup, J. P.; Fitts, R. H.

    1982-01-01

    The effect of hind-limb immobilization on selected Iysosomal enzyme activities was studied in rat hing-limb muscles composed primarily of type 1. 2A, or 2B fibers. Following immobilization, acid protease and acid phosphatase both exhibited signifcant increases in their activity per unit weight in all three fiber types. Acid phosphatase activity increased at day 14 of immobilization in the three muscles and returned to control levels by day 21. Acid protease activity also changed biphasically, displaying a higher and earlier rise than acid phosphatase. The pattern of change in acid protease, but not acid phosphatase, closely parallels observed muscle wasting. The present data therefore demonstrate enhanced proteolytic capacity of all three fiber types early during muscular atrophy. In addition, the data suggest a dependence of basal hydrolytic and proteolytic activities and their adaptive response to immobilization on muscle fiber composition.

  4. Lipid phosphate phosphatases regulate lysophosphatidic acid production and signaling in platelets: studies using chemical inhibitors of lipid phosphate phosphatase activity.

    Science.gov (United States)

    Smyth, Susan S; Sciorra, Vicki A; Sigal, Yury J; Pamuklar, Zehra; Wang, Zuncai; Xu, Yong; Prestwich, Glenn D; Morris, Andrew J

    2003-10-31

    Blood platelets play an essential role in ischemic heart disease and stroke contributing to acute thrombotic events by release of potent inflammatory agents within the vasculature. Lysophosphatidic acid (LPA) is a bioactive lipid mediator produced by platelets and found in the blood and atherosclerotic plaques. LPA receptors on platelets, leukocytes, endothelial cells, and smooth muscle cells regulate growth, differentiation, survival, motility, and contractile activity. Definition of the opposing pathways of synthesis and degradation that control extracellular LPA levels is critical to understanding how LPA bioactivity is regulated. We show that intact platelets and platelet membranes actively dephosphorylate LPA and identify the major enzyme responsible as lipid phosphate phosphatase 1 (LPP1). Localization of LPP1 to the platelet surface is increased by exposure to LPA. A novel receptor-inactive sn-3-substituted difluoromethylenephosphonate analog of phosphatidic acid that is a potent competitive inhibitor of LPP1 activity potentiates platelet aggregation and shape change responses to LPA and amplifies LPA production by agonist-stimulated platelets. Our results identify LPP1 as a pivotal regulator of LPA signaling in the cardiovascular system. These findings are consistent with genetic and cell biological evidence implicating LPPs as negative regulators of lysophospholipid signaling and suggest that the mechanisms involve both attenuation of lysophospholipid actions at cell surface receptors and opposition of lysophospholipid production. PMID:12909631

  5. Low molecular weight protein tyrosine phosphatase (LMWPTP) upregulation mediates malignant potential in colorectal cancer

    NARCIS (Netherlands)

    E. Hoekstra (Elmer); L.L. Kodach (Liudmila L.); A. Mooppilmadham Das (Asha); R.R. Ruela-de-Sousa (Roberta); C.V. Ferreira (Carmen); J.C. Hardwick (James); C.J. van der Woude (Janneke); M.P. Peppelenbosch (Maikel); T.L.M. ten Hagen (Timo); G.M. Fuhler (Gwenny)

    2015-01-01

    textabstractPhosphatases have long been regarded as tumor suppressors, however there is emerging evidence for a tumor initiating role for some phosphatases in several forms of cancer. Low Molecular Weight Protein Tyrosine Phosphatase (LMWPTP; acid phosphatase 1 [ACP1]) is an 18 kDa enzyme that influ

  6. Phosphorylcholine Phosphatase: A Peculiar Enzyme of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Domenech

    2011-01-01

    Full Text Available Pseudomonas aeruginosa synthesizes phosphorylcholine phosphatase (PchP when grown on choline, betaine, dimethylglycine or carnitine. In the presence of Mg2+ or Zn2+, PchP catalyzes the hydrolysis of p-nitrophenylphosphate (p-NPP or phosphorylcholine (Pcho. The regulation of pchP gene expression is under the control of GbdR and NtrC; dimethylglycine is likely the metabolite directly involved in the induction of PchP. Therefore, the regulation of choline metabolism and consequently PchP synthesis may reflect an adaptive response of P. aeruginosa to environmental conditions. Bioinformatic and biochemistry studies shown that PchP contains two sites for alkylammonium compounds (AACs: one in the catalytic site near the metal ion-phosphoester pocket, and another in an inhibitory site responsible for the binding of the alkylammonium moiety. Both sites could be close to each other and interact through the residues 42E, 43E and 82YYY84. Zn2+ is better activator than Mg2+ at pH 5.0 and it is more effective at alleviating the inhibition produced by the entry of Pcho or different AACs in the inhibitory site. We postulate that Zn2+ induces at pH 5.0 a conformational change in the active center that is communicated to the inhibitory site, producing a compact or closed structure. However, at pH 7.4, this effect is not observed because to the hydrolysis of the [Zn2+L2−1L20(H2O2] complex, which causes a change from octahedral to tetrahedral in the metal coordination geometry. This enzyme is also present in P. fluorescens, P. putida, P. syringae, and other organisms. We have recently crystallized PchP and solved its structure.

  7. Long-term and seasonal variations in CO2: linkages to catchment alkalinity generation

    Directory of Open Access Journals (Sweden)

    S.A. Norton

    2001-01-01

    Full Text Available As atmospheric emissions of S have declined in the Northern Hemisphere, there has been an expectation of increased pH and alkalinity in streams believed to have been acidified by excess S and N. Many streams and lakes have not recovered. Evidence from East Bear Brook in Maine, USA and modelling with the groundwater acid-base model MAGIC (Cosby et al. 1985a,b indicate that seasonal and yearly variations in soil PCO2 are adequate to enhance or even reverse acid-base (alkalinity changes anticipated from modest decreases of SO4 in surface waters. Alkalinity is generated in the soil by exchange of H+ from dissociation of H2CO3, which in turn is derived from the dissolving of soil CO2. The variation in soil PCO2 produces an alkalinity variation of up to 15 meq L-1 in stream water. Detecting and relating increases in alkalinity to decreases in stream SO4 are significantly more difficult in the short term because of this effect. For example, modelled alkalinity recovery at Bear Brook due to a decline of 20 meq SO4 L-1 in soil solution is compensated by a decline from 0.4 to 0.2% for soil air PCO2. This compensation ability decays over time as base saturation declines. Variable PCO2 has less effect in more acidic soils. Short-term decreases of PCO2 below the long-term average value produce short-term decreases in alkalinity, whereas short-term increases in PCO2 produce short-term alkalization. Trend analysis for detecting recovery of streams and lakes from acidification after reduced atmospheric emissions will require a longer monitoring period for statistical significance than previously appreciated. Keywords: CO2 , alkalinity, acidification, recovery, soils, climate change

  8. Gabbroic xenoliths in alkaline lavas in the region of Sanganguey Volcano, Nayarit, Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Giosa, T.A.; Nelson, S.A.

    1985-01-01

    Gabbroic xenoliths occur in alkaline cinder cones and lava flows erupted from vents along five parallel lines trending through the calc-alkaline volcano, Sanganguey in the northwestern portion of the Mexican Volcanic Belt. The xenoliths consist of varying proportions of olivine, clinopyroxene, orthopyroxene, and plagioclase. The complete lack of hydrous phases indicates that the gabbros crystallized under conditions of low PH/sub 2/O. Many xenoliths show textures indicative of a cumulate origin and others exhibit recrystallization indicative of subsolidus reactions prior to incorporation in the host liquids. Reaction between xenolithic minerals and host liquids are also observed. The range of Mg numbers calculated for liquids that would have been in equilibrium with olivines in the xenoliths suggests that these olivines crystallized from magmas such as those represented by either calc-alkaline basaltic andesites and andesites or the more evolved alkalic rocks which occur throughout the area. Crystal fractionation models show that the xenoliths may be related to such magmas. The fact that xenoliths occur most commonly in the alkaline rocks suggests that alkaline magmas rise to the surface more rapidly than the more chemically evolved calc-alkaline and alkaline magmas. Alternatively the lack of xenoliths in the more evolved magmas produced by high level crystal fractionation may indicate that the xenoliths are derived from zones below that from which the differentiated magmas begin their final ascent to the surface.

  9. Influence of Oil Viscosity on Alkaline Flooding for Enhanced Heavy Oil Recovery

    Directory of Open Access Journals (Sweden)

    Yong Du

    2013-01-01

    Full Text Available Oil viscosity was studied as an important factor for alkaline flooding based on the mechanism of “water drops” flow. Alkaline flooding for two oil samples with different viscosities but similar acid numbers was compared. Besides, series flooding tests for the same oil sample were conducted at different temperatures and permeabilities. The results of flooding tests indicated that a high tertiary oil recovery could be achieved only in the low-permeability (approximately 500 mD sandpacks for the low-viscosity heavy oil (Zhuangxi, 390 mPa·s; however, the high-viscosity heavy oil (Chenzhuang, 3450 mPa·s performed well in both the low- and medium-permeability (approximately 1000 mD sandpacks. In addition, the results of flooding tests for the same oil at different temperatures also indicated that the oil viscosity put a similar effect on alkaline flooding. Therefore, oil with a high-viscosity is favorable for alkaline flooding. The microscopic flooding test indicated that the water drops produced during alkaline flooding for oils with different viscosities differed significantly in their sizes, which might influence the flow behaviors and therefore the sweep efficiencies of alkaline fluids. This study provides an evidence for the feasibility of the development of high-viscosity heavy oil using alkaline flooding.

  10. Structural elucidation of the NADP(H) phosphatase activity of staphylococcal dual-specific IMPase/NADP(H) phosphatase.

    Science.gov (United States)

    Bhattacharyya, Sudipta; Dutta, Anirudha; Dutta, Debajyoti; Ghosh, Ananta Kumar; Das, Amit Kumar

    2016-02-01

    NADP(H)/NAD(H) homeostasis has long been identified to play a pivotal role in the mitigation of reactive oxygen stress (ROS) in the intracellular milieu and is therefore critical for the progression and pathogenesis of many diseases. NAD(H) kinases and NADP(H) phosphatases are two key players in this pathway. Despite structural evidence demonstrating the existence and mode of action of NAD(H) kinases, the specific annotation and the mode of action of NADP(H) phosphatases remains obscure. Here, structural evidence supporting the alternative role of inositol monophosphatase (IMPase) as an NADP(H) phosphatase is reported. Crystal structures of staphylococcal dual-specific IMPase/NADP(H) phosphatase (SaIMPase-I) in complex with the substrates D-myo-inositol-1-phosphate and NADP(+) have been solved. The structure of the SaIMPase-I-Ca(2+)-NADP(+) ternary complex reveals the catalytic mode of action of NADP(H) phosphatase. Moreover, structures of SaIMPase-I-Ca(2+)-substrate complexes have reinforced the earlier proposal that the length of the active-site-distant helix α4 and its preceding loop are the predisposing factors for the promiscuous substrate specificity of SaIMPase-I. Altogether, the evidence presented suggests that IMPase-family enzymes with a shorter α4 helix could be potential candidates for previously unreported NADP(H) phosphatase activity.

  11. Protein phosphatase 1α is a Ras-activated Bad phosphatase that regulates interleukin-2 deprivation-induced apoptosis

    Science.gov (United States)

    Ayllón, Verónica; Martínez-A, Carlos; García, Alphonse; Cayla, Xavier; Rebollo, Angelita

    2000-01-01

    Growth factor deprivation is a physiological mechanism to regulate cell death. We utilize an interleukin-2 (IL-2)-dependent murine T-cell line to identify proteins that interact with Bad upon IL-2 stimulation or deprivation. Using the yeast two-hybrid system, glutathione S-transferase (GST) fusion proteins and co-immunoprecipitation techniques, we found that Bad interacts with protein phosphatase 1α (PP1α). Serine phosphorylation of Bad is induced by IL-2 and its dephosphorylation correlates with appearance of apoptosis. IL-2 deprivation induces Bad dephosphorylation, suggesting the involvement of a serine phosphatase. A serine/threonine phosphatase activity, sensitive to the phosphatase inhibitor okadaic acid, was detected in Bad immunoprecipitates from IL-2-stimulated cells, increasing after IL-2 deprivation. This enzymatic activity also dephosphorylates in vivo 32P-labeled Bad. Treatment of cells with okadaic acid blocks Bad dephosphorylation and prevents cell death. Finally, Ras activation controls the catalytic activity of PP1α. These results strongly suggest that Bad is an in vitro and in vivo substrate for PP1α phosphatase and that IL-2 deprivation-induced apoptosis may operate by regulating Bad phosphorylation through PP1α phosphatase, whose enzymatic activity is regulated by Ras. PMID:10811615

  12. Effects of Rice Straw Returning on Activities of Soil Phosphatase and Available P Values in Soil%水稻秸秆还田对土壤磷酸酶活性及速效磷含量的影响

    Institute of Scientific and Technical Information of China (English)

    战厚强; 颜双双; 王家睿; 马春梅; 龚振平; 董守坤; 张钦文

    2015-01-01

    The experiment focused on the cold rice,which used plot experiment and laboratory simulation experiment,and studied the effects of rice straw returning on activities of soil phosphatase and available P values in soil so as to provide evidence and reference for the rice straw returning.The result showed that activities of soil acid phosphatase were the highest in the soil,followed by activities of soil neutral phosphatase,activities of soil alkaline phosphatase.The activities of acid phosphatase with straw returning was significantly higher than that of no straw returning,the neutral phosphatase and alkaline phosphatase activities with straw returning were higher than that of no straw returning.Both the activities of acid phosphatase and available P values in soil increased with the amount of straw returned,the activities of alkaline phosphatase decreased with the amount of straw returned but the neutral phosphatase activities was relatively stable.There were a positive correlation between the activities of acid phosphatase and available P values in soil,therefore the activities of acid phosphatase could be used to report to the available P in soil,and the activities of acid phosphatase could be used as sensitive indicators of soil phosphorus status.%本试验针对中国东北稻作区,利用室内模拟试验和小区试验,研究了水稻秸秆还田对土壤磷酸酶活性及土壤速效磷含量的影响,为实现寒地水稻秸秆还田生产提供依据和参考.结果表明,土壤中酸性磷酸酶活性最大,碱性磷酸酶活性次之,中性磷酸酶活性最低;秸秆还田处理的酸性磷酸酶活性极显著高于不还田处理,秸秆还田处理的中性磷酸酶活性和碱性磷酸酶活性显著高于不还田处理;在一定范围内,酸性磷酸酶活性和土壤速效磷含量均随秸秆施入量增加而升高;中性磷酸酶活性相对稳定;碱性磷酸酶活性随秸秆施入量增加而降低.土壤酸性磷酸酶活性与土壤速效

  13. Deep-sea fungi as a source of alkaline and cold-tolerant proteases

    Digital Repository Service at National Institute of Oceanography (India)

    Damare, S.; Raghukumar, C.; Muraleedharan, U.; Raghukumar, S.

    C and 1 bar pressure. Aspergillus ustus (NIOCC#20) producing the highest amounts of the enzyme was selected for further studies. The growth yield was substantial at 30 and 5ºC at 1 bar and elevated hydrostatic pressures. The fungus produced alkaline...

  14. 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.

    nearshore to offshore Trivandrum, the pattern is more patchy. However, we could discern an increase in activity at station 5 and beyond, synchronizing with the decrease in Chl a. Access to DOP data would have clarified these aspects better... for their comments and suggestions which significantly improved the content of the manuscript. This is NIO contribution no. is xxxx. References Ahmed, N., Shahab, S., 2011. Phosphate Solubilization: Their Mechanism Genetics and Application. The Internet. J...

  15. 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

  16. Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma

    DEFF Research Database (Denmark)

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

    2006-01-01

    To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC).......To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC)....

  17. Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma

    DEFF Research Database (Denmark)

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

    2006-01-01

    BACKGROUND: To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC). METHODS: The biomarkers were determined by ELISAs in 153 metastatic PC patients before treatment with parenteral estrogen or total androgen...

  18. Identification of an alkaline phosphatase as a putative Cry1Ac binding protein in Ostrinia furnacalis (Guenée).

    Science.gov (United States)

    Jin, Tingting; Duan, Xiaoli; Bravo, Alejandra; Soberón, Mario; Wang, Zhenying; He, Kanglai

    2016-07-01

    Asian corn borer (ACB), Ostrinia furnacalis, is an important insect pest of maize susceptible to different Cry1A toxins. Based on amino acid sequence alignment of ALP sequences from lepidopteran larvae an alp gene was cloned from ACB, named ofalp. Pull dawn assays using biotinylated Cry1Ac and brush border membrane vesicles isolated from second instar ACB larvae showed that four proteins of 50, 65, 68 and 70kDa precipitated with the Cry1Ac. The 65kDa band cross-reacted with the anti-OfALP monoclonal antibody. GalNac was able to release the binding of Cry1Ac to the 65kDa OfALP in pull down assays. A 37kDa fragment from residues D173 to D473 of OfALP was cloned and expressed in Escherichia coli cells. We show that this ALP-fragment was able to bind Cry1Ac in ligand blot analysis. Our data also indicate that different ALP isoforms or variants may be also Cry1Ac binding proteins since more ALP enzymatic activity was pull down with Cry1Ac than with anti-OfALP antibody. We also analyzed the expression levels of ALP throughout the larval development by qPCR and ALP enzymatic activity. Our data indicated that ALP expression in ACB was observed preferentially in young instar larvae. Finally, we show that resistance in O. furnacalis ACB-AcR strain resistant to Cry1Ac did not correlate with changes in expression of this ALP protein since it shows similar gene expression of ofalp than the susceptible insect strain. Identification of Cry1Ac receptors will help to understand mechanism of action of Cry1Ac in O. furnacalis and to understand mechanism of Cry toxin resistance. Our data indicate that at least one ALP protein is involved in the binding interaction with Cry1Ac in O. furnacalis. PMID:27265829

  19. Alkaline sorbent injection for mercury control

    Science.gov (United States)

    Madden, Deborah A.; Holmes, Michael J.

    2003-01-01

    A mercury removal system for removing mercury from combustion flue gases is provided in which alkaline sorbents at generally extremely low stoichiometric molar ratios of alkaline earth or an alkali metal to sulfur of less than 1.0 are injected into a power plant system at one or more locations to remove at least between about 40% and 60% of the mercury content from combustion flue gases. Small amounts of alkaline sorbents are injected into the flue gas stream at a relatively low rate. A particulate filter is used to remove mercury-containing particles downstream of each injection point used in the power plant system.

  20. Embryonic stem-like cells derived from in vitro produced bovine blastocysts

    Directory of Open Access Journals (Sweden)

    Erika Regina Leal de Freitas

    2011-06-01

    Full Text Available The aim of this work was to study the derivation of bovine embryonic stem-like (ES-like cells from the inner cell mass (ICM of in vitro produced blastocysts. The ICMs were mechanically isolated and six out of seventeen (35% ICMs could attach to a monolayer of murine embryonic fibroblasts (MEF. Ten days after, primary outgrowths were mechanically dissected into several small clumps and transferred to a new MEF layer. Cells were further propagated and passaged by physical dissociation over a 60 days period. The pluripotency of the bovine ES-like cells was confirmed by RT-PCR of Oct-4 and STAT-3 gene markers. The colonies were weakly stained for alkaline phosphatase and the mesoderm and endoderm differentiation gene markers such as GATA-4 and Flk-1, respectively, were not expressed. Embryoid bodies were spontaneously formed at the seventh passage. Results showed that bovine ES-like cells could be obtained and passaged by mechanical procedures from the fresh in vitro produced blastocysts.

  1. Evaluation of biochemical alterations produced by combined exposure of fenvalerate and nitrate in Bubalus bubalis

    Directory of Open Access Journals (Sweden)

    Kamalpreet Kaur Gill

    2014-03-01

    Full Text Available Aim: Evaluation of combined effect of fenvalerate and nitrate on biochemical parameters in buffalo calves. Materials and Methods: Sixteen male buffalo calves were divided into four groups of four calves each. Group I receiving no treatment served as the control. Group II and III animals were orally administered with fenvalerate (1.0 mg/kg/day and sodium nitrate (20 mg/kg/day, respectively, for 21 consecutive days and were kept as positive control. Group IV animals were co-administered with fenvalerate and sodium nitrate at the above dose rates for 21 consecutive days. Biochemical parameters including Aspartate aminotransferase (AST, Alkaline phosphatase (ALP, Gamma-glutamyl transpeptidase (GGT, Lactate dehydrogenase (LDH, Glucose, Total protein, Albumin, Cholesterol, Blood urea nitrogen (BUN and Creatinine were determined on 0, 3, 7, 10, 14, 17 and 21 day of treatment. Estimation of these parameters was also done on 7th day of post-treatment period. Results: Co-administration of fenvalerate and sodium nitrate produced significant increase in the plasma levels of AST, ALP, GGT, LDH, glucose, BUN, cholesterol and creatinine while significant decrease in the plasma levels of total proteins was observed. No significant alteration was observed in albumin levels. Extent of organ damage as evidenced by biochemical alterations was more pronounced in calves exposed to combination of fenvalerate and sodium nitrate as compared to their individual exposures. Conclusion: Fenvalerate and sodium nitrate co-administration potentiates the toxicological injury produced, in comparison to their individual exposure.

  2. 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. PMID:25254238

  3. Production of Alkaline Cellulase by Fungi Isolated from an Undisturbed Rain Forest of Peru

    Directory of Open Access Journals (Sweden)

    Karin Vega

    2012-01-01

    Full Text Available Alkaline cellulase producing fungi were isolated from soils of an undisturbed rain forest of Peru. The soil dilution plate method was used for the enumeration and isolation of fast growing cellulolytic fungi on an enriched selective medium. Eleven out of 50 different morphological colonies were finally selected by using the plate clearing assay with CMC as substrate at different pH values. All 11 strains produced cellulases in liquid culture with activities at alkaline pH values without an apparent decrease of them indicating that they are true alkaline cellulase producers. Aspergillus sp. LM-HP32, Penicillium sp. LM-HP33, and Penicillium sp. LM-HP37 were the best producers of FP cellulase (>3 U mL−1 with higher specific productivities (>30 U g−1 h−1. Three strains have been found suitable for developing processes for alkaline cellulase production. Soils from Amazonian rain forests are good sources of industrial fungi with particular characteristics. The results of the present study are of commercial and biological interest. Alkaline cellulases may be used in the polishing and washing of denim processing of the textile industry.

  4. Protein phosphatase 2A regulatory subunit B56α limits phosphatase activity in the heart.

    Science.gov (United States)

    Little, Sean C; Curran, Jerry; Makara, Michael A; Kline, Crystal F; Ho, Hsiang-Ting; Xu, Zhaobin; Wu, Xiangqiong; Polina, Iuliia; Musa, Hassan; Meadows, Allison M; Carnes, Cynthia A; Biesiadecki, Brandon J; Davis, Jonathan P; Weisleder, Noah; Györke, Sandor; Wehrens, Xander H; Hund, Thomas J; Mohler, Peter J

    2015-07-21

    Protein phosphatase 2A (PP2A) is a serine/threonine-selective holoenzyme composed of a catalytic, scaffolding, and regulatory subunit. In the heart, PP2A activity is requisite for cardiac excitation-contraction coupling and central in adrenergic signaling. We found that mice deficient in the PP2A regulatory subunit B56α (1 of 13 regulatory subunits) had altered PP2A signaling in the heart that was associated with changes in cardiac physiology, suggesting that the B56α regulatory subunit had an autoinhibitory role that suppressed excess PP2A activity. The increase in PP2A activity in the mice with reduced B56α expression resulted in slower heart rates and increased heart rate variability, conduction defects, and increased sensitivity of heart rate to parasympathetic agonists. Increased PP2A activity in B56α(+/-) myocytes resulted in reduced Ca(2+) waves and sparks, which was associated with decreased phosphorylation (and thus decreased activation) of the ryanodine receptor RyR2, an ion channel on intracellular membranes that is involved in Ca(2+) regulation in cardiomyocytes. In line with an autoinhibitory role for B56α, in vivo expression of B56α in the absence of altered abundance of other PP2A subunits decreased basal phosphatase activity. Consequently, in vivo expression of B56α suppressed parasympathetic regulation of heart rate and increased RyR2 phosphorylation in cardiomyocytes. These data show that an integral component of the PP2A holoenzyme has an important inhibitory role in controlling PP2A enzyme activity in the heart.

  5. Method of producing hydrogen, and rendering a contaminated biomass inert

    Science.gov (United States)

    Bingham, Dennis N [Idaho Falls, ID; Klingler, Kerry M [Idaho Falls, ID; Wilding, Bruce M [Idaho Falls, ID

    2010-02-23

    A method for rendering a contaminated biomass inert includes providing a first composition, providing a second composition, reacting the first and second compositions together to form an alkaline hydroxide, providing a contaminated biomass feedstock and reacting the alkaline hydroxide with the contaminated biomass feedstock to render the contaminated biomass feedstock inert and further producing hydrogen gas, and a byproduct that includes the first composition.

  6. Surfactant and adhesive formulations from alkaline biomass extracts

    Science.gov (United States)

    Baxter, Matthew

    This work studies the ability to produce effective surfactant and adhesive formulations using surface active biological material extracted from different biomass sources using alkaline extraction methods. Two urban waste biomass sources were used to produce surfactants, Return Activated Sludge (RAS), and solid Urban Refuse (UR). The third biomass source investigated was isolated mustard protein (MP). RAS and MP extracts were investigated for adhesive production. The results indicate that extracts from the waste biomass sources, RAS and UR, can be combined with a commercial surfactant, sodium dioctyl sulfosuccinate (AOT), to produce surfactants with low interfacial tensions against various oils. These highly surface-active formulations were shown to be useful in the removal of bitumen from contaminated sand. RAS and MP showed potential as protein-based wood adhesives. These sources were used in adhesive formulations to produce a strong bond strength under low-pressure, ambient pressing conditions.

  7. Electrolytic denitrification of alkaline nitrate and nitrite solution

    International Nuclear Information System (INIS)

    Processing of high-level waste at the Savannah River Plant (SRP) will produce a low-level alkaline salt solution, containing approximately 17% sodium nitrate and sodium nitrite. This solution will be incorporated into a cement wasteform, saltstone, and placed in an engineered landfill. Laboratory experiments have demonstrated the technical feasibility of electrochemically reducing the nitrate and nitrite in a synthetic, nonradioactive salt solution similar in composition to that expected to be produced at SRP. Greater than ninety-five percent of the sodium nitrate and sodium nitrite can be reduced electrolytically, producing ammonia, nitrogen, oxygen, and sodium hydroxide. Reduction of the nitrate and nitrite will reduce the leaching of nitrate and nitrite from the saltstone monolith. In addition, significant reductions in the volume of saltstone may be realized if the sodium hydroxide produced by electrolysis can be recycled

  8. Electrochemical processing of alkaline nitrate and nitrite solutions

    International Nuclear Information System (INIS)

    Processing of high-level waste at the Savannah River Plant (SRP) will produce, as a by product, a low- level, alkaline salt solution containing approximately 17% sodium nitrate and sodium nitrite. This solution will be incorporated into a cement formulation, saltstone, and placed in an engineered landfill. Electrochemical methods have been investigated to decrease the nitrate and nitrite in this solution in order to lower the leaching of nitrate and nitrite from saltstone and to reduce the volume of saltstone. Laboratory experiments have demonstrated the technical feasibility of electrolytically reducing the nitrate and nitrite in a synthetic salt solution similar in composition to that expected to be produced at SRP. Greater than 99% of the sodium nitrate and sodium nitrite can be reduced, producing ammonia, nitrogen, oxygen, and sodium hydroxide. In addition, significant reductions in the volume of saltstone may be realized if the sodium hydroxide produced by electrolysis can be recycled

  9. Acid phosphatase and lipid peroxidation in human cataractous lens epithelium

    Directory of Open Access Journals (Sweden)

    Vasavada Abhay

    1993-01-01

    Full Text Available The anterior lens epithelial cells undergo a variety of degenerative and proliferative changes during cataract formation. Acid phosphatase is primarily responsible for tissue regeneration and tissue repair. The lipid hydroperoxides that are obtained by lipid peroxidation of polysaturated or unsaturated fatty acids bring about deterioration of biological membranes at cellular and tissue levels. Acid phosphatase and lipid peroxidation activities were studied on the lens epithelial cells of nuclear cataract, posterior subcapsular cataract, mature cataract, and mixed cataract. Of these, mature cataractous lens epithelium showed maximum activity for acid phosphatase (516.83 moles of p-nitrophenol released/g lens epithelium and maximum levels of lipid peroxidation (86.29 O.D./min/g lens epithelium. In contrast, mixed cataractous lens epithelium showed minimum activity of acid phosphatase (222.61 moles of p-nitrophenol released/g lens epithelium and minimum levels of lipid peroxidation (54.23 O.D./min/g lens epithelium. From our study, we correlated the maximum activity of acid phosphatase in mature cataractous lens epithelium with the increased areas of superimposed cells associated with the formation of mature cataract. Likewise, the maximum levels of lipid peroxidation in mature cataractous lens epithelium was correlated with increased permeability of the plasma membrane. Conversely, the minimum levels of lipid peroxidation in mixed cataractous lens epithelium makes us presume that factors other than lipid peroxidation may also account for the formation of mixed type of cataract.

  10. Mechanism of gold dissolving in alkaline thiourea solution

    Institute of Scientific and Technical Information of China (English)

    CHAI Li-yuan; WANG Yun-yan

    2007-01-01

    Reaction mechanism of gold dissolving in alkaline thiourea solution was studied by electrochemical methods, such as cyclic voltammetry, chronopotentiometry, AC impedance, linear sweep voltammetry. Apparent activation energy of anodic process of gold electrode dissolving in alkaline thiourea solution is 14.91 kJ/mol. Rate determining step is the process of gold thiourea complex diffusing away from electrode surface to solution. The results of AC impedance and chronopotentiometry indicate that thiourea adsorbs on gold electrode surface before dissolving in solution. There does not exist proceeding chemical reactions. Formamidine disulfide, the decomposed product of thiourea, does not participate the process of gold dissolution and thiourea complex. Species with electro-activity produced in the process of electrode reaction adsorbs on the electrode surface. In alkaline thiourea solution, gold dissolving mechanism undergoes the following courses: adsorption of thiourea on electrode surface; charge transfer from gold atom to thiourea molecule; Au[SC(NH2)2]ads+ receiving a thiourea molecule and forming stable Au[SC(NH2)2]2+; and then Au[SC(NH2)2]2+diffusing away from the electrode surface to solution, the last step is the rate-determining one.

  11. Alkaline protease from Thermoactinomyces sp. RS1 mitigates industrial pollution.

    Science.gov (United States)

    Verma, Amit; Ansari, Mohammad W; Anwar, Mohmmad S; Agrawal, Ruchi; Agrawal, Sanjeev

    2014-05-01

    Proteases have found a wide application in the several industrial processes, such as laundry detergents, protein recovery or solubilization, prion degradation, meat tenderizations, and in bating of hides and skins in leather industries. But the main hurdle in industrial application of proteases is their economical production on a large scale. The present investigation aimed to exploit the locally available inexpensive agricultural and household wastes for alkaline protease production using Thermoactinomyces sp. RS1 via solid-state fermentation (SSF) technique. The alkaline enzyme is potentially useful as an additive in commercial detergents to mitigate pollution load due to extensive use of caustic soda-based detergents. Thermoactinomyces sp. RS1 showed good protease production under SSF conditions of 55 °C, pH 9, and 50 % moisture content with potato peels as solid substrate. The presented findings revealed that crude alkaline protease produced by Thermoactinomyces sp. RS1 via SSF is of potential application in silver recovery from used X-ray films. PMID:24122212

  12. Coupling the Alkaline-Surfactant-Polymer Technology and The Gelation Technology to Maximize Oil Production

    Energy Technology Data Exchange (ETDEWEB)

    Malcolm Pitts; Jie Qi; Dan Wilson; Phil Dowling; David Stewart; Bill Jones

    2005-12-01

    the fluid-fluid analyses with the exception of the xanthan gum-chromium acetate gels. Aluminum-polyacrylamide flowing gels are not stable to alkaline-surfactant-polymer solutions of either pH 10.5 or 12.9, either in linear corefloods or in dual separate radial core, common manifold corefloods. Chromium acetate-polyacrylamide flowing and rigid tonguing gels are stable to subsequent alkaline-surfactant-polymer solution injection. Rigid tonguing chromium acetate-polyacrylamide gels maintained permeability reduction better than flowing chromium acetate-polyacrylamide gels. Chromium acetate gels were stable to injection of alkaline-surfactant-polymer solutions at 72 F, 125 F and 175 F in linear corefloods. Chromium acetate-polyacrylamide gels maintained diversion capability after injection of an alkaline-surfactant-polymer solution in stacked; radial coreflood with a common well bore. Chromium acetate-polyacrylamide gel used to seal fractured core maintain fracture closure if followed by an alkaline-surfactant-polymer solution. Chromium acetatexanthan gum rigid gels are not stable to subsequent alkaline-surfactant-polymer solution injection at 72, 125, and 175 F. Silicate-polyacrylamide gels are not stable with subsequent injection of either a pH 10.5 or a 12.9 alkaline-surfactant-polymer solution. Resorcinol-formaldehyde gels were stable to subsequent alkaline-surfactant-polymer solution injection. When evaluated in a dual core configuration, injected fluid flows into the core with the greatest effective permeability to the injected fluid. The same gel stability trends to subsequent alkaline-surfactant-polymer injected solution were observed. Aluminum citrate-polyacrylamide, resorcinol-formaldehyde, and the silicate-polyacrylamide gel systems did not produce significant incremental oil in linear corefloods. Both flowing and rigid tonguing chromium acetate-polyacrylamide gels and the xanthan gum-chromium acetate gel system produced incremental oil with the rigid tonguing

  13. Technetium recovery from high alkaline solution

    Science.gov (United States)

    Nash, Charles A.

    2016-07-12

    Disclosed are methods for recovering technetium from a highly alkaline solution. The highly alkaline solution can be a liquid waste solution from a nuclear waste processing system. Methods can include combining the solution with a reductant capable of reducing technetium at the high pH of the solution and adding to or forming in the solution an adsorbent capable of adsorbing the precipitated technetium at the high pH of the solution.

  14. The Origin of Life in Alkaline Hydrothermal Vents.

    Science.gov (United States)

    Sojo, Victor; Herschy, Barry; Whicher, Alexandra; Camprubí, Eloi; Lane, Nick

    2016-02-01

    Over the last 70 years, prebiotic chemists have been very successful in synthesizing the molecules of life, from amino acids to nucleotides. Yet there is strikingly little resemblance between much of this chemistry and the metabolic pathways of cells, in terms of substrates, catalysts, and synthetic pathways. In contrast, alkaline hydrothermal vents offer conditions similar to those harnessed by modern autotrophs, but there has been limited experimental evidence that such conditions could drive prebiotic chemistry. In the Hadean, in the absence of oxygen, alkaline vents are proposed to have acted as electrochemical flow reactors, in which alkaline fluids saturated in H2 mixed with relatively acidic ocean waters rich in CO2, through a labyrinth of interconnected micropores with thin inorganic walls containing catalytic Fe(Ni)S minerals. The difference in pH across these thin barriers produced natural proton gradients with equivalent magnitude and polarity to the proton-motive force required for carbon fixation in extant bacteria and archaea. How such gradients could have powered carbon reduction or energy flux before the advent of organic protocells with genes and proteins is unknown. Work over the last decade suggests several possible hypotheses that are currently being tested in laboratory experiments, field observations, and phylogenetic reconstructions of ancestral metabolism. We analyze the perplexing differences in carbon and energy metabolism in methanogenic archaea and acetogenic bacteria to propose a possible ancestral mechanism of CO2 reduction in alkaline hydrothermal vents. Based on this mechanism, we show that the evolution of active ion pumping could have driven the deep divergence of bacteria and archaea.

  15. The Origin of Life in Alkaline Hydrothermal Vents.

    Science.gov (United States)

    Sojo, Victor; Herschy, Barry; Whicher, Alexandra; Camprubí, Eloi; Lane, Nick

    2016-02-01

    Over the last 70 years, prebiotic chemists have been very successful in synthesizing the molecules of life, from amino acids to nucleotides. Yet there is strikingly little resemblance between much of this chemistry and the metabolic pathways of cells, in terms of substrates, catalysts, and synthetic pathways. In contrast, alkaline hydrothermal vents offer conditions similar to those harnessed by modern autotrophs, but there has been limited experimental evidence that such conditions could drive prebiotic chemistry. In the Hadean, in the absence of oxygen, alkaline vents are proposed to have acted as electrochemical flow reactors, in which alkaline fluids saturated in H2 mixed with relatively acidic ocean waters rich in CO2, through a labyrinth of interconnected micropores with thin inorganic walls containing catalytic Fe(Ni)S minerals. The difference in pH across these thin barriers produced natural proton gradients with equivalent magnitude and polarity to the proton-motive force required for carbon fixation in extant bacteria and archaea. How such gradients could have powered carbon reduction or energy flux before the advent of organic protocells with genes and proteins is unknown. Work over the last decade suggests several possible hypotheses that are currently being tested in laboratory experiments, field observations, and phylogenetic reconstructions of ancestral metabolism. We analyze the perplexing differences in carbon and energy metabolism in methanogenic archaea and acetogenic bacteria to propose a possible ancestral mechanism of CO2 reduction in alkaline hydrothermal vents. Based on this mechanism, we show that the evolution of active ion pumping could have driven the deep divergence of bacteria and archaea. PMID:26841066

  16. Ocean Acidification: Coccolithophore's Light Controlled Effect on Alkalinity

    Science.gov (United States)

    Dobbins, W.

    2015-12-01

    Coccolithophorids, which play a significant role in the flux of calcite and organic carbon from the photic region to deeper pelagic and benthic zones, are potentially far more useful than siliceous phytoplankton for ocean fertilization projects designed to sequester CO2. However, the production of H+ ions during calcification (HCO3 + Ca+ —> CaCO3 + H+) has resulted in localized acidification around coccolithophore blooms. It has been hypothesized that under the correct light conditions photosynthesis could proceed at a rate such that CO2 is removed in amounts equimolar or greater than the H+ produced by calcification, allowing stable or increasing alkalinity despite ongoing calcification. Previously, this effect had not been demonstrated under laboratory conditions. Fifteen Emiliania huxleyi cultures were separated into equal groups with each receiving: 0, 6, 12, 18, or 24 hours of light each day for 24 days. Daily pH, cell density, and temperature measurements revealed a strong positive correlation between light exposure and pH, and no significant decline in pH in any of the cultures. Alkalinity increases were temperature independent and not strongly correlated with cell density, implying photosynthetic removal of carbon dioxide as the root cause. The average pH across living cultures increased from 7.9 to 8.3 over the first week and changed little for the reminder of the 24-day period. The results demonstrate coccolithophorids can increase alkalinity across a broad range of cell densities, despite the acidification inherent to the calcification process. If the light-alkalinity effect reported here proves scalable to larger cultures, Emiliania huxleyi are a strong candidate for carbon sequestration via targeted ocean fertilization.

  17. Metals in the active site of native protein phosphatase-1.

    Science.gov (United States)

    Heroes, Ewald; Rip, Jens; Beullens, Monique; Van Meervelt, Luc; De Gendt, Stefan; Bollen, Mathieu

    2015-08-01

    Protein phosphatase-1 (PP1) is a major protein Ser/Thr phosphatase in eukaryotic cells. Its activity depends on two metal ions in the catalytic site, which were identified as manganese in the bacterially expressed phosphatase. However, the identity of the metal ions in native PP1 is unknown. In this study, total reflection X-ray fluorescence (TXRF) was used to detect iron and zinc in PP1 that was purified from rabbit skeletal muscle. Metal exchange experiments confirmed that the distinct substrate specificity of recombinant and native PP1 is determined by the nature of their associated metals. We also found that the iron level associated with native PP1 is decreased by incubation with inhibitor-2, consistent with a function of inhibitor-2 as a PP1 chaperone. PMID:25890482

  18. Prostatic acid phosphatase, purification and iodination using Iodogen

    International Nuclear Information System (INIS)

    Prostatic acid phosphatase was purified from prostatic adenomas. The procedure involved chromatography on Concanavalin A-Sepharose, DEAE-cellulose, Bio-Gel P-150 and L-tartrate-Sepharose. The purified phosphatase hydrolyzed p-nitrophenyl phosphate at a rate of 270 μmol.mg-1.min-1 (250C) and showed homogeneity upon polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The final prostatic acid phosphatase preparation was pure and the antisera were monospecific as judged by the highly-sensitive technique of crossed immunoelectrophoresis. Of the procedures evaluated for the radioiodination of the purified enzyme with iodine 125, oxidation with Iodogen was found to give the best radioiodinated product, to be used in radioimmunoassay. (Auth.)

  19. Association of erythrocyte acid phosphatase phenotypes with myopia

    Directory of Open Access Journals (Sweden)

    Himabindu P

    2005-01-01

    Full Text Available Acid phosphatase is a polymorphic nonspecific orthophosphate monoesterase which catalyses the cleaving of phosphoric acid and subsequent breakdown of several monophosphoric esters under acidic pH conditions. Acid phosphatase has a physiologic function as a flavin mononucleotide phosphatase (FMN and regulates the intracellular concentrations of flavin coenzymes that are electron carriers in the oxidative phosphorylation pathway. Myopia or nearsightedness is caused by both environmental and genetic factors. Myopic eyes when subjected to excessive oxidative stress results in retinal detachments .In the present study there is a significant elevation of AA phenotype in myopes when compared to controls. The AA phenotype is more susceptible to oxidative stress and its lower enzyme activity is known to be associated with increased intrauterine growth that further results in increased axial length in progressive myopia. The AA phenotype also confers risk for myopia development in males, early age group and cases with parental consanguinity.

  20. Ultrastructural localization of acid phosphatase in nonhuman primate vaginal epithelium.

    Science.gov (United States)

    King, B F

    1985-01-01

    The vagina of the rhesus monkey is lined by a stratified squamous epithelium. However, little is known regarding the cytochemical composition of its cell organelles and the substances found in the intercellular spaces. In this study we have examined the ultrastructural distribution of acid phosphatase in the vaginal epithelium. In basal and parabasal cells reaction product was found in some Golgi cisternae and vesicles and in a variety of cytoplasmic granules. Reaction product was also found in some, but not all, membrane-coating granules. In the upper layers of the epithelium, the membrane-coating granules extruded their contents and acid phosphatase was localized in the intercellular spaces. The possible roles of acid phosphatase in keratinization, desquamation, or modification of substances in the intercellular compartment are discussed.